Development of A Real-Time PCR Assay for Plasmodiophora brassicae and Its Detection in Soil Samples

文献类型： 外文期刊

第一作者： Li Jin-ping

作者： Li Jin-ping;Li Yan;Shi Yan-xia;Xie Xue-wen;Chai A-li;Li Bao-ju

作者机构：

关键词： species-specific;rDNA-ITS gene;Plasmodiophora brassicae;real-time quantitative PCR;SYBR Green I dye

期刊名称：JOURNAL OF INTEGRATIVE AGRICULTURE （ 影响因子：2.848； 五年影响因子：2.979 ）

ISSN： 2095-3119

年卷期： 2013 年 12 卷 10 期

页码：

收录情况： SCI

摘要： A SYBR Green I real-time PCR assay was developed to detect and quantify Plasmodiophora brassicae ribosomal DNA (rDNA) and internal transcribed spacer (ITS). A pair of primers PBF1/PBR1 was designed based on the conservative region of rDNA-ITS of P. brassicae. The positive plasmid pB12 was obtained and used as the template to create standard curve. The specificity, sensitivity, and reproducibility of real-time PCR were evaluated respectively. Naturally and artificially infested soil samples containing different concentrations of P. brassicae were detected. The results demonstrated that standard curve established by recombinant plasmid was shown a fine linear relationship between threshold cycle and template concentration. The melting curve was specific with the correlation coefficient of 0.995 and that the amplification efficiency was 93.8%. The detection limit of P. brassicae genomic DNA was approximately 40 copies per 25 mu L. The sensitivity of the assay was at least 100-fold higher than conventional PCR. Only DNA from P. brassicae could be amplified and detected using this assay, suggesting the highly specific of this assay. The coefficient of variation was less than 3%, indicating the PCR method revealed high reproducibility. The detection limit in soil samples corresponded to 1000 resting spores g(-1) soil. Bait plants were used to validate the real-time PCR assay. This developed real-time PCR assay allows for fast and sensitive detection of P. brassicae in soil and should be useful in disease management and pest interception so as to prevent further spread of P. brassicae.

分类号：

  • 相关文献

[1]Special Issue: Research status of clubroot (Plasmodiophora brassicae) on cruciferous crops in China. Chai, A. L.,Xie, X. W.,Shi, Y. X.,Li, B. J.. 2014

[2]Comparative Transcriptome Analysis between Broccoli (Brassica oleracea var. italica) and Wild Cabbage(Brassica macrocarpa Guss.) in Response to Plasmodiophorabrassicae during Different Infection Stages. Zhang, Xiaoli,Liu, Yumei,Fang, Zhiyuan,Li, Zhansheng,Yang, Limei,Zhuang, Mu,Zhang, Yangyong,Lv, Honghao. 2016

[3]Estimating Plasmodiophora brassicae gene expression in lines of B. rapa by RT-PCR. Wu, Li-yan,Li, Shi-kai,Gong, Ya-ju,Zhong, Li,He, Jiang-Ming,Siemens, Johannes,Ludwig-Mueller, Jutta. 2012

[4]A Genome-Wide Association Study Reveals New Loci for Resistance to Clubroot Disease in Brassica napus. Li, Lixia,Luo, Yujie,Chen, Biyun,Xu, Kun,Zhang, Fugui,Li, Hao,Huang, Qian,Xiao, Xin,Zhang, Tianyao,Hu, Jihong,Li, Feng,Wu, Xiaoming. 2016

[5]Transcriptome Analysis of Brassica rapa Near-lsogenic Lines Carrying Clubroot-Resistant and -Susceptible Alleles in Response to Plasmodiophora brassicae during Early Infection. Chen, Jingjing,Pang, Wenxing,Piao, Zhongyun,Chen, Bing,Zhang, Chunyu,Zhang, Chunyu. 2016

[6]Dissemination of Plasmodiophora brassicae in livestock manure detected by qPCR. Chai, A. L.,Li, J. P.,Xie, X. W.,Shi, Y. X.,Li, B. J..

[7]Putative role of IAA during the early response of Brassica napus L. to Plasmodiophora brassicae. Xu, Li,Ren, Li,Chen, Kunrong,Liu, Fan,Fang, Xiaoping,Xu, Li,Ren, Li,Chen, Kunrong,Liu, Fan,Fang, Xiaoping.

[8]Quantitative Detection of Chinese Cabbage Clubroot Based on FTIR Spectroscopy. Wang Wei-ping,Wang Wei-ping,Chai A-li,Shi Yan-xia,Xie Xue-wen,Li Bao-ju. 2015

[9]Isolation and characterization of Zhihengliuella aestuarii B18 suppressing clubroot on Brassica juncea var. tumida Tsen. Luo, Yuanli,Zhou, Changyong,Luo, Yuanli,Dong, Daiwen,Gou, Zhiqin,Wang, Xuyi,Jiang, Huan,Yan, Yufang,Wu, Chaojun. 2018

[10]Draft genome sequence of Bacillus amyloliquefaciens HB-26. Liu, Xiao-Yan,Min, Yong,Wang, Kai-Mei,Wan, Zhong-Yi,Zhang, Zhi-Gang,Cao, Chun-Xia,Zhou, Rong-Hua,Jiang, Ai-Bing,Liu, Cui-Jun,Zhang, Guang-Yang,Cheng, Xian-Liang,Yang, Zi-Wen,Zhang, Wei. 2014

[11]Root RNA-seq analysis reveals a distinct transcriptome landscape between clubroot-susceptible and clubroot-resistant Chinese cabbage lines after Plasmodiophora brassicae infection. Jia, Hao,Yang, Yan,Wei, Fang,Tian, Baoming,Jia, Hao,Wei, Xiaochun,Yuan, Yuxiang,Zhao, Yanyan,Yang, Shuangjuan,Yao, Qiuju,Wang, Zhiyong,Zhang, Xiaowei.

[12]Molecular Detection and Pathogenicity Assay of Plasmodiophora brassicae in Chicken Manure. Huang, Rui-rong,Huang, Rong,Hu, Jian-kun,Ho, Hon-hing,He, Yue-qiu.

[13]Identification of chilling-responsive transcripts in peanut (Arachis hypogaea L.). Wang, Chuan Tang,Wang, Xiu Zhen,Chi, Xiao Yuan,Wu, Qi,Chen, Dian Xu,Tang, Yue Yi,Yang, Guan Pin,Feng, Tong,Gao, Hua Yuan,Xu, Ya Long. 2011

[14]Different Tolerance in Bread Wheat, Durum Wheat and Barley to Fusarium Crown Rot Disease Caused by Fusarium pseudograminearum. Liu, Yaxi,Wei, Yuming,Zheng, Youliang,Liu, Yaxi,Ma, Jun,Yan, Wei,Liu, Chunji,Ma, Jun,Yan, Guijun,Yan, Wei,Zhou, Meixue,Zhou, Meixue. 2012

[15]Analysis on Differentially Expressed Genes of Muscle Tissues in Rugao Chicken at the Ages of 2 and 12 Weeks by Microarray. Luan, De Qin,Bin Chang, Guo,Zhen, Ting,Huang, Zheng Yang,Zhou, Wei,Chen, Guo Hong,Liu, Yan. 2011

[16]Molecular identification and interaction assay of the gene (OsUbc13) encoding a ubiquitin-conjugating enzyme in rice. Wang, Ya,Xu, Meng-yun,Liu, Jian-ping,Wang, Mu-gui,Tu, Ju-min,Wang, Ya,Yin, Hai-qing. 2014

[17]Identification of a trypsin gene from Scylla paramamosain and its expression profiling during larval development. Jiang, Keji,Zhang, Fengying,Zhang, Dan,Zhang, Yong,Qiao, Zhenguo,Ma, Lingbo,Tao, Qichang,Pi, Yan. 2011

[18]Ontogeny expression of ghrelin, neuropeptide Y and cholecystokinin in blunt snout bream, Megalobrama amblycephala. Ping, H. -C.,Feng, K.,Zhang, G. -R.,Wei, K. -J.,Wang, W. -M.,Zou, G. -W.. 2014

[19]Expression Pattern Analysis of Four Mlo Genes from Rose. Qiu, Xianqin,Jian, Hongying,Wang, Qigang,Tang, Kaixue,Qiu, Xianqin,Bao, Manzhu.

[20]Large proportion of genes in one cryptic WO prophage genome are actively and sex-specifically transcribed in a fig wasp species. Wang, Guan-Hong,Xiao, Jin-Hua,Huang, Da-Wei,Wang, Guan-Hong,Niu, Li-Ming,Ma, Guang-Chang,Huang, Da-Wei. 2014

作者其他论文 更多>>

微信小程序