菊花去饱和酶CmSAD基因的克隆与表达分析

发布时间：2018-05-11 08:34

  本文选题：菊花 + 低温　； 参考：《河南农业大学学报》2017年03期

【摘要】：以秋菊(Chrysanthemum morifolium Ramat.)抗寒品种‘星光灿烂’为材料,利用RT-PCR和RACE方法从叶片中克隆△9硬脂酰-ACP去饱和酶(Stearoyl-ACP desaturase,SAD)基因,命名为CmSAD,Gen Bank登录号为KC529335。该基因cDNA全长1 203 bp,编码401个氨基酸,相对分子质量为45.57 k D,等电点为6.48,编码的氨基酸序列与新疆雪莲同源性最高(80.29%)。通过对该蛋白进行生物信息学分析得出其没有跨膜结构,主要存在于叶绿体基质中,N端含有一段包含60个氨基酸的叶绿体转运肽。通过实时荧光定量PCR研究低温胁迫下叶片和根系中CmSAD基因的表达量变化,该基因在叶片和根系中均有表达。根系中CmSAD基因的表达量随着温度的降低而降低,16℃时表达量最高;叶片中CmSAD表达量随着温度的降低先升高再降低,5℃时最高。随着温度的降低,菊花叶片和根系中CmSAD基因的表达情况表现出一定的差异。CmSAD基因的表达变化与温度有关,为低温胁迫下菊花脂肪酸代谢的分子机理研究提供了基础。
[Abstract]:Chrysanthemum morifolium Ramat. The gene of 9 stearoyl-ACP desaturase (Stearoyl-ACP desaturase) was cloned from the leaves by RT-PCR and RACE, and named as CmSADGen Bank accession number KC529335. The total length of cDNA is 1 203 BP, encoding 401 amino acids, the relative molecular weight is 45.57 KD, and the isoelectric point is 6.48. The amino acid sequence encoded by this gene has the highest homology with that of Saussurea Sinkiang (80.29). By bioinformatics analysis, it was found that the protein had no transmembrane structure and mainly existed in the chloroplast matrix containing a chloroplast transport peptide containing 60 amino acids. The changes of CmSAD gene expression in leaves and roots under low temperature stress were studied by real-time fluorescence quantitative PCR. The CmSAD gene was expressed in both leaves and roots. The expression of CmSAD gene in roots decreased with the decrease of temperature at 16 鈩，

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