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Transcriptomic changes in the pre-parasitic juveniles of Meloidogyne incognita induced by silencing of effectors Mi-msp-1 and Mi-msp-20

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Abstract

Plant–parasitic root-knot nematode Meloidogyne incognita uses an array of effector proteins to establish successful plant infections. Mi-msp-1 and Mi-msp-20 are two known effectors secreted from nematode subventral oesophageal glands; Mi-msp-1 being a putative secretory venom allergen AG5-like protein, whereas Mi-msp-20 is a pioneer gene with a coiled-coil motif. Expression of specific effector is known to cause disturbances in the expression of other effectors. Here, we used RNA-Seq to investigate the pleiotropic effects of silencing Mi-msp-1 and Mi-msp-20. A total of 25.1–51.9 million HQ reads generated from Mi-msp-1 and Mi-msp-20 silenced second-stage juveniles (J2s) along with freshly hatched J2s were mapped to an already annotated M. incognita proteome to understand the impact on various nematode pathways. As compared to control, silencing of Mi-msp-1 caused differential expression of 29 transcripts, while Mi-msp-20 silencing resulted in differential expression of a broader set of 409 transcripts. In the Mi-msp-1 silenced J2s, cytoplasm (GO:0005737) was the most enriched gene ontology (GO) term, whereas in the Mi-msp-20 silenced worms, embryo development (GO:0009792), reproduction (GO:0000003) and nematode larval development (GO:0002119) were the most enriched terms. Limited crosstalk was observed between these two effectors as a sheer 5.9% of the up-regulated transcripts were common between Mi-msp-1 and Mi-msp-20 silenced nematodes. Our results suggest that in addition to the direct knock-down caused by silencing of Mi-msp-1 and Mi-msp-20, the cascading effect on other genes might also be contributing to a reduction in nematode's parasitic abilities.

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Acknowledgements

Funding from the Department of Biotechnology, Government of India to UR through Grant no. BT/PR5908/AGR/36/727/2012 is acknowledged. The authors thank the Director and the Joint Director (Research), ICAR- Indian Agricultural Research Institute, New Delhi, for extending all the support and facilities to complete the study.

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Authors and Affiliations

  1. Division of Nematology, ICAR-Indian Agricultural Research Institute, New Delhi, India

    Vishal Singh Somvanshi, Madhurima Chatterjee & Uma Rao

  2. Department of Agricultural Entomology, College of Agriculture, Uttar Banga Krishi Viswavidyalaya, Majhian, Dakshin Dinajpur, West Bengal, India

    Victor Phani

  3. Department of Nematology, CCS Haryana Agricultural University, Hisar, Haryana, India

    Prakash Banakar

  4. Bionivid Technology Private Limited, 209, 4th Cross, Kasturi Nagar, Bangalore, Karnataka, India

    Roli Budhwar & Rohit Nandan Shukla

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  1. Vishal Singh Somvanshi
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  2. Victor Phani
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  3. Prakash Banakar
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  4. Madhurima Chatterjee
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Contributions

UR conceptualized the study and received the funding for the study; VSS, UR, VP wrote the manuscript; PB collected the biological material for the study; MC performed the validation experiments; VSS, RB, and RNS carried out the bioinformatic analysis of the data and created representations in consultation with UR.

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Correspondence to Uma Rao.

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This article does not contain any studies with human participants, and no animals were harmed for this study by any of the authors.

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Supplementary file 1 (DOCX 15 kb)

13205_2020_2353_MOESM2_ESM.docx

Supplementary file 2 - Supplementary Table 2. Correlation coefficient values of the two biological replicates of Mi-msp-1 and Mi-msp-20 silenced, and control M. incognita J2 samples used for RNA-Seq experiment in this study. (DOCX 14 kb)

13205_2020_2353_MOESM3_ESM.xlsx

Supplementary file 3 - Supplementary Table 3. A list of the up-regulated and down-regulated transcripts in both the Mi-msp-1 and Mi-msp-20 silenced M. incognita J2s as compared to controls along with significant identified pathways. (XLSX 208 kb)

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Somvanshi, V.S., Phani, V., Banakar, P. et al. Transcriptomic changes in the pre-parasitic juveniles of Meloidogyne incognita induced by silencing of effectors Mi-msp-1 and Mi-msp-20. 3 Biotech 10, 360 (2020). https://doi.org/10.1007/s13205-020-02353-8

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