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易变山羊草抗禾谷孢囊线虫候选基因克隆及鉴定

机译:易变山羊草抗禾谷孢囊线虫候选基因克隆及鉴定

摘要

Cereal cyst nematode (CCN) (Heterodera avenae Woll.) is one of the most economically damaging endoparasite pests of wheat worldwide. We isolated and characterized a novel cereal CCN resistance candidate gene, CreV8, from Aegilops variabilis (2n = 28, UUSvSv). The gene was 3,568 bp long and showed high nucleotide similarity with the known nematode resistance genes Cre3, CreZ, and go35. CreV8 consisted of two exons, one intron and a complete ORF of 2,763 bp. The deduced peptide sequence consisted of 920 amino acid residues and shared 97.83% amino acid identity with Cre3. The sequence harbored a signal peptide domain, a nucleotide-binding ARC (NB-ARC) domain, and a leucine-rich repeat (LRR) domain, all of which are typical characteristics of resistance genes. We proposed the resistance mechanism of CreV8 based on functional analysis and predictions from its conserved domains and tertiary structure. Real-time polymerase chain reaction (PCR) showed that CreV8 was expressed in CCN-inoculated roots of resistant plants, and its expression levels were 2.80 and 27.45 times higher than that in the untreated control at 3 and 11 days post-inoculation, respectively. These results indicate that CreV8 belongs to the nucleotide binding site (NBS)-LRR resistance gene family and that it is a candidate CCN-resistance gene.
机译:谷物囊肿线虫(CCN)(Heterodera avenae Woll。)是全球范围内最具经济破坏力的小麦内寄生虫害之一。我们分离并鉴定了一种新的谷物抗CCN候选基因CreV8,该基因来自变异山羊草(Aegilops variabilis)(2n = 28,UUSvSv)。该基因长3568 bp,与已知的线虫抗性基因Cre3,CreZ和go35表现出高度的核苷酸相似性。 CreV8由两个外显子,一个内含子和一个2763 bp的完整ORF组成。推导的肽序列由920个氨基酸残基组成,与Cre3具有97.83%的氨基酸同一性。该序列包含一个信号肽域,一个核苷酸结合ARC(NB-ARC)域和一个富含亮氨酸的重复(LRR)域,所有这些都是抗性基因的典型特征。基于功能分析和保守域和三级结构的预测,我们提出了CreV8的抗性机制。实时聚合酶链反应(PCR)显示CreV8在接种CCN的抗性植物根中表达,在接种后3天和11天其表达水平分别比未处理的对照高2.80和27.45倍。这些结果表明CreV8属于核苷酸结合位点(NBS)-LRR抗性基因家族,并且它是候选的CCN抗性基因。

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