知识与财富的链接
为探究精胺合成酶（spermine synthase,SPMS）与茶树（Camellia sinensis）耐寒性的关系,以茶树品种‘迎霜’为试验材料,利用简并引物PCR扩增技术结合5′/3′RACE方法,获得与低温胁迫相关的CsSPMS片段cDNA全长序列（GenBank登录号为KJ580429）。该序列全长1 374 bp,包含1 113 bp的完整开放阅读框（ORF）,编码371个氨基酸,预测分子量41.28 kD;构建亚细胞定位载体pJIT166-GFP/SPMS,亚细胞定位结果显示CsSPMS定位在细胞核上。荧光定量PCR分析表明CsSPMS在根、茎、叶、芽、花和果中都有表达,在根、茎中表达量较高;低温胁迫处理下不同组织CsSPMS的表达量,在叶片中2 h达到峰值,而在根中12 h达到高峰;在低温条件下4个茶树品种的耐寒性与CsSPMS表达量密切相关。
In order to explore the relationship between spermine synthase（SPMS）and cold resistance of tea plant（Camellia sinensis）,the full-length cDNA sequence of SPMS gene was cloned using reverse transcription-PCR（RT-PCR）combined with RACE techniques from tea plant cultivar‘Yingshuang’. This gene was named CsSPMS with GenBank accession number KJ580429. It was 1 374 bp in length,containing a 1 113 bp open reading frame（ORF）which encoded 371 amino acid residues of 41.28 kD molecular weight. The localization assays indicated that CsSPMS protein localized in the cell nucleus,after observing its subcellular localization by the fusion expression vector pJIT166-GFP/SPMS. CsSPMS gene expressed in all the tissues,including root,stem,leaf,flower and fruit analyzed by quantitative real-time PCR（qRT-PCR）,the highest expression level were in root and stem. The qRT-PCR analysis also indicated that the expression level of CsSPMS was up-regulated in roots,stems and leaves by cold treatment. The top expression level of CsSPMS in leaves was after 2 h treatment,while after 12 h treatmentin roots. Furthermore,cold resistance of four tea plant cultivars is closely related to expression levels of CsSPMS under low temperature condition.
