Duarte A, Maleita C, Tiago I, Curtis R, Abrantes I
IMAR-CMA, Department of Life Sciences,University of Coimbra,P 3004 517,Coimbra,Portugal.
CIEPQPF, Department of Chemical Engineering,University of Coimbra,P 3030 790Coimbra,Portugal.
J Helminthol. 2016 Jan;90(1):28-38. doi: 10.1017/S0022149X1400073X. Epub 2014 Oct 16.
Meloidogyne hispanica (Mhi) is a difficult-to-control polyphagous root-knot nematode (RKN) species of emerging importance for economically valuable crops. Nematode secretions are likely to be the first signals perceived by the plant and are thought to be involved in various aspects of the plant-nematode interaction. The aims of this work were to identify and characterize M. hispanica parasitism genes: cathepsin L cysteine protease (cpl-1), calreticulin (crt-1), β-1,4-endoglucanase-1 (eng-1) and manganese superoxide dismutase (mnsod). As there are no genomic data available for M. hispanica, primers were designed from the conserved regions of the putative parasitism genes in M. incognita and M. hapla and used to amplify the genes in M. hispanica, which led to the successful amplification of these genes in M. hispanica. Partial gene sequences were also obtained for M. arenaria, M. hapla, M. hispanica, M. incognita and M. javanica cpl-1, crt-1, eng-1 and mnsod genes, and their phylogenetic relationship analysed. In order to determine whether these genes are differentially expressed during M. hispanica development, cDNA was amplified from mRNA isolated from eggs, second-stage juveniles (J2) and females. Amplification products were observed from cDNA of all developmental stages for the Mhi-cpl-1 and Mhi-crt-1 genes. However, the gene Mhi-crt-1 exhibited intense amplification bands in females, while the Mhi-eng-1 gene was equally amplified in eggs and J2 and the Mhi-mnsod gene was only expressed in eggs. In comparison to the other RKN species, the genes Mhi-eng-1 and Mhi-mnsod showed transcription in different nematode developmental stages.
西班牙根结线虫（Mhi）是一种难以控制的多食性根结线虫（RKN），对经济价值作物的重要性日益凸显。线虫分泌物可能是植物感知到的首个信号，并被认为参与了植物与线虫相互作用的各个方面。这项工作的目的是鉴定和表征西班牙根结线虫的寄生基因：组织蛋白酶L半胱氨酸蛋白酶（cpl-1）、钙网蛋白（crt-1）、β-1,4-内切葡聚糖酶-1（eng-1）和锰超氧化物歧化酶（mnsod）。由于没有西班牙根结线虫的基因组数据，根据南方根结线虫和北方根结线虫假定寄生基因的保守区域设计引物，并用于扩增西班牙根结线虫中的这些基因，从而成功在西班牙根结线虫中扩增出这些基因。还获得了花生根结线虫、北方根结线虫、西班牙根结线虫、南方根结线虫和爪哇根结线虫cpl-1、crt-1、eng-1和mnsod基因的部分基因序列，并分析了它们的系统发育关系。为了确定这些基因在西班牙根结线虫发育过程中是否差异表达，从卵、二龄幼虫（J2）和雌虫分离的mRNA中扩增cDNA。在所有发育阶段的cDNA中均观察到Mhi-cpl-1和Mhi-crt-1基因的扩增产物。然而，Mhi-crt-1基因在雌虫中表现出强烈的扩增条带，而Mhi-eng-1基因在卵和J2中扩增程度相同，Mhi-mnsod基因仅在卵中表达。与其他根结线虫物种相比，Mhi-eng-1和Mhi-mnsod基因在不同线虫发育阶段表现出转录情况。
