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PHENOTYPIC AND GENOTYPIC EVALUATIONS FOR CBB
   RESISTANCE IN COMMON BEAN POPULATIONS




       L. Kachulu, M.S. Mwala, R. Chirwa and L. Madubanya

  10th African Crop Science Society Conference 10-13 Oct, 2011, Maputo,
                              Mozambique
Bean Production areas in
        Africa
                       Expanding to
                       tropical
                       lowland areas
                       of western
                       Africa




                      Source: Bean
                      Atlas, 1998
Major production biotic constraints of
beans in Africa (Wortmann, et al. 1998)


       Constraint      Yield loss ton (p.a.)
  Angular leaf spot          384,200
  Anthracnose                328,000
  Bean stem maggot           297,100
  Root rots                  221,100
  CBB                        220,400
Common bacterial blight (CBB)


   CBB –a worldwide seed-
    borne disease of common
    bean- yield loss of up to
    40%.
   Contaminated seed (internal
    or external) acts as primary
    source of inoculum can
    survive for extended period
    10 years
   Quantitative trait controlled
    by more than one gene of
    QTLs
Breeding for CBB resistance

   •   CBB resistance breeding -constrained by the
       instability of resistance because of its
       quantitative nature
   •   Despite the instability-resistance QTLs have
       been introgressed into breeding lines
   •   Markers (SCARs) for these QTLs are
       available and employed in MAS as a way to
       improve the selection of cultivars with CBB
       resistance
Breeding for CBB resistance

   SCAR markers BC420, SU91, and SAP6 are tightly-
    linked with three major QTLs on chromosomes, B6, B8,
    and B10 respectively.

   Different chromosomal positions of these SCAR
    markers makes them attractive sources for introgressing
    independent QTLs conditioning resistance to CBB into
    susceptible bean lines
Objective of the study

   The study was carried out to evaluate the
    reaction of F4.6 lines to CBB and validate the
    presence of SCAR markers SU91700,
    SAP6820 and BC420900
Materials and Methods

    The crosses were made from CBB resistant
    sources of Meso (VAX3, VAX6 ) and Andean
    (RMX2, RMX19 & RMX20), and susceptible
    recipient parents of Andean origin

   Exp1: Phenotypic screening for CBB
    resistance- Conducted in the G/house using
    isolates Xf260 and Xf410. Fully expanded
    trifoliate leaves were inoculated using multiple
    needle method and scored at 1-9 CIAT scale after
Materials and Methods

   Exp2: Genotypic screening for CBB
    resistance-DNA was extracted from the
    greenhouse plants, template DNA was used in
    PCR reaction to amplify SCAR markers SU91,
    SAP6 and BC420.

   Presence or absence of the resulting fragments
    for each marker was determined using agarose
    gel electrophoresis
Results :
                            EXP1:Phenotypic screening-
Level of resistance for CBB in the populations were intermediate.
Average population scores of 5 and 6- due to segregation
Results :
CBB scores for the advanced lines within populations
revealed presence of some resistant (1-3) genotypes,
as well as intermediate and susceptible
Population    Number of lines and reaction to CBB
              Resistant (1-3)   Intermediate (4-6) Susceptible (7-9)
BRB211/VAX3          3                 19                  4
BRB214/VAX3          10                17                  9
BRB215/VAX6          0                  7                  3
BRB265/VAX6          6                  8                  6
CMB107/RMX2          0                  6                 10
RMA70/RMX20          0                 13                  6
RMA72/VAX6           21                45                 22
BRB264/VAX3          3                  5                 10
RMA72/RMX19          0                  9                 18
Results from Exp. 2

Parent   SU91   SAP6   BC420
VAX 3      +      +       _
VAX6       +      +       _
                               • SU91 marker was
                                 present in the VAX
RMX2        _     +       _
                                 parents,
RMX19       _     +       _
                               • Andean parents
BRB211      _     +       _      possessed SAP6.
BRB214      _     _       _    • SAP6 was also
BRB215      _             _      present in some
BRB265      _      +      _      susceptible
                                 cultivars.
CMB107      _      _      _
                               • None of the
RMA70       _      _      _      parents carried
RMA72       _      +      _      the BC420 marker
BRB264      _      _      _
The SCAR markers were easily scored as present (+)
   or absent (-) of a single band on agarose gel.

Population     -SAP6/-   SAP6   SU91   +SAP6/
               SU91                    +SU91
                                                Some resistant genotypes
*BRB211/VAX3
                  1               2
                                                lacked both SCAR markers
BRB214/VAX3                      10
                                                SU91 marker was associated
BRB264/VAX3       1               2             with genotypes that showed
RMA72/VAX6                        8      13     higher levels of phenotypic
                                                resistance to CBB.
BRB265/VAX6                       6

*BRB264/VAX3      2        7              1     SAP6 was found in
RMA70/RMX20       3        3                    susceptible genotypes
CMB107/RMX2       3        7
                                                *Resistant genotypes
RMA72/RMX19       9        9
                                                *Susceptible genotypes
RMA72/VAX6        8        7
Conclusion...

   The findings of this study show that, 43 F4.6 lines
    were resistant to CBB. These will further be
    used as donor parental lines in the breeding
    programmes or evaluated for yield perfomance

   28 of the resistant lines possessed SU91 , 2
    lacked both markers and 13 had a combination
    of the two markers i.e. SU91 and SAP6.
Conclusion...
   The findings of the study provide a basis for
    advancing lines with improved levels of
    resistance to CBB and reducing the number of
    lines that need to be verified in the field
   Given the absence of BC420 marker in the
    study populations, resistance could be
    enhanced further by crossing with resistance
    sources carrying the BC420marker
   Need to find other markers for resistance to
    CBB other than SU91, SAP6 or BC420
THANK YOU FOR
LISTENING

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Evaluating CBB Resistance in Common Bean Using Markers

  • 1. PHENOTYPIC AND GENOTYPIC EVALUATIONS FOR CBB RESISTANCE IN COMMON BEAN POPULATIONS L. Kachulu, M.S. Mwala, R. Chirwa and L. Madubanya 10th African Crop Science Society Conference 10-13 Oct, 2011, Maputo, Mozambique
  • 2. Bean Production areas in Africa Expanding to tropical lowland areas of western Africa Source: Bean Atlas, 1998
  • 3. Major production biotic constraints of beans in Africa (Wortmann, et al. 1998) Constraint Yield loss ton (p.a.) Angular leaf spot 384,200 Anthracnose 328,000 Bean stem maggot 297,100 Root rots 221,100 CBB 220,400
  • 4. Common bacterial blight (CBB)  CBB –a worldwide seed- borne disease of common bean- yield loss of up to 40%.  Contaminated seed (internal or external) acts as primary source of inoculum can survive for extended period 10 years  Quantitative trait controlled by more than one gene of QTLs
  • 5. Breeding for CBB resistance • CBB resistance breeding -constrained by the instability of resistance because of its quantitative nature • Despite the instability-resistance QTLs have been introgressed into breeding lines • Markers (SCARs) for these QTLs are available and employed in MAS as a way to improve the selection of cultivars with CBB resistance
  • 6. Breeding for CBB resistance  SCAR markers BC420, SU91, and SAP6 are tightly- linked with three major QTLs on chromosomes, B6, B8, and B10 respectively.  Different chromosomal positions of these SCAR markers makes them attractive sources for introgressing independent QTLs conditioning resistance to CBB into susceptible bean lines
  • 7. Objective of the study  The study was carried out to evaluate the reaction of F4.6 lines to CBB and validate the presence of SCAR markers SU91700, SAP6820 and BC420900
  • 8. Materials and Methods  The crosses were made from CBB resistant sources of Meso (VAX3, VAX6 ) and Andean (RMX2, RMX19 & RMX20), and susceptible recipient parents of Andean origin  Exp1: Phenotypic screening for CBB resistance- Conducted in the G/house using isolates Xf260 and Xf410. Fully expanded trifoliate leaves were inoculated using multiple needle method and scored at 1-9 CIAT scale after
  • 9. Materials and Methods  Exp2: Genotypic screening for CBB resistance-DNA was extracted from the greenhouse plants, template DNA was used in PCR reaction to amplify SCAR markers SU91, SAP6 and BC420.  Presence or absence of the resulting fragments for each marker was determined using agarose gel electrophoresis
  • 10. Results : EXP1:Phenotypic screening- Level of resistance for CBB in the populations were intermediate. Average population scores of 5 and 6- due to segregation
  • 11. Results : CBB scores for the advanced lines within populations revealed presence of some resistant (1-3) genotypes, as well as intermediate and susceptible Population Number of lines and reaction to CBB Resistant (1-3) Intermediate (4-6) Susceptible (7-9) BRB211/VAX3 3 19 4 BRB214/VAX3 10 17 9 BRB215/VAX6 0 7 3 BRB265/VAX6 6 8 6 CMB107/RMX2 0 6 10 RMA70/RMX20 0 13 6 RMA72/VAX6 21 45 22 BRB264/VAX3 3 5 10 RMA72/RMX19 0 9 18
  • 12. Results from Exp. 2 Parent SU91 SAP6 BC420 VAX 3 + + _ VAX6 + + _ • SU91 marker was present in the VAX RMX2 _ + _ parents, RMX19 _ + _ • Andean parents BRB211 _ + _ possessed SAP6. BRB214 _ _ _ • SAP6 was also BRB215 _ _ present in some BRB265 _ + _ susceptible cultivars. CMB107 _ _ _ • None of the RMA70 _ _ _ parents carried RMA72 _ + _ the BC420 marker BRB264 _ _ _
  • 13. The SCAR markers were easily scored as present (+) or absent (-) of a single band on agarose gel. Population -SAP6/- SAP6 SU91 +SAP6/ SU91 +SU91 Some resistant genotypes *BRB211/VAX3 1 2 lacked both SCAR markers BRB214/VAX3 10 SU91 marker was associated BRB264/VAX3 1 2 with genotypes that showed RMA72/VAX6 8 13 higher levels of phenotypic resistance to CBB. BRB265/VAX6 6 *BRB264/VAX3 2 7 1 SAP6 was found in RMA70/RMX20 3 3 susceptible genotypes CMB107/RMX2 3 7 *Resistant genotypes RMA72/RMX19 9 9 *Susceptible genotypes RMA72/VAX6 8 7
  • 14. Conclusion...  The findings of this study show that, 43 F4.6 lines were resistant to CBB. These will further be used as donor parental lines in the breeding programmes or evaluated for yield perfomance  28 of the resistant lines possessed SU91 , 2 lacked both markers and 13 had a combination of the two markers i.e. SU91 and SAP6.
  • 15. Conclusion...  The findings of the study provide a basis for advancing lines with improved levels of resistance to CBB and reducing the number of lines that need to be verified in the field  Given the absence of BC420 marker in the study populations, resistance could be enhanced further by crossing with resistance sources carrying the BC420marker  Need to find other markers for resistance to CBB other than SU91, SAP6 or BC420