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Genetic transformation studies on sorghum (Sorghum bicolor) for sucrose isomerase

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Relation http://oar.icrisat.org/9121/
 
Title Genetic transformation studies on sorghum
(Sorghum bicolor) for sucrose isomerase
 
Creator Shyam sunder, M J
 
Subject Sorghum
 
Description Seeds of sweet sorghum cultivars SSV74 and SSV84 were the primary explants
used for the introduction pdSI genes(expression of Sucrose Isomerase enzyme)
which converts into Isomaltulose(palatinose,α-D-glycopyranosyl-1,6-Dfructofuranose).
Isomaltulose is an isomer of sucrose which has greater health
advantages for diabetics and used as low calories sweetener, it is also a renewable
starting material in industrial productions. The pdSI gene was under regulation of
35S promoter cloned into binary vector PMDC99. Both the vectors also contain
hygromycin phosphotransferase gene (hptII) as plant selection marker against
hygromycin A kill-curve experiment was performed to determine the optimal
concentration at which the control explants is able to survive rate was high in 2.5
& 0.5 mg/L concentration whereas 7.5 mg/L, the survival rate of approximately
12-15%.
Agrobacterium cloned with pdSI gene was used for transfer of the genes into
sweet sorghum cultivar. Pre-soaked imbibed seeds of sweet sorghum were
punctured using sterilized needle and dipped in Agro culture (with pdSI gene) in
YEB media for 15-20 minutes. The seeds were co-cultivated on MS media 48-72
hours. The explants were then transferred in to MS media augmented with
cefotaxime and hygromycin. The selection marker strategy is as follows, During
germination stage hygromycin concentration was 2.5mg/L, in elongation stage,5mg/L and rooting was done at 7.5mg/L. plantlets which are showed rooting and
survival at selection pressure of 7.5mg/L were then transferred to soil for
acclimatization. Leaf sample of healthy, putative plants were collected and DNA
was extracted using CTAB method for further molecular characterization.
 
Date 2013
 
Type Thesis
NonPeerReviewed
 
Format application/pdf
 
Language en
 
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Identifier http://oar.icrisat.org/9121/1/M%20Shyam%20Sunder.pdf
Shyam sunder, M J (2013) Genetic transformation studies on sorghum (Sorghum bicolor) for sucrose isomerase. Masters thesis, Jawaharlal Nehru Technological University.