Forward screening for“broad” leaf mutants · • Backcross candidates to cv. Sebastian •...
Transcript of Forward screening for“broad” leaf mutants · • Backcross candidates to cv. Sebastian •...
![Page 1: Forward screening for“broad” leaf mutants · • Backcross candidates to cv. Sebastian • Generate Mapping populations by crossing candidates to different Barley cultivars .](https://reader034.fdocuments.in/reader034/viewer/2022050312/5f73f745e06eba0db72a52f7/html5/thumbnails/1.jpg)
Forward screening for“broad” leaf mutants
Moritz Jöst Lenhard Lab
University Potsdam
![Page 2: Forward screening for“broad” leaf mutants · • Backcross candidates to cv. Sebastian • Generate Mapping populations by crossing candidates to different Barley cultivars .](https://reader034.fdocuments.in/reader034/viewer/2022050312/5f73f745e06eba0db72a52f7/html5/thumbnails/2.jpg)
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Leaf size candidates HorTILLUS in the field in Katowice 2016
![Page 3: Forward screening for“broad” leaf mutants · • Backcross candidates to cv. Sebastian • Generate Mapping populations by crossing candidates to different Barley cultivars .](https://reader034.fdocuments.in/reader034/viewer/2022050312/5f73f745e06eba0db72a52f7/html5/thumbnails/3.jpg)
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Leaf size candidates HorTILLUS in the Greenhouse in Potsdam 2017
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![Page 4: Forward screening for“broad” leaf mutants · • Backcross candidates to cv. Sebastian • Generate Mapping populations by crossing candidates to different Barley cultivars .](https://reader034.fdocuments.in/reader034/viewer/2022050312/5f73f745e06eba0db72a52f7/html5/thumbnails/4.jpg)
Outlook
• Grow leaf size candidates from HorTILLUS under highly controlled conditions (Percival)
• If Leaf length difference can be verified: –> detailed growth analysis
• Backcross candidates to cv. Sebastian • Generate Mapping populations by crossing
candidates to different Barley cultivars
![Page 5: Forward screening for“broad” leaf mutants · • Backcross candidates to cv. Sebastian • Generate Mapping populations by crossing candidates to different Barley cultivars .](https://reader034.fdocuments.in/reader034/viewer/2022050312/5f73f745e06eba0db72a52f7/html5/thumbnails/5.jpg)
Phenotypic characterization of “broad” leaf mutants and identification of candidate
mutations
Moritz Jöst Lenhard Lab
University Potsdam
![Page 6: Forward screening for“broad” leaf mutants · • Backcross candidates to cv. Sebastian • Generate Mapping populations by crossing candidates to different Barley cultivars .](https://reader034.fdocuments.in/reader034/viewer/2022050312/5f73f745e06eba0db72a52f7/html5/thumbnails/6.jpg)
“Broad Leaf” mutants from the Nordgen collection Name „Parent cultivar“ Genotyping Mutagen
blf 1 Bonus Bonus OBS
blf2 Bonus Bonus colchicine, X-rays
blf3 Bonus Bonus X-rays
blf5 Bonus Bonus ethylene imine
blf6 Foma Bonus neutrons
blf7 Bonus Bonus gamma-rays
blf8 Foma Bonus ethylene imine
blf10 Foma Foma neutrons
blf11 Foma Foma ethylene imine
blf12 Foma Foma fast neutrons
blf13 Foma Bonus n-butyl methanesulfonate
blf14 Foma Kristina N-ethyl-N-nitrosourea
blf15 Foma Kristina N-ethyl-N-nitrosourea
blf16 Kristina Kristina neutrons
blf17 Kristina Kristina ethyl methanesulfonate
blf18 Kristina Kristina ethyl methanesulfonate
blf19 Kristina Kristina ethyl methanesulfonate
broad leaf 1-6
broad leaf 1-1
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“Broad Leaf” mutants from the Nordgen collection - Phenotyping in the phytotron
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Field trial in Katowice 2016 D
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Kristina Foma Bonus Kristina Foma Bonus
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Task: select the “best” 3 “Broad Leaf” mutants for further analysis:
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NG_blf13 NG_blf2 NG_blf11
WT mutant
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Select the 3 best “Broad leaf”mutants and…:
• Mapping by sequencing: • Select F2 populations for mapping:
• X cv. Bowman • X cv. Barke • X cv. Golden Promise
• Exome capture + sequencing of WT and mutant
pools • Further fine mapping
![Page 13: Forward screening for“broad” leaf mutants · • Backcross candidates to cv. Sebastian • Generate Mapping populations by crossing candidates to different Barley cultivars .](https://reader034.fdocuments.in/reader034/viewer/2022050312/5f73f745e06eba0db72a52f7/html5/thumbnails/13.jpg)
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Selecting a mapping population: NG_blf13
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Selecting a mapping population: NG_blf2
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cv. Golden Promise
![Page 15: Forward screening for“broad” leaf mutants · • Backcross candidates to cv. Sebastian • Generate Mapping populations by crossing candidates to different Barley cultivars .](https://reader034.fdocuments.in/reader034/viewer/2022050312/5f73f745e06eba0db72a52f7/html5/thumbnails/15.jpg)
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F2 NG_blf11 x cv. Barke
Selecting a mapping population: NG_blf11
NG_blf11
cv. Bowman
cv. Golden Promise
cv. Barke
![Page 16: Forward screening for“broad” leaf mutants · • Backcross candidates to cv. Sebastian • Generate Mapping populations by crossing candidates to different Barley cultivars .](https://reader034.fdocuments.in/reader034/viewer/2022050312/5f73f745e06eba0db72a52f7/html5/thumbnails/16.jpg)
Mapping by sequencing: • About 400 plants per F2 population were
phenotyped • Per population 50 phenotypically “WT” and 50
“mutant” plants were selected: • Leaf material sampled and pooled • DNA extraction -> Material will be send to Agnieszka
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NG_blf2 x Bowman
![Page 17: Forward screening for“broad” leaf mutants · • Backcross candidates to cv. Sebastian • Generate Mapping populations by crossing candidates to different Barley cultivars .](https://reader034.fdocuments.in/reader034/viewer/2022050312/5f73f745e06eba0db72a52f7/html5/thumbnails/17.jpg)
Outlook
• DNA pools will be subjected to exome capture and sequenced at the University of Silesia
-> Candidate mutations • Fine mapping using KASP Markers • Detailed phenotypic analysis
• Sections of mature leaf blades • 3D reconstructions of shoot apices
![Page 18: Forward screening for“broad” leaf mutants · • Backcross candidates to cv. Sebastian • Generate Mapping populations by crossing candidates to different Barley cultivars .](https://reader034.fdocuments.in/reader034/viewer/2022050312/5f73f745e06eba0db72a52f7/html5/thumbnails/18.jpg)
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