USDA-ARS Research Geneticist and Professor of … NY Expo...Onion Breeding Goals • For Growers:...
Transcript of USDA-ARS Research Geneticist and Professor of … NY Expo...Onion Breeding Goals • For Growers:...
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USDA-ARS Research Geneticistand
Professor of HorticultureUniversity of Wisconsin-Madison
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Onion Breeding and Genetics
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Onion Breeding and GeneticsBiennial (2 years per generation)
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Onion Breeding and Genetics
Year 1: Seed to Bulb in Field
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Onion Breeding and Genetics
Year 1: Seed to Bulb in Field
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Onion Breeding and Genetics
Bulb storage and vernalization
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Onion Breeding and GeneticsYear 2: Bulb selection for seed production
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Onion Breeding and GeneticsClassical crossing and seed production
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Onion Breeding Goals• For Growers:
• High quality seed• Multiple pest resistances• High yields• Low post-harvest losses
• For Consumers:• Attractive bulbs• Good flavor• Heath-enhancing attributes
• For Processors:• Uniformity• Single centers• High dry matter
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USDA Hybrids
• ‘Granex’ (1952)o YB986A x TEG951Co Short-day onion (Vidalia)o Flat shape with low pungencyo Short storage ability
• ‘Spartan Banner 80’ (1979)o MSU(611-1Ax611B) x 2399Bo Long-day pungent oniono Good storage and pink
root resistance
http://southeastagnet.com
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Inbred Releases
• B8667 A&Bo F1MSM2SM2
o CMS Line is BC6
o Deep red color to internal ringso Medium pungency
• Ski-1 A&Bo F1MSM2SM2
o CMS Line is BC5
o Early maturity, good storage
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Development of Value-Added Onions
Traits recognized by the consumer
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#1 Consumer Complaint:Onions Make You Cry
Development of Value-Added Onions
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LF = Lachrymatory Factor
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Onion ThiosulfinatesSO4 Cysteine
Nature 419:685
Glutathione
“Flavor Precursors”
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Onion ThiosulfinatesSO4 Cysteine
Nature 419:685
Glutathione
“Flavor Precursors”
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Onion ThiosulfinatesSO4 Cysteine
Nature 419:685
Glutathione
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Onion ThiosulfinatesSO4 Cysteine Glutathione
Pungent Flavor
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Onion ThiosulfinatesSO4 Cysteine Glutathione
Pungent Flavor
Reduce Aggregation of Blood Platelets
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Onion ThiosulfinatesSO4 Cysteine Glutathione
Pungent Flavor
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Onion ThiosulfinatesSO4 Cysteine Glutathione
Pungent Flavor
Nature 419:685
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‘Tearless Onion’SO4 Cysteine Glutathione
Pungent Flavor
Health Benefits
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Onions Grown on Low Sulfur SoilsSO4 Cysteine GlutathioneX
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Onions Grown on Low Sulfur SoilsSO4 Cysteine GlutathioneX
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Genetic Variation for Low PungencySO4 Cysteine GlutathioneX
MGG 265:543-551TAG 114:815-822
SiRATPS
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SO4 Cysteine GlutathioneX
MGG 265:543-551TAG 114:815-822
SiRATPS
Genetic Variation for Low Pungency
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Mutation Breeding for Low TearingSO4 Cysteine
X
Sci. Rep. 6:23779
Glutathione
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Mutation Breeding for Low TearingSO4 Cysteine
X
Sci. Rep. 6:23779
Glutathione
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Reduced Tearing and ThiosulfinatesSO4 Cysteine GlutathioneX X
SiRATPS
X
• Low sulfur soil• Low sulfur uptake• Alliinase mutation
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SO4 Cysteine Glutathione
X
Targeted Mutation of LFS
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LFS Maps to Chromosome 5
G3 2:643-651
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Tandem Duplication of LFS
G3 2:643-651
• Two copies of LFS are tandemly linked
LFS LFS
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Tandem Duplication of LFS• Two copies of LFS are tandemly linked• Low probability for mutating BOTH copies
G3 2:643-651
LFS LFSX
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Tandem Duplication of LFS• Two copies of LFS are tandemly linked• Low probability for mutating BOTH copies
• Targeted knock-out of both copies of LFS with interfering RNA or gene editing
LFS LFSX X
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Interfering (i) RNA
http://upload.wikimedia.org/wikipedia/commons/6/6c/RNAi.jpg
NO PROTEIN
X
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RNAi for LFS Produced First ‘Tearless’ Onion
Plant Physiol 147:2096
SO4 Cysteine Glutathione
X
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RNAi for LFS Produced First ‘Tearless’ Onion
Plant Physiol 147:2096
SO4 Cysteine Glutathione
NO LFS PROTEIN
X
LFS
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RNAi for LFS Produced First ‘Tearless’ Onion
Plant Physiol 147:2096
SO4 Cysteine Glutathione
X
RNAi plants
Protein Gel
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RNAi for LFS Produced First ‘Tearless’ Onion
Plant Physiol 147:2096
SO4 Cysteine Glutathione
X
RNAi plants
Protein Gel
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RNAi for LFS Produced First ‘Tearless’ Onion
Plant Physiol 147:2096
SO4 Cysteine Glutathione
X
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‘Tearless Onion’RNAi knock-down does NOT cause tearing AND
accumulates MORE healthy thiosulfinates!!
• Transgenic onion• No commercialization
– Consumer reluctance for GMOs– Regulatory and licensing costs
• Great product; large market; No go!
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CRISPR/Cas9 Targeted Gene Mutation
https://www.frontiersin.org/files/Articles/162559/fgene-06-00300-HTML/image_m/fgene-06-00300-g001.jpg
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CRISPR/Cas9 Targeted Gene Mutation
https://www.frontiersin.org/files/Articles/162559/fgene-06-00300-HTML/image_m/fgene-06-00300-g001.jpg
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LFS LFSX X
Knock-0ut BOTHcopies of LFS
CRISPR/Cas9 Targeted Gene Mutation
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CRISPR/Cas9 Targeted Gene Mutation
SO4 Cysteine Glutathione
X
Knock-out BOTH copies of LFS and
NON-TRANSGENIC
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• Knock-out native gene– Targeted mutation– Presently not regulated
• Change protein– Same gene– May not exist naturally– Regulated?
• Build a new gene– Similar to transgenics– Regulated?
Regulation of CRISPR/CAS
Polyphenol OxidaseX
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• Natural genetic variation– Basis of breeding for 1000s years
• Mutation breeding– Random approach– Few successes relative to effort
• Transgenics– Powerful approach and highly successful– Public opposition; high regulation costs
• Gene editing (CRISPR/Cas)– Non-random approach– Enormous potential for new variation– Regulation and public acceptance?
Onion Breeding Approaches
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Excellent Research By• Japan
• House Foods• New Zealand
• Plant and Food Research• USA
• USDA and University of Wisconsin• J. Craig Venter Institute• Funding
• USDA ARS, NRI, FRA, IFAFS, and SCRI programs
• Seed companies