辣椒ATP硫酸化酶基因(CaATPS1)的克隆与逆境表达

Cloning of a Pepper CaATPS1 Gene and Its Expression in Response to Stresses

采用RT-PCR和RACE方法,克隆了叶绿体ATP硫酸化酶基因的全长序列,命名为Ca ATPS1,Gen Bank登录号为KX009745。该基因包含1个长为1 377 bp的完整开放阅读框,编码458个氨基酸,预测分子量51.11 k D,等电点6.74。氨基酸同源性分析表明,该基因与Gen Bank中登录的茄科植物烟草、马铃薯及其它物种中的ATPS1氨基酸序列具有较高的同源性;进化树分析表明,辣椒Ca ATPS1与同为茄科属的烟草、马铃薯和胡麻属的芝麻处于同一进化分枝上,而与十字花科植物进化关系较远;荧光定量PCR分析显示,Ca ATPS1能被低温、干旱、高盐、机械损伤、过氧化氢、水杨酸、乙烯利以及DC3000侵染等各种胁迫和信号分子诱导。

The full-length c DNA sequences of an ATP sulfurylase gene（ ATPS） isolated from leaf tissues of pepper（ Capsicum annuum L.） were analyzed using reverse transcription PCR（ RT-PCR） and rapid amplification of c DNA ends（ RACE） technology. The gene,designated as Ca ATPS1 with Gen Bank accession number AJY78079,contained an open reading frame of 1 377 bp encoding a polypeptide of 458 amino acid residues with molecular mass of 51. 11 k D and p I value of 6. 74. The protein sequence homology comparison showed that the Ca ATPS1 exhibited a high homology with the ATPS1 of Solanum tuberosum,Nicotiana sylvestris and other plants. A phylogeny tree showed that the Ca ATPS1 had a relatively close evolutionary relationship with Solanum tuberosum, Nicotiana sylvestris and Sesamum indicum,and a distant relationship with Brassicaceae plants. Quantitative RT-PCR analysis showed that the expression of Ca ATPS1 could be significantly induced by cold,drought,high salt,mechanical damage,H2O2,salicylic acid（ SA）,ethephon,and DC3000 infection.