| 松材线虫rDNA的测序和PCR-SSCP分析 |
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| 引用本文： | 张立海,廖金铃,冯志新.松材线虫rDNA的测序和PCR-SSCP分析[J].植物病理学报,2001,31(1):84-89. |
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| 作者姓名： | 张立海 廖金铃 冯志新 |
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| 作者单位： | 华南农业大学植物线虫研究室, 广州 510642 |
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| 基金项目： | 国家林业局松材线虫治理工程项目[财农字(1998)84号]；广东省“九五”重点项目资助(98278079) |
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| 摘 要： | 本文为克服形态鉴定的不足,用分子生物学的方法鉴别松材线虫。线虫核糖体DNA(r DNA)的内部转录间隔区(ITS1)区(约308bp)的测序结果显示:松材线虫种内区别很小,不超过1bp;拟松材线虫种内区别较大,最大达7bp;这2种线虫的种间区别为32~39bp。根据以上测序结果,本文结合单条线虫DNA的提取技术,对14个松材线虫和拟松材线虫样本进行了单链构象多态性(PCR-SSCP)分析,结果表明PCR-SSCP分析技术可明确区分这2种线虫,该技术可为单条松材线虫的鉴定提供一套灵敏而可靠的方法。
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| 关 键 词： | 松材线虫 测序 rDNA ITS PCR-SSCP |
| 文章编号： | 0412—0914(2001)01—0084-06 |
| 修稿时间： | 1999年6月2日 |
SEQUENCING AND PCR-SSCP ANALYSIS OF RIBOSOMAL DNA OF Bursaphelenchus NEMATODES |
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| ZHANG Li-hai,LIAO Jin-ling,FENG Zhi-xin.SEQUENCING AND PCR-SSCP ANALYSIS OF RIBOSOMAL DNA OF Bursaphelenchus NEMATODES[J].Acta Phytopathologica Sinica,2001,31(1):84-89. |
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| Authors: | ZHANG Li-hai LIAO Jin-ling FENG Zhi-xin |
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| Institution: | Plant Nematode Research Lab, South China Agricultural University, Guangzhou 510642, China |
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| Abstract: | To overcome limitations in the morphological identification of Bursaphelenchus species, a molecular approach was used. The internal transcribed spacer 1 (ITS1) of the nuclear ribosomal DNA (rDNA) region was amplified and sequenced. A pairwise comparison of this sequence data revealed that the differences in the ITS1 region (approximately 308 bp) between four B. xylophilus isolates were slight (only 1 bp), but there are marked differences (up to 7 bp) in this region between the B. mucronatus isolates from China (Guangdong) and France. The data also revealed that the differences in the ITS1 region between the two Bursaphelenchus species range from 32 39 bp. Based on these data, the method of extracting the genomic DNA of the individual pinewood nematode and PCR SSCP (single strand conformation polymorphism) analysis were employed for the unequivocal differentiation of the Bursaphelenchus variant from B. xylophilus and B. mucronatus. These methods should provide a valuable tool for sensitive and accurate identification of individual worm of B. xylophilus. |
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| Keywords: | Bursaphelenchus xylophilus sequencing rDNA ITS PCR- SSCP |
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