GENETIC MODIFICATION OF TOMATO (Solanum lycopersicum) TO INCREASE THE UPTAKE AND TRANSLOCATION OF ZINC

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Date
2015-09-09
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UNIVERSITY OF AGRICULTURAL SCIENCES GKVK, BENGALURU
Abstract
Zinc (Zn) is one of the essential micronutrient with major biological roles both in plants and humans. Deficiency of Zn is by far the most ubiquitous problem in the world affecting the plant and human nutrition. Zn deficiency in humans can be addressed through dietary diversification, mineral supplementation, food fortification and biofortification. Among these, biofortification of food crops either through genetic engineering or molecular breeding approaches is considered to be promising and sustainable. To achieve this better understanding of molecular mechanisms regulating the Zn uptake and transport in plant is most crucial. In the present investigation an attempt was made to identify the candidate Zn transporter gene in tomato responsible for high fruit Zn content. Initially 41 tomato genotypes were screened for Zn content. The contrast lines identified based on the fruit Zn content were subjected for Zn deficient and sufficient conditions and analysed for Zn transporter expression through RT- PCR. The candidate Zn transporter genes up-regulated in the high Zn type germplasm lines, SlZIPL and SlZIP4 were identified and cloned. Further the genes were sub-cloned to pBINPLUS binary vector driven by constitutive promoter (CaMV35S) and the two independent gene constructs PCaMV35S: SlZIPL: TpolyA and PCaMV35S: SlZIP4: TpolyA were used for plant transformation. To develop transgenic tobacco and tomato plants, Agrobacterium mediated tissue culture technique was followed. Few tobacco transgenic lines over expressing SlZIPL and SlZIP4 showed high leaf Zn and chlorophyll content. Similarly, tomato SlZIPL and SlZIP4 transgenics also showed high fruit, leaf and petiole Zn content compared to wild type. Further the RT-PCR expression analysis of tomato transgenics showed the higher expression of SlZIPL and SlZIP4. These results imply that, SlZIPL and SlZIP4 are candidate Zn transporters in tomato involved in better Zn transport and hence high fruit Zn content
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