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GSR Breeding Strategy – Exploiting the
  Hidden Reservoir of Diversity for
  Improving Complex Traits in Rice
          Zhi-Kang Li/CAAS

                                GA biosynthetic pathway
                          Geranylgeranyl
                          diphospate                             AS
                                       CPS         GA12 -aldehyde
                                                                 GA7ox
                     ent - Copalyl diphosphate
                                                         GA 12
                                          KS
                                         KS                 GA13ox
                         ent -Kaurene                     GA 53
                         ent -Kaurenol           GA 15            GA 44
                                      KO
                                       KO                  GA
                        ent -Kaurenal                    C20ox GA 19
                                                 GA 24    sd-1
                      ent -Kaurenoic acid        GA 9             GA 20
                                        KAO
                                       KAO
                                                         GA3ox
                        ent -7a-hydroxy
                         kaurenoic acid          GA4              GA 1
Subjects

Introduction

Concept of GSR

GSR Breeding Strategy and
Technology
Perspectives
The impact of ‘Green Revolution’ in China

        60000
                 Impact of ‘Green Revolution’                                                Impact of hybrid rice
        50000

        40000

        30000
Yield




        20000

        10000

          0
                1965
                       1967
                              1969
                                     1971
                                            1973
                                                   1975
                                                          1977
                                                                 1979
                                                                        1981
                                                                               1983
                                                                                      1985
                                                                                             1988
                                                                                                    1990
                                                                                                           1992
                                                                                                                  1994
                                                                                                                         1996
                                                                                                                                1998
                                                                                                                                       2000
                                                                                                                                              2002
                                                                  Year

                                                                                                                           Yield/unit area
                                                                                                                           Total production
Important problems in rice
       production worldwide

• Increasing problems in abiotic and biotic
  stresses
• Over-use of pesticides
• Abuse of chemical fertilizers
• Shortage of water
• Ever increasing demands for yield increase
• Urgent need for improved quality
In China
Current agricultural practices: higher inputs-for high yields- polluted
  environments
China consumes ~1/3 of the global production of chemical fertilizers
  and pesticides annually on only 7% of the world’s cultivated lands

                                     Fertilizer application and grain production in China




                                                                                                                       Total grain production(in
                                                                                                                       10000 t)、unit area yield
                                   4500                                                                        75000




                                                                                                                                   production(
                                                                                                               70000
                                   4000
                                                       Fertilizer use                                          65000
      Fertilizer use (in 10000t)




                                   3500                Grain production
                                                       Grain yield per unit area                               60000




                                                                                                                               (kg/10ha)
                                                                                                                                       )
                                   3000                                                                        55000




                                                                                                                               )、unit
                                                                                                               50000
                                   2500
                                                                                                               45000




                                                                                                                               )、
                                   2000
                                                                                                               40000
                                   1500                                                                        35000
                                                                                                               30000
                                   1000
                                                                                                               25000
                                    500
                                                                                                               20000
                                     0                                                                         15000
                                         1975   1978     1982    1985    1988      1991   1994   1997   2000
Trends of rice production and uses of pesticides
                  and fertilizers in China in the last 30 years
                       40                                                 150                          50




                                                                                                            Fertilizers (million ton)
                                                                                                                                    )
                7      35   单产                                                                         45
                                                                          130




                                                                               Pesticides(10000 ton)
                                                                                                   )
                            农药                                                                         40
Yield(ton/ha)
     (ton/ha)




                       30   化肥                                            110
                6                                                                                      35
                       25
                                                                          90                           30
                       20




                                                                                         (
                5                                                         70                           25
                       15
                                                                                                       20
                                                                          50
                4      10                                                                              15
                        5                                                 30
                                                                                                       10
                3       0                                                 10                           5
                    1978 1981 1984 1981 1984 1987 1990 1993 1996 1999 2002 2005
                          1975 1978 1987 1990 1993 1996 1999 2002 2005
5



              Grain yield(ton/hm2)
                                                                      Grain yield per unit area
                                                                     Grain yield/ha
                                          4
                           /


                                          3


                                          2


                                          1
                                                    1961     1966     1971      1976           1981      1986          1991          1996          2001         2006
                                          350                                                                                                                        350
                                          350                                                                                                              350




                                                                                                                                                                         Nitrogen use efficiency (kg/kg )
          Nitrogen consumption(kg/hm2 )




                                          300                                                 氮肥用量                                                                   300
                                          300                                                                                                              300




                                                                                                                                                                                                            )
公顷)




                                                                                       PFP
                                          250                                     N consumption                                                                      250




                                                                                                                                                                                                              kg/kg
                                          250                                                                                                              250
 /




                                          200
                                                                                             NUE                                                                     200
                                          200                                                                                                              200




                                                                                                                                                                                                            氮肥生产效率(
氮肥用量(公斤




                                          150                                                                                                                        150
                                          150                                                                                                              150
                                          100                                                                                                                        100
                                          100                                                                                                              100
                                           50                                                                                                                       50
                                           50                                                                                                              50
                                                0                                                                                                               0
                                                0                                                                                                          0
                                                      1961    1966     1971     1976          1981     1986     1991          1996          2001      2006
                                                     1961    1966     1971    1976           1981     1986    1991       1996          2001         2006
Annual yield losses

~20% from abiotic stress: drought,
       problem soils, etc)

~15% from biotic stress: diseases
and insects, even with the heavy
       uses of pesticides.
Water crisis and
                                drought in China




Fresh water resources per capita in China is less than a
quarter of the world average;
Agriculture uses ~70% of the fresh water in China, and
rice uses ~70% of the fresh water used in agriculture;
Drought is occurring more frequently than ever before;
Great yield loss of rice to drought in major rice areas.
Which places have been left out by Green
                         Revolution: Rice Yield Gaps in Irrigated and
                            Rainfed Ecosystems in Asia, 1967-97
                   6.0
                                                                                    Irrigated     Rainfed

                   5.0                Irrigated                               100
                                        (40%)

                   4.0                                                        80
Y ie ld (t/h a )




                                               Largely Irrigated
                                                    (30%)                     60
                   3.0

                                                                              40
                   2.0

                                                                              20
                                                       Rainfed
                   1.0                                  (30%)
                                                                               0
                                                                                    MC              TC
                   0.0
                     1967 1970 1973 1976 1979 1982 1985 1988 1991 1994 1997
                                                                                     Adoption percentage
                                             Year
                                                                                     of modern cultivars
 Source: M. Hossain, IRRI
Consequences of the
 Green Revolution
               Productivity




                  Diversity
Chinese scientists are calling
    “Second Green Revolution”
   Less inputs, more production, and
     environmental sustainability
    少投入、多产出、保护环境

- to develop and widely adopted “Green Super
 Rice” that can produce high and stable yields
     under less inputs (chemical fertilizers,
   pesticides and water, and stress resilient).
What are “GSR”?
High yielding cultivars with multiple “Green”
traits:
Resistances/ tolerances to:
Abiotic stresses: Drought, salinity, alkalinity, etc.
Diseases: Blast, bacterial blight, sheath blight,
  viruses, and false smut etc
Insects: Brown plant hopper, stem borer, etc
High resource-use efficiencies: Water and
nutrients (N, P)
To develop GSR rice varieties, breeders
are facing the following challenges:

    Many target traits:
    Yield and its related traits
    Stability
     - Resistances to biotic stresses
     - Tolerances to abiotic stresses

    Quality
     - Eating, cooking, and milling
     - Micronutrients
Where are the sources
 of genetic variation
  for improving the
    ‘green’ traits?
Rice Germplasm Collections in
   Genebanks Worldwide: ~ 215,000 entries

10% of 22 wild species


                                     27% of
                                    modern
                                    varieties


        90% of the                               73% of
       Cultivated rice                          landraces



                     Everson et al. 1998
Status of the International Rice
   Genebank Collection at IRRI
                             Incoming
                Accessions               Total
                              samples

O. sativa       85,999       15,784     101,783

O. glaberrima    1,333          288       1,621

Wild species     3,970          495       4,465

Total           91,302       16,567     107,869
Characterization of O. sativa
% accessions scored
                        accessions (n=79,925)
                100
                 90
                 80
                 70
                 60
                 50
                 40
                 30
                 20
                 10
                  0
                             44 morpho-agronomic traits
No. accessions




                20,000
                10,000
                30,000
                40,000
        Blast   50,000
    Bacterial
     blight
      Sheath
      blight
                             Screened




 Rice tungro

       BPH 1

       BPH 2
                             Resistant




       BPH 3
     Green
                                             resistance/tolerances at IRRI




  leafhopper
Whitebacked
planthopper
                                         Evaluation of rice germplasm for stress




      Cold
    tolerance
Cold tolerance (1-3 = tolerant; 5 =
    intermediate; 7-9 = susceptible)
2,500
            n=6,625
2,000

1,500

1,000

 500

   0
        1         3   5     7      9
Yield responses of the 193 parental lines of IRMBN
     to the terminal drought under the lowland condition

25
                                  ±
                             -19.1±44.0%
20


15


10


5


0
 -100 -80 -60 -40 -20   0   20   40   60   80 100 120 140 160

                  Yield reduction (in %)
Establishment of the core collections for
              major crops in the CAAS’s genebank
      Core collections of rice, wheat, corn and                                                       Indica




      soybean were established with only 5% of
      the accessions representing over 90%                                                                         Jap M.
                                                                                                                     .

                                                                                                                   Jap U.
                                                                                                                     .

                                                                                                                   Jap L.
                                                                                                                     .



      genetic diversity of the whole collections.                                               0.1
                                                                                                0.1
                                                                                                        Japonica
                                                                                                                   Ind EM.
                                                                                                                      .
                                                                                                                   Ind M.
                                                                                                                      .

                                                                                                                   Ind M
                                                                                                                     . L




              Basic Collections             Core Collection                 Mini-core

           Accessions   Entries sampling % Representation   Entries sampling % Representation


    Rice      61479 3074             5%         89.9%             300 0.5%            66.6%
  Wheat         23135 1160            5%          90.1%            231 1.0%             69.1%
  Soybean 28809 1439                  5%          91.0%            280       1.0% 71.0%
Comparison of genetic diversity among core collections
with different sampling ratio                                            (Jia jizheng,2004)
Current Status of Characterization and Utilization of
 Germplasm Collections in Genebanks Worldwide

  Collection and conservation largely completed

  Characterization – very superficial
   - Phenotypic evaluation and description
   To identify accessions with desirable (often extreme)
   phenotypes to be used as parents in breeding programs
   - Few accessions with desirable phenotypes for most traits
   - No accessions with desirable phenotypes for certain traits

  How much valuable genetic variation for target traits in
  the primary gene pool for breeding remains unclear!
Utilization – very poorly
- Less than 5% of the collected germplasm in the Genebanks
  have been utilized in the worldwide breeding programs

Reasons for Poor Utilization of Germplasm
 Outstanding commercial genotypes are commonly
 destroyed by crosses with unimproved exotic germplasm
 (Duvick 1984)
 Slow but consistent genetic improvement can still be
 achieved even within a narrow base in many breeding
 programs
 Selection of parental lines in breeding programs are largely
 based on phenotype and very few accessions appear to have
 “desirable” phenotype for complex traits
The GSR Breeding Strategy: Exploiting
 the maximum genetic diversity in the
      primary gene pool of rice
Objectives
 To exam if there is sufficient (novel) genetic
 variation for target traits in the primary gene
 pool for most complex traits
 If yes, to develop an effective and efficient strategy
 to exploit the genetic diversity for complex traits
  - Integration with the molecular tools
  - Integration with gene discovery
  - Generation of information and training personnel
The Basic Idea
Full Exploitation of The Genetic Diversity in the
          Primary Gene Pool of Rice
The GSR Breeding Strategy
                          Recipients                            Donors
                        (46 best commercial         X     (203 WMCC, 300 CMC +
                    Varieties and hybrid parents)          20 wild rice accessions)
                                                                                Genotyping by re-
                     BC breeding to introgress                                  sequencing of all
                   superior alleles from MCC into                                   parents
                      elite genetic backgrounds

Parents for next                          Trait-specific                   Molecular database
 round of DQP                          IL sets in elite GBs                  of the parents

                                                                    Genotyping &
                                                                    phenotyping


Development of
                                                                 Genetic characterization
 new cultivars                                                            of ILs
                                Breeding by MRS
  and PL sets                       or DQP
                                                                                  -omics and
                                                                                 bioiformatics
  Release to              Theory and
                         technology of
   farmers                breeding by
                                                            Gene networks and metabolic
                        molecular design                   pathways of the target traits and
                                                        functional diversity of alleles at the loci
The Chinese GSR research institutions - China
 National Rice Molecular Breeding Network
Goal

The goal of NCMBN was to develop
superior inbred and hybrid crop
cultivars with significantly improved
yield stability, yield potential, and
grain quality for the major rice
growing areas in China.
Specific objectives
To broaden the genetic base of crop cultivars in major crop
growing areas of China by maximizing the gene flow from the
primary rice gene pool into elite genetic backgrounds through
backcross breeding;
To exploit the hidden diversity of the primary rice gene pool
for improving complex target traits;
To develop IL sets for important traits in elite rice genetic
backgrounds;
To discover and characterize large numbers of QTLs and
QTL networks underlying important traits, and to mine
allelic diversity at important QTLs;
To establish genetic/phenotypic database for the ILs;
To train a new generation of molecular rice breeders in China.
Proof of the Concept
Two Fundamental Questions:

• How much useful genetic diversity,
  particular for the complex
  phenotypes, within the primary
  gene pool of rice?
• Can we combine the process of
  breeding with gene discovery
  (gene/QTL discovery and allelic
  mining)?
Part I: Introgression breeding
        and mass selection

Recurrent Parents
IR64 - Indica, high yield/widely adaptable

New Plant Type - Japonica, high yield potential

Teqing - Indica, high yield/widely adaptable
The “value” added and base broadening
   approach – Introgression breeding


    Widely         Add new genes/traits          IR64 introgression
                   by backcross breeding
  adaptable                                     lines with improved
  high yield                                        target traits
varieties (IR64)


 IR64 lines with improved              Discovery of desirable QTLs using
target traits and the “same”              DNA markers and MAS for
                                              pyramiding QTLs
 yield potential and quality
Procedure of the backcross breeding for development of ILs for
               gene/QTL identification and cultivar development

       RP x donors                 F1s x RP                25 BC1F1s x RP
                                                       X
                                                       Self and bulk
                                                          harvest
                         ~25 BC1F2s x RP                                           ~25 BC2F1s x RP
                                                                                           X    Self and bulk
                                                                                                   harvest
                       Bulk BC1F2 populations                                    Bulk BC2F2 populations

 Screen for
target traits:    1,      2,     3,     4,    5,                  n         1,      2,     3,     4,     5,               n


                  Replicated progeny testing for the introgression lines (ILs) for the selected target traits (tolerances to
                 drought, salinity, submergence, BPH, etc), and for yield performances and other agronomic traits under
                                                          non-stress conditions


                               Large numbers of ILs with selected target traits in the elite RP genetic background

                                                                                           Genotyped w/ DNA markers to track the gene
                                                                                           flow and to identify genes/QTLs for the target
                                                                                                                traits

                        Promising ILs with selected target                       Best ILs with selected target trait(s)
                       trait(s) and good yield performances                            used as parental lines for
                                nominated for NCTs                                 pyramiding genes/QTLs from
                                                                                           different donors

                          Development and releases of
                          green super rice cultivars for                           To rice farmers in the target
                          specific target environments                                    environments
Four Major Groups of Target Traits

  Tolerances to abiotic stresses (drought,
  salinity, low and high temperatures, N
  and P use efficiency, etc.)

  Resistances to biotic stresses (blast,
  sheath blight, false smut, BPH, stem
  borer, BB, etc.)

  Yield potential and heterosis

  Quality parameters
Donor gene pool (203 accessions from
34 countries)
                  Sub-G1 (54)
                   Sub-G2 (6)
                                     I (indica)
                  Sub-G3 (33)
                  Sub-G4 (46)
                Ba-Bao-Mi (Yunnan)
                   Sub-G5(15)
                                     II (japonica)
                  Sub-G6 (12)
                   Sub-G7 (28)
                   Sub-G8 (9)        III (New group)
                Jalmagna (India)
Gene diversity of the donor gene pool
Proportions of accessions Relative genetic diversity of
from different geographic   accessions in different
         regions              geographic regions
Europe America     Others     Europe America Others
15%                          10%
       NA                          NA


                   SA                          SA
    China         33%           China         40%
    22%                         26%
            SEA                         SEA
            30%                         24%
Target traits

• Tolerances to drought, salinity, submergence,
  zinc deficiency, phosphorus deficiency,
  anaerobic germination, etc.

• Resistances to BB, BPH, blast, tungro, sheath
  blight, etc.

• Yield and related traits
• Different maturities
• Different types of grain quality parameters
BC Breeding Procedure
                     RP     X   Donors of diverse origins


                           F1s X        RP


                          BC1F1 X       RP


                                BC2F1
                                    x
                     BC2F2 bulk populations


                    Selection for target traits


Progeny testing           Survival plants                   Genotyping
Major differences of introgression breeding and
             the conventional BC breeding
                          BC breeding       Marker aided    Introgression
                                            BC breeding       breeding
RP selection                 Elite             Same             Same
Donor selection               Yes               Yes          No/diverse

Target traits               Single          Single or few   No limitation
                       monogenic/dominant
Selection                Mass selection        MAS          Mass selection
Progeny testing               yes               yes              yes
Pop. size                    Small             Small           Regular
Selection efficiency         High               High          Depends
Breeding efficiency           Low               Low             High
Selection strategy for target
    traits in random BC
          progenies
Screening of BC2F2 populations for tolerance
     to terminal drought at the reproductive stage
             under the lowland conditions
Stress for 20 days (March 27,2001) Stress for 56 days (May 2, 2001)
Variation in BC2F2 populations for drought
  tolerance under the lowland conditions
Summary of selected drought tolerant BC2F2 plants
          under lowland stress conditions

                         NPT       IR64     Teqing   Total
 Total plants selected    897      2775      489      4161

  # of selected plants     8.5      22.4      6.3     13.2
    per population       (3.8%)   (10.0%)   (3.0%)   (6.8%)
        Range            0 - 85   0 - 100   0 - 30   0 - 100
  No. of populations      113      124       105      320
   No. of I donors        59        67       59       185
   No. of J donors        32        45       19        96

Contributing donor (%)    80.5     90.3      74.3     87.8
The Screening of BC2F2 populations
 under upland drought conditions




                          The RP, IR64
Summary of selected drought tolerant BC2F2 plants
  under lowland stress conditions

                           IR64         Teqing         Total
Total plants selected       192          334           526
 Ave. selected plants    9.6 (4.6%)   10.8 (5.2%)   10.3 (5.0%)
   Per population
       Range               4 - 20       3 - 15        4 - 30
 No. of populations         20            31            51
   No. of I donors          12            23            35

  No. of J donors            8            8             16

Contributing donor (%)      100          100           100
Parental performances and 442 selected DT BC2F2
plants for drought tolerance from 19 BC populations
VG Donors           Origin       IR64 (S) Teqing (M) NPT(SS)
                                                                Total
                                 LL (UL)     LL (UL)   LL
 I   BR24 (S)       Bangladesh    14 (27)     12 (7)   3       29 (34)
 I   STYH (S)       Myanmar       20 (26)       -      4       24 (26)
 I   OM1723 (S)     Vietnam       7 (17)      7 (6)    0       14 (23)
 J   FR13A (SS)     India         15 (16)    17 (15)   0       32 (31)
 J   Type3 (SS)     India         23 (15)    10 (12)   0       33 (27)
 J   Binam (M)      Iran          20 (19)    14 (13)   1       35 (32)
 J   HAN (M)        China         11 (13)       -      3       14 (13)
 I   Zihui100 (S)   China           8           -      9         17
 J   Khazar (S)     Iran            58          -                58
     Total                       176 (133)   60 (53)   22      256 (186)
Screening of BC2F2 populations for salinity
        tolerance at the seedling stage




Young seedlings were subjected to:
                                               IR64   ST CK
6 dSm-1 for 3 days, 12 dSm-1 for 2 weeks, 18
                                                      Pokali
dSm-1 for 1 week, and 24 dSm-1 for 1 week
Confirmation by progeny testing




Seedling screening at EC 24 dSm-1 for 3 weeks
Table 1. Summary results of BC populations for screening salinity tolerance


Details                                   BC2F2 screening                     BC2F3 reconfirmation
                                 IR64     Teqing    NPT      Total   IR64        Teqing    NPT       Total

Total BC2F2 populations           62        58       55      175      24           34       10        68
Single plant selections per BC
population                       4 - 12    4 - 13   1 - 14           0 - 43      0 - 49   0 - 11
Total selected BC2F3 lines        490       428      374     1292     448         392       21       861
Selection intensity (%)          3.95      3.69     3.40     3.69
Number of indica donors           47        47       42      136      20           27       7         54
Selected lines                    369       345      289     1003     372         269       21       662
Selection intensity (%)          4.39      3.67     3.44     3.69
Number of japonica donors          9         9        7       25       3           6        1         10
Selected lines                    70        66       44      180      43          123       0        166
Selection intensity (%)          3.89      3.67     3.14     3.60
Number of intermediate donors      4         1        3       8        1           1        2         4
Selected lines                    35         5       19       59      33           0        0         33
Selection intensity (%)          4.38      2.50     3.16     3.69
Summary results of BC populations
             for screening salinity tolerance
                                    BC2F2 screening                 BC2F3 reconfirmation

                           IR64      Teqing   NPT      Total    IR64     Teqing   NPT      Total

# of BC2F2 pop.             62         58      55       175      24       34       10       68

# of selections per pop.   4 - 12    4 - 13   1 - 14   1 - 14   0 - 43   0 - 49   0 - 11

Total selected lines        490       428      374     1292      448      392      21      861

# of contributing
donors                      60         57      52       169

SI (%)                     3.95       3.69    3.40     3.69

 For individual BC populations of 200 plants, a difference of 4% between two populations in
 selection intensity (survival rate) is statistically significant at P < 0.05 when the selection
 intensity is between 0.1 and 0.5.
Screening of BC2F2 populations for
  submergence tolerance in a deep-water pond




Thirty-five-day old seedlings were submerged under deep water
for two weeks, then allowed to recover
Summary results of BC populations
          for screening anaerobic germination
               and submergence tolerance
                              Anaerobic germination                  Submergence

                           IR64   Teqing   NPT      Total   IR64     Teqing    NPT Total

# of BC pop.                47     47       36      130      60        57       62      179

# of selections per pop.   0-5     0-6     0 - 14           0 - 15    3 - 13   0 - 12

Total selected lines        47     81       215     343      652       483      530     1665

# of contributing
donors                      46     46       35      127      59        57       60      176

SI (%)                     0.52    0.93    3.11     1.32    1.08      0.85     0.85     0.93
Screening of BC2F2 populations for
           anaerobic germination




Direct seeding then submerged under 10 cm-deep water
Summary results of BC populations screened for anaerobic
germination and submergence tolerance

 Details                                 Anaerobic germination                    Submergence
                                  IR64      Teqing    NPT        Total   IR64     Teqing   NPT      Total
 Total BC2F2 populations           47         47       36        130      60       57       62      179
 Single plant selections per BC
 population                       0-5        0-6     0 - 14              0 - 15   3 - 13   0 - 12
 Total selected BC2F3 lines        47         81      215        343      652      483      530     1665
 Selection intensity (%)          0.52       0.93     3.11       1.32    1.08      0.85    0.85     0.93
 Number of indica donors           37         38       29        104      47       50       50      147
 Selected lines                    31         67       77        175      538      431      451     1411
 Selection intensity (%)          0.43       0.90     3.10       0.84    1.15      0.86    0.90     0.96
 Number of japonica donors         6          8        5          19       8        7        7       22
 Selected lines                    11         14       31         56      83       52       63      198
 Selection intensity (%)          0.91       1.03     3.10       1.47    1.00      0.74    0.90     0.90
 Number of intermediate donors     3          0        1          4        4        0        3       7
 Selected lines                    5          0        7          12      40        0       16       56
 Selection intensity (%)          0.83       0.00     3.50       1.50    1.04      0.00    0.53     0.80
Screening for BPH Resistance




       IR64
Summary results of BC populations screened for low temperature
germination and brown planthopper resistance

Details                                  Low temperature germination              Brown planthopper resistance
                                   IR64         TQ       NPT      Total     IR64         TQ       NPT        Total
Total BC2F2 populations             14          15        10           39    64          67        62        193
Single plant selections per BC
population                         0 - 21      0 - 18   0 - 31              0 - 22      0 - 22     0-2
Total selected BC2F3 lines          79          77        51       207       652         255        2        909
Selection intensity (%)             5.64       5.13      5.10      5.31     10.19       3.81       0.03      4.71
Number of indica donors              1           1        0            2     49          60        49        158
Selected lines                       0           3        0            3     565         221        2        788
Selection intensity (%)             0.00        3.0      0.00      1.5      11.53       3.68       0.04      4.99
Number of japonica donors            9          11        7            27     9          11         7            27
Selected lines                      55          52        39       146       54          21         0            75
Selection intensity (%)             6.11       4.73      5.57      5.41     6.00        1.91       0.00      2.78
Number of intermediate donors        4           3        3            10     4           3         3            10
Selected lines                      24          22        12           58    33           2         0            35
Selection intensity (%)             6.00       7.33      4.00      5.80     8.25        0.67       0.00      3.50

 For individual BC populations of 100 plants, a difference of 2.5% between two populations in selection
 intensity (survival rate) is statistically significant at P < 0.05 when the selection intensity is < 0.1.
Summary results of BC populations for screening
  low temperature germination and BPH resistance

                           Low temperature germination                  BHP resistance

                           IR64     Teqing   NPT      Total    IR64      Teqing   NPT     Total

# of BC2F2 pop.             14       15       10       39        64        74      62      200

# of selections per pop.   0 - 21   0 - 18   0 - 31            0 - 22    0 - 22   0-2

Total selected lines        79       77       51      207       652       255       2      909

# of contributing
donors                      14       14       10       38        62        67      59      189

SI (%)                     5.64      5.13    5.10     5.31     10.19      3.81    0.03     4.71

 For individual BC populations of 200 plants, a difference of 4% between two populations in
 selection intensity (survival rate) is statistically significant at P < 0.05 when the selection
 intensity is between 0.1 and 0.5.
Screening for tolerance to phosphorus
                deficiency
Tested in natural conditions in Pangil, Laguna.
Screening for tolerance to zinc deficiency
Summary results of BC populations screened for zinc deficiency tolerance

                                                                               BC2F4 progeny testing under very severe
Details                          BC2F2 screening under moderate field stress                field stress
                                  IR64       Teqing       NPT        Total     IR64      Teqing      NPT         Total
Total number of BC populations     51          42          36         129       11         21          10         42
Single plant selections per BC
population                       4 - 12      2 - 12      0 - 12      0 - 12    0 - 10    0 - 15        0
Total selected BC2F3 lines        454         383         324        1161       72        255          0          327
Selection intensity (%)           7.42        7.60        7.50        7.50     2.14       2.41        0.00       2.01
Number of indica donors            40          33          25          98        9         17          9          35
Selected lines                    370         320         237         927       63        200          0          263
Selection intensity (%)           7.77        8.08        7.90        7.88     2.07       2.31        0.00       1.91
Number of japonica donors           7          6           7           20        1          4          1           6
Selected lines                     63          63          84         210        0         55          0          55
Selection intensity (%)           7.50        8.75        7.86        8.75     0.00       2.86        0.00       2.29
Number of intermediate donors       3           -          1           4         1          0          0           1
Selected lines                     21           -          9           30        9          0          0           9
Selection intensity (%)           5.83          -         7.50        6.25     5.63       0.00        0.00       5.63
Selection for changed grain
       type of Teqing
Selection for high
       yield
High tiller number
Screening for blast resistance
Low N stress
experiment
under rain-off
shelter
Improving japonica for cold
tolerance at the reproductive stage
Screening results of 11 CY1 (recurrent parent) BC2F4 backcross populations for cold
tolerance at the booting stage in 2008 (1 SI = selection intensity, SF = spikelet
fertility; 2 Different letters indicate statistical significance at P < 0.05, based on the
Duncan testing)

                                                   No. of                       SF (%)
                                     Population              SI 1
 Donor parent    Subspecies   Code                selected
                                        size
                                                   plants
                                                             (%)     Mean ± SD2           Range

    Bg90-2         Indica      A        450         41       9.1      63.3±8.4D          50.3-86.5
     X21           Indica      B        450         29       6.4     64.3±9.9CD          50.6-87.1
     X22           Indica      C        450         28       6.2    65.6±10.1BCD         50.7-87.3
      Q5           Indica      D        450         31       6.9    71.1±11.1ABC         50.9-91.4
  Chhomrong       Japonica     E        450         24       5.3        75.6±            51.4-87.8
    Doddi          Indica      F        450         25       5.6    71.0±10.0ABC         50.2-90.0
  Fengaizhan       Indica      G        450         44       9.8        74.1±            52.2-98.5
 Shennong265      Japonica     H        450         21       4.7    69.2±11.4ABCD        50.7-93.8
   Yuanjing7      Japonica     I        450         41       9.1     71.1±9.8ABC         50.0-90.1
    OM997          Indica      J        450         21       4.7     72.1±9.1AB          54.9-89.6
     Cs94          Indica      K        450         19       4.2     64.2±8.6CD          51.4-86.0
     Mean                               450        29.5      6.5       69.2±4.2
     CY1          Japonica                          324               24.8±4.3E          19.0-30.0
Screening of the BC2F4 bulk populations for cold tolerance
                    (CT) at the booting stage:

Seeds of the BC2F4 bulk populations were sown in the seedling nursery on April
15, 2008, and 450 40-day old seedlings of each BC2F4 bulk population were
transplanted into a 45-row plot with 10 plants in each row and a spacing was
25×15cm in the sheltered water pond of JAAS on May 25. Two rows of CY1
were also transplanted on both sides of each plot as the checks. The water-pond
were irrigated with water of normal temperature (25~28℃). When CY1 entered
the stage of panicle initiation, the LT treatment was initiated by irrigation of
flowing cold water (19±0.5℃), which was adjusted in a nearby water pool by
                              ℃
mixing cold underground water (9℃) with the river water. The depth of the
                                       ℃
cold water in the pond was 20cm and the treatment was maintained for ~30
days until panicles of almost all plants exerted completely. Then, irrigation
with normal temperature water was resumed until the maturity. At the
maturity, all plants except those with >3 days earlier heading or those with
>3 days delayed heading than CY1 were harvested for measuring the
spikelet fertility. Under this LT treatment, CY1 had a spikelet fertility of
     ±
24.8±4.3%, then, any plant with spikelet fertility >50% were selected.
Evaluation of 324 BC2F6 introgression lines and their recurrent parent, CY1 for cold
 tolerance of at the seedling and booting stages in 2009 (SNP = spikelet number per panicle,
 FGN = filled grain number per panicle, SF = spikelet fertility)

                          Cold stress at the seedling
                No. of              stage                                  Cold stress at the reproductive stage
Donor parent   selected   Survival rate of seedlings
                 lines               (%)                        SNP                          FGN                       SF (%)
                            Mean            Range       Mean            Range         Mean           Range         Mean      Range

  BG90-2         41          19.5          10.0~62.5    114.5         76.3~164.2      55.0          22.1~95.7      48.0     18.5~71.3

    X21          29          19.4          11.1~40.3    99.7          73.3~125.0      50.5          29.5~90.3      50.3     30.9~74.9

    X22          28          19.6          12.5~40.7    113.2         70.3~139.0      58.5          4.5~94.9       51.4      5.6~79.4

    Q5           31          18.3          15.0~33.8    114.6         68.3~185.4      56.5       15.3~113.6        49.2     14.6~83.6

Chhomromg        24          17.0          15.0~32.5    109.8         77.6~175.2      74.0       47.0~104.8        66.9     45.6~83.4

   Doddi         25          17.3          12.5~25.0    112.9         89.4~182.6      73.9       51.5~108.5        65.4     48.7~80.6

 Fengaizhan      44          16.7          12.5~25.0    96.5          70.2~115.1      67.1          14.3~96.0      70.1     12.7~86.1

Shennong265      21          15.6          0.0~23.3     96.9          65.0~144.8      62.0          36.9~99.2      63.0     40.0~79.9

 Yuanjing7       41          18.5          15.0~45.0    116.0         93.5~151.0      80.6       30.4~106.4        69.2     25.5~84.2

  OM997          21          37.1          15.0~82.4    81.6          56.2~108.1      42.0          14.9~64.0      51.1     23.3~72.0

   Cs94          19          22.3          15.0~42.5    113.3         82.0~137.9      69.5          31.8~99.4      60.0     34.2~75.5

   CY1                       14.8          10.8~20.0    106.6         98.6~114.6      36.7          32.7~40.8      35.1      33.7~36.5
  LSD0.05                            4.5                        8.3                           8.2                         6.2
Comparison of selection efficiencies of 11 CY1 BC2F6 populations for cold
                      tolerances at the seedling and reproductive stages

                                                                            The reproductive stage                                    The seedling stage
           Population
                                            N1                FGN                        SNP                      SF (%)                        SR (%)
    Code             Donor                               N1            N2           N1            N2           N1           N2             N1               N2
     A             BG90-2                   41           18             0           13             6           17            1              5                0
     B                X21                   29            7             0            0             3           15            0              4                0
     C                X22                   28           17             2            8             1           17            3              5                0
     D                 Q5                   31           15             2           10             4           20            3              1                0
     E          Chhomromg                   24           17             0            7             3           22            0              2                0
     F               Doddi                  25           23             0            1             0           24            0              0                0
     G           Fengaizhan                 44           35             1            0             7           42            1              0                0
     H         Shennong265                  21           12             0            2             9           18            0              0                1
     I            Yuanjing7                 41           40             0            9             0           40            0              3                0
     J              OM997                   21            2             1            0            13           13            0             14                0
     K                Cs94                  19           14             0            5             1           14            0              6                0
                      Total                324          200             6           55            47          242            8             40                1
1N is the total number of selections based on single plant spikelet fertility (SF) in BC2F4 populations from Table 1, N1 and N2 are the numbers of the BC2F6 lines
showing significantly higher or significantly lower than CY1 for the measured traits. FGN, SNP, SF and SR are filled grain number per panicle, spikelet number
per panicle, spikelet fertility and survival rate of seedlings.
Mean performances of 116 ILs of five populations for 11 traits evaluated under
                normal and cold water stress conditions in 2010
       Trait 1            Chhomrong         Doddi      Fengaizhan   Shennong265   Yuanjing7    Mean    CK
         N                       24          24           24            20           24        116
Under the normal conditions
      PH (cm)                      ±
                              135.2±4.4        ±
                                          130.6±5.8         ±
                                                       125.7±5.4          ±
                                                                     127.2±3.7         ±
                                                                                  132.3±4.5    130.3   117.1
    BM (g/plant)               37.1±3.3    36.5±3.4     38.1±3.5      33.5±3.3     35.1±2.4    36.2    34.7
        FGN               143.4±15.0      143.7±17.2   148.1±12.3   135.1±15.6         ±
                                                                                  156.3±19.4   145.7   138.9

        SNP               176.6±20.8      179.4±20.9   171.2±17.4   172.9±17.7         ±
                                                                                  197.2±21.8   179.7   168.6
       SF (%)                 81.6±6.6     80.4±8.1         ±
                                                        86.9±5.8     78.4±7.2      79.2±5.3    81.4    82.4
   GY (g/plant)                   ±
                              19.4±2.94    20.5-3.0         ±
                                                        20.9±2.5         ±
                                                                     18.7±2.5          ±
                                                                                   18.7±2.7    19.6    19.7
       HI (%)                 55.0±5.2     59.1±4.9     57.9±3.7     59.1±4.4      56.0±4.8    57.3    60.3
         PN                    5.4±0.6     5.7±0.4      5.7±0.6       5.6±0.7         ±
                                                                                   4.8±0.5      5.5     5.7
      GW (g)                  24.3±1.5        ±
                                          26.3±1.6          ±
                                                        22.0±2.1     24.5±1.1          ±
                                                                                   26.3±1.4    24.7    24.9
       HD (d)                 113.9±3.3   113.0±4.1    116.3±6.0     112.5±4.7    113.6±3.9    113.9   111.1
Under the cold water stress
      PH (cm)                      ±
                              118.1±7.9        ±
                                          119.9±7.7         ±
                                                       119.8±7.7          ±
                                                                     122.4±6.6         ±
                                                                                  125.2±5.7    121.0   107.8
    BM (g/plant)               16.2±3.2    16.1±2.7     16.8±2.5     15.3±3.6      17.5±3.6    16.4    12.0
        FGN                   46.8±28.4   39.1±20.2        ±
                                                       63.8±14.3         ±
                                                                     22.9±14.1    46.4±19.8    44.5    12.8

        SNP               118.9±23.9      121.7±13.8   110.4±12.9   117.7±24.1    139.6±19.6   121.8   121.5

       SF (%)                 38.8±18.3   32.3±14.5        ±
                                                       57.3±10.3         ±
                                                                     19.7±12.1    33.3±14.6    36.9    10.5
   GY (g/plant)                   ±
                               3.5±2.2        ±
                                           3.0±1.7         ±
                                                        5.0±1.3          ±
                                                                      1.9±1.2          ±
                                                                                   3.6±1.9      3.5     1.0
       HI (%)                 33.3±7.6     30.7±6.7         ±
                                                        40.9±4.4         ±
                                                                     25.8±7.2      30.8±5.1    32.5    25.1
         PN                    3.3±0.5     3.4±0.6      3.3±0.5       3.7±0.9      3.2±0.5      3.4     3.5
      GW (g)                  17.9±1.5     18.0±1.6     17.5±2.7     17.1±0.9      17.9±1.3    17.7    16.4
       HD (d)                 131.3±2.2   128.2±3.3    132.0±4.1     126.9±4.1    128.6±3.0    129.5   125.2
The numbers of ILs from the 5 populations that deviated significantly CY1 for 11
measured traits evaluated under cold water stress (S) and normal (N) conditions in 2010
                                                            GY            GW            HD             HI         PH                         SF
                              BM (g/plant)     FGN                                                                              PN                   SNP
     Donor      N1    Treat                              (g/plant)        (g)           (d)           (%)        (cm)                        (%)

                               N1     N2     N1    N2    N1    N2    N1     N2      N1    N2      N1    N2      N1    N2    N1   N2      N1    N2   N1   N2
Chhomrong       24             18      0      18    0     14    0     15        0   24        0   14        0   19      2   3        8   21    0    6    12
     Doddi      24             18      0      19    0     15    0     16        1   16        2   14        2   21      0   5        7   19    0    4    4

Fengaizhan      24     S       22      0      24    0     23    0     6         3   22        1   24        0   20      0   2        6   24    0    0    12

Shennong26
    5      20                  12      1      7     0     6     0     5         2   10        8   7         6   18      0   9        5   10    0    6    8
    Yuanjing7   24             21      0      21    0     19    0     17        0   16        0   15        1   24      0   3        8   20    0    12   1


      Total     116            91      1      89    0     77    0     59        6   88    11      74        9   102     2   22   34      94    0    28   38
Chhomrong       24             15      2      11    5     9     8     4     13      13        2   1     16      24      0   3        9   7     9    11   4
     Doddi      24             11      4      10    5     9     5     17        4   11        3   6         7   24      0   4        4   8     12   13   5
Fengaizhan      24             16      0      14    2     9     1     2     20      19        3   3     15      21      0   5        7   15    4    8    8
                       N
Shennong26
    5      20                  3       8      5     7     2     9     5     10      8         7   5         7   20      0   8    10      5     13   9    7
    Yuanjing7   24             5       3      17    2     5     14    18        2   11        3   3     18      24      0   0    21      4     12   20   1


      Total     116            50     17      57    21    34    37    46    49      62    18      18    63      113     0   20   51      39    50   61   25

1N is the total number of ILs with CT selected from each population; N1 and N2 are the numbers of the ILs showing significantly higher and lower trait
values than CY1.
GY = grain yield, BM = biomass, PH = plant height, PL = panicle length, PN = panicle number per plant, FGN = filled grain number per panicle, SNP =
spikelet number per panicle, SF = spikelet fertility, HD = heading date, GW = 1000-grain weight, HI = harvest index.
Mean performances of 19 promising ILs under cold water stress and
                 normal conditions in 2009 and 2010 (Meng et al. 2012)

 Line#    Pop.1                   2010 under the normal condition                                    2010 under cold water stress                     2009 under stress
                   PH     BM     SNP    SF       GY      HI     PN   GW     HD       PH     BM     SNP     SF    GY      HI      PN   GW     HD      SNP     SF      SR

CK(CY1)           117.1   34.7   168.6   82.4   19.7   60.3    5.7   24.9   111.1   107.8   12.0   121.5   10.5   1.0   25.1    3.5   16.4   125.2   106.6   35.1   14.8
LW213      G      134.2   48.5   192.3   91.1   28.1   60.1    6.4   22.8   103.0   129.2   19.5   131.2   69.1   7.2   46.8    3.5   15.7   128.0   101.7   81.2   12.5
LW164      F      126.5   45.7   204.5   87.8   28.5   65.2    6.4   23.5   117.0   118.8   17.2   120.0   27.3   2.5   26.0    3.3   16.1   129.0   113.2   56.9   15.0
LW180      F      131.7   38.8   182.0   89.8   23.4   63.1    5.7   25.6   101.0   131.7   18.0   138.0   39.6   4.9   36.3    3.3   19.1   123.0   107.3   68.0   17.5
LW157      F      133.4   41.0   218.7   71.9   23.4   59.9    6.0   25.2   116.0   114.8   20.9   132.8   40.1   5.0   33.8    4.3   17.1   131.0   110.6   60.3   17.5
LW170      F      129.3   40.0   183.3   83.7   22.8   59.7    5.9   24.0   117.0   117.8   19.6   119.5   45.5   5.3   38.4    4.8   16.7   131.0   118.4   71.9   17.5
LW214      G      122.0   38.0   159.8   85.6   22.9   63.5    6.7   17.9   120.0   108.5   14.3   103.7   54.9   4.5   44.4    3.3   16.3   137.0   86.7    84.5   15.0
LW188      G      121.0   40.1   164.3   84.2   23.1   60.6    6.7   21.0   121.0   130.0   16.6   119.7   66.9   5.8   46.2    3.2   17.0   138.0   90.9    82.5   15.0
LW142      E      136.7   39.3   162.7   79.2   21.5   57.7    6.7   23.9   112.0   121.5   16.7   119.8   61.6   5.2   43.1    3.2   18.1   134.0   91.2    60.0   15.0
LW154      E      137.8   39.9   198.3   82.9   21.9   58.0    5.4   23.4   112.0   128.5   19.7   160.7   67.4   7.4   47.7    3.0   17.4   130.0   121.6   71.3   15.0
LW174      F      129.5   37.6   151.3   86.6   21.7   61.0    6.7   25.9   115.0   120.3   17.7   122.0   48.2   5.3   41.4    4.2   17.8   128.0   126.2   80.3   25.0
LW207      G      132.6   40.0   192.8   79.9   22.1   58.0    5.8   21.2   121.0   127.0   20.1   118.3   66.6   6.7   43.1    3.8   17.5   132.0   115.1   83.7   20.0

LW264       I     138.8   40.7   221.5   77.7   22.6   58.0    5.3   24.9   107.0   130.0   17.2   169.2   26.3   3.1   29.1    2.8   17.9   125.0   120.3   79.1   42.5
LW250       I     130.7   39.2   226.8   79.9   23.3   62.6    5.2   26.0   110.0   136.8   20.1   136.8   65.1   5.8   38.4    2.7   18.0   132.0   108.4   76.1   20.0
LW156      E      138.9   40.3   187.8   78.9   21.0   55.0    5.7   23.0   114.0   127.0   20.2   129.7   57.9   6.8   43.0    4.0   16.3   129.0   131.8   74.0   15.0

LW151      E      131.9   37.7   168.3   82.9   19.2   53.9    5.6   24.2   114.0   122.3   19.3   123.5   67.2   6.6   43.3    3.3   16.6   130.0   122.4   82.9   16.3
LW189      G      132.4   37.9   198.3   84.4   20.1   55.9    4.8   21.7   120.0   133.2   19.0   114.7   59.5   6.2   41.1    3.5   17.0   137.0   103.4   75.7   20.0

LW216      G      129.1   37.3   170.8   87.1   20.1   56.9    5.4   20.7   123.0   111.8   17.1   99.8    51.7   4.3   34.4    2.8   15.9   139.0   99.9    86.1   16.9

LW200      G      127.7   38.6   167.8   88.8   20.5   56.1    5.6   20.3   115.0   122.0   16.4   100.2   66.2   4.9   41.9    3.3   16.5   130.0   93.7    84.2   17.5

LW291       I     121.1   33.1   181.5   84.8   17.5   56.5    4.7   25.1   120.0   126.3   24.9   156.3   39.1   6.6   32.2    4.2   17.7   131.0   143.6   74.5   45.0
LSD0.05            1.9     2.1    8.2     2.6   1.6    1.8     0.3   0.5     1.9     3.7    1.8    11.1    8.1    1.2   4.3     0.4   1.1     1.6     8.2    6.2     4.5
The hidden diversity for highly
heritable traits – BLB resistance
Reactions (lesion lengths) of HHZ and two donors to 14
  tropical races of BLB caused by Xanthomonas oryzae pv oryzae


          P1    P2     P3b    P3c    P4     P5     P6     P7    P8    P9a    P10   P9c    P9b    P9d


          PXO   PXO    PXO    PXO    PXO    PXO    PXO    PXO   PXO   PXO    PXO   PXO    PXO    PXO    aver
           61    86     79    340     71     112    99    145   280   339    341   347    349    363    age


HHZ       9.8   21.2   13.1   25.7   10.4   2.4    29.6   5.0   8.6   28.8   8.4   26.6   15.2   24.6   16.4


PSBRC66   6.4   18.5   16.4   21.4   11.6   0.7    13.0   2.6   8.8   4.1    7.0   12.0   3.4    17.3   10.2


PSBRC28   2.8   20.3   21.6   24.0   11.2   3.9    22.4   4.7   9.2   26.0   8.5   23.7   22.9   21.9   15.9
8个黄华占群体对15个Xoo菌株产生反应类型
                              (2012年8月,北京)




F1-F5
F1-     F6-
        F6-F10   F11-
                 F11-F15   F1-
                           F1-F5   F6-
                                   F6-F10   F11-
                                            F11-F15   F1-F5
                                                      F1-     F6-
                                                              F6-F10   F11-F15
                                                                       F11-



对部分小种抗病的株系                 对全部15个小种高抗的株系              对全部15个小种感病的株系
Blast evaluation of virulent strains   Evaluation of BB resistance of >500
                                       lines (HHZ background) against 14
                                        strains of 10 Xoo races, 2010 WS




                                                           HHZ is
                                                        susceptible to
                                                         most tropic
                                                         BLB races


    Vera Cruz et al
Ten HHZ ILs with broad spectrum resistance to all 14 races
      of bacterial blight pathogen, Xanthomonas oryzae pv oryzae

                                      PX            PX           PX    PX    PX     PX    PX     PX     PX
                  PX    PX     PX     O34    PX     O11   PX     O14   O28   O33    O34   O34    O34    O36    aver
                  O61   O86    O79     0     O71     2    O99     5     0     9      1     7      9      3     age

HHZ               9.8   21.2   13.1   25.7   10.4   2.4   29.6   5.0   8.6   28.8   8.4   26.6   15.2   24.6   16.4

PSBRC66           6.4   18.5   16.4   21.4   11.6   0.7   13.0   2.6   8.8   4.1    7.0   12.0   3.4    17.3   10.2
HHZ15-SAL13-Y2    0.2   0.2    0.2    0.2    0.2    0.2   0.2    0.2   0.2   0.2    0.2   0.2    0.2    0.2    0.2

HHZ15-SAL-13-Y3   0.2   0.2    0.2    0.2    0.2    0.2   0.2    0.2   0.2   0.2    0.2   0.2    0.2    0.2    0.2

HHZ15-DT7-SAL1    0.2   0.2    0.2    0.2    0.2    0.2   0.2    0.2   0.2   0.2    0.2   0.2    0.2    0.2    0.2

HHZ15-DT7-SAL3    0.2   0.2    0.2    0.2    0.2    0.2   0.2    0.2   0.2   0.2    0.2   0.2    0.2    0.2    0.2

HHZ15-DT7-SAL6    0.2   0.2    0.2    0.2    9.3    0.2   0.2    0.2   0.2   0.2    0.2   0.2    0.2    0.2    0.9

PSBRC28           2.8   20.3   21.6   24.0   11.2   3.9   22.4   4.7   9.2   26.0   8.5   23.7   22.9   21.9   15.9

HHZ19-SAL-14-Y3   0.2   0.2    0.2    0.2    0.2    0.2   0.2    0.2   0.2   0.2    0.2   0.2    0.2    0.2    0.2

HHZ19-DT8-SAL2    0.2   0.2    0.2    0.2    0.2    0.2   0.2    0.2   0.2   0.2    0.2   0.2    0.2    0.2    0.2
HHZ19-SAL12-
SAL4              0.2   0.7    0.9    0.5    0.2    0.2   0.2    0.2   0.4   0.2    0.3   0.9    0.2    0.2    0.4
HHZ19-SAL14-
SAL4              0.2   1.0    0.6    0.3    0.2    0.2   0.2    0.2   0.3   0.2    0.7   0.2    0.2    0.2    0.3
HHZ19-SAL15-
SAL2              0.2   3.8    2.2    0.6    0.4    0.4   0.8    0.5   0.3   0.3    0.3   0.6    0.4    0.4    0.8
Reactions of 512 HHZ ILs to 14 tropical Xoo races

            Race           R% (LL <3.0 cm)
             P1                 76.4
             P2                  4.7
             P3b                 4.9
             P3c                 4.9
             P4                 22.4
             P5                 78.4
             P6                  5.1
             P7                 46.6
             P8                 31.0
             P9a                12.3
             P10                12.1
             P9c                 4.7
             P9b                 5.1
             P9d                50.8
Classification of 14 Xoo races into two major groups (virulent
     and less virulent ones) based on the reactions of the 512 HHZ ILs

                 P1
                 P5
                                                                       Weak virulence group
                 P4
                 P7
Name Xoo races




                 P8
                 P10
                 P2
                 P9c
                 P6
                 P3c
                                                                        High virulence group
                 P3b
                 P9a
                 P9b

                  0.0   0.1   0.2      0.3   0.4    0.5    0.6   0.7     0.8   0.9   1.0   1.1   1.2   1.3
                                    Average Distance Between Clusters
Classification of the 512 ILs based on their resistances to the 14 Xoo Phillipines races
                H9-39
                H5-1
                H8-27
                H9-20
                H17-65
                H9-3
                H9-13
                H5-32
                H8-44
                H15-12
                H17-29
                H5-49
                H15-1
                H8-8
                H8-23
                H9-7
                H8-13
                H8-51
                H9-16
                H9-26
                H5-12
                H5-13
                H8-3
                H19-48
                H17-11
                H12-27
                H8-33
                H5-5
                H15-26
                H9-24
                H5-51
                H8-21
                H8-31
                H5-59
                H8-47
                H9-14
                H9-25
                H5-6
                H8-41
                H9-18
                H8-40
                H8-26
                H12-17
                H19-18
                H9-56
                H19-22
                H19-23
                H5-26
                H17-32
                H11-10
                H12-26
                H9-40
                H8-55
                H12-16
                H11-32
                H11-30
                H11-36
                H11-34
                H11-45
                H11-49
                H12-39
                H19-45
                H12-2
                H11-28
                H9-70
                H15-43
                H17-35
                H19-55
                H19-13
                H11-31
                H19-15
                H12-24
                H9-41
                H12-20
                H19-46
                H12-18
                H19-29
                H9-51
                H11-16
                H19-14
                H17-34
                H9-9
                H8-38
                H11-50
                H12-40
                H12-61
                H12-55
                H17-5
                H15-19
                H12-64
                H15-21
                H19-57
                H19-50
                H9-66
                H19-59
                H19-36
                H15-8
                H15-25
                H12-54
                H15-24
                H19-56
                H17-53
                H9-4
                H15-30
                H5-55
                H8-18
                H15-11
                H12-31
                H5-60
Name of lines




                H8-54
                H11-54
                H12-36
                H19-35
                H11-11
                H11-55
                H12-48
                H11-24
                H12-9
                H12-4
                H12-7
                H12-15
                H19-9
                H8-45
                H11-27
                H9-63
                H12-12
                H12-33
                H12-42
                H11-33
                H17-54
                H9-37
                H11-21
                H9-6
                H11-25
                H19-7
                H12-13
                H19-4
                H8-10
                H8-42
                H12-28
                H12-65
                H15-31
                H17-1
                H8-52
                H12-5
                H5-75
                H11-17
                H11-19
                H12-29
                H5-11
                H5-74
                H5-18
                H5-21
                H17-38
                H8-50
                H9-15
                H8-46
                H12-45
                H17-64
                H5-29
                H19-1
                H8-12
                H11-5
                H12-49
                H19-47
                H8-7
                H12-14
                H19-16
                H5-33
                H8-53
                H9-19
                H9-21
                H5-54
                H17-63
                H8-15
                H8-35
                H5-41
                H11-37
                H11-40
                H9-22
                H11-38
                H17-9
                H17-19
                H17-14
                H17-24
                H17-13
                H17-21
                H17-49
                H17-52
                H17-39
                H17-48
                H17-62
                H9-38
                H17-57
                H17-28
                H17-45
                H15-44
                H5-3
                H5-23
                H9-12
                H8-28
                H5-36
                H5-35
                H8-39
                H15-6
                H15-9
                H5-61
                H17-60
                H5-66
                H5-67
                H17-68
                H17-66
                H5-48
                H5-53
                H15-33
                H17-36
                H5-28
                H5-52
                H8-22
                H5-42
                H9-17
                H5-10
                H8-30
                H5-63
                H5-72
                H11-23
                H9-32
                H5-15
                H17-40
                H5-39
                H15-32
                H17-18
                H5-43
                H8-6
                H5-65
                H8-34
                H8-36
                H12-58
                H19-5
                H15-13
                H9-23
                H15-35
                H19-19
                H15-40
                H15-42
                H15-38
                H19-58
                H19-62

                    0.0       0.2        0.4          0.6          0.8             1.0   1.2     1.4         1.6
                                               Average Distance Between Clusters
Are the parental performances
correlated with the performances
     of their BC progenies?
Genetic background effects in
   introgression breeding
Submergence Tolerance
    Number of submergence tolerant plants in 9 BC2F2
    and 3 BC3F2 populations under the field condition
VG Donors          Origin   TKM9 (SS) Khazar (SS) FR13A (T)   Total
                               (I)        (I)        (J)
I    IR64 (S)      India       12          8         14        34

I    Teqing (SS)   India       10          9          8        27
     NPT (SS)
J                  Iran         6          6          6        18
      BC2F2
     NPT (SS)
J                  Iran        10         71          2        83
      BC3F2
      Total                    38         94         30       162
Yield responses of the 193 parental lines of IRMBP
     to the terminal drought under the lowland condition

25
                                  ±
                             -19.1±44.0%
20


15


10


5


0
 -100 -80 -60 -40 -20   0   20   40   60   80 100 120 140 160

                  Yield reduction (in %)
Summary of selected drought tolerant BC2F2 plants
         under lowland stress conditions

                        NPT       IR64     Teqing   Total
Total plants selected    897      2775      489      4161

 # of selected plants     8.5      22.4      6.3     13.2
   per population       (3.8%)   (10.0%)   (3.0%)   (6.8%)
       Range            0 - 85   0 - 100   0 - 30   0 - 100
 No. of populations      113      124       105      320
  No. of I donors        59        67       59       185
  No. of J donors        32        45       19        96

Contributing donors (%) 80.5      90.3      74.3     87.8
Parental performance and 442 selected DT BC2F2
plants for drought tolerance from 19 BC populations
VG Donors           Origin       IR64 (S) Teqing (M) NPT(SS)
                                                                Total
                                 LL (UL)     LL (UL)   LL
 I   BR24 (MR)      Bangladesh    14 (27)     12 (7)   3       29 (34)
 I   STYH (S)       Myanmar       20 (26)       -      4       24 (26)
 I   OM1723 (S)     Vietnam       7 (17)      7 (6)    0       14 (23)
 J   FR13A (SS)     India         15 (16)    17 (15)   0       32 (31)
 J   Type3 (S)      India         23 (15)    10 (12)   0       33 (27)
 J   Binam (M)      Iran          20 (19)    14 (13)   1       35 (32)
 J   HAN (M)        China         11 (13)       -      3       14 (13)
 I   Zihui100 (S)   China           8           -      9         17
 J   Khazar (MR)    Iran            58          -                58
      Total                      176 (133)   60 (53)   22      256 (186)
Comparison of different IR64 and NPT BC generations
in screening for anaerobic germination (%)

                                        BC2F2 bulks           BC3F2 bulks            BC4F2 bulks
                                      IR64      NPT     IR64         NPT       IR64           NPT
 Total number of populations            9        10       -           10         9             10
 Surviving plants/population         0 – 28    0 - 50     -         33 - 78   17 - 78       97 - 162
 Number of indica donors                7        8        -           8          7             8
 Selected lines (indica)               36       158       -          452        296           1038
 Selection intensity (%)               5.1      19.8      -          28.3       21.1          64.9
 Number of japonica donors              2        2        -           2          2             2
 Selected lines (japonica)              2        8        -           35        44            121
 Selection intensity (%)               2.0       8.0      -          17.5       22.0          60.5
 Mean selection intensity (%)          4.4      13.9                 26.1       21.3          62.7

BC2F2, BC3F2 and BC4F2 bulks all had 200 seeds in 2 replications. For individual BC populations of
200 plants, a difference of 4% between two populations in selection intensity (survival rate) is
statistically significant at P < 0.05 when the selection intensity is between 0.1 and 0.5.
Screening for seedling cold tolerance




Twelve-day old seedlings were subjected to cold temperature for
18 days at the mean daily temperature of 11.8 Co, including 3-day
of low temperature at 8 Co between April 24-26 (LAAS, 2002).
Seedling Cold Tolerance (from NARES)
Selection of 861 C418 plants with seedling cold tolerance
     from 28 C418 BC2F2 populations 2002 (LAAS)
                                                      Non-CT
                             BC2F2     CT donors
                                                      donors
    # of populations           28           2            26
  # of surviving plants
                             10.3%        14%            0.3
     per population
         Range            1.4 – 19.3% 10 – 16%        0 – 3.0%
  # of surviving plants
                             10.3%        7.6%         10.5%
   per BC population
The mean population size was 310, ranging from 196 – 465, the
recipient, C418 (japonica) was killed by the stress.
Genetic background effects on the
   performance of BC progenies
Teqing / Chipda        NPT / Chipda
Donor and recipient effects on the number of BC2F2 plants
selected under lowland drought during the 2002 dry season
 Donor          Recurrent parent       Donor             Recurrent parent

           IR64    Teqing      NPT                    IR64     Teqing   NPT
 ASD 16             10***          0   Moroberekan     13                   11

 ASD18     59***      0        54***   MR 77         31****      0

 B4122     37***      2            1   Palung 2        33                   36

 Budda     75***      11               Pokhreli      119****     7      30***

 Chipda    47***      0       85****   Pusa           15**       4

 Chorofa              1        20***   Rasi          63***                   9

 Dacca 6   20         19               Rusty Late    38****      2           4

 Dhan4                1            0   Sadajira 19   55****      0

 Doddi     81***      2                Shwewartun      5*        0          6**

 Gajale    61***      22               SLG-1         26****                  0

 Giza 14   29***      4                SML242         6**        0          5*
Genetic background effects on the number of survival plants under submergence
      in 33 BC4F2 populations from crosses between 3 RPs and 11 donors
     Cross              Survival (%)   Z value   Cross             Survival(%)   Z value
     IR64/SN89366           3.33        1.87     NPT/C418             0.00
     Teqing/SN89366         6.67        3.75     IR64/CH448           9.33        4.14
     NPT/SN89366            0.00                 Teqing/CH448         7.33        3.25
     IR64/Y134              7.33        3.45     NPT/CH448            0.00
     Teqing/Y134            7.33        3.45     IR64/FR13A           7.67        3.64
     NPT/Y134               0.00                 Teqing/FR13A         6.67        3.17
     IR64/BR24              8.00        3.61     NPT/FR13A            0.00
     Teqing/BR24            8.00        3.61     IR64/Madhukar        7.33        3.60
     NPT/BR24               0.00                 Teqing/Madhukar      6.00        2.95
     IR64/Zihui 100        11.67        4.87     NPT/Madhukar         0.00
     Teqing/Zihui 100       7.33        3.06     IR64/IR50            6.67        3.13
     NPT/Zihui 100          0.00                 Teqing/IR50          8.00        3.76
     IR64/IR72              8.00        0.45     NPT/IR50             0.00
     Teqing/IR72            8.00        0.45     IR64/Jhona349        5.00        2.73
     NPT/IR72               7.00                 Teqing/Jhona349      5.67        3.09
     IR64/C418              7.33        3.19     NPT/Jhona349         0.00
    Teqing/C418            10.00        4.35
Genetic background effects on the number of survival plants under submergence
      in 33 BC4F2 populations from crosses between 3 RPs and 11 donors
     Cross              Survival (%)   Z value   Cross             Survival(%)   Z value
     IR64/SN89366           3.33        1.87     NPT/C418             0.00
     Teqing/SN89366         6.67        3.75     IR64/CH448           9.33        4.14
     NPT/SN89366            0.00                 Teqing/CH448         7.33        3.25
     IR64/Y134              7.33        3.45     NPT/CH448            0.00
     Teqing/Y134            7.33        3.45     IR64/FR13A           7.67        3.64
     NPT/Y134               0.00                 Teqing/FR13A         6.67        3.17
     IR64/BR24              8.00        3.61     NPT/FR13A            0.00
     Teqing/BR24            8.00        3.61     IR64/Madhukar        7.33        3.60
     NPT/BR24               0.00                 Teqing/Madhukar      6.00        2.95
     IR64/Zihui 100        11.67        4.87     NPT/Madhukar         0.00
     Teqing/Zihui 100       7.33        3.06     IR64/IR50            6.67        3.13
     NPT/Zihui 100          0.00                 Teqing/IR50          8.00        3.76
     IR64/IR72              8.00        0.45     NPT/IR50             0.00
     Teqing/IR72            8.00        0.45     IR64/Jhona349        5.00        2.73
     NPT/IR72               7.00                 Teqing/Jhona349      5.67        3.09
     IR64/C418              7.33        3.19     NPT/Jhona349         0.00
    Teqing/C418            10.00        4.35
Can we develop ILs with extreme
 phenotypes for selected target
            traits?
Screening results of 11 BC2F4 backcross populations derived from crosses between a
     japonica variety, Chaoyou 1 (the recurrent parent) and 11 donors for cold tolerance at the
                     booting stage and for heat tolerance at the flowering stage

                                                                                Selection for heat tolerance at the flowering stage
                      Selection for cold tolerance at the booting stage

                                                                                                   Seed set (%)         Spikelets per panicle
                                                  Seed set (%)

      Donor 1            N      SI (%) 2     Mean 3         Range         N      SI (%)       Mean 3        Range      Mean 3        Range

     Bg90-2 (I)         41        9.11       63.3 d       50.3 - 86.5     25      6.25    41.3 abc        23.0–77.0   171.2 abc   124.0–253.1

                                                                          9       2.25    38.1 abcd       19.5–50.0   192.5 ab    143.3–275.5
      X21 (I)           29        6.44       64.3 cd      50.6 - 87.1

      X22 (I)           28        6.22      65.6 bcd      50.7 - 87.3      -        -     -                   -           -             -

       Q5 (I)           31        6.89      71.1 abc      50.9 - 91.4     6       1.50    41.1 abc        19.9–75.4               160.4–274.2

Chhomrong (J)           24        5.33                    51.4 - 87.8     17      4.25    37.7 abcd       19.3–65.5   172.9 abc   115.9–258.8

     Doddi (I)          25        5.56      71.0 abc      50.2 - 90.0      -        -     -                   -           -             -

Feng-Ai-Zhan (I)        44        9.78                    52.2 - 98.5     12      3.00                    31.3–70.1   180.7 abc   138.2–238.0

                                                                          9       2.25    28.9 d          17.0–44.4   171.5 abc   112.0–216.7
Shennong265 (J)         21        4.67     69.2 abcd      50.7 - 93.8
                                                                          25      6.25    45.5 ab         23.9–65.6                83.2–255.4
 Yuangeng7 (J)          41        9.11      71.1 abc      50.0 - 90.1
                                                                          13      3.25    33.0 cd         17.0–48.9   171.9 abc   111.6–230.4
     OM997 (I)          21        4.67       72.1 ab      54.9 - 89.6
                                                                          8       2.00    33.6 abcd       24.9–48.3   175.5 abc   121.4–280.4
      Cs94 (I)          19        4.22       64.2 cd      51.4 - 86.0
                                                                          124     3.44
                                                                                          5.2 e            0.0–7.9    157.4 bc    127.4–178.6
  Chaoyou (J)           324       6.55       24.8 e       19.0 - 30.0

 2N    is the number of cold tolerant or heat tolerant BC plants selected from each population and SI = selection intensity.
 3   Different letters indicate the statistical significance in seed set at P < 0.05, based on the Duncan testing of ANOVA.
Table 6 Performances for AG of 11 promising BC4F3 lines and their donors
    Recipient               Donor             Seedling height (cm)           AG (%)            Emerging at 10 d (%)
   NPT                      Khazar                      33.5                   90.0*                      20.0
   NPT                      Khazar                      32.5                  95.0**                     75.0**
   NPT                      FR13A                       37.7                  95.0**                      30.0
   NPT                      TKM 9                       37.7                100.0***                    95.0***
   NPT                      TKM 9                       36.2                100.0***                      60.0*
   NPT                      TKM 9                       37.9                100.0***                    95.0***
   NPT                      Babaomi                     34.9                100.0***                      50.0
   NPT                      Babaomi                     36.7                100.0***                      50.0
   NPT                      OM1706                      33.9                100.0***                      65.0*
   NPT                      OM1706                      36.1                100.0***                    80.0***
   TKM 9                          I                     31.4                   20.0                       12.7
   Khazar                         J                     30.3                    3.0                        0.0
   Babaomi                        I                     26.3                    5.0                        0.0
   Jiangxi-Si-Miao                I                     31.5                    9.0                        0.0
   OM1706                         I                     29.1                   18.0                        2.0
   IR64                           I                     26.5                   20.0                        2.0
   NPT                            J                     36.2                   68.0                       39.0
I = indica and J = japonica. Traits were measured at 21 d after seeding. All 10 ILs had significantly higher AG than the
recurrent parent, NPT at P < 0.001.
Trait specific introgression lines developed
                        # of BC2F2      No. of selected
   Target traits
                        populations      BC2F3 lines

  Drought tolerance        350             4687
  BPH resistance           203              522
   Salinity tolerance      203             1022
Anaerobic germination      130              368
  Zinc deficiency          129              1211
Submergence tolerance      264              798
   Grain quality           65               580
    Other traits           375            12,000+
      Total                                20,000+
Summary of Selection Experiments

• Most donors contributed performance enhancing
  alleles to their BC progenies regardless of their
  performances;

• Appropriate screening (selection) is the key to
  identify improved target traits in the BC
  progenies;

• More distantly related donors, particularly
  landraces, tend to give more transgressive
  segregations for abiotic tolerance in the BC
  progenies.
Conclusions
There are tremendous amounts of hidden genetic
diversity in the current rice germplasm
collections for genetic improvement of all target
traits we tried, which have not been exploited;
Selection of parental lines for breeding based on
phenotype practiced by most breeders is a poor
way in exploiting novel genetic variation for
complex traits;
Backcross breeding, effective selection (efficient
screening) combined with DNA markers are the
effective way to exploit this hidden diversity;
What are we going to do with
   this large number of ILs?

1. Direct development of new cultivars;
2. As genetic stocks for discovery of DT alleles
  or QTLs and functional genomics of DT;
3. As parents for development of superior rice
  cultivars by QTL pyramiding
Progeny Testing of DT ILs Under Stress
   (no irrigation after transplanting)




      DT C418 ILs        Check (C418)
Progeny Testing Under Stress
(no irrigation after transplanting)




         Promising DT C418 ILs
2001-2002 DS BC2F4 progeny testing




IR64 ILs for quality   IR64 (CK)   DT IR64 ILs
Performance of two DT NPT ILs under severe stress




            DT NPT ILs       NPT CK
Performance of some promising salinity
tolerant IR64 ILs in the field condition in Iloilo
                (EC 18 dSm-1 )




                                      11/17, 2003
Performance of some promising salinity
tolerant IR64 ILs in the field condition in
           Iloilo (EC 18 dSm-1 )




                                 11/17, 2003
Promising varieties
developed in the BC
 breeding program
The Recurrent Parents
  C418
                      Liaojing454
(restorer)
Preliminary Yield Trials of Promising
            DT/WUE ILs
                             30%
                            water
                            saving




                              70%
      50%                    water
     water                   saving
     saving
Replicated Preliminary yield trial of DT/WUE ILs
                (Shengyang/2006.9)
Completely               Water
 rainfed                 saving
                          70%




Water                     Water
saving                    saving
 50%                       30%
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
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2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
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2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
2012 GSR - breeding technology
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2012 GSR - breeding technology

  • 1. GSR Breeding Strategy – Exploiting the Hidden Reservoir of Diversity for Improving Complex Traits in Rice Zhi-Kang Li/CAAS GA biosynthetic pathway Geranylgeranyl diphospate AS CPS GA12 -aldehyde GA7ox ent - Copalyl diphosphate GA 12 KS KS GA13ox ent -Kaurene GA 53 ent -Kaurenol GA 15 GA 44 KO KO GA ent -Kaurenal C20ox GA 19 GA 24 sd-1 ent -Kaurenoic acid GA 9 GA 20 KAO KAO GA3ox ent -7a-hydroxy kaurenoic acid GA4 GA 1
  • 2. Subjects Introduction Concept of GSR GSR Breeding Strategy and Technology Perspectives
  • 3. The impact of ‘Green Revolution’ in China 60000 Impact of ‘Green Revolution’ Impact of hybrid rice 50000 40000 30000 Yield 20000 10000 0 1965 1967 1969 1971 1973 1975 1977 1979 1981 1983 1985 1988 1990 1992 1994 1996 1998 2000 2002 Year Yield/unit area Total production
  • 4. Important problems in rice production worldwide • Increasing problems in abiotic and biotic stresses • Over-use of pesticides • Abuse of chemical fertilizers • Shortage of water • Ever increasing demands for yield increase • Urgent need for improved quality
  • 5. In China Current agricultural practices: higher inputs-for high yields- polluted environments China consumes ~1/3 of the global production of chemical fertilizers and pesticides annually on only 7% of the world’s cultivated lands Fertilizer application and grain production in China Total grain production(in 10000 t)、unit area yield 4500 75000 production( 70000 4000 Fertilizer use 65000 Fertilizer use (in 10000t) 3500 Grain production Grain yield per unit area 60000 (kg/10ha) ) 3000 55000 )、unit 50000 2500 45000 )、 2000 40000 1500 35000 30000 1000 25000 500 20000 0 15000 1975 1978 1982 1985 1988 1991 1994 1997 2000
  • 6. Trends of rice production and uses of pesticides and fertilizers in China in the last 30 years 40 150 50 Fertilizers (million ton) ) 7 35 单产 45 130 Pesticides(10000 ton) ) 农药 40 Yield(ton/ha) (ton/ha) 30 化肥 110 6 35 25 90 30 20 ( 5 70 25 15 20 50 4 10 15 5 30 10 3 0 10 5 1978 1981 1984 1981 1984 1987 1990 1993 1996 1999 2002 2005 1975 1978 1987 1990 1993 1996 1999 2002 2005
  • 7. 5 Grain yield(ton/hm2) Grain yield per unit area Grain yield/ha 4 / 3 2 1 1961 1966 1971 1976 1981 1986 1991 1996 2001 2006 350 350 350 350 Nitrogen use efficiency (kg/kg ) Nitrogen consumption(kg/hm2 ) 300 氮肥用量 300 300 300 ) 公顷) PFP 250 N consumption 250 kg/kg 250 250 / 200 NUE 200 200 200 氮肥生产效率( 氮肥用量(公斤 150 150 150 150 100 100 100 100 50 50 50 50 0 0 0 0 1961 1966 1971 1976 1981 1986 1991 1996 2001 2006 1961 1966 1971 1976 1981 1986 1991 1996 2001 2006
  • 8. Annual yield losses ~20% from abiotic stress: drought, problem soils, etc) ~15% from biotic stress: diseases and insects, even with the heavy uses of pesticides.
  • 9. Water crisis and drought in China Fresh water resources per capita in China is less than a quarter of the world average; Agriculture uses ~70% of the fresh water in China, and rice uses ~70% of the fresh water used in agriculture; Drought is occurring more frequently than ever before; Great yield loss of rice to drought in major rice areas.
  • 10. Which places have been left out by Green Revolution: Rice Yield Gaps in Irrigated and Rainfed Ecosystems in Asia, 1967-97 6.0 Irrigated Rainfed 5.0 Irrigated 100 (40%) 4.0 80 Y ie ld (t/h a ) Largely Irrigated (30%) 60 3.0 40 2.0 20 Rainfed 1.0 (30%) 0 MC TC 0.0 1967 1970 1973 1976 1979 1982 1985 1988 1991 1994 1997 Adoption percentage Year of modern cultivars Source: M. Hossain, IRRI
  • 11. Consequences of the Green Revolution Productivity Diversity
  • 12. Chinese scientists are calling “Second Green Revolution” Less inputs, more production, and environmental sustainability 少投入、多产出、保护环境 - to develop and widely adopted “Green Super Rice” that can produce high and stable yields under less inputs (chemical fertilizers, pesticides and water, and stress resilient).
  • 13. What are “GSR”? High yielding cultivars with multiple “Green” traits: Resistances/ tolerances to: Abiotic stresses: Drought, salinity, alkalinity, etc. Diseases: Blast, bacterial blight, sheath blight, viruses, and false smut etc Insects: Brown plant hopper, stem borer, etc High resource-use efficiencies: Water and nutrients (N, P)
  • 14. To develop GSR rice varieties, breeders are facing the following challenges: Many target traits: Yield and its related traits Stability - Resistances to biotic stresses - Tolerances to abiotic stresses Quality - Eating, cooking, and milling - Micronutrients
  • 15. Where are the sources of genetic variation for improving the ‘green’ traits?
  • 16. Rice Germplasm Collections in Genebanks Worldwide: ~ 215,000 entries 10% of 22 wild species 27% of modern varieties 90% of the 73% of Cultivated rice landraces Everson et al. 1998
  • 17. Status of the International Rice Genebank Collection at IRRI Incoming Accessions Total samples O. sativa 85,999 15,784 101,783 O. glaberrima 1,333 288 1,621 Wild species 3,970 495 4,465 Total 91,302 16,567 107,869
  • 18. Characterization of O. sativa % accessions scored accessions (n=79,925) 100 90 80 70 60 50 40 30 20 10 0 44 morpho-agronomic traits
  • 19. No. accessions 20,000 10,000 30,000 40,000 Blast 50,000 Bacterial blight Sheath blight Screened Rice tungro BPH 1 BPH 2 Resistant BPH 3 Green resistance/tolerances at IRRI leafhopper Whitebacked planthopper Evaluation of rice germplasm for stress Cold tolerance
  • 20. Cold tolerance (1-3 = tolerant; 5 = intermediate; 7-9 = susceptible) 2,500 n=6,625 2,000 1,500 1,000 500 0 1 3 5 7 9
  • 21. Yield responses of the 193 parental lines of IRMBN to the terminal drought under the lowland condition 25 ± -19.1±44.0% 20 15 10 5 0 -100 -80 -60 -40 -20 0 20 40 60 80 100 120 140 160 Yield reduction (in %)
  • 22. Establishment of the core collections for major crops in the CAAS’s genebank Core collections of rice, wheat, corn and Indica soybean were established with only 5% of the accessions representing over 90% Jap M. . Jap U. . Jap L. . genetic diversity of the whole collections. 0.1 0.1 Japonica Ind EM. . Ind M. . Ind M . L Basic Collections Core Collection Mini-core Accessions Entries sampling % Representation Entries sampling % Representation Rice 61479 3074 5% 89.9% 300 0.5% 66.6% Wheat 23135 1160 5% 90.1% 231 1.0% 69.1% Soybean 28809 1439 5% 91.0% 280 1.0% 71.0% Comparison of genetic diversity among core collections with different sampling ratio (Jia jizheng,2004)
  • 23. Current Status of Characterization and Utilization of Germplasm Collections in Genebanks Worldwide Collection and conservation largely completed Characterization – very superficial - Phenotypic evaluation and description To identify accessions with desirable (often extreme) phenotypes to be used as parents in breeding programs - Few accessions with desirable phenotypes for most traits - No accessions with desirable phenotypes for certain traits How much valuable genetic variation for target traits in the primary gene pool for breeding remains unclear!
  • 24. Utilization – very poorly - Less than 5% of the collected germplasm in the Genebanks have been utilized in the worldwide breeding programs Reasons for Poor Utilization of Germplasm Outstanding commercial genotypes are commonly destroyed by crosses with unimproved exotic germplasm (Duvick 1984) Slow but consistent genetic improvement can still be achieved even within a narrow base in many breeding programs Selection of parental lines in breeding programs are largely based on phenotype and very few accessions appear to have “desirable” phenotype for complex traits
  • 25. The GSR Breeding Strategy: Exploiting the maximum genetic diversity in the primary gene pool of rice Objectives To exam if there is sufficient (novel) genetic variation for target traits in the primary gene pool for most complex traits If yes, to develop an effective and efficient strategy to exploit the genetic diversity for complex traits - Integration with the molecular tools - Integration with gene discovery - Generation of information and training personnel
  • 26. The Basic Idea Full Exploitation of The Genetic Diversity in the Primary Gene Pool of Rice
  • 27. The GSR Breeding Strategy Recipients Donors (46 best commercial X (203 WMCC, 300 CMC + Varieties and hybrid parents) 20 wild rice accessions) Genotyping by re- BC breeding to introgress sequencing of all superior alleles from MCC into parents elite genetic backgrounds Parents for next Trait-specific Molecular database round of DQP IL sets in elite GBs of the parents Genotyping & phenotyping Development of Genetic characterization new cultivars of ILs Breeding by MRS and PL sets or DQP -omics and bioiformatics Release to Theory and technology of farmers breeding by Gene networks and metabolic molecular design pathways of the target traits and functional diversity of alleles at the loci
  • 28. The Chinese GSR research institutions - China National Rice Molecular Breeding Network
  • 29. Goal The goal of NCMBN was to develop superior inbred and hybrid crop cultivars with significantly improved yield stability, yield potential, and grain quality for the major rice growing areas in China.
  • 30. Specific objectives To broaden the genetic base of crop cultivars in major crop growing areas of China by maximizing the gene flow from the primary rice gene pool into elite genetic backgrounds through backcross breeding; To exploit the hidden diversity of the primary rice gene pool for improving complex target traits; To develop IL sets for important traits in elite rice genetic backgrounds; To discover and characterize large numbers of QTLs and QTL networks underlying important traits, and to mine allelic diversity at important QTLs; To establish genetic/phenotypic database for the ILs; To train a new generation of molecular rice breeders in China.
  • 31. Proof of the Concept
  • 32. Two Fundamental Questions: • How much useful genetic diversity, particular for the complex phenotypes, within the primary gene pool of rice? • Can we combine the process of breeding with gene discovery (gene/QTL discovery and allelic mining)?
  • 33. Part I: Introgression breeding and mass selection Recurrent Parents IR64 - Indica, high yield/widely adaptable New Plant Type - Japonica, high yield potential Teqing - Indica, high yield/widely adaptable
  • 34. The “value” added and base broadening approach – Introgression breeding Widely Add new genes/traits IR64 introgression by backcross breeding adaptable lines with improved high yield target traits varieties (IR64) IR64 lines with improved Discovery of desirable QTLs using target traits and the “same” DNA markers and MAS for pyramiding QTLs yield potential and quality
  • 35. Procedure of the backcross breeding for development of ILs for gene/QTL identification and cultivar development RP x donors F1s x RP 25 BC1F1s x RP X Self and bulk harvest ~25 BC1F2s x RP ~25 BC2F1s x RP X Self and bulk harvest Bulk BC1F2 populations Bulk BC2F2 populations Screen for target traits: 1, 2, 3, 4, 5, n 1, 2, 3, 4, 5, n Replicated progeny testing for the introgression lines (ILs) for the selected target traits (tolerances to drought, salinity, submergence, BPH, etc), and for yield performances and other agronomic traits under non-stress conditions Large numbers of ILs with selected target traits in the elite RP genetic background Genotyped w/ DNA markers to track the gene flow and to identify genes/QTLs for the target traits Promising ILs with selected target Best ILs with selected target trait(s) trait(s) and good yield performances used as parental lines for nominated for NCTs pyramiding genes/QTLs from different donors Development and releases of green super rice cultivars for To rice farmers in the target specific target environments environments
  • 36. Four Major Groups of Target Traits Tolerances to abiotic stresses (drought, salinity, low and high temperatures, N and P use efficiency, etc.) Resistances to biotic stresses (blast, sheath blight, false smut, BPH, stem borer, BB, etc.) Yield potential and heterosis Quality parameters
  • 37. Donor gene pool (203 accessions from 34 countries) Sub-G1 (54) Sub-G2 (6) I (indica) Sub-G3 (33) Sub-G4 (46) Ba-Bao-Mi (Yunnan) Sub-G5(15) II (japonica) Sub-G6 (12) Sub-G7 (28) Sub-G8 (9) III (New group) Jalmagna (India)
  • 38. Gene diversity of the donor gene pool Proportions of accessions Relative genetic diversity of from different geographic accessions in different regions geographic regions Europe America Others Europe America Others 15% 10% NA NA SA SA China 33% China 40% 22% 26% SEA SEA 30% 24%
  • 39. Target traits • Tolerances to drought, salinity, submergence, zinc deficiency, phosphorus deficiency, anaerobic germination, etc. • Resistances to BB, BPH, blast, tungro, sheath blight, etc. • Yield and related traits • Different maturities • Different types of grain quality parameters
  • 40. BC Breeding Procedure RP X Donors of diverse origins F1s X RP BC1F1 X RP BC2F1 x BC2F2 bulk populations Selection for target traits Progeny testing Survival plants Genotyping
  • 41. Major differences of introgression breeding and the conventional BC breeding BC breeding Marker aided Introgression BC breeding breeding RP selection Elite Same Same Donor selection Yes Yes No/diverse Target traits Single Single or few No limitation monogenic/dominant Selection Mass selection MAS Mass selection Progeny testing yes yes yes Pop. size Small Small Regular Selection efficiency High High Depends Breeding efficiency Low Low High
  • 42. Selection strategy for target traits in random BC progenies
  • 43. Screening of BC2F2 populations for tolerance to terminal drought at the reproductive stage under the lowland conditions Stress for 20 days (March 27,2001) Stress for 56 days (May 2, 2001)
  • 44. Variation in BC2F2 populations for drought tolerance under the lowland conditions
  • 45. Summary of selected drought tolerant BC2F2 plants under lowland stress conditions NPT IR64 Teqing Total Total plants selected 897 2775 489 4161 # of selected plants 8.5 22.4 6.3 13.2 per population (3.8%) (10.0%) (3.0%) (6.8%) Range 0 - 85 0 - 100 0 - 30 0 - 100 No. of populations 113 124 105 320 No. of I donors 59 67 59 185 No. of J donors 32 45 19 96 Contributing donor (%) 80.5 90.3 74.3 87.8
  • 46. The Screening of BC2F2 populations under upland drought conditions The RP, IR64
  • 47. Summary of selected drought tolerant BC2F2 plants under lowland stress conditions IR64 Teqing Total Total plants selected 192 334 526 Ave. selected plants 9.6 (4.6%) 10.8 (5.2%) 10.3 (5.0%) Per population Range 4 - 20 3 - 15 4 - 30 No. of populations 20 31 51 No. of I donors 12 23 35 No. of J donors 8 8 16 Contributing donor (%) 100 100 100
  • 48. Parental performances and 442 selected DT BC2F2 plants for drought tolerance from 19 BC populations VG Donors Origin IR64 (S) Teqing (M) NPT(SS) Total LL (UL) LL (UL) LL I BR24 (S) Bangladesh 14 (27) 12 (7) 3 29 (34) I STYH (S) Myanmar 20 (26) - 4 24 (26) I OM1723 (S) Vietnam 7 (17) 7 (6) 0 14 (23) J FR13A (SS) India 15 (16) 17 (15) 0 32 (31) J Type3 (SS) India 23 (15) 10 (12) 0 33 (27) J Binam (M) Iran 20 (19) 14 (13) 1 35 (32) J HAN (M) China 11 (13) - 3 14 (13) I Zihui100 (S) China 8 - 9 17 J Khazar (S) Iran 58 - 58 Total 176 (133) 60 (53) 22 256 (186)
  • 49. Screening of BC2F2 populations for salinity tolerance at the seedling stage Young seedlings were subjected to: IR64 ST CK 6 dSm-1 for 3 days, 12 dSm-1 for 2 weeks, 18 Pokali dSm-1 for 1 week, and 24 dSm-1 for 1 week
  • 50. Confirmation by progeny testing Seedling screening at EC 24 dSm-1 for 3 weeks
  • 51. Table 1. Summary results of BC populations for screening salinity tolerance Details BC2F2 screening BC2F3 reconfirmation IR64 Teqing NPT Total IR64 Teqing NPT Total Total BC2F2 populations 62 58 55 175 24 34 10 68 Single plant selections per BC population 4 - 12 4 - 13 1 - 14 0 - 43 0 - 49 0 - 11 Total selected BC2F3 lines 490 428 374 1292 448 392 21 861 Selection intensity (%) 3.95 3.69 3.40 3.69 Number of indica donors 47 47 42 136 20 27 7 54 Selected lines 369 345 289 1003 372 269 21 662 Selection intensity (%) 4.39 3.67 3.44 3.69 Number of japonica donors 9 9 7 25 3 6 1 10 Selected lines 70 66 44 180 43 123 0 166 Selection intensity (%) 3.89 3.67 3.14 3.60 Number of intermediate donors 4 1 3 8 1 1 2 4 Selected lines 35 5 19 59 33 0 0 33 Selection intensity (%) 4.38 2.50 3.16 3.69
  • 52. Summary results of BC populations for screening salinity tolerance BC2F2 screening BC2F3 reconfirmation IR64 Teqing NPT Total IR64 Teqing NPT Total # of BC2F2 pop. 62 58 55 175 24 34 10 68 # of selections per pop. 4 - 12 4 - 13 1 - 14 1 - 14 0 - 43 0 - 49 0 - 11 Total selected lines 490 428 374 1292 448 392 21 861 # of contributing donors 60 57 52 169 SI (%) 3.95 3.69 3.40 3.69 For individual BC populations of 200 plants, a difference of 4% between two populations in selection intensity (survival rate) is statistically significant at P < 0.05 when the selection intensity is between 0.1 and 0.5.
  • 53. Screening of BC2F2 populations for submergence tolerance in a deep-water pond Thirty-five-day old seedlings were submerged under deep water for two weeks, then allowed to recover
  • 54. Summary results of BC populations for screening anaerobic germination and submergence tolerance Anaerobic germination Submergence IR64 Teqing NPT Total IR64 Teqing NPT Total # of BC pop. 47 47 36 130 60 57 62 179 # of selections per pop. 0-5 0-6 0 - 14 0 - 15 3 - 13 0 - 12 Total selected lines 47 81 215 343 652 483 530 1665 # of contributing donors 46 46 35 127 59 57 60 176 SI (%) 0.52 0.93 3.11 1.32 1.08 0.85 0.85 0.93
  • 55. Screening of BC2F2 populations for anaerobic germination Direct seeding then submerged under 10 cm-deep water
  • 56. Summary results of BC populations screened for anaerobic germination and submergence tolerance Details Anaerobic germination Submergence IR64 Teqing NPT Total IR64 Teqing NPT Total Total BC2F2 populations 47 47 36 130 60 57 62 179 Single plant selections per BC population 0-5 0-6 0 - 14 0 - 15 3 - 13 0 - 12 Total selected BC2F3 lines 47 81 215 343 652 483 530 1665 Selection intensity (%) 0.52 0.93 3.11 1.32 1.08 0.85 0.85 0.93 Number of indica donors 37 38 29 104 47 50 50 147 Selected lines 31 67 77 175 538 431 451 1411 Selection intensity (%) 0.43 0.90 3.10 0.84 1.15 0.86 0.90 0.96 Number of japonica donors 6 8 5 19 8 7 7 22 Selected lines 11 14 31 56 83 52 63 198 Selection intensity (%) 0.91 1.03 3.10 1.47 1.00 0.74 0.90 0.90 Number of intermediate donors 3 0 1 4 4 0 3 7 Selected lines 5 0 7 12 40 0 16 56 Selection intensity (%) 0.83 0.00 3.50 1.50 1.04 0.00 0.53 0.80
  • 57. Screening for BPH Resistance IR64
  • 58. Summary results of BC populations screened for low temperature germination and brown planthopper resistance Details Low temperature germination Brown planthopper resistance IR64 TQ NPT Total IR64 TQ NPT Total Total BC2F2 populations 14 15 10 39 64 67 62 193 Single plant selections per BC population 0 - 21 0 - 18 0 - 31 0 - 22 0 - 22 0-2 Total selected BC2F3 lines 79 77 51 207 652 255 2 909 Selection intensity (%) 5.64 5.13 5.10 5.31 10.19 3.81 0.03 4.71 Number of indica donors 1 1 0 2 49 60 49 158 Selected lines 0 3 0 3 565 221 2 788 Selection intensity (%) 0.00 3.0 0.00 1.5 11.53 3.68 0.04 4.99 Number of japonica donors 9 11 7 27 9 11 7 27 Selected lines 55 52 39 146 54 21 0 75 Selection intensity (%) 6.11 4.73 5.57 5.41 6.00 1.91 0.00 2.78 Number of intermediate donors 4 3 3 10 4 3 3 10 Selected lines 24 22 12 58 33 2 0 35 Selection intensity (%) 6.00 7.33 4.00 5.80 8.25 0.67 0.00 3.50 For individual BC populations of 100 plants, a difference of 2.5% between two populations in selection intensity (survival rate) is statistically significant at P < 0.05 when the selection intensity is < 0.1.
  • 59. Summary results of BC populations for screening low temperature germination and BPH resistance Low temperature germination BHP resistance IR64 Teqing NPT Total IR64 Teqing NPT Total # of BC2F2 pop. 14 15 10 39 64 74 62 200 # of selections per pop. 0 - 21 0 - 18 0 - 31 0 - 22 0 - 22 0-2 Total selected lines 79 77 51 207 652 255 2 909 # of contributing donors 14 14 10 38 62 67 59 189 SI (%) 5.64 5.13 5.10 5.31 10.19 3.81 0.03 4.71 For individual BC populations of 200 plants, a difference of 4% between two populations in selection intensity (survival rate) is statistically significant at P < 0.05 when the selection intensity is between 0.1 and 0.5.
  • 60. Screening for tolerance to phosphorus deficiency Tested in natural conditions in Pangil, Laguna.
  • 61. Screening for tolerance to zinc deficiency
  • 62. Summary results of BC populations screened for zinc deficiency tolerance BC2F4 progeny testing under very severe Details BC2F2 screening under moderate field stress field stress IR64 Teqing NPT Total IR64 Teqing NPT Total Total number of BC populations 51 42 36 129 11 21 10 42 Single plant selections per BC population 4 - 12 2 - 12 0 - 12 0 - 12 0 - 10 0 - 15 0 Total selected BC2F3 lines 454 383 324 1161 72 255 0 327 Selection intensity (%) 7.42 7.60 7.50 7.50 2.14 2.41 0.00 2.01 Number of indica donors 40 33 25 98 9 17 9 35 Selected lines 370 320 237 927 63 200 0 263 Selection intensity (%) 7.77 8.08 7.90 7.88 2.07 2.31 0.00 1.91 Number of japonica donors 7 6 7 20 1 4 1 6 Selected lines 63 63 84 210 0 55 0 55 Selection intensity (%) 7.50 8.75 7.86 8.75 0.00 2.86 0.00 2.29 Number of intermediate donors 3 - 1 4 1 0 0 1 Selected lines 21 - 9 30 9 0 0 9 Selection intensity (%) 5.83 - 7.50 6.25 5.63 0.00 0.00 5.63
  • 63. Selection for changed grain type of Teqing
  • 66. Screening for blast resistance
  • 67. Low N stress experiment under rain-off shelter
  • 68. Improving japonica for cold tolerance at the reproductive stage
  • 69. Screening results of 11 CY1 (recurrent parent) BC2F4 backcross populations for cold tolerance at the booting stage in 2008 (1 SI = selection intensity, SF = spikelet fertility; 2 Different letters indicate statistical significance at P < 0.05, based on the Duncan testing) No. of SF (%) Population SI 1 Donor parent Subspecies Code selected size plants (%) Mean ± SD2 Range Bg90-2 Indica A 450 41 9.1 63.3±8.4D 50.3-86.5 X21 Indica B 450 29 6.4 64.3±9.9CD 50.6-87.1 X22 Indica C 450 28 6.2 65.6±10.1BCD 50.7-87.3 Q5 Indica D 450 31 6.9 71.1±11.1ABC 50.9-91.4 Chhomrong Japonica E 450 24 5.3 75.6± 51.4-87.8 Doddi Indica F 450 25 5.6 71.0±10.0ABC 50.2-90.0 Fengaizhan Indica G 450 44 9.8 74.1± 52.2-98.5 Shennong265 Japonica H 450 21 4.7 69.2±11.4ABCD 50.7-93.8 Yuanjing7 Japonica I 450 41 9.1 71.1±9.8ABC 50.0-90.1 OM997 Indica J 450 21 4.7 72.1±9.1AB 54.9-89.6 Cs94 Indica K 450 19 4.2 64.2±8.6CD 51.4-86.0 Mean 450 29.5 6.5 69.2±4.2 CY1 Japonica 324 24.8±4.3E 19.0-30.0
  • 70. Screening of the BC2F4 bulk populations for cold tolerance (CT) at the booting stage: Seeds of the BC2F4 bulk populations were sown in the seedling nursery on April 15, 2008, and 450 40-day old seedlings of each BC2F4 bulk population were transplanted into a 45-row plot with 10 plants in each row and a spacing was 25×15cm in the sheltered water pond of JAAS on May 25. Two rows of CY1 were also transplanted on both sides of each plot as the checks. The water-pond were irrigated with water of normal temperature (25~28℃). When CY1 entered the stage of panicle initiation, the LT treatment was initiated by irrigation of flowing cold water (19±0.5℃), which was adjusted in a nearby water pool by ℃ mixing cold underground water (9℃) with the river water. The depth of the ℃ cold water in the pond was 20cm and the treatment was maintained for ~30 days until panicles of almost all plants exerted completely. Then, irrigation with normal temperature water was resumed until the maturity. At the maturity, all plants except those with >3 days earlier heading or those with >3 days delayed heading than CY1 were harvested for measuring the spikelet fertility. Under this LT treatment, CY1 had a spikelet fertility of ± 24.8±4.3%, then, any plant with spikelet fertility >50% were selected.
  • 71. Evaluation of 324 BC2F6 introgression lines and their recurrent parent, CY1 for cold tolerance of at the seedling and booting stages in 2009 (SNP = spikelet number per panicle, FGN = filled grain number per panicle, SF = spikelet fertility) Cold stress at the seedling No. of stage Cold stress at the reproductive stage Donor parent selected Survival rate of seedlings lines (%) SNP FGN SF (%) Mean Range Mean Range Mean Range Mean Range BG90-2 41 19.5 10.0~62.5 114.5 76.3~164.2 55.0 22.1~95.7 48.0 18.5~71.3 X21 29 19.4 11.1~40.3 99.7 73.3~125.0 50.5 29.5~90.3 50.3 30.9~74.9 X22 28 19.6 12.5~40.7 113.2 70.3~139.0 58.5 4.5~94.9 51.4 5.6~79.4 Q5 31 18.3 15.0~33.8 114.6 68.3~185.4 56.5 15.3~113.6 49.2 14.6~83.6 Chhomromg 24 17.0 15.0~32.5 109.8 77.6~175.2 74.0 47.0~104.8 66.9 45.6~83.4 Doddi 25 17.3 12.5~25.0 112.9 89.4~182.6 73.9 51.5~108.5 65.4 48.7~80.6 Fengaizhan 44 16.7 12.5~25.0 96.5 70.2~115.1 67.1 14.3~96.0 70.1 12.7~86.1 Shennong265 21 15.6 0.0~23.3 96.9 65.0~144.8 62.0 36.9~99.2 63.0 40.0~79.9 Yuanjing7 41 18.5 15.0~45.0 116.0 93.5~151.0 80.6 30.4~106.4 69.2 25.5~84.2 OM997 21 37.1 15.0~82.4 81.6 56.2~108.1 42.0 14.9~64.0 51.1 23.3~72.0 Cs94 19 22.3 15.0~42.5 113.3 82.0~137.9 69.5 31.8~99.4 60.0 34.2~75.5 CY1 14.8 10.8~20.0 106.6 98.6~114.6 36.7 32.7~40.8 35.1 33.7~36.5 LSD0.05 4.5 8.3 8.2 6.2
  • 72. Comparison of selection efficiencies of 11 CY1 BC2F6 populations for cold tolerances at the seedling and reproductive stages The reproductive stage The seedling stage Population N1 FGN SNP SF (%) SR (%) Code Donor N1 N2 N1 N2 N1 N2 N1 N2 A BG90-2 41 18 0 13 6 17 1 5 0 B X21 29 7 0 0 3 15 0 4 0 C X22 28 17 2 8 1 17 3 5 0 D Q5 31 15 2 10 4 20 3 1 0 E Chhomromg 24 17 0 7 3 22 0 2 0 F Doddi 25 23 0 1 0 24 0 0 0 G Fengaizhan 44 35 1 0 7 42 1 0 0 H Shennong265 21 12 0 2 9 18 0 0 1 I Yuanjing7 41 40 0 9 0 40 0 3 0 J OM997 21 2 1 0 13 13 0 14 0 K Cs94 19 14 0 5 1 14 0 6 0 Total 324 200 6 55 47 242 8 40 1 1N is the total number of selections based on single plant spikelet fertility (SF) in BC2F4 populations from Table 1, N1 and N2 are the numbers of the BC2F6 lines showing significantly higher or significantly lower than CY1 for the measured traits. FGN, SNP, SF and SR are filled grain number per panicle, spikelet number per panicle, spikelet fertility and survival rate of seedlings.
  • 73. Mean performances of 116 ILs of five populations for 11 traits evaluated under normal and cold water stress conditions in 2010 Trait 1 Chhomrong Doddi Fengaizhan Shennong265 Yuanjing7 Mean CK N 24 24 24 20 24 116 Under the normal conditions PH (cm) ± 135.2±4.4 ± 130.6±5.8 ± 125.7±5.4 ± 127.2±3.7 ± 132.3±4.5 130.3 117.1 BM (g/plant) 37.1±3.3 36.5±3.4 38.1±3.5 33.5±3.3 35.1±2.4 36.2 34.7 FGN 143.4±15.0 143.7±17.2 148.1±12.3 135.1±15.6 ± 156.3±19.4 145.7 138.9 SNP 176.6±20.8 179.4±20.9 171.2±17.4 172.9±17.7 ± 197.2±21.8 179.7 168.6 SF (%) 81.6±6.6 80.4±8.1 ± 86.9±5.8 78.4±7.2 79.2±5.3 81.4 82.4 GY (g/plant) ± 19.4±2.94 20.5-3.0 ± 20.9±2.5 ± 18.7±2.5 ± 18.7±2.7 19.6 19.7 HI (%) 55.0±5.2 59.1±4.9 57.9±3.7 59.1±4.4 56.0±4.8 57.3 60.3 PN 5.4±0.6 5.7±0.4 5.7±0.6 5.6±0.7 ± 4.8±0.5 5.5 5.7 GW (g) 24.3±1.5 ± 26.3±1.6 ± 22.0±2.1 24.5±1.1 ± 26.3±1.4 24.7 24.9 HD (d) 113.9±3.3 113.0±4.1 116.3±6.0 112.5±4.7 113.6±3.9 113.9 111.1 Under the cold water stress PH (cm) ± 118.1±7.9 ± 119.9±7.7 ± 119.8±7.7 ± 122.4±6.6 ± 125.2±5.7 121.0 107.8 BM (g/plant) 16.2±3.2 16.1±2.7 16.8±2.5 15.3±3.6 17.5±3.6 16.4 12.0 FGN 46.8±28.4 39.1±20.2 ± 63.8±14.3 ± 22.9±14.1 46.4±19.8 44.5 12.8 SNP 118.9±23.9 121.7±13.8 110.4±12.9 117.7±24.1 139.6±19.6 121.8 121.5 SF (%) 38.8±18.3 32.3±14.5 ± 57.3±10.3 ± 19.7±12.1 33.3±14.6 36.9 10.5 GY (g/plant) ± 3.5±2.2 ± 3.0±1.7 ± 5.0±1.3 ± 1.9±1.2 ± 3.6±1.9 3.5 1.0 HI (%) 33.3±7.6 30.7±6.7 ± 40.9±4.4 ± 25.8±7.2 30.8±5.1 32.5 25.1 PN 3.3±0.5 3.4±0.6 3.3±0.5 3.7±0.9 3.2±0.5 3.4 3.5 GW (g) 17.9±1.5 18.0±1.6 17.5±2.7 17.1±0.9 17.9±1.3 17.7 16.4 HD (d) 131.3±2.2 128.2±3.3 132.0±4.1 126.9±4.1 128.6±3.0 129.5 125.2
  • 74. The numbers of ILs from the 5 populations that deviated significantly CY1 for 11 measured traits evaluated under cold water stress (S) and normal (N) conditions in 2010 GY GW HD HI PH SF BM (g/plant) FGN PN SNP Donor N1 Treat (g/plant) (g) (d) (%) (cm) (%) N1 N2 N1 N2 N1 N2 N1 N2 N1 N2 N1 N2 N1 N2 N1 N2 N1 N2 N1 N2 Chhomrong 24 18 0 18 0 14 0 15 0 24 0 14 0 19 2 3 8 21 0 6 12 Doddi 24 18 0 19 0 15 0 16 1 16 2 14 2 21 0 5 7 19 0 4 4 Fengaizhan 24 S 22 0 24 0 23 0 6 3 22 1 24 0 20 0 2 6 24 0 0 12 Shennong26 5 20 12 1 7 0 6 0 5 2 10 8 7 6 18 0 9 5 10 0 6 8 Yuanjing7 24 21 0 21 0 19 0 17 0 16 0 15 1 24 0 3 8 20 0 12 1 Total 116 91 1 89 0 77 0 59 6 88 11 74 9 102 2 22 34 94 0 28 38 Chhomrong 24 15 2 11 5 9 8 4 13 13 2 1 16 24 0 3 9 7 9 11 4 Doddi 24 11 4 10 5 9 5 17 4 11 3 6 7 24 0 4 4 8 12 13 5 Fengaizhan 24 16 0 14 2 9 1 2 20 19 3 3 15 21 0 5 7 15 4 8 8 N Shennong26 5 20 3 8 5 7 2 9 5 10 8 7 5 7 20 0 8 10 5 13 9 7 Yuanjing7 24 5 3 17 2 5 14 18 2 11 3 3 18 24 0 0 21 4 12 20 1 Total 116 50 17 57 21 34 37 46 49 62 18 18 63 113 0 20 51 39 50 61 25 1N is the total number of ILs with CT selected from each population; N1 and N2 are the numbers of the ILs showing significantly higher and lower trait values than CY1. GY = grain yield, BM = biomass, PH = plant height, PL = panicle length, PN = panicle number per plant, FGN = filled grain number per panicle, SNP = spikelet number per panicle, SF = spikelet fertility, HD = heading date, GW = 1000-grain weight, HI = harvest index.
  • 75. Mean performances of 19 promising ILs under cold water stress and normal conditions in 2009 and 2010 (Meng et al. 2012) Line# Pop.1 2010 under the normal condition 2010 under cold water stress 2009 under stress PH BM SNP SF GY HI PN GW HD PH BM SNP SF GY HI PN GW HD SNP SF SR CK(CY1) 117.1 34.7 168.6 82.4 19.7 60.3 5.7 24.9 111.1 107.8 12.0 121.5 10.5 1.0 25.1 3.5 16.4 125.2 106.6 35.1 14.8 LW213 G 134.2 48.5 192.3 91.1 28.1 60.1 6.4 22.8 103.0 129.2 19.5 131.2 69.1 7.2 46.8 3.5 15.7 128.0 101.7 81.2 12.5 LW164 F 126.5 45.7 204.5 87.8 28.5 65.2 6.4 23.5 117.0 118.8 17.2 120.0 27.3 2.5 26.0 3.3 16.1 129.0 113.2 56.9 15.0 LW180 F 131.7 38.8 182.0 89.8 23.4 63.1 5.7 25.6 101.0 131.7 18.0 138.0 39.6 4.9 36.3 3.3 19.1 123.0 107.3 68.0 17.5 LW157 F 133.4 41.0 218.7 71.9 23.4 59.9 6.0 25.2 116.0 114.8 20.9 132.8 40.1 5.0 33.8 4.3 17.1 131.0 110.6 60.3 17.5 LW170 F 129.3 40.0 183.3 83.7 22.8 59.7 5.9 24.0 117.0 117.8 19.6 119.5 45.5 5.3 38.4 4.8 16.7 131.0 118.4 71.9 17.5 LW214 G 122.0 38.0 159.8 85.6 22.9 63.5 6.7 17.9 120.0 108.5 14.3 103.7 54.9 4.5 44.4 3.3 16.3 137.0 86.7 84.5 15.0 LW188 G 121.0 40.1 164.3 84.2 23.1 60.6 6.7 21.0 121.0 130.0 16.6 119.7 66.9 5.8 46.2 3.2 17.0 138.0 90.9 82.5 15.0 LW142 E 136.7 39.3 162.7 79.2 21.5 57.7 6.7 23.9 112.0 121.5 16.7 119.8 61.6 5.2 43.1 3.2 18.1 134.0 91.2 60.0 15.0 LW154 E 137.8 39.9 198.3 82.9 21.9 58.0 5.4 23.4 112.0 128.5 19.7 160.7 67.4 7.4 47.7 3.0 17.4 130.0 121.6 71.3 15.0 LW174 F 129.5 37.6 151.3 86.6 21.7 61.0 6.7 25.9 115.0 120.3 17.7 122.0 48.2 5.3 41.4 4.2 17.8 128.0 126.2 80.3 25.0 LW207 G 132.6 40.0 192.8 79.9 22.1 58.0 5.8 21.2 121.0 127.0 20.1 118.3 66.6 6.7 43.1 3.8 17.5 132.0 115.1 83.7 20.0 LW264 I 138.8 40.7 221.5 77.7 22.6 58.0 5.3 24.9 107.0 130.0 17.2 169.2 26.3 3.1 29.1 2.8 17.9 125.0 120.3 79.1 42.5 LW250 I 130.7 39.2 226.8 79.9 23.3 62.6 5.2 26.0 110.0 136.8 20.1 136.8 65.1 5.8 38.4 2.7 18.0 132.0 108.4 76.1 20.0 LW156 E 138.9 40.3 187.8 78.9 21.0 55.0 5.7 23.0 114.0 127.0 20.2 129.7 57.9 6.8 43.0 4.0 16.3 129.0 131.8 74.0 15.0 LW151 E 131.9 37.7 168.3 82.9 19.2 53.9 5.6 24.2 114.0 122.3 19.3 123.5 67.2 6.6 43.3 3.3 16.6 130.0 122.4 82.9 16.3 LW189 G 132.4 37.9 198.3 84.4 20.1 55.9 4.8 21.7 120.0 133.2 19.0 114.7 59.5 6.2 41.1 3.5 17.0 137.0 103.4 75.7 20.0 LW216 G 129.1 37.3 170.8 87.1 20.1 56.9 5.4 20.7 123.0 111.8 17.1 99.8 51.7 4.3 34.4 2.8 15.9 139.0 99.9 86.1 16.9 LW200 G 127.7 38.6 167.8 88.8 20.5 56.1 5.6 20.3 115.0 122.0 16.4 100.2 66.2 4.9 41.9 3.3 16.5 130.0 93.7 84.2 17.5 LW291 I 121.1 33.1 181.5 84.8 17.5 56.5 4.7 25.1 120.0 126.3 24.9 156.3 39.1 6.6 32.2 4.2 17.7 131.0 143.6 74.5 45.0 LSD0.05 1.9 2.1 8.2 2.6 1.6 1.8 0.3 0.5 1.9 3.7 1.8 11.1 8.1 1.2 4.3 0.4 1.1 1.6 8.2 6.2 4.5
  • 76. The hidden diversity for highly heritable traits – BLB resistance
  • 77. Reactions (lesion lengths) of HHZ and two donors to 14 tropical races of BLB caused by Xanthomonas oryzae pv oryzae P1 P2 P3b P3c P4 P5 P6 P7 P8 P9a P10 P9c P9b P9d PXO PXO PXO PXO PXO PXO PXO PXO PXO PXO PXO PXO PXO PXO aver 61 86 79 340 71 112 99 145 280 339 341 347 349 363 age HHZ 9.8 21.2 13.1 25.7 10.4 2.4 29.6 5.0 8.6 28.8 8.4 26.6 15.2 24.6 16.4 PSBRC66 6.4 18.5 16.4 21.4 11.6 0.7 13.0 2.6 8.8 4.1 7.0 12.0 3.4 17.3 10.2 PSBRC28 2.8 20.3 21.6 24.0 11.2 3.9 22.4 4.7 9.2 26.0 8.5 23.7 22.9 21.9 15.9
  • 78. 8个黄华占群体对15个Xoo菌株产生反应类型 (2012年8月,北京) F1-F5 F1- F6- F6-F10 F11- F11-F15 F1- F1-F5 F6- F6-F10 F11- F11-F15 F1-F5 F1- F6- F6-F10 F11-F15 F11- 对部分小种抗病的株系 对全部15个小种高抗的株系 对全部15个小种感病的株系
  • 79. Blast evaluation of virulent strains Evaluation of BB resistance of >500 lines (HHZ background) against 14 strains of 10 Xoo races, 2010 WS HHZ is susceptible to most tropic BLB races Vera Cruz et al
  • 80. Ten HHZ ILs with broad spectrum resistance to all 14 races of bacterial blight pathogen, Xanthomonas oryzae pv oryzae PX PX PX PX PX PX PX PX PX PX PX PX O34 PX O11 PX O14 O28 O33 O34 O34 O34 O36 aver O61 O86 O79 0 O71 2 O99 5 0 9 1 7 9 3 age HHZ 9.8 21.2 13.1 25.7 10.4 2.4 29.6 5.0 8.6 28.8 8.4 26.6 15.2 24.6 16.4 PSBRC66 6.4 18.5 16.4 21.4 11.6 0.7 13.0 2.6 8.8 4.1 7.0 12.0 3.4 17.3 10.2 HHZ15-SAL13-Y2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 HHZ15-SAL-13-Y3 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 HHZ15-DT7-SAL1 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 HHZ15-DT7-SAL3 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 HHZ15-DT7-SAL6 0.2 0.2 0.2 0.2 9.3 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.9 PSBRC28 2.8 20.3 21.6 24.0 11.2 3.9 22.4 4.7 9.2 26.0 8.5 23.7 22.9 21.9 15.9 HHZ19-SAL-14-Y3 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 HHZ19-DT8-SAL2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 HHZ19-SAL12- SAL4 0.2 0.7 0.9 0.5 0.2 0.2 0.2 0.2 0.4 0.2 0.3 0.9 0.2 0.2 0.4 HHZ19-SAL14- SAL4 0.2 1.0 0.6 0.3 0.2 0.2 0.2 0.2 0.3 0.2 0.7 0.2 0.2 0.2 0.3 HHZ19-SAL15- SAL2 0.2 3.8 2.2 0.6 0.4 0.4 0.8 0.5 0.3 0.3 0.3 0.6 0.4 0.4 0.8
  • 81. Reactions of 512 HHZ ILs to 14 tropical Xoo races Race R% (LL <3.0 cm) P1 76.4 P2 4.7 P3b 4.9 P3c 4.9 P4 22.4 P5 78.4 P6 5.1 P7 46.6 P8 31.0 P9a 12.3 P10 12.1 P9c 4.7 P9b 5.1 P9d 50.8
  • 82. Classification of 14 Xoo races into two major groups (virulent and less virulent ones) based on the reactions of the 512 HHZ ILs P1 P5 Weak virulence group P4 P7 Name Xoo races P8 P10 P2 P9c P6 P3c High virulence group P3b P9a P9b 0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1.0 1.1 1.2 1.3 Average Distance Between Clusters
  • 83. Classification of the 512 ILs based on their resistances to the 14 Xoo Phillipines races H9-39 H5-1 H8-27 H9-20 H17-65 H9-3 H9-13 H5-32 H8-44 H15-12 H17-29 H5-49 H15-1 H8-8 H8-23 H9-7 H8-13 H8-51 H9-16 H9-26 H5-12 H5-13 H8-3 H19-48 H17-11 H12-27 H8-33 H5-5 H15-26 H9-24 H5-51 H8-21 H8-31 H5-59 H8-47 H9-14 H9-25 H5-6 H8-41 H9-18 H8-40 H8-26 H12-17 H19-18 H9-56 H19-22 H19-23 H5-26 H17-32 H11-10 H12-26 H9-40 H8-55 H12-16 H11-32 H11-30 H11-36 H11-34 H11-45 H11-49 H12-39 H19-45 H12-2 H11-28 H9-70 H15-43 H17-35 H19-55 H19-13 H11-31 H19-15 H12-24 H9-41 H12-20 H19-46 H12-18 H19-29 H9-51 H11-16 H19-14 H17-34 H9-9 H8-38 H11-50 H12-40 H12-61 H12-55 H17-5 H15-19 H12-64 H15-21 H19-57 H19-50 H9-66 H19-59 H19-36 H15-8 H15-25 H12-54 H15-24 H19-56 H17-53 H9-4 H15-30 H5-55 H8-18 H15-11 H12-31 H5-60 Name of lines H8-54 H11-54 H12-36 H19-35 H11-11 H11-55 H12-48 H11-24 H12-9 H12-4 H12-7 H12-15 H19-9 H8-45 H11-27 H9-63 H12-12 H12-33 H12-42 H11-33 H17-54 H9-37 H11-21 H9-6 H11-25 H19-7 H12-13 H19-4 H8-10 H8-42 H12-28 H12-65 H15-31 H17-1 H8-52 H12-5 H5-75 H11-17 H11-19 H12-29 H5-11 H5-74 H5-18 H5-21 H17-38 H8-50 H9-15 H8-46 H12-45 H17-64 H5-29 H19-1 H8-12 H11-5 H12-49 H19-47 H8-7 H12-14 H19-16 H5-33 H8-53 H9-19 H9-21 H5-54 H17-63 H8-15 H8-35 H5-41 H11-37 H11-40 H9-22 H11-38 H17-9 H17-19 H17-14 H17-24 H17-13 H17-21 H17-49 H17-52 H17-39 H17-48 H17-62 H9-38 H17-57 H17-28 H17-45 H15-44 H5-3 H5-23 H9-12 H8-28 H5-36 H5-35 H8-39 H15-6 H15-9 H5-61 H17-60 H5-66 H5-67 H17-68 H17-66 H5-48 H5-53 H15-33 H17-36 H5-28 H5-52 H8-22 H5-42 H9-17 H5-10 H8-30 H5-63 H5-72 H11-23 H9-32 H5-15 H17-40 H5-39 H15-32 H17-18 H5-43 H8-6 H5-65 H8-34 H8-36 H12-58 H19-5 H15-13 H9-23 H15-35 H19-19 H15-40 H15-42 H15-38 H19-58 H19-62 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 Average Distance Between Clusters
  • 84. Are the parental performances correlated with the performances of their BC progenies?
  • 85. Genetic background effects in introgression breeding
  • 86. Submergence Tolerance Number of submergence tolerant plants in 9 BC2F2 and 3 BC3F2 populations under the field condition VG Donors Origin TKM9 (SS) Khazar (SS) FR13A (T) Total (I) (I) (J) I IR64 (S) India 12 8 14 34 I Teqing (SS) India 10 9 8 27 NPT (SS) J Iran 6 6 6 18 BC2F2 NPT (SS) J Iran 10 71 2 83 BC3F2 Total 38 94 30 162
  • 87. Yield responses of the 193 parental lines of IRMBP to the terminal drought under the lowland condition 25 ± -19.1±44.0% 20 15 10 5 0 -100 -80 -60 -40 -20 0 20 40 60 80 100 120 140 160 Yield reduction (in %)
  • 88. Summary of selected drought tolerant BC2F2 plants under lowland stress conditions NPT IR64 Teqing Total Total plants selected 897 2775 489 4161 # of selected plants 8.5 22.4 6.3 13.2 per population (3.8%) (10.0%) (3.0%) (6.8%) Range 0 - 85 0 - 100 0 - 30 0 - 100 No. of populations 113 124 105 320 No. of I donors 59 67 59 185 No. of J donors 32 45 19 96 Contributing donors (%) 80.5 90.3 74.3 87.8
  • 89. Parental performance and 442 selected DT BC2F2 plants for drought tolerance from 19 BC populations VG Donors Origin IR64 (S) Teqing (M) NPT(SS) Total LL (UL) LL (UL) LL I BR24 (MR) Bangladesh 14 (27) 12 (7) 3 29 (34) I STYH (S) Myanmar 20 (26) - 4 24 (26) I OM1723 (S) Vietnam 7 (17) 7 (6) 0 14 (23) J FR13A (SS) India 15 (16) 17 (15) 0 32 (31) J Type3 (S) India 23 (15) 10 (12) 0 33 (27) J Binam (M) Iran 20 (19) 14 (13) 1 35 (32) J HAN (M) China 11 (13) - 3 14 (13) I Zihui100 (S) China 8 - 9 17 J Khazar (MR) Iran 58 - 58 Total 176 (133) 60 (53) 22 256 (186)
  • 90. Comparison of different IR64 and NPT BC generations in screening for anaerobic germination (%) BC2F2 bulks BC3F2 bulks BC4F2 bulks IR64 NPT IR64 NPT IR64 NPT Total number of populations 9 10 - 10 9 10 Surviving plants/population 0 – 28 0 - 50 - 33 - 78 17 - 78 97 - 162 Number of indica donors 7 8 - 8 7 8 Selected lines (indica) 36 158 - 452 296 1038 Selection intensity (%) 5.1 19.8 - 28.3 21.1 64.9 Number of japonica donors 2 2 - 2 2 2 Selected lines (japonica) 2 8 - 35 44 121 Selection intensity (%) 2.0 8.0 - 17.5 22.0 60.5 Mean selection intensity (%) 4.4 13.9 26.1 21.3 62.7 BC2F2, BC3F2 and BC4F2 bulks all had 200 seeds in 2 replications. For individual BC populations of 200 plants, a difference of 4% between two populations in selection intensity (survival rate) is statistically significant at P < 0.05 when the selection intensity is between 0.1 and 0.5.
  • 91. Screening for seedling cold tolerance Twelve-day old seedlings were subjected to cold temperature for 18 days at the mean daily temperature of 11.8 Co, including 3-day of low temperature at 8 Co between April 24-26 (LAAS, 2002).
  • 92. Seedling Cold Tolerance (from NARES) Selection of 861 C418 plants with seedling cold tolerance from 28 C418 BC2F2 populations 2002 (LAAS) Non-CT BC2F2 CT donors donors # of populations 28 2 26 # of surviving plants 10.3% 14% 0.3 per population Range 1.4 – 19.3% 10 – 16% 0 – 3.0% # of surviving plants 10.3% 7.6% 10.5% per BC population The mean population size was 310, ranging from 196 – 465, the recipient, C418 (japonica) was killed by the stress.
  • 93. Genetic background effects on the performance of BC progenies Teqing / Chipda NPT / Chipda
  • 94. Donor and recipient effects on the number of BC2F2 plants selected under lowland drought during the 2002 dry season Donor Recurrent parent Donor Recurrent parent IR64 Teqing NPT IR64 Teqing NPT ASD 16 10*** 0 Moroberekan 13 11 ASD18 59*** 0 54*** MR 77 31**** 0 B4122 37*** 2 1 Palung 2 33 36 Budda 75*** 11 Pokhreli 119**** 7 30*** Chipda 47*** 0 85**** Pusa 15** 4 Chorofa 1 20*** Rasi 63*** 9 Dacca 6 20 19 Rusty Late 38**** 2 4 Dhan4 1 0 Sadajira 19 55**** 0 Doddi 81*** 2 Shwewartun 5* 0 6** Gajale 61*** 22 SLG-1 26**** 0 Giza 14 29*** 4 SML242 6** 0 5*
  • 95. Genetic background effects on the number of survival plants under submergence in 33 BC4F2 populations from crosses between 3 RPs and 11 donors Cross Survival (%) Z value Cross Survival(%) Z value IR64/SN89366 3.33 1.87 NPT/C418 0.00 Teqing/SN89366 6.67 3.75 IR64/CH448 9.33 4.14 NPT/SN89366 0.00 Teqing/CH448 7.33 3.25 IR64/Y134 7.33 3.45 NPT/CH448 0.00 Teqing/Y134 7.33 3.45 IR64/FR13A 7.67 3.64 NPT/Y134 0.00 Teqing/FR13A 6.67 3.17 IR64/BR24 8.00 3.61 NPT/FR13A 0.00 Teqing/BR24 8.00 3.61 IR64/Madhukar 7.33 3.60 NPT/BR24 0.00 Teqing/Madhukar 6.00 2.95 IR64/Zihui 100 11.67 4.87 NPT/Madhukar 0.00 Teqing/Zihui 100 7.33 3.06 IR64/IR50 6.67 3.13 NPT/Zihui 100 0.00 Teqing/IR50 8.00 3.76 IR64/IR72 8.00 0.45 NPT/IR50 0.00 Teqing/IR72 8.00 0.45 IR64/Jhona349 5.00 2.73 NPT/IR72 7.00 Teqing/Jhona349 5.67 3.09 IR64/C418 7.33 3.19 NPT/Jhona349 0.00 Teqing/C418 10.00 4.35
  • 96. Genetic background effects on the number of survival plants under submergence in 33 BC4F2 populations from crosses between 3 RPs and 11 donors Cross Survival (%) Z value Cross Survival(%) Z value IR64/SN89366 3.33 1.87 NPT/C418 0.00 Teqing/SN89366 6.67 3.75 IR64/CH448 9.33 4.14 NPT/SN89366 0.00 Teqing/CH448 7.33 3.25 IR64/Y134 7.33 3.45 NPT/CH448 0.00 Teqing/Y134 7.33 3.45 IR64/FR13A 7.67 3.64 NPT/Y134 0.00 Teqing/FR13A 6.67 3.17 IR64/BR24 8.00 3.61 NPT/FR13A 0.00 Teqing/BR24 8.00 3.61 IR64/Madhukar 7.33 3.60 NPT/BR24 0.00 Teqing/Madhukar 6.00 2.95 IR64/Zihui 100 11.67 4.87 NPT/Madhukar 0.00 Teqing/Zihui 100 7.33 3.06 IR64/IR50 6.67 3.13 NPT/Zihui 100 0.00 Teqing/IR50 8.00 3.76 IR64/IR72 8.00 0.45 NPT/IR50 0.00 Teqing/IR72 8.00 0.45 IR64/Jhona349 5.00 2.73 NPT/IR72 7.00 Teqing/Jhona349 5.67 3.09 IR64/C418 7.33 3.19 NPT/Jhona349 0.00 Teqing/C418 10.00 4.35
  • 97. Can we develop ILs with extreme phenotypes for selected target traits?
  • 98. Screening results of 11 BC2F4 backcross populations derived from crosses between a japonica variety, Chaoyou 1 (the recurrent parent) and 11 donors for cold tolerance at the booting stage and for heat tolerance at the flowering stage Selection for heat tolerance at the flowering stage Selection for cold tolerance at the booting stage Seed set (%) Spikelets per panicle Seed set (%) Donor 1 N SI (%) 2 Mean 3 Range N SI (%) Mean 3 Range Mean 3 Range Bg90-2 (I) 41 9.11 63.3 d 50.3 - 86.5 25 6.25 41.3 abc 23.0–77.0 171.2 abc 124.0–253.1 9 2.25 38.1 abcd 19.5–50.0 192.5 ab 143.3–275.5 X21 (I) 29 6.44 64.3 cd 50.6 - 87.1 X22 (I) 28 6.22 65.6 bcd 50.7 - 87.3 - - - - - - Q5 (I) 31 6.89 71.1 abc 50.9 - 91.4 6 1.50 41.1 abc 19.9–75.4 160.4–274.2 Chhomrong (J) 24 5.33 51.4 - 87.8 17 4.25 37.7 abcd 19.3–65.5 172.9 abc 115.9–258.8 Doddi (I) 25 5.56 71.0 abc 50.2 - 90.0 - - - - - - Feng-Ai-Zhan (I) 44 9.78 52.2 - 98.5 12 3.00 31.3–70.1 180.7 abc 138.2–238.0 9 2.25 28.9 d 17.0–44.4 171.5 abc 112.0–216.7 Shennong265 (J) 21 4.67 69.2 abcd 50.7 - 93.8 25 6.25 45.5 ab 23.9–65.6 83.2–255.4 Yuangeng7 (J) 41 9.11 71.1 abc 50.0 - 90.1 13 3.25 33.0 cd 17.0–48.9 171.9 abc 111.6–230.4 OM997 (I) 21 4.67 72.1 ab 54.9 - 89.6 8 2.00 33.6 abcd 24.9–48.3 175.5 abc 121.4–280.4 Cs94 (I) 19 4.22 64.2 cd 51.4 - 86.0 124 3.44 5.2 e 0.0–7.9 157.4 bc 127.4–178.6 Chaoyou (J) 324 6.55 24.8 e 19.0 - 30.0 2N is the number of cold tolerant or heat tolerant BC plants selected from each population and SI = selection intensity. 3 Different letters indicate the statistical significance in seed set at P < 0.05, based on the Duncan testing of ANOVA.
  • 99. Table 6 Performances for AG of 11 promising BC4F3 lines and their donors Recipient Donor Seedling height (cm) AG (%) Emerging at 10 d (%) NPT Khazar 33.5 90.0* 20.0 NPT Khazar 32.5 95.0** 75.0** NPT FR13A 37.7 95.0** 30.0 NPT TKM 9 37.7 100.0*** 95.0*** NPT TKM 9 36.2 100.0*** 60.0* NPT TKM 9 37.9 100.0*** 95.0*** NPT Babaomi 34.9 100.0*** 50.0 NPT Babaomi 36.7 100.0*** 50.0 NPT OM1706 33.9 100.0*** 65.0* NPT OM1706 36.1 100.0*** 80.0*** TKM 9 I 31.4 20.0 12.7 Khazar J 30.3 3.0 0.0 Babaomi I 26.3 5.0 0.0 Jiangxi-Si-Miao I 31.5 9.0 0.0 OM1706 I 29.1 18.0 2.0 IR64 I 26.5 20.0 2.0 NPT J 36.2 68.0 39.0 I = indica and J = japonica. Traits were measured at 21 d after seeding. All 10 ILs had significantly higher AG than the recurrent parent, NPT at P < 0.001.
  • 100. Trait specific introgression lines developed # of BC2F2 No. of selected Target traits populations BC2F3 lines Drought tolerance 350 4687 BPH resistance 203 522 Salinity tolerance 203 1022 Anaerobic germination 130 368 Zinc deficiency 129 1211 Submergence tolerance 264 798 Grain quality 65 580 Other traits 375 12,000+ Total 20,000+
  • 101. Summary of Selection Experiments • Most donors contributed performance enhancing alleles to their BC progenies regardless of their performances; • Appropriate screening (selection) is the key to identify improved target traits in the BC progenies; • More distantly related donors, particularly landraces, tend to give more transgressive segregations for abiotic tolerance in the BC progenies.
  • 102. Conclusions There are tremendous amounts of hidden genetic diversity in the current rice germplasm collections for genetic improvement of all target traits we tried, which have not been exploited; Selection of parental lines for breeding based on phenotype practiced by most breeders is a poor way in exploiting novel genetic variation for complex traits; Backcross breeding, effective selection (efficient screening) combined with DNA markers are the effective way to exploit this hidden diversity;
  • 103. What are we going to do with this large number of ILs? 1. Direct development of new cultivars; 2. As genetic stocks for discovery of DT alleles or QTLs and functional genomics of DT; 3. As parents for development of superior rice cultivars by QTL pyramiding
  • 104. Progeny Testing of DT ILs Under Stress (no irrigation after transplanting) DT C418 ILs Check (C418)
  • 105. Progeny Testing Under Stress (no irrigation after transplanting) Promising DT C418 ILs
  • 106. 2001-2002 DS BC2F4 progeny testing IR64 ILs for quality IR64 (CK) DT IR64 ILs
  • 107. Performance of two DT NPT ILs under severe stress DT NPT ILs NPT CK
  • 108. Performance of some promising salinity tolerant IR64 ILs in the field condition in Iloilo (EC 18 dSm-1 ) 11/17, 2003
  • 109. Performance of some promising salinity tolerant IR64 ILs in the field condition in Iloilo (EC 18 dSm-1 ) 11/17, 2003
  • 110. Promising varieties developed in the BC breeding program
  • 111. The Recurrent Parents C418 Liaojing454 (restorer)
  • 112. Preliminary Yield Trials of Promising DT/WUE ILs 30% water saving 70% 50% water water saving saving
  • 113. Replicated Preliminary yield trial of DT/WUE ILs (Shengyang/2006.9) Completely Water rainfed saving 70% Water Water saving saving 50% 30%