Differential expression of Rs-eng-1b in two populations of Radopholus similis (Tylenchida: Pratylecnchidae) and its relationship to pathogenicity
Differential expression of Rs-eng-1b in two populations of Radopholus similis (Tylenchida: Pratylecnchidae) and its relationship to pathogenicity
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Rs-eng-1b在Radopholus similis（垫刃纲：短体纲）两个群体中的差异表达及其与致病性的关系
DOI：
10.1007/s10658-012-0015-4
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发表时间：
2012-08-01
影响因子：
1.8
通讯作者：
Li, Yu
中科院分区：
文献类型：
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作者：
Zhang, Chao;Xie, Hui;Li, Yu
以来源于不同寄主植物的相似穿孔线虫Rs-C和Rs-P群体为材料，构建了对香蕉和红掌致病力差异较大的相似穿孔线虫SSH文库。为了从SSH文库中筛选具有显著表达差异的克隆，随机选择总共2，400个克隆，并对它们进行反向北方印迹。在2，400个克隆中，89个克隆显示出显著的表达差异。在对这89个克隆进行测序之后，获得了来自87个克隆的不同序列。将87个不同的序列与NCBI中的非冗余核苷酸数据库（nr）进行比对，我们发现5个序列与Rs-eng-1b高度保守。再用其中的两个467 bp（GW 395922）和742 bp（GW 395923）的序列分别进行5′ RACE-PCR和3′ RACE-PCR。随后，获得了Rs-eng-1b（EU 414839）的全长（1，427 bp）。qPCR结果表明，Rs-eng-1b在对寄主植物致病性强的群体Rs-C中的表达量是对寄主植物致病性弱的群体Rs-P中的2.7倍。此外，来自Rs-C群体的基因Rs-eng-1b在四个不同发育阶段也表现出表达差异。Rs-eng-1b相对表达丰度从高到低依次为雌虫、幼虫、雄虫、卵。我们进一步利用RNAi技术对Rs-eng-1b的致病性进行了研究，这是第一次利用RNAi技术对Rs-eng-1b进行研究。RNAi结果表明，Rs-eng-1b基因的表达量与致病性呈正相关。RNAi处理时间越长，线虫群体的致病性越低。非内源性gfp dsRNA对Rs-eng-1b的表达和致病性无明显影响。similis Rs-C群体。综上所述，Rs-eng-1b可能是一个重要的致病相关基因。相似的
We constructed a SSH (suppression subtractive hybridization) library based on two populations (Rs-C and Rs-P) of Radopholus similis from different host plants and exhibiting differences in pathogenicity on Musa paradisiaca and Anthurium andraeanum plants. In order to screen the clones with significant expression differences from the SSH library, a total of 2,400 clones was randomly selected and reverse northern blotting was performed on them. Out of the 2,400 clones, 89 clones showed significant expression differences. Out of sequencing these 89 clones, distinct sequences from 87 clones were obtained. Aligning the 87 distinct sequences against the non-redundant nucleotide database (nr) in NCBI, we found that five sequences were highly conserved with Rs-eng-1b. Two of five sequences with lengths of 467 base pairs (bp) (GW395922) and 742 bp (GW395923) were further employed to perform 5′ RACE-PCR and 3′ RACE-PCR, respectively. Subsequently, the complete length of Rs-eng-1b (EU414839) was obtained (1,427 bp). Our qPCR result showed that expression of Rs-eng-1b in the population Rs-C with high pathogenicity on host plants was approximately 2.7 times as much as the expression of Rs-eng-1b in the population Rs-P with low pathogenicity on host plants. Furthermore, the gene Rs-eng-1b from the Rs-C population also showed expression differences amongst four different development stages. The order of Rs-eng-1b relative expression abundance from high to low was females, juveniles, males, and eggs. We further used RNAi to test whether Rs-eng-1b of Rs-C population was responsible for pathogenicity which was the first RNAi work about Rs-eng-1b. The RNAi results showed that Rs-eng-1b expression had a positive correlation to pathogenicity of the population. The longer the RNAi treatment, the less pathogenic the nematode population was. Non-endogenous gfp dsRNA had no significant influence on the expression of Rs-eng-1b and pathogenicity of R. similis Rs-C population. In conclusion, all our evidence indicated Rs-eng-1b might be a crucial pathogenicity-related gene in R. similis.