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硝酸盐转运蛋白(NRT)是植物吸收和利用硝态氮的一种关键蛋白。运用RACE技术从茶树中扩增出NRT基因的cDNA,并利用实时荧光定量PCR检测了CsNRT基因在不同茶树器官与品种之间的差异表达。结果表明:CsNRT基因的cDNA全长2 061 bp,开放阅读框为1 818 bp,编码含由605个氨基酸组成的蛋白质,GenBank登录号为KJ160503,属于NRT2基因家族。CsNRT为组成型基因,对不同处理的水培茶苗进行定量表达分析显示,该基因在根、茎、叶中都有表达,其中在根部的表达水平最高,1.0 mmol·L-1的NO3-可诱导其表达量上升7.53倍。不同茶树品种中CsNRT基因的表达也有较大差异,‘龙井长叶’和‘凫早2号’的表达量较高,前者强烈响应0.5和1.0 mmol·L-1 NO3-的诱导,后者的响应浓度为1.0和2.0mmol·L-1,而‘舒茶早’在各浓度下的表达差异不明显。
Nitrate transporter (NRT) is a key protein that plants absorb and utilize nitrate nitrogen. The RACE technique was used to amplify the cDNA of NRT gene from tea plant, and the differential expression of CsNRT gene in different tea plant organs and cultivars was detected by real-time fluorescence quantitative PCR. The results showed that the cDNA of CsNRT gene was 2 061 bp in length and 1 818 bp in open reading frame, encoding a protein consisting of 605 amino acids. GenBank accession No. KJ160503 belongs to NRT2 gene family. CsNRT is a constitutive gene. Quantitative expression analysis of hydroponics cultivars with different treatments showed that the gene was expressed in roots, stems and leaves, and the highest expression level was in the roots. The expression level of NO3- Can induce its expression increased by 7.53 times. The expression of CsNRT gene was also different in different tea varieties. The expression of CsNRT gene in Longjing Longye and Longzao 2 was high, the former strongly responded to the induction of 0.5 and 1.0 mmol·L-1 NO3- The response concentrations were 1.0 and 2.0 mmol·L-1, while the expression of ’Shu Chazao’ at each concentration was not significantly different.