R de Prado

University
of
Córdoba

Roundup Ready ® Maize symposium: The
European Perspective

Weed Resistance to Glyphosate in Southern Europe:
Present and Future Perspective.

Dr. Rafael De Prado and Fidel González-Torralva

University of Cordoba, Spain

Main crops produced in the Mediterranean Region

Roundup Ready® Maize symposium: The European Perspective

University
of SUSTAINABLE AGRICULTURE
Córdoba

Is that in which the production systems permit the

obtainment of a continuous benefit from the use of

water, soil, genetic resources, etc. to meet the

current needs of the population without destroying

the basic natural resources for future generations.

Castillo, 1992


University
of
Córdoba
Evolution of herbicides use in Spain

70

60

50

40 1999
2004
30
2009

20

10

0
Wheat Olive Stone fruit Vineyard Citrus


University
of
Córdoba RESISTANCE

The Weed Science Society of America (WSSA)
defines herbicide resistance as

“the inherited ability of a plant to survive and
reproduce following exposure to a dose of
herbicide normally lethal to the wild type”.


Distribution of Resistant Weed Biotypes in Southern Europe


Resistance mechanisms in weeds
University Non target site
of
Córdoba
Foliar surface
Absorption

Cuticule

Apoplast

Systematization
Translocation

Metabolization
Herbicide
application Resistant Plant


Resistance mechanisms in weeds
University Target site
of Córdoba

Target site
modification

Overexpression

Herbicide
application Resistant Plant

University
of
MULTIPLE RESISTANCE
Córdoba

ALS INHIBITORS ACCase INHIBITORS PSII INHIBITORS

SUSCEPTIBLE
BIOTYPE

DETOXIFICATION SYSTEM

RESISTANT
BIOTYPE
STOP ACCase STOP PSII


Evolution of ALS Resistance in Southern Europe

30
University
of 25

Number of resistant species
Córdoba
20

15

10
Evolution of PS II (C1) Resistance in Southern Europe
5 World: 107
70 Southern Europe : 26
0
60
1990 1995 2000 2005 2010

50 Year

40

30
Evolution of ACCase Resistance in Southern Europe
20 30
World: 68
10 Southern Europe : 59 25

0
20
1975 1980 1985 1990 1995 2000
Year
15

10

5 World: 38
Southern Europe : 26
0
1990 1995 2000 2005 2010
Year


Evolution of Glyphosate Resistance in Southern Europe
University
of
Córdoba

6

5
RESISTANT SPECIES

4

3

2

1

0
2002 2003 2004 2005 2006 2007 2008 2009 2010 2011
YEAR

World: 18
Southern Europe : 5

WORST WEEDS IN THE MEDITERRANEAN REGION: ORCHARDS AND OLIVE TREES

Ecbalium elaterium
Conium maculatum
Conyza spp

Cuscuta campestris Euphorbia helioscopia Malva parviflora

WORST WEEDS IN THE MEDITERRANEAN REGION: ORCHARDS AND OLIVE TREES
REGION:

Avena sterilis Paspalum distichum

Lolium multiflorum Lolium rigidum Cyperus rotundus

Evolution of Glyphosate Resistance in Southern Europe
University
of
Córdoba

Main differences between glyphosate R and S Lolium
multiflorum biotypes.

R/S factor depends of growth stage.

Shikimate does not accumulate in R biotypes.

Absorption and translocation of 14C-glyphosate in S is higher than R
biotypes.

Spray retention is higher in S biotypes.

Mutations in EPSPS have been founded in R biotypes.


University
of
Córdoba

ED50 Evolution of Lolium multiflorum
treated with glyphosate
1500 R
S
1000
-1
g ai ha

500

0
13 21 55-55 65
Growth Stage (BBCH)


Shikimic Acid (Shikimate) Pathway
University
of
Córdoba

AShikimate accumulation in L.
multiflorum biotypes

1,6
Shikimic acid (mg g-1 fresh weight)
)

1,4

1,2

1

0,8
R Biotype
0,6
S Biotype
0,4

0,2

0
0 24 48 72 96
Hours after treatment

Retention and contact angle in L. multiflorum treated with
University glyphosate
of
Córdoba

Adaxial Surface
µL spraying solution
g-1 dry matter Angle (degrees)
S R S R
1,763b 1,265a
62,4 (2,83) 112,9 (5,00)
Major of 90º Minor of 90º
θ = 180 – tan-1[(x.b2)/(y.a2)] θ = 2.tan-1(h/x)

a

b
x

y
θ

Unpublished data, 2010


Wax differences between R and S L. multiflorum biotypes
University
of
Córdoba

2µm²

CAD software quantification

Area Covered by Wax (%)
S R
42,9 (0,66) 102,6 (2,38)


Absorption and translocation of 14C-Glyphosate in R and S biotypes of
University glyphosate resistant L. multiflorum.
of
Córdoba

48 hours after 14C-Glyphosate
application
R S

Protein Alignment of EPSPS Glyphosate Resistant
University
of
Lolium spp.
Córdoba

*Arrows indicate amino acid mutations sites

Glyphosate (2250 g ai ha-1) treatment in olive orchard
University with glyphosate resistant L. multiflorum
of
Córdoba

0 DAT 15 DAT

30 DAT 75 DAT

University
of
Efficacy (%) of herbicides mixture with glyphosate applied in
Córdoba
glyphosate-resistant L. multiflorum.

DAT
Herbicide
15 30 45 60 75
Untreated 0,0±0,0c 0,0±0,0c 0,0±0,0c 0,0±0,0c 0,0±0,0c
G1125 54,0±12,3b 47,4±9,2b 47,3±7,3b 42,1±6,2b 40,5±6,4b
G2250 61,2±8,7bc 67,4±6,1bc 62,7±10,2bc 60,2±7,1bc 57,2±7,3b
G1125+O 58,2±9,2b 62,8±8,0bc 53,1±9,3b 52,1±6,8b 52,2±10,4b
G1125+F 49,1±8,3b 99,1±2,7a 100,0±0,0a 100,0±0,0a 100,0±0,0a
G1125+C 79,1±6,3ab 100,0±0,0a 100,0±0,0a 100,0±0,0a 100,0±0,0a
G1125+Q 71,1±4,0ab 100,0±0,0a 100,0±0,0a 100,0±0,0a 100,0±0,0a

G=glyphosate (g ae ha-1); O=orosorb (0,8% v/v); F=flazasulfuron (50 g ai ha-1);
C=clethodim (120 g ai ha-1) and Q=quizalofop (50 g ai ha-1).

Glyphosate at 2160 g ai ha-1

30 DAT 45 DAT
0 DAT

60 DAT Control 60 DAT

Visual assessment of different herbicide treatments on Conyza spp.

DAYS AFTER TREATMENT
TREATMENT 15 30 45 60
A 0,0 ± 0,0 0,0 ± 0,0 0,0 ± 0,0 0,0 ± 0,0
B 71,7 ± 12,6 80,0 ± 5,0 80,0 ± 5,0 85,0 ± 13,2
C 85,0 ± 8,7 100,0 ± 0,0 100,0 ± 0,0 100,0 ± 0,0
D 18,3 ± 10,4 30,0 ± 5,0 41,7 ± 7,6 56,7 ± 5,8
E 26,7 ± 5,8 70,0 ± 5,0 80,0 ± 10,0 93,3 ± 2,9
F 18,3 ± 2,9 20,0 ± 10,0 20,0 ± 5,0 31,7 ± 7,6
G 60,0 ± 5,0 60,0 ± 10,0 65,0 ± 5,0 70,0 ± 10,0
H 55,0 ± 10,0 81,7 ± 10,4 85,0 ± 5,0 88,3 ± 7,6
I 40,0 ± 0,0 70,0 ± 10,0 76,7 ± 2,9 96,7 ± 2,9
J 68,3 ± 7,6 88,3 ± 7,6 96,7 ± 2,9 100,0 ± 0,0
K 60,0 ± 5,0 68,3 ± 2,9 71,7 ± 2,9 70,0 ± 17,3
A Control
B Glyphosate 2160 g ai ha-1
C Glyphosate 4320 g ai ha-1
D Flazasulfuron 200 g ha-1
E Glyphosate 2160 g ai ha-1+ Flazasulfuron 200 g ha-1
F Saflufenacil 20 g ha-1+ Dash 0,75%
G Glyphosate 2160 g ai ha-1+ Saflufenacil 20 g ha-1 + Dash 0,75%
Glyphosate 2160 g ai ha-1 + Saflufenacil 20 g ha-1 + Dash 0,75%
H
** Saflufenacil 20 g ha-1 + Dash 0,75%
I Fusta 8 L ha-1
J Fusta 8 L ha-1 + glyphosate 720 g ai ha-1
K Glyphosate 2160 g ai ha-1 + Saflufenacil 20 g ha-1 + Aceite 1 %

University “Would it be possible to have a sustainable crop
of
Córdoba
management in the future using herbicides?”

Good agricultural …We are
more than
6,500
practices… millions of
people

University
of
Córdoba Innovation in Agro-Technologies are
Key-Factors to Satisfy Future Food Demand

Trillion Kilocalories /Year
7.37 2.13 11.28
10.6 0.52 Reserve

10.0
2.29

1.72

5.1 5.17 1.23
5.0 -0.13
Improvements
-1.12

Demand Supply Demand Arable Conversion Gap Fertilizer Irrigation Traditional Bio- Supply
1995 2025 Land Loss Crop technology 2025
Protection and
Seeds
Source: Kern, M., 1998

Future of Agriculture fig. 2

Summarize
University
of
Córdoba

- Resistance to herbicides is wide spread in Europe as well as globally ( see www.weedscience.org ).
www.weedscience.

- Confirmed glyphosate resistant weed biotypes of Lolium rigidum, Lolium multiflorum, Conyza canadensis
rigidum, multiflorum,
and Conyza bonariensis are known in Europe, these resistant biotypes are exclusively spread in perennial
crops like vineyards, citrus plantations and olive groves. They were selected because there was a sole
groves.
reliance on glyphosate in these crops to control weeds.
weeds.

- Glyphosate resistant weed biotypes can be managed by: by:
•glyphosate applications when the plants are most susceptible
•complementary cultural practices, like tillage or mowing
•herbicide sequences and/or tank-mixes
tank-

-Mixture of auxinic herbicides alone and/or with glyphosate are a good solution to control dicotiledonea
weeds.
weeds.

-ALS and ACCase inhibitor herbicides mixture with glyphosate must be managed with care. In terms of
care.
resistance development they can be named “High Risk Herbicides.
Herbicides.

THANK YOU FOR YOUR
ATTENTION

R de Prado

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