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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Cs SPMS片段cDNA全长序列(Gen Bank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C SPMS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 tolerance of Camellia sinensis, tea varieties Ying Yinguang was used as experimental material and degenerated primers PCR amplification combined with 5 ’/ 3’RACE The full-length cDNA sequence of Cs SPMS fragment (GenBank accession number KJ580429) was obtained. The full length cDNA was 1 374 bp in length and contained a 1 113 bp open reading frame (ORF) encoding 371 amino acids with a predicted molecular mass of 41.28 kD. The subcellular localization vector pJIT166-GFP / SPMS was constructed. On the nucleus. Quantitative real-time PCR analysis showed that CSPMS was expressed in roots, stems, leaves, buds, flowers and fruit, and higher in roots and stems. The expression of CsSPMS in different tissues under low temperature stress was significantly lower in 2 h Reached the peak value, and peaked at 12 h in roots. The cold tolerance of 4 tea cultivars at low temperature was closely related to the expression of CsSPMS.