期刊文献+
任意字段
题名或关键词
题名
关键词
文摘
作者
第一作者
机构
刊名
分类号
参考文献
作者简介
基金资助
栏目信息

油菜茎基溃疡病菌的实时荧光PCR检测 被引量：8

Detection of Leptosphaeria maculans by real-time PCR
原文传递
导出
摘要 根据油菜茎基溃疡病菌Leptosphaeria maculans与其近似种ITS序列的差异,设计了检测L.maculans的引物Lmb3/R2和探针Probe-M,建立了L.maculans的实时荧光PCR检测方法。试验结果表明,来自加拿大、澳大利亚和乌克兰等国的22株L.maculans菌株都能得到阳性扩增,而供试的30株L.biglobosa菌株和6株其他菌株以及空白对照没有荧光信号的增加。该检测方法的灵敏度达到4 pg菌丝DNA,整个检测过程控制在4 h内,其快速、特异和灵敏的特点可以满足进境油菜籽样品的快速初检以及病菌分离物的快速鉴定。 Real-time PCR was used for detection of Leptosphaeria maculans, the causative agent of oilseed phoma stem canker. Specific primers Lmb3/R2 and a TaqMan-MGB probe （Probe-M） specific to L. macu- lans were designed from the differentiation of ITS sequences between L. maculans and related species in the GenBank database. The results showed that all 22 isolates of L. maculans from oilseed samples from Austra- lia, Ukraine and Canada could get positive amplification with the limit of 4 pg DNA, and negative results for 30 isolates of L. biglobosa and the other 6 isolates of related species from oilseed samples. Real-time PCR could be especially useful for mass screening of oilseed sample and diagnosis of suspected isolate.
作者 周国梁 尚琳琳 林泓 印丽萍 徐殿胜 易建平
机构地区 上海出入境检验检疫局 华东理工大学生物工程学院
出处 《植物病理学报》 CAS CSCD 北大核心 2011年第1期10-17,共8页 Acta Phytopathologica Sinica
基金 上海出入境检验检疫局科研项目(HK069-2010) 上海市科委项目(09DZ0580201)
关键词 油菜茎基溃疡病菌 实时荧光PCR Leptosphaeria maculans real-time PCR
分类号 S41-31 [农业科学—植物保护] S435.654 [农业科学—农业昆虫与害虫防治]
  • 相关文献

参考文献12

  • 1Williams R H, Fitt B D L. Differentiating A and B groups of Leptosphaeria maculans, causal agent of stem canker (blackleg) of oilseed rape [ J ]. Plant Pathology, 1999, 48:161 - 175.
  • 2Plummer K M, Dunse K, Howlett B J, Nonaggressive strains of the blackleg fungus, Leptosphaeria maeulans, are present in Australia and can be distinguished from aggressive strains by molecular analysis [J].Australian Journal of Botany, 1994, 42 : 1 - 8.
  • 3Shoemaker R A, Brun H. The teleomorph of the weakly aggressive segregate of Leptosphaeria rnaculans [J]. Canadian Journal of Botany, 2001, 79:412 - 419.
  • 4West J S, Balesdent M H, Rouxel T, et al. Colonization of winter oilseed rape tissues by A/Tox + and B/ Tox0 Leptosphaeria maculans (Phoma stern canker) in France and England [J]. Plant Pathology, 2002, 51, 311 -321.
  • 5West J S, Kharbanda P D, Barbetti M J, et al. Epidemiology and management of Leptosphaeria maculans (Phoma stem canker) on oilseed rape in Australia, Canada and Europe [J].Plant Pathology, 2001, 50 : 10 -27.
  • 6Fitt B B H, Brun H, Barbetti M, et al. World-wide importance of phoma stem canker ( Leptosphaeria maculans and L. biglobosa ) on oilseed rape (Brassica hapus) [ J ]. European Journal of Plant Pathology, 2006, 114. 3-15.
  • 7Zhou Y, Fitt B D L, Welham S J, et al. Effects of severity and timing of stem canker (Leptosphaeria maculans) symptoms on yield of winter oilseed rape (Brassica napus) in the UK[J]. European Journal of Plant Pathology, 1999, 105.715-728.
  • 8Taylor J L. A simple, sensitive, and rapid method for detecting seed contaminated with highly virulent Leptosphaeria maculans[J]. Applied and Environmental Microbiology, 1993, 59(11 ) : 3681 -3685.
  • 9Mahuku G S, Goodwin P H, Hall R. A competitive polymerase chain reaction to quantify DNA of Leptosphaeria maculans during blackleg development in oilseed rape [J]. Molecular Plant-Microbe Interactions, 1995, 8 : 761 - 767.
  • 10Liu S Y, Liu Z, Fitt B D L, et al. Resistance to Leptosphaeria maculans ( Phoma stem canker) in Brassica napus (oilseed rape ) induced by L. biglobosa and chemical defense activators in field and controlled environments[J]. Plant Pathology, 2006, 55:401 -412.

同被引文献94

  • 1刘刚.质检总局决定对进口油菜籽实施紧急检疫措施[J].农药市场信息,2009(23):45-45. 被引量：1
  • 2吴元华,王左斌,刘志恒,赵秀香,粱景颐.我国烟草新病害—靶斑病[J].中国烟草学报,2006,12(6):22-22. 被引量：59
  • 3ByrneM,Fraulino D,DiLorenzo J,et al.Cloning,sequencing and e valu-ation of the 13-tubulin gene from Leishmania donotmni as a referenee gene for RT-qPCR [ J].The Journal of Young Investigators,2011,21(3):41-50.
  • 4StuderD,LiseherS,Jochum W,et al.Ribosomal protein L13a as a refer-enee gene for human bone marrow-derived mesenehymal stromal eells dur-ing expansion,adipo-,ehondro-,and osteogenesis [ J].Tissue Engineer-ing Part C:Melhods,2012,18(10):761-771.
  • 5Gu C S,Chen S M,Liu Z L,et al.Reference gene selection for quantita-tive real-time PCR in chrysanthemum subjected to biotic and abiotic stress [ J ].Molecular Biotechnology,2011,49(2):192-197.
  • 6Biederman J,Yee J,Cortes P.Validation of internal control genes for gene expression analysis in diabetic glomerulosclerosis [J].Kidney Inter-national,2004,66:2308-2314.
  • 7Changseo L,Seungyong L,Seung GS,et al.Real-time PCR determi-nation of rRNA gone copy number:absolute and relative quantification as-says with Escherichia coli [ J].Applied Microbiology and Biotechnology,2008,78:371-376.
  • 8Xu ML,Melchinger AE,Iathberstedt T.Species-specific detection of the maize pathogens Sporisorium reiliana and Ustilago maydis by dot blot hybridization and PCR-based assays [J].Plant Disease,1999,83(4):390-395.
  • 9Margnez-Espinoza AD,Leon-Ramlr CG,Singh N,et al.Use of PCR to detect infection of differentially susceptible maize cultivars using Ustilago maydis strains of variable vindence [ J].International Microbiol-ogy,2003,6:117-120.
  • 10Kumar B,Kumar P,Rajput,et al.Comparative reproducibility of SYBR Green Ⅰ and TaqMan real-time PCR chemistries for the analysis of matrix and hemagglutinin genes of Influenza A viruses [ J ].hlternational Journal of Collaborative Research on Internal Medicine & Public Health,2012,4(7):1346-1352.

引证文献8

二级引证文献26

植物病理学报
2011年 第1期

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部