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RNAi silencing of the Meloidogyne incognita Rpn7 gene reduces nematode parasitic success

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Abstract

RNA interference (RNAi) techniques provide a major breakthrough in functional analysis for plant parasitic nematodes (PPNs). It offers the possibility of identifying new essential targets and consequently developing new resistance transgenes. To validate the potential of Mi-Rpn7 as a target for controlling root knot nematode Meloidogyne incognita and to evaluate the feasibility of our modified platform for assessing silencing phenotypes, we knocked down the Rpn7 gene of M. incognita using RNAi in vitro and in vivo. After soaking with 408-bp Rpn7 dsRNA, pre-parasitic second-stage juvenile (J2) nematodes showed specific transcript knockdown, resulting in an interrupted locomotion in an attraction assay with Pluronic gel medium, and consequently in a reduction of nematode infection ranging from 55.2% to 66.5%. With in vivo expression of Rpn7 dsRNA in transformed composite plants, the amount of egg mass per gram root tissue was reduced by 34% (P < 0.05) and the number of eggs per gram root tissue was reduced by 50.8% (P < 0.05). Our results demonstrated that the silencing of the Rpn7 gene in M. incognita J2s significantly reduced motility and infectivity. Although it does not confer complete resistance, Mi-Rpn7 RNAi in hairy roots produced significant negative impacts on reproduction and motility of M. incognita. In addition, the presented modified procedure provides technique reference for PPN genes functional analysis or target screening.

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Acknowledgments

The authors would like to thank Professors Tevi Tzfira and Dr. Peter M Gresshoff, the University of Queensland, Australia, for providing pSAT6 RNAi vector, pCAMBIA3301 vector and A. rhizogenes strain K599 kindly. This study was financially supported by Program for Changjiang Scholars and Innovative Research Team in University (IRT1042), and the Special Fund for Agro-scientific Research in the Public Interest, China (No. 20113018) and the National Foundation of Natural Sciences, China (No. 30971901), and project 948 from the Ministry of Agriculture of China (No. 2011-G4)

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Authors and Affiliations

  1. Key Laboratory of Plant Pathology, Ministry of Agriculture of China, Department of Plant Pathology, China Agricultural University, Beijing, 100193, China

    Junhai Niu, Heng Jian, Jianmei Xu, Changlong Chen, Quanxin Guo & Qian Liu

  2. Tropical Crops Genetic Resources Institute, CATAS, Danzhou, Hainan, 571737, China

    Junhai Niu

  3. Department of Vegetable Science, China Agricultural University, Beijing, 100193, China

    Junhai Niu & Yangdong Guo

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  1. Junhai Niu
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  2. Heng Jian
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  3. Jianmei Xu
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  4. Changlong Chen
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Correspondence to Heng Jian.

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Niu, J., Jian, H., Xu, J. et al. RNAi silencing of the Meloidogyne incognita Rpn7 gene reduces nematode parasitic success. Eur J Plant Pathol 134, 131–144 (2012). https://doi.org/10.1007/s10658-012-9971-y

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