刺梨果实半乳糖内酯脱氢酶基因cDNA片段的克隆及在不同器官的表达.pdfVIP

中国农业科学 2005, 38(3):571-575 Scientia Agriultura Sinica 刺梨果实半乳糖内酯脱氢酶基因cDNA片段的 克隆及在不同器官的表达 1,2 1 2 安华明 , 陈力耕 , 樊卫国 1 2 （浙江大学园艺系，杭州 310029 ； 贵州大学园艺系，贵阳 550025 ） 摘要：根据GenBank中登录的拟南芥、花椰菜、甘薯和草莓等植物的半乳糖内酯脱氢酶基因(GalLDH)保守序 列设计引物，采用RT-PCR法，从刺梨果实中扩增出该基因的cDNA片段855 bp并测序。序列分析表明，刺梨GalLDH 与草莓 GalLDH 的相似性达 92%，与其它植物同源基因的相似性也在 80%左右；该片段编码 285 个氨基酸，推导的 氨基酸序列与其它植物同源基因的相似性高达75%～87%。Northern杂交分析表明，该基因在刺梨不同器官中的表 达差异明显：在果实内有很高的表达水平，叶内较弱，而在茎和根内只能检测到微弱的信号。这种差异与不同器 官中维生素C 的积累量高度一致。 关键词：刺梨；半乳糖内酯脱氢酶(GalLDH)；维生素C；克隆；基因表达 Cloning of cDNA Fragment of L-Galactono-1,4-Lactone Dehydrogenase and Its Expression in Different Organs of Rosa roxburghii Tratt AN Hua-ming ，CHEN Li-geng ，FAN Wei-guo 1 ( Department of Horticulture, Zhejiang University, Hangzhou 310029； 2Department of Horticulture, Guizhou University, Guiyang 550025) Abstract ：855 bp cDNA encoding L-galactono-1,4-lactone dehydrogenase (GalLDH) fragment was cloned from fruit of Rosa. roxburghii Tratt by the method of RT-PCR on the basis of the homologous genes of Arabidopsis thaliana , cauliflower, sweet potato, strawberry, etc. in GenBank. Sequensing analysis shows that 79%-92% identity in nucleotide sequence and 75%-87% identity in amino acid sequence to that of strawberry and Arabidopsis thaliana , etc. was observed. Northern blot showed that the expression of GalLDH was significantly different in different organs. The transcription level of GalLDH in fruit was significantly higher than that in leaf, stem and root, respectively. Furthermor