Skip to main content
SpringerLink
Log in

Molecular cloning and virus-induced gene silencing of MiASB in the southern root-knot nematode, Meloidogyne incognita

  • Original Research
  • Published:
European Journal of Plant Pathology Aims and scope Submit manuscript

Abstract

Root-knot nematodes (Meloidogyne spp.), cause serious damage to agricultural production worldwide. In this study, we designed special primers based on the predicted Mitochondrial ATP synthase b subunit gene (ASB) sequence to clone the same gene in M. incognita (MiASB). The identity between the cloned MiASB and the predicted MiASB was as high as 100 %. Using the tobacco rattle virus (TRV)-mediated virus-induced gene silencing (VIGS) system, we delivered MiASB RNAi triggers to the M. incognita feeding site on tomato seedlings, resulting in significantly fewer galls on the seedlings. Sixty days after inoculation with M. incognita, the number of root galls induced on the MiASB silence-treated seedlings was reduced by 64.3 % compared to that on the control seedlings, and reduced by 64.1 % compared to that on untreated control seedlings. This study revealed the MiASB silencing had a positive effect on the control of root-knot nematodes, and MiASB may be associated with the formation of galls caused by the nematode.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10

Similar content being viewed by others

References

  • Abad, P., Gouzy, J., Aury, J. M., Castagnone-Sereno, P., Danchin, E. G. J., Deleury, E., et al. (2008). Genome sequence of the metazoan plant-parasitic nematode Meloidogyne incognita. Nature Biotechnology, 26, 909–915.

    Article  CAS  PubMed  Google Scholar 

  • Becker, A., & Lange, M. (2010). VIGS–genomics goes functional. Trends in plant science, 15, 1–4.

    Article  CAS  PubMed  Google Scholar 

  • Bonora, M., Patergnani, S., Rimessi, A., De Marchi, E., Suski, J. M., Bononi, A., et al. (2012). ATP synthesis and storage. Purinergic Signalling, 8, 343–357.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Caboni, P., Ntalli, N. G., Aissani, N., Cavoski, I., & Angioni, A. (2012). Nematicidal activity of (E, E)-2, 4-Decadienal and (E)-2-Decenal from Ailanthus altissima against Meloidogyne javanica. Journal of Agricultural and Food Chemistry, 60, 1146–1151.

    Article  CAS  PubMed  Google Scholar 

  • Dang, Q. L., Kim, W. K., Nguyen, C. M., Choi, Y. H., Choi, G. J., Jang, K. S., et al. (2011). Nematicidal and Antifungal Activities of Annonaceous Acetogenins from Annona squamosa against Various Plant Pathogens. Journal of Agricultural and Food Chemistry, 59, 11160–11167.

    Article  CAS  PubMed  Google Scholar 

  • Di Stilio, V. S., Kumar, R. A., Oddone, A. M., Tolkin, T. R., Salles, P., & McCarty, K. (2010). Virus-induced gene silencing as a tool for comparative functional studies in thalictrum. PLoS ONE, 5(8), e12064.

    Article  PubMed Central  PubMed  Google Scholar 

  • Dubreuil, G., Magliano, M., Dubrana, M. P., Lozano, J., Lecomte, P., Favery, B., et al. (2009). Tobacco rattle virus mediates gene silencing in a plant parasitic root-knot nematode. Journal of Experimental Botany, 60, 4041–4050.

    Article  CAS  PubMed  Google Scholar 

  • Dudkina, N. V., Kouril, R., Peters, K., Braun, H., & Boekema, E. J. (2010). Structure and function of mitochondrial supercomplexes. Biochimica et Biophysica Acta, 1797, 664–670.

    Article  CAS  PubMed  Google Scholar 

  • Gruber, G., & Marshansky, V. (2008). New insights into structure-function relationships between archeal ATP synthase (A1A0) and vacuolar type ATPase (V1V0). BioEssays, 30, 1096–1109.

    Article  CAS  PubMed  Google Scholar 

  • Hakulinen, J.K., Klyszejko, A.L., Hoffmann, J., Eckhardt-Strelau, L., Brutschy, B., Vonck, J., & Meier, T. (2012). Structural study on the architecture of the bacterial ATP synthase F0 motor. PNAS, 2012, E2050–E2056. Doi:10.1073/pnas.1203971109.

  • Huang, G., Allen, R., Davis, E., Baum, T., & Hussey, R. (2006). Engineering broad root-knot resistance in transgenic plants by RNAi silencing of a conserved and essential root-knot nematode parasitism gene. Proceedings of the National Academy of Sciences of the United States of America, 103, 14302–14306.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Jablonska, B., Ammiraju, J. S., Bhattarai, K. K., Mantelin, S., Martinez de Ilarduya, O., Roberts, P. A., et al. (2007). The Mi-9 gene from Solanum arcanum conferring heat-stable resistance to root-knot nematodes is a homolog of Mi-1. Plant Physiology, 143, 1044–1054.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Kościańska, E., Kalantidis, K., Wypijewski, K., Sadowski, J., & Tabler, M. (2005). Analysis of RNA silencing in agroinfiltrated leaves of Nicotiana benthamiana and Nicotiana tabacum. Plant Molecular Biology, 59, 647–661.

    Article  PubMed  Google Scholar 

  • Kourmpetis, Y. A. I., van Dijk, A. D. J., van Ham, R. C. H. J., & ter Braak, C. J. F. (2011). Genome-wide computational function prediction of arabidopsis proteins by integration of multiple data sources. Plant Physiology, 155, 271–281.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Leal-Bertioli, S. C. M., José, A. C. V. F., Alves-Freitas, D. M. T., Moretzsohn, M. C., Guimaraes, P. M., Nielen, S., et al. (2009). Identification of candidate genome regions controlling disease resistance in Arachis. BMC Plant Biology, 9, 112.

    Article  PubMed Central  PubMed  Google Scholar 

  • Li, H. Q., Bai, C. Q., Chu, S. S., Zhou, L., Du, S. S., Li, Z. L., et al. (2011). Chemical composition and toxicities of the essential oil derived from Kadsura heteroclita stems against Sitophilus zeamais and Meloidogyne incognita. Journal of Medicinal Plants Research, 5, 4943–4948.

    CAS  Google Scholar 

  • Lin, B., Zhuo, K., Wu, P., Cui, R., Zhang, L. H., & Liao, J. (2013). A novel effector protein, MJ-NULG1a, targeted to giant cell nuclei plays a role in Meloidogyne javanica parasitism. Molecular Plant - Microbe Interactions, 26, 55–66.

    Article  CAS  PubMed  Google Scholar 

  • Livak, K. J., & Schmittgen, T. D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods, 25, 402–408.

    Article  CAS  PubMed  Google Scholar 

  • Moyroud, E., Minguet, E. G., Ott, F., Yant, L., Pose, D., Monniaux, M., et al. (2011). Prediction of regulatory interactions from genome sequences using a biophysical model for the arabidopsis LEAFY transcription factor. The Plant Cell, 23, 1293–1306.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Naz, I., Palomares-Rius, J. E., Saifullah, Blok, V., Khan, M. R., Ali, S., et al. (2012). In vitro and in planta nematicidal activity of Fumaria parviflora (Fumariaceae) against the southern root-knot nematode Meloidogyne incognita. Plant Pathology. doi:10.1111/j.1365-3059.2012.02682.x.

    Google Scholar 

  • Ntalli, N. G., Ferrari, F., Giannakou, I., & Menkissoglu-Spiroudi, U. (2010). Phytochemistry and nematicidal activity of the essential oils from 8 greek lamiaceae aromatic plants and 13 terpene components. Journal of Agricultural and Food Chemistry, 58, 7856–7863.

    Article  CAS  PubMed  Google Scholar 

  • Ntalli, N. G., Manconi, F., Leonti, M., Maxia, A., & Caboni, P. (2011). Aliphatic ketones from ruta chalepensis (rutaceae) induce paralysis on root knot nematodes. Journal of Agricultural and Food Chemistry, 59, 7098–7103.

    Article  CAS  PubMed  Google Scholar 

  • Oka, Y., Nacar, S., Putievsky, E., Ravid, U., Yaniv, Z., & Spiegel, Y. (2000). Nematicidal activity of essential oils and their components against the root-knot nematode. Phytopathology, 90, 710–715.

    Article  CAS  PubMed  Google Scholar 

  • Opperman, C. H., Bird, D. M., Williamson, V. M., Rokhsar, D. S., Burke, M., Cohn, J., et al. (2008). Sequence and genetic map of Meloidogyne hapla: a compact nematode genome for plant parasitism. Proceedings of the National Academy of Sciences of the United States of America, 105, 14802–14807.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Paumard, P., Vaillier, J., Coulary, B., Schaeffer, J., Soubannier, V., Mueller, D. M., et al. (2002). The ATP synthase is involved in generating mitochondrial cristae morphology. The EMBO Journal, 21, 221–230.

    Article  CAS  PubMed  Google Scholar 

  • Ratcliff, F., Martin-Hernandez, A. M., & Baulcombe, D. C. (2001). Tobacco rattle virus as a vector for analysis of gene function by silencing. Plant Journal, 25, 237–245.

    Article  CAS  PubMed  Google Scholar 

  • Rosso, M. N., Dubrana, M. P., Cimbolini, N., Jaubert, S., & Abad, P. (2005). Application of RNA interference to root–knot nematode genes encoding esophageal gland proteins. Molecular Plant - Microbe Interactions, 18, 615–620.

    Article  CAS  PubMed  Google Scholar 

  • Rosso, M. N., Jones, J. T., & Abad, P. (2009). RNAi and functional genomics in plant parasitic nematodes. Annual Review of Phytopathology, 47, 207–232.

    Article  CAS  PubMed  Google Scholar 

  • Runo, S. (2011). Engineering host-derived resistance against plant parasites through RNA interference: challenges and opportunities. Bioeng Bugs, 2, 208–213.

    Article  PubMed  Google Scholar 

  • Sahu, P. P., Puranik, S., Khan, M., & Prasad, M. (2012). Recent advances in tomato functional genomics: utilization of VIGS. Protoplasma, 249, 1017–1027.

    Article  CAS  PubMed  Google Scholar 

  • Sambrook, J., & Russell, D. W. (2001). Molecular cloning: a laboratory manual (3rd E). New York: Cold Spring Harbor Laboratory Press.

  • Senthil-Kumar, M., & Mysore, K. S. (2011). New dimensions for VIGS in plant functional genomics. Trends in Plant Science, 16, 656–665.

    Article  CAS  PubMed  Google Scholar 

  • The C. elegans Sequencing Consortium. (1998). Genome sequence of the nematode C. elegans: a platform for investigating biology. Science, 282, 2012–2018.

    Article  Google Scholar 

  • Valentine, T. A., Randall, E., Wypijewski, K., Chapman, S., Jones, J., & Oparka, K. J. (2007). Delivery of macromolecules to plant parasitic nematodes using a tobacco rattle virus vector. Plant Biotechnology Journal, 5, 827–834.

    Article  CAS  PubMed  Google Scholar 

  • Valentine, T., Shaw, J., Blok, V. C., Phillips, M. S., Oparka, K. J., & Lacomme, C. (2004). Efficient virus-induced gene silencing in roots using a modified tobacco rattle virus vector. Plant Physiology, 136, 3999–4009.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Velásquez, A. C., Chakravarthy, S., & Martin, G. B. (2009). Virus-induced gene silencing (VIGS) in Nicotiana benthamiana and tomato. JoVE, 28, 1292. doi:10.3791/1292.

    PubMed  Google Scholar 

  • Wang, Q., Lei, Y., Xu, X., Wang, G., & Chen, L. (2012). Theoretical prediction and experimental verification of protein-coding genes in plant pathogen genome Agrobacterium tumefaciens strain C58. PLoS ONE, 7, e43176.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Watt, I. N., Montgomery, M. G., Runswick, M. J., Leslie, A. G. W., & Walker, J. E. (2010). Bioenergetic cost of making an adenosine triphosphate molecule in animal mitochondria. PNAS, 107, 16823–16827.

    Article  CAS  PubMed  Google Scholar 

  • Wittig, I., & Schägger, H. (2009). Supramolecular organization of ATP synthase and respiratory chain in mitochondrial membranes. Biochimica et Biophysica Acta, 1787, 672–680.

    Article  CAS  PubMed  Google Scholar 

  • Yadav, B. C., Veluthambib, K., & Subramaniam, K. (2006). Host-generated double stranded RNA induces RNAi in plant-parasitic nematodes and protects the host from infection. Molecular & Biochemical Parasitology, 148, 219–222.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Key Laboratory of Biology and Genetic Resource Utilization of Fruit Tree in South Subtropics, Ministry of Agriculture/Fruit Research Institute, Guangdong Academy of Agricultural Science, Guangzhou, 510640, People’s Republic of China

    Yonghong Huang

  2. Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, 100081, People’s Republic of China

    Yonghong Huang, Mei Mei, Zhenchuan Mao & Bingyan Xie

  3. College of Horticulture and Landscape, Hunan Agriculture University, Changsha, 410128, People’s Republic of China

    Mei Mei

  4. Dongguan Institute of Banana and Vegatable Sciences, Dongguan, Guangdong, 523061, People’s Republic of China

    Shun Lv, Jiankun Zhou & Shi Chen

Authors
  1. Yonghong Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mei Mei
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Zhenchuan Mao
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shun Lv
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jiankun Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Shi Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Bingyan Xie
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Bingyan Xie.

Additional information

Dr. Mei Mei and Dr. Yonghong Huang contributed equally to this research.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Huang, Y., Mei, M., Mao, Z. et al. Molecular cloning and virus-induced gene silencing of MiASB in the southern root-knot nematode, Meloidogyne incognita . Eur J Plant Pathol 138, 181–193 (2014). https://doi.org/10.1007/s10658-013-0321-5

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10658-013-0321-5

Keywords

Search

Navigation