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蔗糖磷酸合成酶(Sucrose phosphate synthase,SPS)是蔗糖合成途径的关键限速酶,在蔗糖积累和碳分配中具有重要作用.在茎组织中SPSⅡ表达量占SPS转录总量的40%,表明SPSⅡ cDNA的克隆与表达对蔗茎中蔗糖的积累有着重要的影响.通过RT-PCR技术克隆分离SPSⅡ基因cDNA片段.序列分析表明该cDNA片段包含1个3 183 bp的开放读码框,可编码1 060个氨基酸.GenBank登录号为EU269038.通过实时荧光定量PCR检测SPSⅡ基因在甘蔗糖分积累的不同时期、不同组织部位的相对表达量.结果表明,在糖分积累初期蔗茎中的SPSⅡ相对表达量最大,在糖分积累中期蔗叶中的SPSⅡ相对表达量达到最高峰;同组织部位中,糖分积累初期SPSⅡ相对表达量高于糖分积累中期、后期.图6参11
Sucrose phosphate synthase (SPS) is a key rate-limiting enzyme in sucrose synthesis pathway, which plays an important role in sucrose accumulation and carbon partitioning. SPSII expression in stem tissues accounted for 40% of total SPS transcripts, indicating that SPSII The cloning and expression of cDNA have an important effect on the accumulation of sucrose in cane stems.The cDNA fragment of SPSⅡ gene was cloned and isolated by RT-PCR.The sequence analysis showed that the cDNA fragment contained a 3 183 bp open reading frame encoding 1 060 amino acids.The GenBank accession number was EU269038.The relative expression level of SPSII gene in different tissues and parts of cane sugar accumulation was detected by real-time fluorescence quantitative PCR.The results showed that the relative expression level of SPSII in sugarcane stem during the early stage of sugar accumulation was the largest , The relative expression level of SPSII in sugarcane leaf reached the peak in the middle stage of sugar accumulation.The relative expression level of SPSII in early stage of sugar accumulation in the same tissues was higher than that in middle and late stages of sugar accumulation