大豆GmLecRlk基因耐盐功能分析

Analysis of salt tolerance function of GmLecRlk gene in soybean

凝集素类受体蛋白激酶属于类受体蛋白激酶(RLKs)家族,在植物的抗病防御反应、生长发育、胞内信号传导以及非生物胁迫反应过程中发挥重要作用。实验室前期对过表达抗逆转录因子GmNFYB1大豆进行转录组测序,获得了差异表达基因GmLecRlk(Glyma.07G005700),其开放阅读框长度为546 bp,编码181个氨基酸,蛋白结构域分析显示其含有两个丝氨酸/苏氨酸激酶结构域,属于一种G型凝集素类受体蛋白激酶。实时荧光定量PCR结果显示,GmLecRlk基因在大豆的根、茎、叶、荚中均有不同程度的表达,在根中表达量最高;200 mmol/LNaCl处理下,GmLecRlk的mRNA丰度先降低后升高,在12 h时达到最高值,表明该基因参与大豆对盐胁迫的响应。利用发根农杆菌K599获得GmLecRlk过表达转基因发状根复合植株,在盐胁迫处理下,其存活率高于对照;在拟南芥中异源表达GmLecRlk基因,转基因拟南芥在盐胁迫处理下的萌发率、绿化率和根长均高于野生型拟南芥。综上所述,GmLecRlk参与大豆对盐胁迫的反应,过量表达基因能提高大豆和拟南芥的耐盐性,为培育和改良抗盐大豆新品种提供新的途径和理论指导。

Lectin receptor like protein kinases belong to the receptor like protein kinases(RLKs)family,which play an important role in plant disease resistance and defense response, growth and development, intracellular signal transduction and abiotic stress response. Transcriptome sequencing was performed on soybean GmNFYB1, and the differentially expressed gene GmLecRlk(Glyma.07G005700)was obtained. Its length of open reading frame was 546 bp, encoding 181 amino acids. Protein domain analysis showed that GmLecRlk contained two serine/threonine kinase domains. It belongs to a G type lectin receptor protein kinase. The results of real-time quantitative PCR showed that GmLecRlk gene was expressed in root, stem, leaf and pod of soybean, and the highest expression level was found in root. Under 200 mmol/L NaCl treatment, the mRNA abundance of GmLecRlk decreased first and then increased, and reached the maximum value at 12 h, indicating that the gene was involved in the response of soybean to salt stress. Agrobacterium tumefaciens K599 was used to obtain GmLecRlk overexpressed transgenic hair-root complex plants. Under salt stress, the survival rate of overexpressed soybean hair-root complex plants was higher than that of control plants. GmLecRlk gene was heterologous expressed in Arabidopsis thaliana, and the germination rate, greening rate and root length of transgenic Arabidopsis thaliana under salt stress were higher than those of wild-type Arabidopsis. In conclusion, GmLecRlk is involved in the response of soybean to salt stress, and the overexpression of GmLecRlk gene can improve the salt tolerance of soybean and Arabidopsis thaliana, providing a new approach and theoretical guidance for breeding and improving salt-resistant soybean varieties.