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利用系统发生分析对印度尼西亚可可中的 sp. 进行分子鉴定。

Molecular Identification of sp. From Indonesian Cocoa Using Phylogenetic Analysis.

出版信息

Pak J Biol Sci. 2022 Jan;25(3):245-253. doi: 10.3923/pjbs.2022.245.253.

Abstract

<b>Background and Objective:</b> Diseases caused by <i>Phytophthora</i> species cause widespread damage worldwide and are troubling cocoa farmers in Indonesia. The specific species causing disease in an area can be ascertained by characterizing its rDNA fragments. This study aimed to identify <i>Phytophthora</i> sp., samples from cocoa plantations in Southeast Sulawesi, Indonesia, based on phylogenetic analysis of rDNA fragments. <b>Materials and Methods:</b> Identification of rDNA fragments of <i>Phytophthora</i> sp., done by amplifying rDNA fragments using PCR (Polymerase Chain Reactions) techniques with the specific primer of <i>Phytophthora</i> (Phy-F and Phy-R) which can amplify regions of ITS1, 5.8S rRNA and ITS2. The rDNA fragments are then sequenced and analyzed using: The BLAST (Basic Local Alignment Search Tools) provided by NCBI (National Center for Biotechnology Information) via (www.ncbi.nlm.nih.gov/blast) to analyze the local alignment of DNA sequences with Genbank DNA data and Mega 7.0.26 software is used to construct the phylogenetic tree. <b>Results:</b> The DNA sequencing results showed the rDNA measuring 786 bp consisted of complete sequences of ITS 1 (210 bp), 5.8S rRNA (162 bp) and ITS 2 (414 bp). Based on phylogenetic tree analysis using the maximum likelihood method with 1000 bootstrap replications showed that the rDNA of <i>Phytophthora</i> sp., isolates and 29 comparator isolates formed 2 large groups. <i>Phytophthora</i> sp., formed a subgroup with <i>Phytophthora palmivora</i> with a bootstrap value of 99%. <b>Conclusion:</b> The type of <i>Phytophthora</i> spreading in cocoa plantations in Southeast Sulawesi, Indonesia, is 1 group with <i>Phytophthora palmivora</i>.

摘要

<b>背景与目的:</b>由<i>Phytophthora</i>物种引起的疾病在全球范围内造成广泛破坏,困扰着印度尼西亚的可可种植者。通过对 rDNA 片段的特征分析,可以确定导致特定地区疾病的特定物种。本研究旨在根据 rDNA 片段的系统发育分析,从印度尼西亚东南苏拉威西的可可种植园中鉴定<i>Phytophthora</i>sp。<b>材料与方法:</b>使用 PCR(聚合酶链反应)技术,用特定的<i>Phytophthora</i>引物(Phy-F 和 Phy-R)扩增 rDNA 片段,对<i>Phytophthora</i>sp.rDNA 片段进行鉴定,该引物可扩增 ITS1、5.8S rRNA 和 ITS2 区域。然后对 rDNA 片段进行测序和分析,使用:NCBI(美国国家生物技术信息中心)提供的 BLAST(基本局部比对搜索工具)(通过 www.ncbi.nlm.nih.gov/blast)分析 DNA 序列与 Genbank DNA 数据的局部比对,使用 Mega 7.0.26 软件构建系统发育树。<b>结果:</b>DNA 测序结果显示,rDNA 长 786bp,由 ITS1(210bp)、5.8S rRNA(162bp)和 ITS2(414bp)的完整序列组成。使用最大似然法进行系统发育树分析,1000 次自举复制,结果显示,<i>Phytophthora</i>sp.rDNA 的分离株和 29 个比较分离株形成了 2 个大组。<i>Phytophthora</i>sp.与<i>Phytophthora palmivora</i>形成一个亚群,自举值为 99%。<b>结论:</b>印度尼西亚东南苏拉威西可可种植园中传播的<i>Phytophthora</i>类型是与<i>Phytophthora palmivora</i>属于 1 组。

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