爪哇根结线虫Mj-1-1基因的克隆、鉴定及RNAi表型分析  

Cloning,identification and RNA interference analysis of Mj-1-1 gene from Meloidogyne javanica

作　　者：扈丽丽[1,2] 卓侃[1,2] 林柏荣[1,2] 陈海燕[1] 廖金铃[1,2,3] HU Li-li ZHUO Kan LIN Bo-rong CHEN Hai-yan LIAO Jin-ling(Laboratory of Plant Nematology, South China Agricultural University, Guangzhou 510642, China = Guangdong Province Key Laboratory of Microbial Signals and Disease Control, Guangzhou 510642, China Guangdong Vocational College of Ecological Engineering, Guangzhou 510520, China)

机构地区：[1]华南农业大学植物线虫研究室,广州510642 [2]广东省微生物信号与作物病害防控重点实验室,广州510642 [3]广东生态工程职业学院,广州510520

出　　处：《植物病理学报》2017年第1期75-81,共7页Acta Phytopathologica Sinica

基　　金：国家重点基础研究发展计划(973计划)(2013CB127501);国家自然科学基金资助项目(31401716)

摘　　要：根据爪哇根结线虫食道腺内表达的EST序列,结合反式剪接序列SL1,从爪哇根结线虫中克隆了一个假定寄生基因Mj-1-1(KU358725),该基因的c DNA全长为573 bp,包含450 bp的开放阅读框(ORF),编码149个氨基酸。DNA全长为740 bp,包含2个内含子,长度分别为20 bp和111 bp。NCBI BLASTn比对表明,Mj-1-1与南方根结线虫的M.incognita zk1236.5(JQ284068)基因相似性最高,为97%。原位杂交表明Mj-1-1基因在爪哇根结线虫的背食道腺中表达,qRT-PCR结果表明Mj-1-1基因在爪哇根结线虫寄生性3龄幼虫阶段表达量最高。对爪哇根结线虫侵染前2龄幼虫的Mj-1-1基因进行沉默,调查发现,沉默Mj-1-1后,爪哇根结线虫对番茄的侵染力显著下降,表明Mj-1-1对爪哇根结线虫侵染和寄生具有重要的调控作用。A putative parasitism gene named Mj-1-1（KU358725） was cloned based on the EST library of Meloidogyne javanica.The full length cDNA of Mj-1-1 is 573 bp,containing a 450 bp open reading frame（ORF）.The ORF encodes 149 amino acids.Two introns（20 bp and 111 bp） exist in the 740 bp DNA sequence.NCBI Blastn showed that Mj-1-1 had a 97%similarity with a sequence（JQ284068） of Meloidogyne incognita.In situ hybridization showed that Mj-1-1 was expressed in the dorsal gland cell of M.javanica.The highest expression level was detected in the third stage juveniles by qRT-PCR.Infection ability of nematode was significantly deceased after soaking the pre-parasitic juveniles in the dsRNA of Mj-1-1.The results showed that Mj-1-1 could play an important regulatory role in the parasitism of M.javanica.

关 键 词：爪哇根结线虫 Mj-1-1基因 克隆 鉴定 RNAI 

分 类 号：S432.45[农业科学—植物病理学]