甘蓝型油菜类甜蛋白激酶BnTLK1与真菌病害抗性相关  

Thaumatin-like protein kinase BnTLK1 from Brassica napus related to fungal disease resistance

作　　者：钟雪 李祥[1] 左蓉 刘杰 刘胜毅[1] 白泽涛[1] 童超波[1] ZHONG Xue;LI Xiang;ZUO Rong;LIU Jie;LIU Sheng-yi;BAI Ze-tao;TONG Chao-bo(Key Laboratory of Biology and Genetic Improvement of Oil Crops,the Ministry of Agriculture and Rural Affairs,Oil Crops Research Institute,Chinese Academy of Agricultural Sciences,Wuhan 430062,China)

机构地区：[1]中国农业科学院油料作物研究所,农业农村部油料作物生物学与遗传育种重点实验室,湖北武汉430062

出　　处：《中国油料作物学报》2021年第5期743-751,共9页Chinese Journal of Oil Crop Sciences

基　　金：国家自然科学基金(31770250);国家重点研发计划项目(2018YFE0108000、2016YFD0100305)。

摘　　要：类甜蛋白(thaumatin-likeprotein,TLP)是病程相关蛋白第5家族蛋白,在部分物种中发现有的TLP蛋白C端融合了激酶结构域。为探讨该类融合基因在植物与病原菌互作中的作用,通过生物信息学方法在甘蓝型油菜中鉴定到一个N端具有TLP结构域,C端为富含丝氨酸苏氨酸受体类激酶结构域的基因(BnTLK1)并成功克隆该基因。序列分析表明BnTLK1第254至276个氨基酸具有典型的跨膜结构,三级结构形成两个明显独立的TLP和激酶结构域。转录组数据显示,该基因在多个组织中表达丰度较低,此外,相比于菌核病感病品种Westar,高抗品种ZY821中其表达丰度较高,且核盘菌诱导后表达明显上调,表明该基因可能参与油菜抗菌核病过程。为了验证BnTLK1对核盘菌等真菌的影响,通过原核表达系统成功表达BnTLK1蛋白,在0.4 mmol/LIPTG和17℃低温条件下诱导培养,最终获得BnTLK1可溶性蛋白,抑菌实验发现BnTLK1能够抑制核盘菌和灰霉菌丝的生长。As the fifth family of pathogenesis-related proteins,TLP(thaumatin-like protein)plays an important defensive role in the interaction between plant and pathogen.In some species,the C-terminal of TLP fused kinase domain,but the fusion gene function required to be discussed.In Brassica napus,we identified and cloned a gene named BnTLK1,which contained a TLP domain at N termini and serine/threonine-rich receptor-like kinase domain at C termini.Sequence characterization showed that BnTLK1 had typical transmembrane structure and formed two independent units at 3D structure level.The expression of BnTLK1 was low in different tissues,high expressed in the cultivar′ZY821′of high resistance to Sclerotinia sclerotiorum,and induced by S.sclerotiorum based on public transcriptome data analysis.To investigate the effect of BnTLK1 on S.sclerotiorum and other fungus,BnTLK1 were expressed in prokaryotic system and get the soluble protein BnTLK1 after overnight induction at 0.4 mmol/L IPTG and 17℃.Bacteriostatic experiment further confirmed BnTLK1 could indeed inhibit the mycelium growth of Sclerotinia sclerotiorum and Botrytis cinerea.

关 键 词：甘蓝型油菜 类甜蛋白 激酶 核盘菌 原核表达 抑菌活性 

分 类 号：Q789[生物学—分子生物学]