5030
DOAA HASSAN ALI SOLIMAN
IN VITRO STUDIES ON DROUGHT TOLERANCE USING GENE TRANSFER ON TOMATO
Tomatoes- MM- SSB- RG - Agrobacterium tumefaciensis- PtdIns-PLC2- Gene transfer- Drought- PCR- GUS assay
The aim of this study is to investigate best conditions of previously published protocols on Agrobacterium-mediated transformation of tomato, to deliver PtdIns-PLC2 gene, which renders the trait of tolerance to drought, to Money Maker (MM), Super Strain B (SSB) and Rio Grande (RG) tomato cultivars. The obtained results indicated that the efficiency of Agrobacterium-mediated gene transfer in tomato tissues is influenced by many factors; the most important are cultivars, explants type and bacterial optical density. The highest transformation efficiency was 68% with Money Maker flamingo bill like explant. Whereas the transformation efficiency with Super Strain B and Rio Grande cotyledons were 48% and 40%, respectively. On contrary, the transformation efficiency of Rio Grande and Super Strain B hypocotyls were 20% and 16 %, respectively. The best optical density for cotyledons and hypocotyls of the SSB and RG was 0.05 followed by 0.5 and 1.0. The presence of transgenes and their expression were confirmed by molecular analysis (PCR, RT PCR and Real Time PCR) and histochemical analysis (GUS assay). In conclusion, GUS assays and PCR affirmed the successful delivery and expression of PtdIns-PLC2 gene in Money Maker, Super Strain B and Rio Grande tomato plants regenerated, elongated and rooted on selective media. The obtained Money Maker transformants were exposed to drought stress in vitro using different concentrations of Mannitol to assess the activity of thegene and estimate the occurred physiological variations
2015
M.Sc
Cairo
Agriculture