凹叶厚朴低温胁迫SSH文库的构建及差异基因的表达分析

Construction of Magnolia officinalis Low Temperature Stress SSH Library and Differential Gene Expression Analysis

[目的]通过凹叶厚朴差减文库的构建及序列分析,找出其主要抗寒相关功能基因。[方法]随机挑选单克隆菌落进行PCR检测,对837个阳性克隆测序后进行拼接得到97个unigenes,将97个unigenes在NCBI进行BLAST比对,后进行BLAST2GO功能分类,推定出11个相关基因进行荧光定量PCR。[结果]CHS、APX、TRX、ERF、DREB、LHCB、WRKY、HSP、PP2C、GAPDH 10个基因在凹叶厚朴受到低温侵害时出现差异表达,推断这10个基因在低温时可能表现出防御机制。[结论]该研究为与凹叶厚朴亲缘关系相近的植物抗低温特性遗传育种提供理论基础。

Objective] The aim was to obtain cold resistance relevant genes through construction of Magnolia officinalis subtractive library and sequence analysis.[Method] PCR detection was conducted on randomly selected monoclonal colony, after 837 positive clone sequencing, 97 unigenes were obtained, BLAST comparison was conducted on NCBI, then BLAST2GO functional classification was carried out.[Result] Fluo-rescence quantitative PCR was conducted on 11 relevant genes.CHS, APX, TRX, ERF, DREB, LHCB, WRKY, HSP, PP2C, GAPDH exhibited the defense mechanism when Magnolia officinalis was damaged by low temperature.[Conclusion] The study can provide theoretical basis for cold resistance genetic breeding of plants with similar affinities to Magnolia officinalis.