【作者】 李娜娜； 邵文韵； 刘畅； 陆建良； 郑新强； 梁月荣；

【Author】 LI Nana;SHAO Wenyun;LIU Chang;LU Jianliang;ZHENG Xinqiang;LIANG Yuerong;Tea Research Institute of Zhejiang University;

【机构】 浙江大学茶叶研究所；

【摘要】 八氢番茄红素脱氢酶是类胡萝卜素生物合成途径的关键酶之一,本实验采用3’/5’RACE和RT-PCR技术成功扩增出茶树八氢番茄红素脱氢酶基因(PDS)的3’端和5’端序列,序列拼接获得全长cDNA序列,命名为CsPDS,并将其登录至GenBank,登录号KF646537。经生物信息学分析,所得基因cDNA序列全长为2295bp,开放阅读框(ORF)1749 bp,编码582个氨基酸,预测分子量约为64.86 kDa,理论等电点(PI)6.77,亲水性蛋白。该基因编码的氨基酸序列与柿树PDS序列的同源性达到87%,多序列比对表明茶树PDS具有高度保守区域,基于邻接法的进化树显示与柿树、葡萄的亲缘关系最近。实时荧光定量PCR分析结果显示,CsPDS在"黄金芽"体内表达没有受抑制,表明"黄金芽"黄色白化可能不是在CsPDS基因转录水平异常而引起。更多还原

【Abstract】 Phytoene desaturase is one of the key enzymes involving in carotenoid biosynthetic pathway. 3’/5’ RACE and RT-PCR technique were used to amplify the 3’ and 5’ end sequence of phytoenedesaturase gene( PDS) from tea plant, then a full-length cDNA sequence was obtained after sequence splicing, which named as CsPDS, GenBank accession number KF646537. The bioinformatic characterization indicated that the full length cDNA of CsPDS was 2295 bp, which contained 1749 bp open reading frame and encoded 582 amino acid residues with a putative molecular mass of 64.86 kDa and a theoretical isoelectric point of 6.77. It belongs to a hydrophilic protein. The deduced amino acid was most closely to Diospyros kaki with 87% sequence homology, multiple sequence alignment showed that many highly conserved regions of CsPDS, phylogenetic analysis based on the neighbor-joining method displayed that the closest evolutionary relationship with Diospyros kaki andVitisvinifera. Quantitative real-time PCR analysis showed that the expression of CsPDSgene was not inhibited in ‘Huangjinya’, it indicates thatyellow-albinism of ‘Huangjinya’ may not be caused by CsPDS gene atthe transcriptional level.更多还原