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目的研究天麻晚期胚胎富集蛋白(LEA2)与抗寒性的关系。方法根据天麻种子转录组测序结果,得到天麻晚期胚胎富集蛋白基因序列,获得重组质粒后导入大肠杆菌进行低温实验。结果获得两条天麻LEA2基因c DNA序列。GeLEA2-1和GeLEA2-2全长分别为477和942 bp,编码158和313个氨基酸,其编码蛋白理论相对分子质量为17.30×103和34.85×103。系统发育分析显示,天麻LEA2基因分别与拟南芥和水稻LEA2基因具有较高相似性。此外,天麻LEA2基因与已知的多种植物的LEA2蛋白具有相似的功能结构域。低温实验显示,与对照组相比,含有GeLEA2重组质粒的大肠杆菌的抗寒性较差。结论推测天麻LEA2为抗寒相关的负向调节基因。
Objective To study the relationship between late embryogenic enriched protein (LEA2) and cold resistance in Gastrodia elata Blume. Methods According to the sequencing results of Gastrodia seed transcriptome, the gene sequence of late embryogenesis enriched protein was obtained. The recombinant plasmids were introduced into Escherichia coli for hypothermia test. Results Two DNA sequences of LEA2 gene from Gastrodia elata Blume were obtained. The full length of GeLEA2-1 and GeLEA2-2 were 477 bp and 942 bp, encoding 158 and 313 amino acids, respectively. The theoretical molecular weights of GeLEA2-1 and GeLEA2-2 were 17.30 × 103 and 34.85 × 103, respectively. Phylogenetic analysis showed that the LEA2 gene of Gastrodia elata was highly similar to the LEA2 gene of Arabidopsis and rice, respectively. In addition, the LEA2 gene of Gastrodiale has a similar functional domain as the LEA2 protein of many plants known. Low-temperature experiments showed that compared with the control group, Escherichia coli containing GeLEA2 recombinant plasmid has poor cold resistance. Conclusions Gastrodia elata Blume LEA2 is a negative regulator of cold-resistance.