黑果枸杞外整流钾离子通道SKOR基因的克隆及表达分析

摘要

To determine the molecular mechanism of Lycium ruthenicum, a perennial shrub species that has at-tracted great interest in recent years due to its nutritional value and abiotic stress resistance, with its high salt tolerance capability, a potential outward-rectifying potassium channel gene was cloned based on a transcriptome sequencing database analysis. Analyses also included real time (RT)-polymerase chain reaction (PCR), a ho-mology comparison analysis, and a gene expression analysis. Results of the RT-PCR indicated that the length of this potential L. ruthenicum SKOR (LrSKOR) gene was 2448 bp encoding 815 amino acids. The homology comparison analysis showed that the SKOR gene shared more than a 60％ amino acid sequence similarity with reported plants, such as Arabidopsis thaliana, and had over 90％ similarity with other Solanaceae plants. Gene expression analysis with RT-PCR showed that the LrSKOR gene was strongly induced by NaHCO3 stress and weakly induced by NaCl stress; its expression level with NaHCO3 stress was 6.81 times higher than NaCl stress and 23.79 times higher than the control. Thus, expression of LrSKOR, regulated by both saline and saline-alka-li stress, with pH imposed a strong influence on its expression. The results could help in LrSKOR gene function prediction as well as lay a foundation for further research of high salinity stress response in L. ruthenicum.[Ch, 4 fig. 27 ref.]%黑果枸杞Lycium ruthenicum为多年生野生灌木,由于其珍贵的营养价值和强非生物胁迫抗性受到广泛关注.为研究黑果枸杞钾离子(K+)/钠离子(Na+)选择性运输的分子机制,从转录组数据中筛选出拟外整流K+通道基因,提取黑果枸杞根系总RNA为模板,采用反转录聚合酶链式反应(RT-PCR)方法分离出该拟似SKOR基因.序列分析显示:该基因序列长2448 bp,编码815个氨基酸.通过比对发现:该基因与拟南芥Arabidopsis thaliana等植物中已报道的SKOR基因编码的氨基酸序列同源性在60％以上,与几种茄科Solanaceae植物的SKOR同源性高达90％以上.进一步对该基因进行表达分析,结果显示:LrSKOR被盐碱处理强烈诱导而被高盐处理微弱诱导,盐碱处理是高盐处理的6.81倍,是对照的23.79倍,说明LrSKOR的表达受到盐以及盐碱胁迫调控,并且pH值对其表达水平有显著影响.图4参27