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University

of

Córdoba

Weed Resistance to Glyphosate in Southern Europe:

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

SUSTAINABLE AGRICULTURE

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

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of

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

current needs of the population without destroying

the basic natural resources for future generations.

Castillo, 1992


Evolution of herbicides use in Spain

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of

Córdoba

50

60

70

0

10

20

30

40

Wheat Olive Stone fruit Vineyard Citrus

1999

2004

2009


RESISTANCE

The Weed Science Society of America (WSSA)

defines herbicide resistance as

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of

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“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


Systematization

Foliar surface

Cuticule

Apoplast

Absorption

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of

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Resistance mechanisms in weeds

Non target site

Herbicide

applicationResistant Plant

Metabolization

Translocation

Systematization


Target site

modification

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of Córdoba

Resistance mechanisms in weeds

Target site

Overexpression

Herbicide

application Resistant Plant

SUSCEPTIBLE

PSII INHIBITORSALS INHIBITORS ACCase INHIBITORS

MULTIPLE RESISTANCEUniversity

of

Córdoba

SUSCEPTIBLE

BIOTYPE

DETOXIFICATION SYSTEM

STOP ACCase STOP PSII

RESISTANT

BIOTYPE


Nu

mb

er

of

res

ista

nt

sp

ec

ies

40

50

60

70

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of

Córdoba

Evolution of PS II (C1) Resistance in Southern Europe

Evolution of ALS Resistance in Southern Europe

Year

1990 1995 2000 2005 2010

Nu

mb

er

of

res

ista

nt

sp

ec

ies

0

5

10

15

20

25

30

World: 107Southern Europe : 26

Year

1975 1980 1985 1990 1995 2000

Nu

mb

er

of

res

ista

nt

sp

ec

ies

0

10

20

30

40



Evolution of ACCase Resistance in Southern Europe

Year

1990 1995 2000 2005 2010

Nu

mb

er

of

res

ista

nt

sp

ec

ies

0

5

10

15

20

25

30


Evolution of Glyphosate Resistance in Southern EuropeUniversity

of

Córdoba

3

4

5

6

RESIS

TAN

T SPEC

IES

World: 18

Southern Europe : 5


0

1

2

3

2002 2003 2004 2005 2006 2007 2008 2009 2010 2011

RESIS

TAN

T SPEC

IES

YEAR

Ecbalium elateriumConium maculatum

WORST WEEDS IN THE MEDITERRANEAN REGION: ORCHARDS AND OLIVE TREES

Cuscuta campestris Malva parviflora

Conyza spp

Conium maculatum

Euphorbia helioscopia

Avena sterilis Paspalum distichum

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

Lolium multiflorum Cyperus rotundusLolium rigidum

Evolution of Glyphosate Resistance in Southern Europe

� R/S factor depends of growth stage.

�Shikimate does not accumulate in R biotypes.

Main differences between glyphosate R and S Lolium

multiflorum biotypes.

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of

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�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.


ED50 Evolution of Lolium multiflorum

treated with glyphosate

1500 R

S

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of

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0

500

1000

13 21 55-55 65

Growth Stage (BBCH)

g a

i h

a-1

S


Shikimic Acid (Shikimate) Pathway

1,6)

AShikimate accumulation in L.

multiflorum biotypes

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of

Córdoba

0

0,2

0,4

0,6

0,8

1

1,2

1,4

1,6

0 24 48 72 96

Sh

ikim

ic a

cid

(mg

g-1

fres

hw

eig

ht)

Hours after treatment

R Biotype

S Biotype

S R

62,4 (2,83) 112,9 (5,00)

Angle (degrees)

Adaxial SurfaceAdaxial Surface

Retention and contact angle in L. multiflorum treated with glyphosate

µL spraying solution

g-1 dry matter

S R

1,763b 1,265a

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of

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MinorMinor of 90ºof 90º

θθ = 2.tan= 2.tan--11(h/x)(h/x)

Major of 90º

θ = 180 – tan-1[(x.b2)/(y.a2)]

62,4 (2,83) 112,9 (5,00)

θ

bx

y

a

Unpublished data, 2010


1,763b 1,265a

2µm²

CAD software quantification

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of

Córdoba

Wax differences between R and S L. multiflorum biotypes

CAD software quantification

S R

42,9 (0,66) 102,6 (2,38)

Area Covered by Wax (%)


R S

48 hours after 14C-Glyphosate

application

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of

Córdoba

Absorption and translocation of 14C-Glyphosate in R and S biotypes of

glyphosate resistant L. multiflorum.

Protein Alignment of EPSPS Glyphosate Resistant Lolium spp.University

of

Córdoba

*Arrows indicate amino acid mutations sites


Glyphosate (2250 g ai ha-1) treatment in olive orchard with glyphosate resistant L. multiflorumUniversity

of

Córdoba

0 DAT

30 DAT 75 DAT

15 DAT

HerbicideDAT

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

Efficacy (%) of herbicides mixture with glyphosate applied in

glyphosate-resistant L. multiflorum.

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of

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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).

0 DAT

Glyphosate at 2160 g ai ha-1

30 DAT 45 DAT0 DAT 30 DAT 45 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 ± 10,0G 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%

HGlyphosate 2160 g ai ha-1 + Saflufenacil 20 g ha-1 + Dash 0,75%

** 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 %

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

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of

Córdoba

…We are

more than

6,500

millions of

people

Good agricultural

practices…

10.0

Trillion Kilocalories /Year

10.6

11.28

2.29

2.137.37

Reserve0.52

1.72

Innovation in Agro-Technologies are

Key-Factors to Satisfy Future Food Demand

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of

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5.0

Fertilizer TraditionalCrop

Protection and Seeds

Irrigation Bio-technology

5.1 5.17

ArableLand

-0.13

Conversion Loss

-1.12

Improvements

1995Demand Supply

2025

Demand Gap

1.23

2025

Supply

Source: Kern, M., 1998

Future of Agriculture fig. 2

Summarize

-- ResistanceResistance toto herbicidesherbicides isis widewide spreadspread inin EuropeEurope asas wellwell asas globallyglobally (( seesee wwwwww..weedscienceweedscience..orgorg ))..

-- ConfirmedConfirmed glyphosateglyphosate resistantresistant weedweed biotypesbiotypes ofof LoliumLolium rigidumrigidum,, LoliumLolium multiflorummultiflorum,, ConyzaConyza canadensiscanadensisandand ConyzaConyza bonariensisbonariensis areare knownknown inin Europe,Europe, thesethese resistantresistant biotypesbiotypes areare exclusivelyexclusively spreadspread inin perennialperennialcropscrops likelike vineyards,vineyards, citruscitrus plantationsplantations andand oliveolive grovesgroves.. TheyThey werewere selectedselected becausebecause therethere waswas aa solesolereliancereliance onon glyphosateglyphosate inin thesethese cropscrops toto controlcontrol weedsweeds..

-- GlyphosateGlyphosate resistantresistant weedweed biotypesbiotypes cancan bebe managedmanaged byby::••glyphosateglyphosate applicationsapplications whenwhen thethe plantsplants areare mostmost susceptiblesusceptible••complementarycomplementary culturalcultural practices,practices, likelike tillagetillage oror mowingmowing

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••complementarycomplementary culturalcultural practices,practices, likelike tillagetillage oror mowingmowing••herbicideherbicide sequencessequences and/orand/or tanktank--mixesmixes

--MixtureMixture ofof auxinicauxinic herbicidesherbicides alonealone and/orand/or withwith glyphosateglyphosate areare aa goodgood solutionsolution toto controlcontrol dicotiledoneadicotiledoneaweedsweeds..

--ALSALS andand ACCaseACCase inhibitorinhibitor herbicidesherbicides mixturemixture withwith glyphosateglyphosate mustmust bebe managedmanaged withwith carecare.. InIn termsterms ofofresistanceresistance developmentdevelopment theythey cancan bebe namednamed “High“High RiskRisk HerbicidesHerbicides..

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