Cloning And Analysis Oil HPL And GDS Of Jasminum Sambac

The jasmine [Jasminum sambac (Linn.) Aiton] is the important raw materials of spiceindustry and Tea industry, is an important resource of the flavoring substances ofplants.The formation of flower aroma volatile molecules are a class of low-molecular,low-boiling, low-polarity compounds. In this study, the materials is double-petal jasmine'sleaves and flower buds that collected from Fujian Agriculture and Forestry University.Using RT-PCR and RACE techniques to obtain the fatty acid hydroperoxide lyase (HPL),and the germacrene D, synthase(GDS) cDNA, using semi-quantitative RT-PCR analysis ofthe HPL and GDS expression levels of different periods in the leaves and buds, in order toclone and analyse the jasmine' aroma synthesis enzyme gene.1Cloning Jasmine's HPL full-length cDNA and analysis the expression of HPL bysemi-quantitative RT-PCRObtained HPL genes in GenBank, and design degenerate primers to amplifyconserved region fragments. Obtain5'and3' sequence by the RACE technique, and obtainthe total length of1703bp HPL gene what containing1479bp ORF. It is encoding492amino acids(Mw=55372.8, pI=8.39). There is a non-coding region with80bp in the end of5ËŠ-region.3'end contains134bp non-coding sequences. The amino acid sequencestranslated from ORF is compared with those such as olive, tomato, potato and so on. Andfinally construct the phylogenetic tree.Take Jasmine Actin gene as a positive control to detect the expression on leaves, budsof period1-3and florescence. At last, the results show that the jasmine HPL geneexpression of the5period, and the expression levels is following the developmentalspecificity of HPL: the expression of HPL puts up a high level when flowers growing, andwhen the flowers matured, the level turns lower; and old leaves' expression is lower thannew leaves', too.2Cloning Jasmine's GDS full-length cDNA and analysis the expression of GDS bysemi-quantitative RT-PCRObtained GDS genes in GenBank, and design degenerate primers to amplify conserved region fragments. Obtain5'and3' sequence by the RACE technique, and obtainthe total length of1863bp HPL gene what containing1674bp ORF. It is encoding552amino acids(Mw=64016.4, pI=5.26). There is a non-coding region with90bp in the end of5ËŠ-region.3'end contains104bp non-coding sequences. The amino acid sequencestranslated from ORF is compared with those such as basil, kiwi fruit, grapes and so on.And finally construct the phylogenetic tree.Take Jasmine Actin gene as a positive control to detect the expression on leaves, budsof period1-3and florescence. At last, it results that the the jasmine GDS gene merelyexpression in the Period3of bud and florescence. In line with the expressed in specificorgans, tissues and developmental stages. And From the point of genetics, it supports thetemperament to spend.