Abstract
Plant-parasitic nematodes (PPNs) are known to be devastating pests of agricultural crops; the rice root knot nematode Meloidogyne graminicola being the most damaging in rice. Variability in response to any trait is important not only to understand plant-stress interactions, but also its utility in stress mitigation. In this direction, the focus of the study was to exploit activation tagging, a well-established functional genomics approach in rice as a plausible strategy to assess variability in the phenotypic and molecular response to M. graminicola infection. T-DNA insertional mutants were developed in rice (acc. JBT 36/14) using an apical meristem-targeted in planta transformation strategy. A panel of 30 mutants that demonstrated molecular evidences for T-DNA integration and portraying a robust phenotype were chosen for the study. Vigorous phenotyping of these mutants against M. graminicola second stage juveniles both under in vitro pluronic gel medium and soil conditions depicted variable response based on four disease scoring parameters and the derived nematode reproductive factor. Among 30 mutants that were evaluated, five showed a resistant phenotype while 25 depicted a susceptible reaction. The resistant plants clearly demonstrated a reduction in all the disease parameters both under in vitro as well as greenhouse conditions. Quantification of transcripts of 25 genes associated with host defence responses belonging to salicylic acid (SA), jasmonic acid (JA) and ethylene (ET) pathways also indicated corroboration with the phenotype exhibited by the mutants. Distinct pattern of gene expression profiles was observed not only between the susceptible and resistant mutants, but also amongst the resistant ones, demonstrating the possibility of lesser known defense pathways being operated in these plants, apparently depending upon the T-DNA integration loci. The variability in the response to nematode challenge demonstrated in this study can be a step towards deciphering of compatible/incompatible relationships that can form a knowledge base towards not only understanding plant-nematode interactions but also extrapolation of their utility in rice improvement programmes.
Similar content being viewed by others
Abbreviations
- PPNs:
-
Plant-parasitic nematodes
- RRKN:
-
Rice root knot nematode
- WT:
-
Wild type JBT 36/14
- PB 1121:
-
Pusa basmati 1121
- J2s:
-
Juveniles
- SA:
-
Salicylic acid
- JA:
-
Jasmonic acid
- ET:
-
Ethylene
References
Bridge J, Plowright RA, Peng D (2005) Nematode parasites of rice. In: Luc M, Sikora RA, Bridge J (eds) Plant parasitic nematodes in subtropical and tropical agriculture, 2nd edn. CABI Publishing, Wallingford, pp 87–130
Byrd DW, Kirkpatrick T, Barker KR (1983) An improved technique for clearing and staining plant tissues for detection of nematodes. J Nematol 15:142–143
Chang TT (1976) The origin, evolution, cultivation, dissemination, and diversification of Asian and African rice. Euphytica 25(1):425–441. https://doi.org/10.1007/BF00041576
Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissues. Focus 12:13–15
Dutta TK, Ganguly AK, Gaur HS (2012) Global status of rice root-knot nematode, Meloidogyne graminicola. Afr J Microbiol Res 6:6016–6021. https://doi.org/10.5897/AJMR12.707
Gheysen G, Mitchum MG (2011) How nematodes manipulate plant development pathways for infection. Curr Opin Plant Biol 14:415–421. https://doi.org/10.1016/j.pbi.2011.03.012
Glazebrook J (2005) Contrasting mechanisms of defense against biotrophic and necrotrophic pathogens. Annu Rev Phytopathol 43:205–227. https://doi.org/10.1146/annurev.phyto.43.040204.135923
Golden AN, Birchfield W (1965) Meloidogyne graminicola (Heteroderidae) a new species of root-knot nematode from grass. Proc Helminthol Soc Wash 32:228–231
Jain RK, Khan MR, Kumar V (2012) Rice root-knot nematode (Meloidogyne graminicola) infestation in rice. Arch Phytopathol Plant Protect 45(6):635–645. https://doi.org/10.1080/03235408.2011.588059
Jeong DH, An S, Kang HG, Moon S, Han JJ, Park S, Lee HS, An K, An G (2002) T-DNA insertional mutagenesis for activation tagging in rice. Plant Physiol 130(4):1636–1644. https://doi.org/10.1104/pp.014357
Ji H, Gheysen G, Denil S, Lindsey K, Topping JF, Nahar K, Haegeman A, De Vos WH, Trooskens G, Van Criekinge W, De Meyer T, Kyndt T (2013) Transcriptional analysis through RNA sequencing of giant cells induced by Meloidogyne graminicola in rice roots. J Exp Bot 64:3885–3898. https://doi.org/10.1093/jxb/ert219
Ji H, Gheysen G, Ullah C, Verbeek R, Shang C, De Vleesschauwer D, Hofte M, Kyndt T (2015a) The role of thionins in rice defence against root pathogens. Mol Plant Pathol 16:870–881. https://doi.org/10.1111/mpp.12246
Ji H, Kyndt T, He W, Vanholme B, Gheysen G (2015b) b-Aminobutyric acid-induced resistance against root-knot nematodes in rice is based on increased basal defense. Mol Plant-Microbe Interact 28:519–533. https://doi.org/10.1094/MPMI-09-14-0260-R
Kandoth PK, Mitchum MG (2013) War of the worms: how plants fight underground attacks. Curr Opin Plant Biol 16:457–463. https://doi.org/10.1016/j.pbi.2013.07.001
Kumari C, Dutta TK, Banakar P, Rao U (2016) Comparing the defence-related gene expression changes upon root-knot nematode attack in susceptible versus resistant cultivars of rice. Sci Rep 6:22846. https://doi.org/10.1038/srep22846
Kyndt T, Denil S, Haegeman A, Trooskens G, Bauters L, Criekinge WV, Meyer TD, Gheysen G (2012a) Transcriptional reprogramming by root knot and migratory nematode infection in rice. New Phytol 196:887–900. https://doi.org/10.1111/j.1469-8137.2012.04311
Kyndt T, Nahar K, Haegeman A, De Vleesschauwer D, Hofte M, Gheysen G (2012b) Comparing systemic defence-related gene expression changes upon migratory and sedentary nematode attack in rice. Plant Biol 14:73–82. https://doi.org/10.1111/j.1438-8677.2011.00524.x
Kyndt T, Vieira P, Gheysen G, de Almeida-Engler J (2013) Nematode feeding sites: unique organs in plant roots. Planta 238(5):807–818. https://doi.org/10.1007/s00425-013-1923-z
Kyndt T, Fernandez D, Gheysen G (2014) Plant-parasitic nematode infection in rice: molecular and cellular insights. Annu Rev Phytopathol 52:7.1–7.19. https://doi.org/10.1146/annurev-phyto-102313-050111
Lin WC, Lu CF, Wu JW, Cheng ML, Lin YM, Yang NS, Black L, Green SK, Wang JF, Cheng CP (2004) Transgenic tomato plants expressing the Arabidopsis NPR1 gene display enhanced resistance to a spectrum of fungal and bacterial diseases. Transgenic Res 13:567–581
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real time quantitative PCR and the 2 –ΔΔCT method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
Manimaran P, Reddy S, Moin M, Reddy MR, Yugandhar P, Mohanraj SS, Balachandran SM, Kirti PB (2017) Activation-tagging in indica rice identifies a novel transcription factor subunit, NF-YC13 associated with salt tolerance. Sci Rep 7:9341. https://doi.org/10.1038/s41598-017-10022-9
Mantelin S, Bellafiore S, Kyndt T (2017) Meloidogyne graminicola: a major threat to rice agriculture. Mol Plant Pathol 18(1):3–15. https://doi.org/10.1111/mpp.12394
Moin M, Bakshi A, Saha A, Udayakumar M, Reddy AR, Rao KV, Siddiq EA, Kirti PB (2016) Activation tagging in indica rice identifies ribosomal proteins as potential targets for manipulation of water-use efficiency and abiotic stress tolerance in plants. Plant Cell Environ 39:2440–2459. https://doi.org/10.1111/pce.12796
Nagaveni V, Sashidhar V, Sreevathsa R (2011) Overexpression of phytochelatin synthase (AtPCS) in rice for tolerance to cadmium stress. Biologia 66(6):1060–1073. https://doi.org/10.2478/s11756-011-0135-x
Nahar K, Kyndt T, De Vleesschauwer D, Hofte M, Gheysen G (2011) The jasmonate pathway is a key player in systemically induced defence against root-knot nematodes in rice. Plant Physiol 157:305–316. https://doi.org/10.1104/pp.111.177576
Nahar K, Kyndt T, Nzogela YB, Gheysen G (2012) Abscisic acid interacts antagonistically with classical defence pathways in rice-migratory nematode interaction. New Phytol 196:901–913. https://doi.org/10.1111/j.1469-8137.2012.04310.x
Nguyen PV, Bellafiore S, Petitot AN, Haidar R, Bak A, Abed A, Gantet P, Mezzalira I, Engler J, Fernandez D (2014) Meloidogyne incognita-rice (Oryza sativa) interaction: a new model system to study plant-root-knot nematode interactions in monocotyledons. Rice 7:23. https://doi.org/10.1186/s12284-014-0023-4
Padgham JL, Duxbury JM, Mazid AM, Abawi GS, Hossain H (2004) Yield loss caused by Meloidogyne graminicola on lowland rainfed rice in Bangladesh. J Nematol 36:42–48
Pieterse CMJ, Leon-Reyes A, Van der Ent S, Van Wees SCM (2009) Networking by small-molecule hormones in plant immunity. Nat Chem Biol 5(5):308–316. https://doi.org/10.1038/nchembio.164
Plowright RA, Coyne DL, Nash P, Jones MP (1999) Resistance to the rice nematodes Heterodera sacchari, Meloidogyne graminicola and M. incognita in Oryza glaberrima and O. glaberrima x O. sativa interspecific hybrids. Nematology 1:745–751. https://doi.org/10.1093/aob/mcw256
Pokharel RR, Duxbury JM, Abawai G (2012) Evaluation of protocol for assessing the reaction of rice and wheat germplasm to infection by Meloidogyne graminicola. J Nematol 44(3):274–283
Roberts PA, Mathews WC, Veremis JC (1998) Genetic mechanisms of host plant resistance to nematodes. In: Baker KR, Peterson GA, Windham GL (eds) Plant nematode interactions. Amer Soci Agro, Madison, pp 209–238
Rohde RA (1965) The nature of resistance in plants to nematodes. Phytopathology 55:1159–1162. https://doi.org/10.1007/978-1-4615-9305-8_16
Shivakumara TN, Sreevathsa R, Dash PK, Sheshshayee MS, Papolu PK, Rao U, Tuteja N, Udayakumar M (2017) Overexpression of Pea DNA Helicase 45 (PDH45) imparts tolerance to multiple abiotic stresses in chilli (Capsicum annuum L.). Sci Rep 7:2760. https://doi.org/10.1038/s41598-017-02589-0
Soriano IR, Schmit V, Brar DS, Prot JC, Reversat G (1999) Resistance to the rice root-knot nematode Meloidogyne graminicola identified in Oryza longistaminata and O. glaberrima. Nematology 1(4):395–398. https://doi.org/10.1163/156854199508397
Spoel SH, Johnson JS, Dong X (2007) Regulation of tradeoffs between plant defences against pathogens with different lifestyles. Proc Natl Acad Sci U S A 104:18842–18847. https://doi.org/10.1073/pnas.0708139104
Vlot AC, Dempsey DA, Klessig DF (2009) Salicylic acid, a multifaceted hormone to combat disease. Annu Rev Phytopathol 47:177–206. https://doi.org/10.1146/annurev.phyto.050908.135202
Weige lD, Ahn JH, Blazquez MA, Borevitz JO, Christensen SK, Fankhauser C, Ferrandiz C, Kardailsky I, Malancharuvil EJ, Neff MM, Nguyen JT, Sato S, Wang ZY, Xia Y, Dixon RA, Harrison MJ, Lamb CJ, Yanofsky MF, Chory J (2000) Activation tagging in Arabidopsis. Plant Physiol 122:1003–1013
Acknowledgments
The Ph.D student BH is very grateful and acknowledges the Post Graduate School, Indian Council of Agricultural Research-Indian Agricultural Research Institute, New Delhi for financial assistance of the present research work. The authors acknowledge financial assistance by DBT funded project (BT/PR-18924/COE/34/48/2017).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they do not have any conflict of interest.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Hatzade, B., Sreevathsa, R., Makarla, U. et al. T-DNA activation tagging in rice results in a variable response to Meloidogyne graminicola infection. Biologia 74, 1197–1217 (2019). https://doi.org/10.2478/s11756-019-00281-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.2478/s11756-019-00281-4