Hira Abbas, Nazia Nahid, Muhammad Shah Nawaz Ul Rehman, Tayyaba Shaheen, Sadia Liaquat
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Alt a 1 gene was amplified using gene specific primers. The isolated pathogen produced similar symptoms when inoculated on mungbean and tobacco. The sequence analysis of the internal transcribed spacer (ITS) region, a 600 bp fragment amplified using specific primers, ITS1 and ITS2 showed 100% identity with A. alternata. Potato virus X (PVX) -based silencing vector expressing Alt a 1 gene was constructed to control this pathogen through RNA interference in tobacco. Out of 50 inoculated plants, 9 showed delayed onset of disease. Furthermore, to confirm our findings at molecular level semi-quantitative reverse transcriptase polymerase chain reaction was used. Both phenotypic and molecular investigation indicated that RNAi induced through the VIGS vector was efficacious in resisting the pathogen in the model host, Tobacco (Nicotiana tabacum). 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Diversity of 50 genotypes of mungbean was assessed against A. alternata and data on pathological traits was subjected to cluster analysis. The results showed that genotypes of mungbean were grouped into four clusters based on resistance parameters under the influence of disease. The principal component biplot demonstrated that all the disease-related parameters (% disease incidence, % disease intensity, lesion area, and % of infection) were strongly correlated with each other. Alt a 1 gene that is precisely found in Alternaria species and is responsible for virulence and pathogenicity. Alt a 1 gene was amplified using gene specific primers. The isolated pathogen produced similar symptoms when inoculated on mungbean and tobacco. The sequence analysis of the internal transcribed spacer (ITS) region, a 600 bp fragment amplified using specific primers, ITS1 and ITS2 showed 100% identity with A. alternata. 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引用次数: 0
摘要
对绿豆种植区进行了一次全面调查,以观察由交替丝核菌(Alternaria alternata)引起的叶斑病。在叶片上观察到了交替孢霉叶斑病的症状。评估了 50 种绿豆基因型对交替孢霉的多样性,并对病理特征数据进行了聚类分析。结果表明,在病害影响下,绿豆基因型根据抗性参数被分为四个群组。主成分双平面图表明,所有与病害相关的参数(病害发生率、病害强度、病害面积和感染率)之间都有很强的相关性。Alt a 1 基因恰恰存在于 Alternaria 物种中,是毒力和致病性的决定因素。Alt a 1 基因是利用基因特异性引物扩增的。分离出的病原体接种到绿豆和烟草上会产生相似的症状。使用特异引物 ITS1 和 ITS2 扩增的内部转录间隔区(ITS)600 bp 片段的序列分析表明,该病原体与交替马铃薯病毒(A. alternata)100% 相同。构建了表达 Alt a 1 基因的基于马铃薯病毒 X(PVX)的沉默载体,通过 RNA 干扰烟草来控制这种病原体。在 50 株接种植株中,有 9 株表现出延迟发病。此外,为了在分子水平上证实我们的发现,我们使用了半定量逆转录酶聚合酶链反应。表型和分子研究都表明,通过 VIGS 载体诱导的 RNAi 能有效抵抗模式宿主烟草(Nicotiana tabacum)中的病原体。据我们所知,这项研究尚属首次报道。
Assessment of Resistance Induction in Mungbean against Alternaria alternata through RNA Interference.
A comprehensive survey of mungbean-growing areas was conducted to observe leaf spot disease caused by Alternaria alternata. Alternaria leaf spot symptoms were observed on the leaves. Diversity of 50 genotypes of mungbean was assessed against A. alternata and data on pathological traits was subjected to cluster analysis. The results showed that genotypes of mungbean were grouped into four clusters based on resistance parameters under the influence of disease. The principal component biplot demonstrated that all the disease-related parameters (% disease incidence, % disease intensity, lesion area, and % of infection) were strongly correlated with each other. Alt a 1 gene that is precisely found in Alternaria species and is responsible for virulence and pathogenicity. Alt a 1 gene was amplified using gene specific primers. The isolated pathogen produced similar symptoms when inoculated on mungbean and tobacco. The sequence analysis of the internal transcribed spacer (ITS) region, a 600 bp fragment amplified using specific primers, ITS1 and ITS2 showed 100% identity with A. alternata. Potato virus X (PVX) -based silencing vector expressing Alt a 1 gene was constructed to control this pathogen through RNA interference in tobacco. Out of 50 inoculated plants, 9 showed delayed onset of disease. Furthermore, to confirm our findings at molecular level semi-quantitative reverse transcriptase polymerase chain reaction was used. Both phenotypic and molecular investigation indicated that RNAi induced through the VIGS vector was efficacious in resisting the pathogen in the model host, Tobacco (Nicotiana tabacum). To the best of our knowledge, this study has been reported for the first time.