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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。
According to the EST sequences expressed in the esophageal glands of Meloidogyne javanica and the trans-splicing sequence SL1, a putative Mj-1-1 (KU358725) was cloned from the root-knot nematode of Java. The full-length cDNA of this gene was 573 bp, containing 450 bp open reading frame (ORF) encoding 149 amino acids. The full-length DNA is 740 bp in length and contains two introns, 20 bp in length and 111 bp in length. The NCBI BLASTn alignment showed that Mj-1-1 had the highest similarity to M. incognita zk1236.5 (JQ284068) gene of Meloidogyne incognita, 97%. In situ hybridization showed that the Mj-1-1 gene was expressed in the dorsal esophageal glands of Meloidogyne javanica, and the qRT-PCR results showed that the Mj-1-1 gene was most expressed in the 3rd instar larvae of M.javi. The Mj-1-1 gene of 2nd instar larvae before root-knot nematode infection was silenced. It was found that the infection of Mj-1 against Mj-1 was significantly decreased after Mj-1-1 silencing, indicating that Mj-1- 1 has an important regulatory effect on root knot nematode infection and parasitism.