PCR-RFLP技术鉴定龙目岛辣椒黄曲病病毒

M. Windarningsih
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This research conducted with this following step: field observation of yellow leaf curl symptoms, DNA extraction, amplification the coat protein with spesific primer PYLCV, and RFLP analyses. Field result that conducted in three regency of Lombok island (East Lombok, West Lombok and Central Lombok) shown that infection symptoms in chili pepper such as various yellow-green spot in leaves, leaf curl, become small and wrapped on top, and in plant that have severe infection the plant become stunted. Results of molecular identification using PCR in sample plant aredetected the Begomovirus existence with amplification products sized 840 bp. The analysis results of PCR-RFLP using restriction enzymes HaeIII have shown the genetic variability between isolate from Lombok island, Centre of Java, and DIY. The result of this research has proved the ability of PYLCV primer in PCR to detect Begomovirus obtained from different places. This research proved that the universal primer ability in PCR for Begomovirus detection from different area and as the first report there are infections of Pepper Yellow Leaf Curl Virus in Lombok island NTB.Yellow leaf curl virus causes pepper yellow leaf curl diseases that caused by Geminiviruses (Begomovirus) became epidemic in Indonesia on 2000. This disease is widespread at several chili pepper production centers in Java, Sumatera, Bali, Kalimantan, and Sulawesi. There is no report about existence of it in Lombok island, West Nusa Tenggara Province, one of chili pepper production centre in Indonesia. The aim of this research is to identify Begomovirus which infect chili pepper with molecular approach and to find out the variabilities genetic of Begomovirus from Lombok island, Central Jawa and DIY with DNA fingerprint pattern based on Polymerase Chain Reaction (PCR) and Reaction Fragment Length Polymorphisme (RFLP). 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引用次数: 3

摘要

黄卷叶病毒引起辣椒黄卷叶病,由双病毒(begomvirus)引起,于2000年在印度尼西亚流行。该病在爪哇、苏门答腊、巴厘岛、加里曼丹和苏拉威西的几个辣椒生产中心广泛传播。在印度尼西亚的辣椒生产中心之一的西努沙登加拉省龙目岛,没有关于它存在的报道。本研究采用分子方法鉴定侵染辣椒的贝古莫病毒,并利用聚合酶链式反应(PCR)和反应片段长度多态性(RFLP)的DNA指纹图谱对龙目岛、中爪哇和DIY的贝古莫病毒进行变异遗传分析。本研究进行了以下步骤:黄卷叶症状的田间观察,DNA提取,特异性引物PYLCV扩增外壳蛋白,RFLP分析。在龙目岛东龙目岛、西龙目岛和中龙目岛3个县进行的田间试验结果表明,辣椒感染后表现为叶片出现各种黄绿色斑点、卷曲、叶片变小、顶部缠绕等症状,严重感染后植株发育迟缓。PCR分子鉴定结果显示,样品植物中存在begomavirus,扩增产物大小为840 bp。利用限制性内切酶HaeIII的PCR-RFLP分析结果显示,龙目岛、爪哇中心和DIY分离株之间存在遗传变异。本研究结果证实了PCR中PYLCV引物对不同来源的begomavirus的检测能力。本研究证实了该引物对不同地区的Begomovirus检测具有普遍的引物能力,并首次报道了辣椒黄卷叶病毒在龙目岛NTB的感染。黄卷叶病毒引起辣椒黄卷叶病,由双病毒(begomvirus)引起,于2000年在印度尼西亚流行。该病在爪哇、苏门答腊、巴厘岛、加里曼丹和苏拉威西的几个辣椒生产中心广泛传播。在印度尼西亚的辣椒生产中心之一的西努沙登加拉省龙目岛,没有关于它存在的报道。本研究采用分子方法鉴定侵染辣椒的贝古莫病毒,并利用聚合酶链式反应(PCR)和反应片段长度多态性(RFLP)的DNA指纹图谱对龙目岛、中爪哇和DIY的贝古莫病毒进行变异遗传分析。本研究进行了以下步骤:黄卷叶症状的田间观察,DNA提取,特异性引物PYLCV扩增外壳蛋白,RFLP分析。在龙目岛三个县(东龙目岛、西龙目岛和中龙目岛)进行的实地调查结果表明:
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Identification of virus causing the yellow leaf curl diseases on chili pepper in Lombok Island by PCR-RFLP technique
Yellow leaf curl virus causes pepper yellow leaf curl diseases that caused by Geminiviruses (Begomovirus) became epidemic in Indonesia on 2000. This disease is widespread at several chili pepper production centers in Java, Sumatera, Bali, Kalimantan, and Sulawesi. There is no report about existence of it in Lombok island, West Nusa Tenggara Province, one of chili pepper production centre in Indonesia. The aim of this research is to identify Begomovirus which infect chili pepper with molecular approach and to find out the variabilities genetic of Begomovirus from Lombok island, Central Jawa and DIY with DNA fingerprint pattern based on Polymerase Chain Reaction (PCR) and Reaction Fragment Length Polymorphisme (RFLP). This research conducted with this following step: field observation of yellow leaf curl symptoms, DNA extraction, amplification the coat protein with spesific primer PYLCV, and RFLP analyses. Field result that conducted in three regency of Lombok island (East Lombok, West Lombok and Central Lombok) shown that infection symptoms in chili pepper such as various yellow-green spot in leaves, leaf curl, become small and wrapped on top, and in plant that have severe infection the plant become stunted. Results of molecular identification using PCR in sample plant aredetected the Begomovirus existence with amplification products sized 840 bp. The analysis results of PCR-RFLP using restriction enzymes HaeIII have shown the genetic variability between isolate from Lombok island, Centre of Java, and DIY. The result of this research has proved the ability of PYLCV primer in PCR to detect Begomovirus obtained from different places. This research proved that the universal primer ability in PCR for Begomovirus detection from different area and as the first report there are infections of Pepper Yellow Leaf Curl Virus in Lombok island NTB.Yellow leaf curl virus causes pepper yellow leaf curl diseases that caused by Geminiviruses (Begomovirus) became epidemic in Indonesia on 2000. This disease is widespread at several chili pepper production centers in Java, Sumatera, Bali, Kalimantan, and Sulawesi. There is no report about existence of it in Lombok island, West Nusa Tenggara Province, one of chili pepper production centre in Indonesia. The aim of this research is to identify Begomovirus which infect chili pepper with molecular approach and to find out the variabilities genetic of Begomovirus from Lombok island, Central Jawa and DIY with DNA fingerprint pattern based on Polymerase Chain Reaction (PCR) and Reaction Fragment Length Polymorphisme (RFLP). This research conducted with this following step: field observation of yellow leaf curl symptoms, DNA extraction, amplification the coat protein with spesific primer PYLCV, and RFLP analyses. Field result that conducted in three regency of Lombok island (East Lombok, West Lombok and Central Lo...
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