油菜抗菌肽基因BnCRP1的克隆表达和活性分析  

Antimicrobial Activity Detection of Antimicrobial Peptide Gene BnCRP1 from Brassica napus

作　　者：曹慧慧[1,2] 王磊 闫艳华 林田 柯涛 黄军艳[1] 程晓晖[1] 刘胜毅[1] Cao Huihui;Wang Lei;Yan Yanhua;Lin Tian;Ke Tao;Huang Junyan;Cheng Xiaohui;Liu Shengyi(Oil Crops Research Institute,Chinese Academy of Agriculture Science,Key Laboratory of Biology and Genetic Improvement of Oil Crops,Ministry of Agriculture,Wuhan 430062;Tangshan Food and Drug Comprehensive Testing Center,Tangshan 063000;Department of Life Science and Technology,Nanyang Normal University,Nanyang 473061)

机构地区：[1]中国农业科学院油料作物研究所,农业部油料作物生物学与遗传育种重点实验室,武汉430062 [2]唐山市食品药品综合检验检测中心,唐山063000 [3]南阳师范学院生命科学与技术学院,南阳473061

出　　处：《中国粮油学报》2020年第2期91-97,共7页Journal of the Chinese Cereals and Oils Association

基　　金：国家重点研发计划(2018YFD0200904、2016YFD 0100602、2016YFD0101007);国家油菜产业技术体系(CARS-12)

摘　　要：通过生物信息学的方法,充分利用甘蓝型油菜组织转录组测序数据和已有的抗菌肽数据库,筛选获得一个核苷酸长度为102 bp,编码34个氨基酸长度的新型抗菌肽基因BnCRP1,BnCRP1抗菌肽序列中有9个半胱氨酸,半胱氨酸含量超过27%,是一种新型的植物来源的半胱氨酸富集类抗菌肽(Cystin rich protein CRPs),在抗菌肽分类中属于打结素类抗菌肽(Knottin-like Peptides)。抑菌活性检测结果表明抗菌肽BnCRP1对细菌和真菌都具有较强的抑菌活性。拟南芥离体叶片涂抹接菌实验证实,BnCRP1能够显著增强植物对腐生营养型真菌核盘菌的抗性,能够作为生防制剂用于菌核病的防治中,还可以作为一个新基因源用于植物基因工程,以培育抗病转基因植物。本研究为利用生物信息手段和基因工程技术大规模鉴定新植物来源抗菌肽提供参考。Through the method of bioinformatics,in this study,the sequencing data of the tissue transcription group of Brassica napus and the database of antimicrobial peptides have been obtained,and a new antimicrobial peptide gene BnCRP1 which harbored 102 bp encoding 34 amino acids has been got.There were 9 cysteines in the BnCRP1 sequence,and the cysteine content exceeded 27%.It was a new type of cysteine-enriched antimicrobial peptide(Cystine rich peptide,CRP)which derived from plants belonging to the Knottin-like peptides in the classification of antimicrobial peptides.Antimicrobial activity demonstrated that BnCRP1 exhibited strong antimicrobial activity against Gram-positive bacteria,Gram-negative bacteria and yeast.BnCRP1 also had strong antifungal activity against Sclerotinia sclerotiorum.BnCRP1 has been proved to significantly enhance the resistance of plants to saprotrophic fungal Sclerotinia sclerotiorum in vitro leaf grafting experiments of Arabidopsis.It can be used as a biocontrol preparation for the prevention and treatment of sclerotinia,and can also be used as a new gene source for plant genetic engineering to cultivate disease-resistant transgenic plants.This study laid a foundation for the large-scale identification of antimicrobial peptides from new plants by bioinformatics and genetic engineering.

关 键 词：油菜 抗菌肽 半胱氨酸富集类抗菌肽 融合表达 活性检测 

分 类 号：S33[农业科学—作物遗传育种]