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

Shyam sunder, M J (2013) Genetic transformation studies on sorghum (Sorghum bicolor) for sucrose isomerase. Masters thesis, Jawaharlal Nehru Technological University.

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Supervisors

Supervisors NameSupervisors ID
Pooja Bhatnagar Mathur, AICRISAT (Patancheru)
Saida Naik, LJNTU (Hyderabad)

Abstract

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.

Item Type: Thesis (Masters)
Divisions: RP-Dryland Cereals
CRPS: CGIAR Research Program on Dryland Cereals
Uncontrolled Keywords: Molecular Characterization, Genetic Transformation, Sucrose Isomerase
Subjects: Mandate crops > Sorghum
Depositing User: Mr T L Gautham
Date Deposited: 09 Nov 2015 09:15
Last Modified: 18 Nov 2015 10:42
URI: http://oar.icrisat.org/id/eprint/9121
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