Hostname: page-component-cd9895bd7-jn8rn Total loading time: 0 Render date: 2024-12-18T13:56:00.588Z Has data issue: false hasContentIssue false
  • English
  • Français

Host-mediated RNAi of a Notch-like receptor gene in Meloidogyne incognita induces nematode resistance

Published online by Cambridge University Press:  25 April 2018

Deshika Kohli ,
Parameswaran Chidambaranathan ,
J. Prasanth Tej Kumar ,
Ashish Kumar Singh ,
Anil Kumar ,
Anil Sirohi ,
K. Subramaniam ,
Ramamurthy Srinivasan ,
Navneeta Bharadvaja  and
Pradeep K. Jain Open the ORCID record for Pradeep K. Jain [Opens in a new window]
Deshika Kohli
Affiliation:
ICAR-NRC on Plant Biotechnology, Pusa Campus, New Delhi 110012, India Delhi Technological University, Shahbad Daulatpur, Delhi-110042, India
Parameswaran Chidambaranathan
Affiliation:
ICAR-NRC on Plant Biotechnology, Pusa Campus, New Delhi 110012, India
J. Prasanth Tej Kumar
Affiliation:
ICAR-NRC on Plant Biotechnology, Pusa Campus, New Delhi 110012, India
Ashish Kumar Singh
Affiliation:
Division of Nematology, ICAR-Indian Agricultural Research Institute, Pusa Campus, New Delhi 110012, India
Anil Kumar
Affiliation:
ICAR-NRC on Plant Biotechnology, Pusa Campus, New Delhi 110012, India
Anil Sirohi
Affiliation:
Division of Nematology, ICAR-Indian Agricultural Research Institute, Pusa Campus, New Delhi 110012, India
K. Subramaniam
Affiliation:
Department of Biotechnology, Indian Institute of Technology, Madras, Chennai 600036, India
Ramamurthy Srinivasan
Affiliation:
ICAR-NRC on Plant Biotechnology, Pusa Campus, New Delhi 110012, India
Navneeta Bharadvaja
Affiliation:
Delhi Technological University, Shahbad Daulatpur, Delhi-110042, India
Pradeep K. Jain*
Affiliation:
ICAR-NRC on Plant Biotechnology, Pusa Campus, New Delhi 110012, India
*
Author for correspondence: Pradeep K. Jain, E-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

Abstract

GLP-1 (abnormal germline proliferation) is a Notch-like receptor protein that plays an essential role in pharyngeal development. In this study, an orthologue of Caenorhabditis elegans glp-1 was identified in Meloidogyne incognita. A computational analysis revealed that the orthologue contained almost all the domains present in the C. elegans gene: specifically, the LIN-12/Notch repeat, the ankyrin repeat, a transmembrane domain and different ligand-binding motifs were present in orthologue, but the epidermal growth factor-like motif was not observed. An expression analysis showed differential expression of glp-1 throughout the life cycle of M. incognita, with relatively higher expression in the egg stage. To evaluate the silencing efficacy of Mi-glp-1, transgenic Arabidopsis plants carrying double-stranded RNA constructs of glp-1 were generated, and infection of these plants with M. incognita resulted in a 47–50% reduction in the numbers of galls, females and egg masses. Females obtained from the transgenic RNAi lines exhibited 40–60% reductions in the transcript levels of the targeted glp-1 gene compared with females isolated from the control plants. Second-generation juveniles (J2s), which were descendants of the infected females from the transgenic lines, showed aberrant phenotypes. These J2s exhibited a significant decrease in the overall distance from the stylet to the metacorpus region, and this effect was accompanied by disruption around the metacorporeal bulb of the pharynx. The present study suggests a role for this gene in organ (pharynx) development during embryogenesis in M. incognita and its potential use as a target in the management of nematode infestations in plants.

Keywords

Arabidopsisdouble-stranded RNA (dsRNA)glp-1 genein vivo RNAiMeloidogyne incognitapharynx development
Type
Research Article
Information
Parasitology , Volume 145 , Issue 14 , December 2018 , pp. 1896 - 1906
Check for updates
Copyright
Copyright © Cambridge University Press 2018 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Abad, P, et al. (2008) Genome sequence of the metazoan plant-parasitic nematode Meloidogyne incognita. Nature Biotechnology 26, 909–915.Google Scholar
Agrawal, N, et al. (2003) RNA interference: biology, mechanism, and applications. Microbiology and Molecular Biology Reviews 67, 657–685.Google Scholar
Artavanis-Tsakonas, S, Rand, MD and Lake, RJ (1999) Notch signaling: cell fate control and signal integration in development. Science 284, 770–776.Google Scholar
Austin, J and Kimble, J (1989) Transcript analysis of glp-1and lin-12, homologous genes required for cell interactions during development of C. elegans. Cell 58, 565–571.Google Scholar
Bailey, TL, et al. (2015) The MEME Suite. Nucleic Acids Research 43, W39–W49.Google Scholar
Banerjee, S, et al. (2017) RNA interference: a novel source of resistance to combat plant parasitic nematodes. Frontiers in Plant Science 8, 834.Google Scholar
Bass, BL (2000) Double-stranded RNA minireview as a template for gene silencing. Cell 101, 235–238.Google Scholar
Bird, DM, et al. (2009) The genomes of root-knot nematodes. Annual Review of Phytopathology 47, 333–351.Google Scholar
Bybd, DW Jr., Kirkpatrick, T and Barker, KR (1983) An improved technique for clearing and staining plant tissues for detection of nematodes. Journal of Nematology 15, 142–143.Google Scholar
Calderón-Urrea, A, et al. (2016) Early development of the root-knot nematode Meloidogyne incognita. BMC Developmental Biology 16, 10.Google Scholar
Castagnone-Sereno, P, et al. (2010) Genome-wide survey and analysis of microsatellites in nematodes, with a focus on the plant-parasitic species Meloidogyne incognita. BMC Genomics 11, 598.Google Scholar
Christensen, S, et al. (1996) lag-1, a gene required for lin-12 and glp-1 signaling in Caenorhabditis elegans, is homologous to human CBF1 and Drosophila Su(H). Development 122, 1373–1383.Google Scholar
Clough, SJ and Bent, AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. The Plant Journal 16, 735–743.Google Scholar
Cotton, JA, et al. (2014) The genome and life-stage specific transcriptomes of Globodera pallida elucidate key aspects of plant parasitism by a cyst nematode. Genome Biology 15, R43.Google Scholar
Crittenden, SL, et al. (1997) Genes required for GLP-1 asymmetry in the early Caenorhabditis elegans embryo. Developmental Biology 181, 36–46.Google Scholar
Diederich, RJ, et al. (1994) Cytosolic interaction between deltex and Notch ankyrin repeats implicates deltex in the Notch signaling pathway. Development 120, 473–481.Google Scholar
Dinh, PTY, Brown, CR and Elling, AA (2014) RNA interference of effector gene Mc16D10L confers resistance against Meloidogyne chitwoodi in Arabidopsis and Potato. Phytopathology 104, 1098–1106.Google Scholar
Edgar, RC (2004) MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Research 32, 1792–1797.Google Scholar
Eisenback, JD and Hunt, DJ (2009) General morphology. In Perry, RN, Moens, M and Starr, JL (eds). Root Knot Nematode. Oxfordshire, UK: CAB International 2, pp. 18–50.Google Scholar
Elling, AA (2013) Major emerging problems with minor Meloidogyne species. Phytopathology 103, 1092–1102.Google Scholar
Elsworth, B, Wasmuth, J and Blaxter, M (2011) NEMBASE4: the nematode transcriptome resource. International Journal of Parasitology 41, 881–894.Google Scholar
Evans, TC, et al. (1994) Translational control of maternal glp-1 mRNA establishes an asymmetry in the C. elegans embryo. Cell 77, 183–194.Google Scholar
Fanelli, E, et al. (2005) Analysis of chitin synthase function in a plant parasitic nematode, Meloidogyne artiellia, using RNAi. Gene 349, 87–95.Google Scholar
Finn, RD, et al. (2016) The Pfam protein families database: towards a more sustainable future. Nucleic Acids Research 44, D279–D285.Google Scholar
García, LE and Sánchez-Puerta, MV (2012) Characterization of a root-knot nematode population of Meloidogyne arenaria from Tupungato (Mendoza, Argentina). Journal of Nematology 44(3), 291–301.Google Scholar
Gasteiger, E, et al. (2005) Protein identification and analysis tools on the ExPASy server. In Walker, JM (ed). The Proteomics Protocols Handbook. Totowa, NJ: Humana Press, pp. 571–607.Google Scholar
Geng, C, et al. (2016) A novel serine protease, Sep1, from Bacillus firmus DS-1 has nematicidal activity and degrades multiple intestinal-associated nematode proteins. Scientific Reports 6, 25012.Google Scholar
Gleason, CA, Liu, QL and Williamson, VM (2008) Silencing a candidate nematode effector gene corresponding to the tomato resistance gene Mi-1 leads to acquisition of virulence. Molecular Plant-Microbe Interactions 21(5), 576–585.Google Scholar
Gray, LJ, Curtis, RH and Jones, JT (2001) Characterisation of a collagen gene subfamily from the potato cyst nematode Globodera pallida. Gene 263, 67–75.Google Scholar
Heitzler, P and Simpson, P (1993) Altered epidermal growth factor-like sequences provide evidence for a role of Notch as a receptor in cell fate decisions. Development 117, 1113–1123.Google Scholar
Hu, B, et al. (2015) GSDS 2.0: an upgraded gene feature visualization server. Bioinformatics (Oxford, England) 31(8), 1296–1297.Google Scholar
Iberkleid, I, Sela, N and Brown Miyara, S (2015) Meloidogyne javanica fatty acid- and retinol-binding protein (Mj-FAR-1) regulates expression of lipid-, cell wall-, stress- and phenyl propanoid-related genes during nematode infection of tomato. BMC Genomics 16, 272.Google Scholar
Kaur, H and Attri, R (2013) Morphological and morphometrical characterization of Meloidogyne incognita from different host plants in four districts of Punjab, India. The Journal of Nematology 45, 122–127.Google Scholar
Kelley, MR, et al. (1987) Mutations altering the structure of epidermal growth factor-like coding sequences at the Drosophila Notch locus. Cell 57, 539–548.Google Scholar
Kumar, A, et al. (2017) Host-delivered RNAi-mediated root-knot nematode resistance in Arabidopsis by targeting splicing factor and integrase genes. Journal of General Plant Pathology 83, 91–97.Google Scholar
Kumar, S, Stecher, G and Tamura, K (2016) MEGA7: Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Molecular Biology and Evolution 33, 1870–1874.Google Scholar
Kyte, J and Doolittle, R (1982) A simple method for displaying the hydropathic character of a protein. Journal of Molecular Biology 157, 105–132.Google Scholar
Livak, KJ and Schmittgen, TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta DeltaC(T)) method. Methods 25(4), 402–408.Google Scholar
Lu, CJ, et al. (2016) Nuclear receptor nhr-48 is required for pathogenicity of the second stage (J2) of the plant parasite Meloidogyne incognita. Scientific Reports 6, 34959.Google Scholar
Lunt, DH, et al. (2014) The complex hybrid origins of the root knot nematodes revealed through comparative genomics. PeerJ 2, e356.Google Scholar
Mango, SE (2007) The C. elegans pharynx: a model for organogenesis. In Wormbook, The C. elegans Research community. WormBook. doi/10.1895/wormbook.1.129.1.Google Scholar
Martin, J, et al. (2015) Helminth.net: expansions to Nematode.net and an introduction to Trematode.net. Nucleic Acids Research 43, D698–D706.Google Scholar
Martinuz, A, Schouten, A and Sikora, RA (2013) Post-infection development of Meloidogyne incognita on tomato treated with the endophytes Fusarium oxysporum strain Fo162 and Rhizobium etli strain G12. BioControl 58, 95–104.Google Scholar
Moens, M, Perry, RN and Starr, JL (2009) Meloidogyne species: a diverse group of novel and important plant parasites. In Perry, RN, Moens, M and Starr, JL (eds). Root-knot nematodes. Wallingford, UK: CABI Publishing, pp. 1–17.Google Scholar
Niu, J, et al. (2012) RNAi silencing of the Meloidogyne incognita Rpn7 gene reduces nematode parasitic success. European Journal of Plant Pathology 134, 131–144.Google Scholar
Nguyễn, PV, et al. (2014) Meloidogyne incognita – rice (Oryza sativa) interaction: a new model system to study plant-root-knot nematode interactions in monocotyledons. Rice 7, 23.Google Scholar
Nsengimana, J, et al. (2013) Silencing of Mg-pat-10 and Mg-unc-87 in the plant parasitic nematode Meloidogyne graminicola using siRNAs. Agriculture 3, 567–678.Google Scholar
Opperman, CH, et al. (2008) Sequence and genetic map of Meloidogyne hapla: a compact nematode genome for plant parasitism. Proceedings of the National Academy of Sciences USA 105, 14802–14807.Google Scholar
Priess, JR, Schnabel, H and Schnabel, R (1987) The glp-1 locus and cellular interactions in early C. elegans embryos. Cell 51, 601–611.Google Scholar
Rastogi, S, et al. (2015) Caenorhabditis elegans glp-4 encodes a valylaminoacyl tRNA synthetase. Genes Genomes Genetics 5, 719–728.Google Scholar
Rebay, I, et al. (1991) Specific EGF repeats of Notch mediate interactions with Delta and Serrate: implications for Notch as a multifunctional receptor. Cell 67, 687–699.Google Scholar
Rehman, S, Gupta, VK and Goyal, AK (2016) Identification and functional analysis of secreted effectors from phytoparasitic nematodes. BMC Microbiology 16, 48.Google Scholar
Roehl, H and Kimble, J (1993) Control of cell fate in C. elegans by a GLP-1 peptide consisting primarily of ankyrin repeats. Nature 364, 632–635.Google Scholar
Roehl, H, et al. (1996) Roles of the RAM and ANK domains in signaling by the C. elegans GLP-1 receptor. The EMBO Journal 15, 7002–7012.Google Scholar
Rudel, D and Kimble, J (2001) Conservation of glp-1 regulation and function in nematodes. Genetics 157, 639–654.Google Scholar
Tytgat, T, et al. (2004) An SXP/RAL-2 protein produced by the subventral pharyngeal glands in the plant parasitic root-knot nematode Meloidogyne incognita. Parasitology Research 95, 50–54.Google Scholar
Veronico, P, et al. (2001) Nematode chitin synthases: gene structure, expression and function in Caenorhabditis elegans and the plant parasitic nematode Meloidogyne artiellia. Molecular Genetics and Genomics 266, 28–34.Google Scholar
Verwoerd, TC, Dekker, BM and Hoekema, A (1989) A small-scale procedure for the rapid isolation of plant RNAs. Nucleic Acids Research 17(6), 2362.Google Scholar
Wang, T, Deom, CM and Hussey, RS (1998) Identification of a Meloidogyne incognita cuticle collagen gene and characterization of the developmental expression of three collagen genes in parasitic stages. Molecular and Biochemical Parasitology 93, 131–134.Google Scholar
Whitehead, AG (1968) Taxonomy of Meloidogyne (Nematodea: Heteroderidae) with descriptions of four new species. Journal of Zoology l31(3), 263–401.Google Scholar
Xie, J, et al. (2016) A novel Meloidogyne incognita effector Misp12 suppresses plant defense response at latter stages of nematode parasitism. Frontiers in Plant Science 7, 964.Google Scholar
Yadav, BC, Veluthambi, K and Subramaniam, K (2006) Host-generated double stranded RNA induces RNAi in plant-parasitic nematodes and protects the host from infection. Molecular and Biochemical Parasitology 148, 219–222.Google Scholar
Ye, W, Zeng, Y and Kerns, J (2015) Molecular characterisation and diagnosis of root-knot nematodes (Meloidogyne spp.) from Turf grasses in North Carolina, USA. PLoS ONE 10(11), e0143556.Google Scholar
Yochem, J and Greenwald, I (1989) glp-1 and lin-12, genes implicated in distinct cell-cell interactions in C. elegans, encode similar transmembrane proteins. Cell 58, 553–563.Google Scholar
Supplementary material: File

Kohli et al. supplementary material

Figure S1

Download Kohli et al. supplementary material(File)
File 45.4 KB
Supplementary material: File

Kohli et al. supplementary material

Tables S1-S4B

Download Kohli et al. supplementary material(File)
File 35.6 KB
Supplementary material: File

Kohli et al. supplementary material

Figures S2-S5B

Download Kohli et al. supplementary material(File)
File 2.9 MB