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茄子蛀果蛾种群的分子遗传多样性及其抗药性 SCAR 标记的开发。

Molecular genotypic diversity of populations of brinjal shoot and fruit borer, Leucinodes orbonalis and development of SCAR marker for pesticide resistance.

机构信息

Department of Zoology, University of Madras, Chennai, 600025, India.

Department of Zoology, N.M.S.S. Vellaichamy Nadar College, Madurai, 625019, India.

出版信息

Mol Biol Rep. 2021 Dec;48(12):7787-7800. doi: 10.1007/s11033-021-06791-2. Epub 2021 Oct 16.

DOI:10.1007/s11033-021-06791-2
PMID:34655405
Abstract

BACKGROUND

The brinjal shoot and fruit borer, Leucinodes orbonalis is a destructive pest of Solanum melongena. The control of L. orbonalis with extensive application of synthetic chemical insecticides resulted in the development of resistance with known genetic heterogeneity among populations. Understanding the genetic diversity of their populations is important in developing strategies for their management. The present investigation was performed to characterize populations of L. orbonalis for their genetic diversity in the entire region of Tamil Nadu, South India using random amplified polymorphic DNA (RAPD) primers as a tool of the molecular marker.

METHODS AND RESULTS

Among 60 random 10-mer primers, only ten primers generated reproducible and scorable banding profile. Among the ten different random primers, the primers namely OPG 7, OPG 8, OPS 2 and OPS 7 generated the highest genetic variation with over 80% genetic polymorphism. Phylogram analysis produced 18 clusters with eight major and ten minor clusters. Cluster analysis, statistical fitness, population structure and analysis of molecular variance confirmed the significant genetic variation among different populations. A trait specific marker obtained through RAPD was cloned, sequenced and used to develop a stable diagnostic SCAR marker for DNA fingerprinting to distinguish the populations. Amplification of this locus in the samples of 20 different populations indicated recognition of the trait for pesticide resistance in 12 populations.

CONCLUSIONS

The results suggest that the biochemical nature of host plant varieties of this insect pest and variation in the application of different insecticides are essential contributing factors for the genotypic variations observed among populations of L. orbonalis.

摘要

背景

茄子蛀果蛾,Leucinodes orbonalis,是茄子的毁灭性害虫。由于广泛使用合成化学杀虫剂来控制 L. orbonalis,导致种群产生了抗性,且已知存在遗传异质性。了解其种群的遗传多样性对于制定管理策略非常重要。本研究旨在利用随机扩增多态性 DNA(RAPD)引物作为分子标记工具,对印度南部泰米尔纳德邦的整个地区的 L. orbonalis 种群进行遗传多样性特征描述。

方法和结果

在 60 个随机的 10 碱基引物中,只有 10 个引物产生了可重复和可评分的带型谱。在这 10 个不同的随机引物中,引物 OPG 7、OPG 8、OPS 2 和 OPS 7 产生了最高的遗传变异,遗传多态性超过 80%。系统发育树分析产生了 18 个聚类,其中包括 8 个主要聚类和 10 个次要聚类。聚类分析、统计拟合度、种群结构和分析分子方差证实了不同种群之间存在显著的遗传变异。通过 RAPD 获得的一个特性特异标记被克隆、测序,并用于开发稳定的 DNA 指纹图谱 SCAR 标记,以区分种群。在 20 个不同种群的样本中扩增该基因座,表明在 12 个种群中识别出了对杀虫剂的抗性特性。

结论

结果表明,这种昆虫的寄主植物品种的生化特性和不同杀虫剂的应用变化是导致 L. orbonalis 种群中观察到的遗传变异的重要因素。

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