Fecha actualización

Transgenic alfalfa tolerant to

herbicides, from Lab to Field

Gabriela C. Soto

Weeds problematic

• 120 species of weeds worldwide.

• Important productive losses (50% -80%).

• Low forage quality.

• No alfalfa selective herbicides.

• No/few commercial tolerant alfalfa cultivars.

What we do propose?

To develop Alfalfa cultivars tolerant to herbicides by combining traditional breeding with biotechnology

High level of biosafety

Alfalfa-BAR plants

Agrobacterium mediated transformation of alfalfa

García et al. 2014 BMC Plant Biology and Jozefkowicz et al., 2016 PCR

Genome walking Assays

Comprehensive molecular evaluation

Integrity and heritability transgen and BAR protein confirmed.

WT Bio10 BioX H2O

Event specific PCR

Southern Blot

Western Blot

PCR

Comprehensive molecular evaluation

Jozefkowicz et al., 2018 TAG

Increased vigor during the breeding program

Introgression, manual crosses

3 years The lights!

Alfalfa-

BAR

14 days old-plants 3 month old-plants mature flowers

Transgenic trait at different growth stages

Alfalfa-BAR elite elite elite

0d

7d

10

0 m

g/l a

mm

on

ium

glu

fosi

nat

e

transgenic wt

100 mg/l herbicide

Alfalfa-BAR plants showed tolerance at all different growth stages

Alfalfa-BAR Alfalfa-BAR

GMO transfer CONABIA PhaseI

Field ammonium glufosinate tolerance assays

IGEAF

EEA Manfredi

CONABIA PhaseI

2 L GA/Ha

+1%SO4(NH4)2

3,5 L GA/Ha

+1%SO4(NH4)2

without herbicide

+1%SO4(NH4)2

4.65+0.12 kg/year.m2

4.59+0.20 kg/year.m2

4.45+0.14 kg/year.m2

Commercial cultivars Alfalfa-BAR.

Alfalfa-Bar plants showed tolerance to glufosinate field doses

Jozefkowicz et al., 2018 TAG

Evaluation of agronomic skills

The introgression of BAR transgene in elite material was successful

WT RR BAR Doble

2 weeks Glufosinate Assays 2 weeks Glyphosate Assays

Second step: Glufosinate+Glyphosate tolerance

Next steps: Glufosinate+Glyphosate+IMI tolerance CRISPR/Cas9

Thank You soto.gabrielacinthia@inta.gob.ar