桔小实蝇是直接危害果蔬生产并对出口贸易形成重要影响的有害生物群,口岸检疫对其鉴定依赖形态观察.本试验利用特异的嵌套式PCR 引物对桔小实蝇rDNA基因片段进行扩增,达到快速、可靠鉴定出柑橘果实中微量桔小实蝇卵,通过进一步序列分析,确定该实蝇与近缘种的关系.本试验建立的分子鉴定体系,可应用于口岸桔小实蝇卵快速、准确检疫,并能确定样本的种属关系.
孔秋莲
,
叶军
,
岳玲
,
陈志军
,
包英姿
,
徐赟
,
郭卡
,
袁忠谊
,
戚文元
. 快速检测水果中桔小实蝇卵的分子诊断方法研究[J]. 华北农学报, 2010
, 25(1)
: 226
-230
.
DOI: 10.7668/hbnxb.2010.01.046
Bactrocera dorsalis(Hendel),as oriental fruit fly,is a serious quarantine pest of fruits in tropical and subtropical countries.Its rDNA gene includes 18S RNA,ITS-1,5.8S RNA,ITS-2,and 28S RNA.In this study,a nested PCR was carried out for amplification of ITS fragments containing 5.8S RNA.The sequence of the PCR fragment was greatly similar with Bactrocera dorsalis(Hendel)′s in NCBI Genbank.The PCR technique can be used to rapidly identify eggs of Bactrocera dorsalis(Hendel) in fruit.
[1] 张彬,刘映红,赵岚岚,等.桔小实蝇研究进展[J].中国农学通报,2008,24(11):391-397.
[2] 宁红,秦蓁.分子生物学技术在检疫性有害生物诊断中的应用[J].植物检疫,2002,16(2):98-100.
[3] Armstrong K F,Cameron C M,Frampton E R.Fruit fly(Diptera:Tephritidae)species identification:A rapid molecular diagnostic technique for quarantine application[J].Bulletin of Entomological Reaeareh,1997,87:111-118.
[4] 王振华,朱堂明,吕志藻,等.用PCR方法鉴定区分柑橘大实蝇与橘小实蝇[J].植物检疫,2008,22(3):155-157.
[5] 潘志萍,曾玲,温硕洋.桔小实蝇抗性品系的微卫星DNA分析[J].昆虫学报,2006,49(5):874-877.
[6] 徐浪,余道坚,张润杰,等.桔小实蝇线粒体基因组全序列及其分析[J].昆虫学报,2007,50(8):755-761.
[7] He M,Haymer D S He M,Polymorphic intron sequences detected within and between populations of the oriental fruit fly(Diptera tephfitidae)[J].Annals of Entormological Society of America,1997,90(6):825-831.
[8] 吴红芝,孔宝华,陈海如,等.RT-PCR检测番茄不孕病毒[J].植物检疫,2002,16(2):66-69.
[9] 易建平,陶庭典,潘良文,等.套式PCR直接检测印度腥黑穗病菌冬孢子[J].植物检疫,2002,16(4):197-200.
[10] 贺水山,闻伟刚,杨兰英,等.松材线虫PCR快速检测方法研究[J].植物检疫,2002,16(6):321-324.
[11] 刘伟.rDNA在昆虫纲系统发育研究中的应用[J].昆虫知识,1998,35(2):123-126.
[12] Smith P T,Kambha(n)ti S,Armstrong K A.Phylogenetic relationships among Bactrocera species(Diptera:Tephritidae) inferred from mitochondrial DNA sequences[J].Mor Phylogenet and Evol,2003,26:8-17.
[13] Muraji M,Nakahara S.Phylogentic rehtitmships among fruit flies,Bactrocera(Diptear:Tephritidae),based on the mitochondrial rDNA sequences[J].Insect Mol Biol,2001,10(6):549-559.
[14] Regier J C,Shultz J W.Molecular phylogeny of the major arthopoda groups indicates polyphyly of crustaceans and a new hypothesis for the origin of hexapods[J].Mol Bio Evol,1997,14:902-913.
[15] Bachmann L,Schihel J M,Rabb M,et al.Satellite DNA as a taxonomic marker[J].Biochem Biophys Acts,1993,1089:396-400.
