摘要
为了研究棉纤维醛酮还原酶基因在参与细胞伸长发育中所起的作用,本研究利用RT-PCR技术从处于快速伸长发育时期的棉花纤维组织中克隆得到棉花醛酮还原酶(aldo/keto reductase,AKR)基因c DNA,该基因全长读码框为1134 bp,编码377个氨基酸。氨基酸序列生物信息学分析表明Gh AKR蛋白具有较高的保守性,包含氧化还原酶、醛酮还原酶结合位点和跨膜信号序列等功能序列和位点;进化树分析显示Gh AKR与可可(Tc AKR)的亲缘关系较近。q RT-PCR和酶活性分析表明Gh AKR基因与纤维细胞的快速伸长发育密切相关,尤其在开花后5-15 d纤维快速伸长发育时期具有较高的表达。构建35S::Gh AKR过量表达载体并转化野生型拟南芥(哥伦比亚生态型),转基因拟南芥的主根伸长发育获得了显著的促进,与野生型相比,转基因拟南芥主根伸长增加了约1.4倍。拟南芥根中甲醇和乙醇含量分析显示,转基因拟南芥根中具有较高的甲醇和乙醇积累。这些结果表明棉纤维Gh AKR基因与纤维细胞伸长发育联系紧密,可能通过增加细胞内的醇含量进一步促进细胞的伸长。本研究可为棉花Gh GAKR基因参与纤维伸长发育分子机制解析,以及利用基因工程培育优质棉花品种提供一定参考。
In order to study the function of cotton fiber aldehyde/ketone reductase（AKR） gene and its role in cell elongation development, the full-length open reading frame（ORF） of Gh AKR c DNA was cloned from fast elongating fiber tissues by RT-PCR, The length of Gh AKR ORF contains 1134 bp nucleotides and codes a protein of 377 amino acids. Sequence analysis indicated that Gh AKR is a conservative protein with a transmembrane signal peptide, functional sites of oxidoreductase and aldehyde/ketone reductase binding sites. Phylogenetic tree analysis showed that Gh AKR has a close evolutionary relationship with Tc AKR. Quantitative real-time PCR（q RT-PCR） and enzyme activity analyses displayed that Gh AKR is closely relevant to fiber fast elongation development with abundant expression at 5-15 day-post anthesis（dpa） wild-type（WT） ovule and fiber tissues. Transgenic Arabidopsis plants expressing 35 S：：Gh AKR vector showed enhanced main root elongation with 1.4 fold increase than control. Methanol and alcohol were accumulated in roots of transgenic Arabidopsis plants. These results revealed that Gh AKR is involved in fiber elongation through increasing the content of methanol and alcohol in the cell. The study provided a basis for further understanding of Gh AKR participating in cell elongation and cotton genetic breeding.
作者
杨洋
李鸿彬
Yang Yang;Li Hongbin(College of Life Sciences, Key laboratory of Agrobiotechnology, Shihezi, Xinjiang 832003, Chin)
出处
《石河子大学学报（自然科学版）》
CAS
2017年第5期612-617,共6页
Journal of Shihezi University(Natural Science)
基金
国家自然科学基金项目(31660408)
新疆兵团科技计划项目(2016AC017)
新疆兵团杰出青年项目(2014CD003)
关键词
棉花纤维
醛酮还原酶
拟南芥
细胞伸长
醇含量
cotton fiber
aldehyde ketone reductase
Arabidopsis thaliana
cell elongation
methol and alcohol
