Genetic engineering of oilseed rape resistant to seed shattering.
2008
Shakib, A`li Mohammad | Shafi`i, A`li Reza | Ahmad Raji, Mana | O`ruj Lu, Mahnaz
In order to study the possibility for production of resistant plants to seed shattering in oilseed rape by genetic engineering, at first two BnShp and BnFUL genes which are involved in silique opening were isolated by RT-PCR and screening of a cDNA library constructed from expressing mRNAs in silique. At the same time in vitro plant regeneration and gene transfer method was optimized for rape culivars. Two strategies including silecing of BnSHP using RNAi technique and over-expression of BnFUL were chosen. For this purpose, molecular recombinant constructs were made by inserting two segments of 470 and 780 bp on pGSA1252 and pCAMBIA3301 vectors for BnSHP and BnFUL , respectively. These constructs were then transferred into rapeseed using Agrobacterium. PCR analysis showed presense of the BnSHP trans gene in rape lines. Although, due sever damage to winter cold during vernalization, only one transgenic line was obtained. T1 seeds were produced for the next analysis. From transferring the BnFUL gene, 8 transgenic lines for Okapi cultivar were also obtained. PCR analysis showed presense of the BnFUL transgene in these lines. Preliminary analysis by Southern blot confirmed the presense of one to 3 copies of BnFUL transgene. As these lines containing the herbicide resistant gene there is a potential for them to be used for weed control using herbicide. T1 seeds from these lines were prepared for further assessment.
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