transgenic alfalfa tolerant to herbicides, from lab to field soto ppt... · gabriela c. soto ....
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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
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