百萨偃麦草(Thinopyrum bessarabicum Lve,2n=2x=14,JJ)是小麦改良的重要亲缘物种。为了解GSK和HKT1基因在百萨偃麦草耐盐胁迫中的作用,本研究采用同源克隆的方法从百萨偃麦草中扩增获得了糖原合成酶激酶基因ThbGSK及高亲和性钾离子转运蛋白基因ThbHKT1的全长cDNA,ThbGSK基因全长1 233bp,与水稻、短柄草、小麦等物种的氨基酸一致性为93.92%~99.02%,表明GSK基因在这些物种间具有较高的保守性;聚类分析表明,ThbGSK与小麦TaGSK关系最近。ThbHKT1基因全长1 602bp,与水稻、小麦、短柄草、大麦等物种的氨基酸一致性为66.17%~92.87%,说明HKT1基因在这些物种间变异较大;聚类分析表明,ThbHKT1与大麦HvHKT1关系最近。RT-PCR和荧光定量PCR分析表明,经250mmol.L-1 NaCl胁迫处理后,GSK基因在百萨偃麦草和中国春诱导0~24h的叶片中表现为组成型表达,而HKT1基因在百萨偃麦草和中国春中均表现为诱导后上调表达。
Thinopyrum bessarabicum Lve(2n = 2x = 14,JJ) represents an important genetic resource for wheat improvement.In this study,two full-length orthologous genes of glycogen synthase kinase(GSK) designated ThbGSK and high-affinity K+ transporters(HKT) designated ThbHKT1 were cloned from Th.bessarabicum leaf cDNA.In comparison with that of rice,Brachypondium distachyon and wheat,the identity of amino acid of the gene varied from 93.92%~92.02%,indicating high conservation of this gene in these species.Cluster analysis showed a closer relationship between ThbGSK and TaGSK1.In comparison with that of rice,wheat,Brachypondium distachyon and barley,the identity of amino acid of the gene ThbHKT1 varied from 66.17%~92.87%,indicating variation of this gene in these species.Cluster analysis showed a closer relationship between ThbGSK and HvGSK1.Stressed with 250 mmol·L-1 NaCl for 0~24 h,GSK showed constitutive expression patterns both in Th.bessarabicum and Chinese Spring while HKT1 showed up-regulated after semi-quantitative and quantitative analysis.