香蕉穿孔线虫双重PCR快速检测技术研究

	香蕉穿孔线虫双重PCR快速检测技术研究
	葛建军 1; 曹爱新 1; 周国梁 2; 赵国柱 3; 谢丙炎 4
刊名	植物病理学报
	2007
卷号	37 期号:5 页码:472-478
关键词	香蕉穿孔线虫分子检测双重PCR
ISSN号	0412-0914
其他题名	Development of duplex PCR assay for detection of burrowing nematode, Radopholus similis
英文摘要	香蕉穿孔线虫（Radopholus similis）是国际上公认的植物检疫性有害生物之一,是热带地区果树上最重要的线虫。通过设计的属水平上的特异性引物R1、R2（R1/2）和种水平上的特异性引物RS1、RS2（RS1/2）进行双重PCR扩增香蕉穿孔线虫,可扩增出长度为362和291 bp的2个特异性片段谱带。同时分别对检测体系中反应参数进行优化,最佳延伸温度为51.4℃,引物R1/2与RS1/2最佳浓度配比为0.2μm o.lL~(-1)/1.2μmol.L~(-1),并测试检测引物的灵敏度,能进行有效检测的最低起始核酸量为100 pg;同时,也能对单条线虫进行检测以及样品中线虫的检测。专化性测试证明,2对引物能有效地进行香蕉穿孔线虫的快速分子诊断。; The burrowing nematode, Radopholus similis（Cobb） Thorne, is one of considerable plant quarantine pests, it may be the most important nematode pest on fruit crops （ especially citrus and banana） in the tropics. Two pairs of primers R1, R2 （R1/2） and RS1, RS2 （RS1/2） were developed to identify the burrowing nematode. The primer R1/2 and RS1/2 could produce 362 bp band for all nematodes of genus Radopholus, but only R. similis produced 291 bp band. At the same time, the parameters of PCR were optimized and sensitivity were tested. The optimized conditions were an extension temperature of 51.4℃ and primers concentrations of R1/2 and RS1/2 at 0.2 μmoL/L and 1.2 μmol/L, individually. The minimum amount of DNA of R. similis could be detected was as low as 100 pg, and it also could detect DNA from individual nematode and nematodes from samples of plant. Thus, it demonstrates that duplex PCR approach will provided an accurate and effective method for quick detection of burrowing nematode.
学科主题	植物保护
语种	中文
内容类型	期刊论文
源URL	[http://111.203.20.206/handle/2HMLN22E/100681]
专题	蔬菜花卉研究所_植物保护研究室
作者单位	1.中国检验检疫科学研究院动植物检疫研究所, 北京, 100029 2.上海出入境检验检疫局, 上海, 200135 3.北京林业大学生命科学与技术学院, 北京, 100083 4.中国农业科学研究院蔬菜研究所, 北京, 100081
推荐引用方式 GB/T 7714	葛建军,曹爱新,周国梁,等. 香蕉穿孔线虫双重PCR快速检测技术研究[J]. 植物病理学报,2007,37(5):472-478.
APA	葛建军,曹爱新,周国梁,赵国柱,&谢丙炎.(2007).香蕉穿孔线虫双重PCR快速检测技术研究.植物病理学报,37(5),472-478.
MLA	葛建军,et al."香蕉穿孔线虫双重PCR快速检测技术研究".植物病理学报 37.5(2007):472-478.