Annual Report Of Research Activity: Construction Of A cDNA Library And Its Screening
2004
Saifi, Ali Reza | Mohammad Shakib, Ali
Rapeseed is one of the most important sources of edible oil in Iran. Its oil quantity and quality and adaptation ability to cold climates are the major advantages of rapeseed. Yield lost because of seed shattering during harvesting is a great problem that may cause seed decrease up to 50%. No satisfying remedy for this disadvantage has been presented via cultural and classical plant breeding methods, so there are some endeavors to adopt genetic engineering techniques to produce shatterproof rapeseed lines in the world. Several regulatory genes in Arabidopsis thaliana, a near model plant to rapessed, have been identified involving in pod opening process. Agl1, Agl5 and Agl8 are the most important ones. The objective of this investigation was to isolate these genes homologues in rapeseed. For this aim pods of rapeseed was harvested and RNA was extracted by Chomczynski method. After mRNA purification, a cDNA library was constructed and screened against Arabidopsis-derived probes. Another strategy was to adopt PCR-based techniques. At first, one pair of primers was designed for BnShp1, a cDNA clone showing high degree of similarity to Agl1 and Agl5, has been isolated from rapeseed. After alignment of several MADS-box genes, a unique sequence was distinguished for Agl8, which was used for primer designing to amplify Agl8-like in rapeseed. Amplification using this primer was very interesting and resulted in isolation of a new clone which shows homology to Agl8 and it is the most important aim of our experiments. More detailed molecular analysis is undergoing.
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