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Developing Wheat Tilling Population

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by

S.m. Shakil

on 15 November 2014

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Transcript of Developing Wheat Tilling Population

Development of a Wheat Tilling Population
Presented By S.M. Shakil
What is Tilling ?
To develop new varieties with improved properties
Combines random mutagenesis with high throughput screening of molecular markers

EMS Treatment
Sterilize approx. 200/tube seeds with Detergent
Washing (2 times)
Treat with EMS for 16 h
Washing (3 times)
Plantation
M2 Generation
718 plants germinated out of 800 Lines
Tissue collection from 250 lines
16 Cholorotic mutations
9 Dwarf, 2 Taller, 4 Broad leaved plants
Estimated frequency approx. 2%
Salt Tolerant Lines
High Protein Lines
Slow Carbohydrate Release Lines
Salinity Test in Belinda
Salinity Test Gom-25
Suggestions ?
In Agar In MS Medium
In Agar In MS Medium
Germination frequency 90%
Total 2674 M2 lines
Ethyl Methanesulfonate (EMS)
Random mutations
Point mutation
Ethyl group of EMS reacts with Guanine resulting O-6-ethylguanine.
Replace T instead of C, resulting A:T pair instead of G:C

Mutation Frequency Analysis
Genomic DNA Extraction
M3 Population
Salinity test in mutant lines
Salinity Test in Belinda
Control Belinda
Agar+150mMNacl
M5 Line:126 (Control)
M5 Line:126 (Agar+150mM NaCl)
Salinity Test in Gom-25
Agar
Agar+ 150mM NaCl
Random primers in Gom-25
Random primer Optimization
Random primer mixture
Random primers in mutants
Amplified genes in Gom-25
HKT8, SOS1 in Gom-25 & Mutants
(Product Size-649bp, 659bp)
HKT8, SOS1 in Gom-25 & Mutants
(Product Size-2141bp, 2012bp)
M2 Plants in Bangladesh
Tilling Method
Conclusion
M4 Generation: 1754 mutant lines
Salt tolerace analysis in Gothenberg.
Genomic DNA Stock: 204 mutants from M2 generation
Tissue material stock: 110 mutants from M3 generation
Mutation frequency analysis
Acknowledgments
Prof. Olof Olsson
Prof. Lutfar Rahman
Dr. Aakash Chawde
A. Prof. Henrik Aronsson

Bindu Ayoor
Svettlana Leonova
Hans-Olof Johansson
EMS Optimization in Gom-25
1% EMS Treated plants
EMS Optimization
Large Scale M1 population
M1 Population development
Rare mutation
Chlorotic Mutation
M2 Population
Genes involves in salt stress
Mutation Frequency
Large Scale M1 population
Chlorotic Mutation in M2 population
M1 Population
Draft & Taller plants in M2 population
Line: 172
Line 391, 421
Line: 233, 245, 256
Full transcript