谷子种子线虫检测及分析

doi:10.7668/hbnxb.201751613

摘要/Abstract

摘要： 为了快速检测谷子种子携带线虫情况并了解不同地区线虫种群的多态性，利用PCR技术对采集自中国谷子主产区不同区域内的99份谷子种子的带线虫情况进行了检测，并分析其不同地区线虫种群的遗传多样性。结果表明，根据贝西滑刃线虫核糖体28S rRNA-D2/D3片段设计的引物对谷子线虫具有高度的特异性和灵敏性，扩增出245 bp的特异性目的片段，对谷子线虫的检测浓度最低为0.125 ng/μL。在99份谷子种子DNA中，有33份种子检测到特异性扩增条带，测序结果表明，所有扩增条带对应序列与线虫28S rRNA序列的相似性达97%以上，进一步说明33份种子携带线虫。重复试验结果表明，阳性种子的带线虫频率介于33.3%~100%，且带线虫的谷子种子主要来自春谷区。分析33份阳性样品的28S rRNA片段序列，存在29个变异位点和22种单倍型，其中单倍型AB1为优势单倍型，河北张家口市和内蒙古松山区两地线虫的单倍型遗传多样性最丰富。本研究建立了一种特异性好、灵敏度高的检测谷子种子带线虫的一步PCR方法；谷子线虫病是夏谷区的主要病害，随着谷子春夏谷区的交流，线虫病成为春谷区病害，种子带病可能是主要初侵染源；不同地理来源的线虫28S rRNA序列存在差异，AB1为优势单倍型。

关键词: 谷子, 贝西滑刃线虫, 种子带菌, 聚合酶链式反应, 单倍型

Abstract: In order to quickly detect the conditions of Aphelenchoides besseyi contamination in millet seeds and understand the population variations of Aphelenchoides besseyi from different regions, a total of 99 millet seed samples collected from different millet production regions of China were detected by PCR technology, and the population variation of Aphelenchoides besseyi was analyzed. The results showed that the specific primers designed according to the 28S rRNA-D2/D3 fragment of Aphelenchoides besseyi had a high specificity and sensitivity to millet nematodes, and could amplify a fragment of 245 bp with the lowest test concentration of 0.125 ng/μL Aphelenchoides besseyi. Among the 99 millet seed samples, 33 samples were detected to be contaminated, in which the contamination rates ranged from 33.3% to 100%, and they mainly distributed in spring millet areas. The sequence analysis of 28S rRNA fragments from 33 positive samples revealed that there were 29 variable sites and 22 haplotypes, of which the haplotype AB1 was the dominant haplotype. The haplotype genetic diversity of Aphelenchoides besseyi was the most abundant in Zhangjiakou, Hebei Province and Songshan District, Inner Mongolia. In this study, a PCR-based detection method was established to detect Aphelenchoides besseyi in millet seeds with a good specificity and high sensitivity, which could be used to detect the millet seeds carrying Aphelenchoides besseyi. The millet nematode is a major disease in summer millet areas, but with the millet exchange between spring and summer millet areas, it has become a disease in spring millet area. The seed carrier may be the main primary infection source. The 28S rRNA sequences of Aphelenchoides besseyi from different geographic origins were diverse, and AB1 was the dominant haplotype.

Key words: Foxtail millet, Aphelenchoides besseyi, Seed-borne, Polymerase chain reaction, Haplotype

中图分类号:

S515

宋振君, 李志勇, 王永芳, 全建章, 马继芳, 白辉, 董志平. 谷子种子线虫检测及分析[J]. 华北农学报, 2019, 34(4): 208-216. doi: 10.7668/hbnxb.201751613.

SONG Zhenjun, LI Zhiyong, WANG Yongfang, QUAN Jianzhang, MA Jifang, BAI Hui, DONG Zhiping. Detection and Analysis of Millet Seeds with Aphelenchoides besseyi[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2019, 34(4): 208-216. doi: 10.7668/hbnxb.201751613.

http://www.hbnxb.net/CN/Y2019/V34/I4/208

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