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Identification of differentially expressed genes in Brassica juncea var. tumida Tsen following infection by Plasmodiophora brassicae

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Abstract

Mustard clubroot, caused by Plasmodiophora brassicae, is one of the most serious diseases affecting Brassica juncea var. tumida Tsen, a mustard plant which is the raw material of a traditional fermented food manufactured in the Chongqing Municipality, P. R. China. We used suppression subtractive hybridization (SSH) to better understand the interaction between B. juncea var. tumida and P. brassicae, and the complex regulation of resistance mechanisms occurring in B. juncea var. tumida after infection by P. brassicae. A total of 1,842 different gene clones were selected from the forward subtracted library (using diseased roots as tester and healthy roots as driver), and 224 positive spots were identified following cDNA array dot blotting. Elimination of polyA tails and sequences shorter than 100 bp generated 196 high-quality gene sequences with an average length of 332 bp. Bioinformatic analysis showed that these 196 sequences represented 173 unigenes, comprising 14 contigs and 159 singlets. Of these, 146 ESTs (84.4 % of the total) were significantly similar to known sequences in plants, the remaining 23 (13.3 % of the total) were of P. brassicae origin. We used quantitative reverse transcription-PCR to analyze the six genes most likely to be involved in disease resistance or the stress response to evaluate the efficiency of SSH, and the results showed that our library data is reliable. Further study of these genes might be helpful for breeding resistance of mustard plants to P. brassicae.

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Acknowledgments

This research was financially supported by the Natural Science Foundation of Chongqing Science and Technology Commission (CSTC, 2008BB1370). We also gratefully appreciate the support from Chongqing Fuling Institute of Agricultural Science.

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Authors and Affiliations

  1. Key Lab of Gene Function and Regulation at Chongqing, Bioengineering College of Chongqing University, Chongqing, 40030, People’s Republic of China

    Yuanli Luo, Youping Yin, Yan Liu, Xiaotu Pei & Zhongkang Wang

  2. Fuling Agricultural Science Institute of Chongqing, Chongqing, 40030, People’s Republic of China

    Yuanli Luo, Yonghong Fan, Xuyi Wang & Rong Leng

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  1. Yuanli Luo
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  2. Youping Yin
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  4. Xiaotu Pei
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  5. Zhongkang Wang
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  6. Yonghong Fan
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Correspondence to Zhongkang Wang.

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Supplementary Table 2

Differentially expressed genes in the SSH library (DOC 169 kb) (DOC 169 kb)

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Luo, Y., Yin, Y., Liu, Y. et al. Identification of differentially expressed genes in Brassica juncea var. tumida Tsen following infection by Plasmodiophora brassicae . Eur J Plant Pathol 137, 43–53 (2013). https://doi.org/10.1007/s10658-013-0215-6

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