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Development of transgenics for fungal resistance and discovery of chemically induced mutations in Pearl Millet (Pennisetum glaucum L.) population by TILLING

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Relation http://oar.icrisat.org/122/
 
Title Development of transgenics for fungal resistance and discovery of chemically induced mutations in Pearl Millet (Pennisetum glaucum L.) population by TILLING
 
Creator Jalaja, N
 
Subject Millets
 
Description Protocols for in vitro plantlet regeneration in pearl millet and genetic transformation using inflorescence
derived callus cultures via Agrobacterium and microprojectile methods were standardized. Transgenic
pearl millet plants were confirmed by gene specific amplification of chitinase and osmotin and also the
bar genes. However, the transgenics turned out to be sterile, though pollen was produced.
� Mutagenized population of pearl millet from inbred line P1449-2-P1 was produced using the chemical
mutagen ethylmethane sulfonate and M2 generation was raised. Phenotypic analysis was carried out in
M2 generation. The mutagenized population was screened for point mutations using the TILLING
technique.
� The mutant lines (9,938 lines) were screened for two candidate genes, one associated with water stress
(dehydration responsive element binding factor, DREB2A) and the second associated with pathogen
resistance (enhanced disease resistance, EDR2).
� Allele mining was carried out for DREB2A gene; amino acid arginine was changed to threonine and
serine to proline in the hot spot region that was predicted by the CODDLE software. It is hoped that these
amino acid changes would bring loss of function of this gene and thus can help to find out the function of
the gene under water stress conditions.
� TILLING was performed for EDR2 gene; but because of non-specificity, the mutations could not be
detected in the LICOR gel.
 
Date 2011
 
Type Thesis
PeerReviewed
 
Format application/pdf
 
Language en
 
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Identifier http://oar.icrisat.org/122/1/Report-var-2011.pdf
Jalaja, N (2011) Development of transgenics for fungal resistance and discovery of chemically induced mutations in Pearl Millet (Pennisetum glaucum L.) population by TILLING. PhD thesis, Osmania University.