基于转录组测序的山药NBS-LRR家族基因鉴定和分析  

Identification and Analysis of NBS-LRR Family Genes in Dioscorea opposita Based on Transcriptome Sequencing

作　　者：李顺朋 李朝闯 王兰宁 张昊[1,2] 王瑞飞 李明军[1,3] 杨清香 LI Shunpeng;LI Chaochuang;WANG Lanning;ZHANG Hao;WANG Ruifei;LI Mingjun;YANG Qingxiang(College of Life Sciences,Henan Normal University,Xinxiang 453007,Henan,China;Henan International Joint Laboratory of Agricultural Microbia l Ecology and Technology,Xinxiang 453007,Henan,China;Engineering Technology Research Center of Nursing and Utilization of Genuine Chinese Crude Drugs of Henan Province,Xinxiang 453007,Henan,China)

机构地区：[1]河南师范大学生命科学学院,河南新乡453007 [2]河南省农业微生物生态与技术国际联合实验室,河南新乡453007 [3]河南省道地药材保育及利用工程技术中心,河南新乡453007

出　　处：《中国蔬菜》2024年第8期47-57,共11页China Vegetables

基　　金：国家中药材产业技术体系项目(CARS-21);国家重点研发计划资助项目(2023YFD1600603);河南省高等学校重点科研项目(24A210016);中原英才计划-科技创新领军人才项目(224200510011)。

摘　　要：为了探究NBS-LRR类抗病基因在铁棍山药响应不同真菌侵染中的作用,通过同源克隆从山药基因组中扩增出NBS类基因DoRGA2和DoRGA4,RT-qPCR分析发现山药黑斑病病原真菌互隔交链孢和内生真菌耐盐青霉侵染会不同程度诱导DoRGA2和DoRGA4上调表达。同时,利用转录组测序分析互隔交链孢和耐盐青霉侵染山药愈伤组织后的NBS-LRR家族基因,结果共鉴定到47个山药NBS-LRR家族基因,包括N、NL、CN、CNL和TN 5种类型,分别有25、15、3、3、1个基因;不同亚家族NBS-LRR蛋白的进化路径存在差异,且这些NBS-LRR类蛋白以酸性为主,普遍具有亲水性,主要定位在细胞膜或细胞质。这些NBS-LRR家族基因中,TRINITY_DN9146_c0_g1、TRINITY_DN8009_c1_g1、TRINITY_DN49830_c0_g1和TRINITY_DN39475_c0_g1在接种互隔交链孢后显著上调表达,而TRINITY_DN18700_c0_g3在接种耐盐青霉菌后显著上调。To explore the role of NBS-LRR resistance genes in response to different fungal infections in‘Tiegun’yam,NBS-L RR genes DoRGA2 and DoRGA4 were amplified from the genome of‘Tiegun’yam by homologous cloning.RT-qPCR analysis revealed that the infection of the black spot disease pathogenic fungus Alternaria alternata and the endophytic fungus Penicillium halotolerans could induce the up-regulated expression of DoRGA2 and DoRGA4 in different degrees.Meanwhile,RNA-seq was performed to analyze the NBS-LRR family genes of‘Tiegun’callus infected by A.alternata and P.halotolerans,the results showed that a total of 47 NBS-LRR family genes were identified,including 5 types of N,NL,CN,CNL and TN,with 25,15,3,3 and 1 gene,respectively.Furthermore,different subfamilies of NBS-LRR proteins had different evolutionary paths,and these proteins were mainly acidic proteins with hydrophilic properties,and were mainly located in the cell membrane or cytoplasm.In addition,among these NBS-LRR family genes,the expressions of TRINITY_DN9146_c0_g1,TRINITY_DN8009_c1_g1,TRINITY_DN49830_c0_g1 and TRINITY_DN39475_c0_g1 were significantly up-regulated after inoculation with A.alternata,while TRINITY_DN18700_c0_g3 was significantly up-regulated after infection with P.halotolerans.

关 键 词：山药 NBS-LRR基因 互隔交链孢 黑斑病 RNA-SEQ 

分 类 号：S632.1[农业科学—蔬菜学]