苜蓿滑刃线虫线粒体基因组及其系统发育研究  

Mitochondrial Genome and Phylogeny of Aphelenchoides medicagus

作　　者：薛清 杜虹锐 薛会英[2] 王译浩 王暄 李红梅 XUE Qing;DU Hong-rui;XUE Hui-ying;WANG Yi-hao;WANG Xuan;LI Hong-mei(College of Plant Protection,Key Laboratory of Integrated Management of Crop Diseases and Pests,Ministry of Education,Nanjing Agricultural University,Nanjing 210095;College of Resources and Environment,Tibet College of Agriculture and Animal Husbandry,Linzhi 860000)

机构地区：[1]南京农业大学植物保护学院,农作物生物灾害综合治理教育部重点实验室,南京210095 [2]西藏农牧学院资源与环境学院,林芝860000

出　　处：《生物技术通报》2021年第7期98-106,共9页Biotechnology Bulletin

基　　金：国家自然科学基金项目(32001876);中央高校基本科研业务费专项资金(KYYZ202109)。

摘　　要：苜蓿滑刃线虫(Aphelenchoides medicagus)是一种兼性植物寄生线虫,可在多种真菌上完成生活史,同时对大豆和苜蓿具有弱寄生性。本研究采用Illumina平台对苜蓿滑刃线虫进行低覆盖全基因组测序,从获得的全基因组序列中提取线粒体基因,并利用“种子”序列进行组装;同时对其中富含AT的非编码部分进行PCR扩增与Sanger测序,将扩增与组装的结果进行拼接,共获得14411 bp的线粒体基因组。基因注释结果显示,苜蓿滑刃线虫共有蛋白编码基因12个,tRNA基因22个,rRNA基因2个,其基因构成和排列与贝西滑刃线虫(A.besseyi)、松材线虫(Bursaphelenchus xylophilus)及拟松材线虫(B.mucronatus)完全相同。基于氨基酸序列的系统发育显示,苜蓿滑刃线虫与贝西滑刃线虫互为姐妹群,且与松材线虫、拟松材线虫处在高度支持的滑刃线虫科单系中。本研究表明,低覆盖全基因组测序法可以成功组装出线粒体基因组的大部分序列,证明了利用该方法获取线虫线粒体全基因组序列的可行性。Aphelenchoides medicagus is a facultative plant-parasitic nematode that may complete its life cycles on various fungi,meanwhile it has weak parasitism on soybean and alfalfa.In presented study,Illumina platform was used to sequence A.medicagus genome in low coverage.Assembly was made by extracting mitochondrial reads and subsequently assembled by the seed sequence.For the AT rich noncoding region,amplification was made using PCR and sequenced by Sanger method.The jointed result from the assembled and amplified sequences revealed the mitochondrial genome of A.medicagus consisted of 14411 bp nucleotides,including 12 protein coding genes,22 tRNA and 2 rRNA genes after gene annotation.These genes are arranged in same way as those in A.besseyi,Bursaphelenchus xylophilus,and B.mucronatus.The phylogeny analysis using amino acid sequences showed that A.medicagus and A.besseyi clustered together as a sister group,which together with B.xylophilus and B.mucronatus to form a highly supported monophyletic Aphelenchoididae clade.This study demonstrates that low-coverage whole-genome sequencing method can be used to assemble the most of mitochondrial genome,and thus support this technique to be a feasible tool in mitochondrial genome sequencing.

关 键 词：苜蓿滑刃线虫 滑刃线虫科 线粒体基因组 Illumina平台 低覆盖全基因组测序 

分 类 号：S432.45[农业科学—植物病理学]