The Expression Pattern And Regulatory Of CBT-ol Synthetase Gene(CYC) In Tobacco

Tobacco CBT-ol synthetase gene(CYC)is the key gene of CBT-ol biosynthesis.Expression level of different tobacco types(Dabai 1,Cigar 1,Basma and K326),different flue-cured tobacco varieties(K326,Zhongyan 100,Honghua Dajinyuan,Cuibi 1 and Yuyan 11),and different tissues of K326(root,stem,upper leaves,middle leaves,lower leaves,glandular hairs,leaves without glandular hairy,flowers)were studied,in order to study the expression rule of CYC gene.Meanwhile,the plant expression vector p C-CYC-Pro-GUS built were transformed into K326 via agrobacterium-mediated transformation under the condition of 30 mg/ml concentration of kana to get positive transgenic plants,analysized specifically expressed in the tissues and the expression pattern under different temperature range Surface exudates compositions of CYC-RNAi transgenic plants were determined with GC/MS method.The main findings are as follows:1.CYC gene expression is different.Expression level of CYC genevaried among different tobacco types with the highest quantity in Basma,followed by K326 and Ciger 1 up to Dabai 1;varied among different flue-cured tobacco varieties with the highest quantity in followed by Honghua Dajinyuan,Yuyan 11,Cuibi 1 up to Zhongyan 100.K326;varied among different tissues of K326 with the highest quantity in glandular trichomes.2.Specific expression of CYC gene promoter:Get the roots,stems and leaves of K326 and transgenic plants respectively,GUS dye solution,dyeing found that the transgenic plants glandular hairs on the leaves and stems were stained with blue.The GUS reaction indicated that CYC gene promoter was consolidated and transcripted in the tobacco K326 genome,and specifically expressed in the tobacco glandular trichomes.Using transgenic plants as the materials,there were three settings in the temperature test,low temperature(4?),normal temperature(28?),high temperature(42?),sampled at 1 hour,6hour,12 hour respectively.Total protein extraction for GUS enzyme activity tests showed that the GUS enzyme activity was higher under 28? than those in the temperature range of 4? and 42?,which indicated that the tobacco CYC gene promoter expression was regulated by the temperature.3.The trichome exudates components of CYC-RNAi transgenic plants: 10 CYC-RNAi transgenic strains under controlled conditions of field experiment were carried out in accordance with the high quality tobacco cultivation management specification.The content of CBT-ols and CBT-diols of six in ten strains were significantly lower than K326,meanwhile the content of hydrocarbons and sucrose ester were the same level.The other two strains of glandular trichome secretion content increased significantly compared with K326.May the dissimilarity associated with specific insertional locus.