Saipul Abbas, Ernawati Djaya, Erwin Najamuddin, Amelia Sebayang, Ayyub Ar Rahman, Aminah Aminah, H. Hasbi, S. Sipi, Nur Fathurahman Ridwan, R. Ismayanti, Elisurya Ibrahim
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White-back planthoppers found on rice plantations showing symptoms of yellowing and stunted leaves were sampled for further analysis, including total DNA isolation of insects, standard PCR amplification for insect and Nested-PCR for phytoplasma identification, gene sequencing for both amplicons, and nucleotide analysis using BLAST method and Mega X program. The PCR with CO1 primer successfully amplified a 700 bp amplicon from insects, whereas nested-PCR using fP1/rP7 primers followed by m23SR/R16F2n amplified phytoplasma supposedly around 1800 bp and 1250 bp of 16S RNA gene, respectively. The DNA sequencing analysis results indicate that the insect samples were identified as 83% Sogatella vibix species based on homology percentage analysis using BLAST and Mega X Program. 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引用次数: 0
摘要
南苏拉威西岛是印度尼西亚最大的水稻生产中心之一。水稻植物的几种重要病害,如病毒和植物病原体引起的病害,可通过昆虫媒介传播,尤其是叶蝉和茎螨。由病毒和植物病原体引起的病害症状多种多样,但视觉效果相似,难以区分。本研究旨在分析与水稻种植园中常见的白背飞虱相关的植原体的存在情况。 研究方法是在 Sidrap 区的六个村庄进行昆虫样本调查和探索。对在水稻种植园中发现的表现为叶片发黄和发育不良症状的白背飞虱进行取样,以进一步分析,包括昆虫的总 DNA 分离、昆虫的标准 PCR 扩增和植原体鉴定的 Nested-PCR、两种扩增子的基因测序,以及使用 BLAST 方法和 Mega X 程序进行核苷酸分析。使用 CO1 引物进行的 PCR 成功地从昆虫身上扩增出 700 bp 的扩增片段,而使用 fP1/rP7 引物和 m23SR/R16F2n 进行的嵌套 PCR 则分别扩增出植物原原体 16S RNA 基因的 1800 bp 和 1250 bp 左右。DNA 测序分析结果表明,根据 BLAST 和 Mega X 程序的同源性百分比分析,昆虫样本被鉴定为 83% 的 Sogatella vibix 物种。至于植原体,则更倾向于 16SrI 组或 Candidatus phytoplasma asteris(翠菊黄斑植原体),同源性百分比为 99%。
Phytoplasma Associated with White-backed Planthopper on Rice Plants in Sidrap Regency, South Sulawesi
South Sulawesi is one of the largest rice production centers in Indonesia. Several important diseases of rice plants, such as those caused by viruses and phytoplasmas, can be transmitted by insect vectors, especially leafhoppers and stem plant. Symptoms of diseases caused by viruses and phytoplasmas are quite diverse but visually similar and difficult to distinguish. This study aims to analyze the presence of phytoplasma associated with white-backed planthopper which are commonly found in rice plantations. The research method used is by conducting surveys and explorations of insect samples in six villages in Sidrap District. White-back planthoppers found on rice plantations showing symptoms of yellowing and stunted leaves were sampled for further analysis, including total DNA isolation of insects, standard PCR amplification for insect and Nested-PCR for phytoplasma identification, gene sequencing for both amplicons, and nucleotide analysis using BLAST method and Mega X program. The PCR with CO1 primer successfully amplified a 700 bp amplicon from insects, whereas nested-PCR using fP1/rP7 primers followed by m23SR/R16F2n amplified phytoplasma supposedly around 1800 bp and 1250 bp of 16S RNA gene, respectively. The DNA sequencing analysis results indicate that the insect samples were identified as 83% Sogatella vibix species based on homology percentage analysis using BLAST and Mega X Program. As for the phytoplasma, it leans more towards the 16SrI group or Candidatus phytoplasma asteris (Aster yellows phytoplasma) with a homology percentage of 99%.