, Volume 250, Issue 5, pp 1505–1520 | Cite as

Concurrent overexpression of rice G-protein β and γ subunits provide enhanced tolerance to sheath blight disease and abiotic stress in rice

  • Durga Madhab Swain
  • Ranjan Kumar Sahoo
  • Ravindra Kumar Chandan
  • Srayan Ghosh
  • Rahul Kumar
  • Gopaljee JhaEmail author
  • Narendra TutejaEmail author
Original Article


Main conclusion

Our study demonstrates that simultaneous overexpression of RGB1 and RGG1 genes provides multiple stress tolerance in rice by inducing stress responsive genes and better management of ROS scavenging/photosynthetic machineries.


The heterotrimeric G-proteins act as signalling molecules and modulate various cellular responses including stress tolerance in eukaryotes. The gamma (γ) subunit of rice G-protein (RGG1) was earlier reported to promote salinity stress tolerance in rice. In the present study, we report that a rice gene-encoding beta (β) subunit of G-protein (RGB1) gets upregulated during both biotic (upon a necrotrophic fungal pathogen, Rhizoctonia solani infection) and drought stresses. Marker-free transgenic IR64 rice lines that simultaneously overexpress both RGB1 and RGG1 genes under CaMV35S promoter were raised. The overexpressing (OE) lines showed enhanced tolerance to R. solani infection and salinity/drought stresses. Several defense marker genes including OsMPK3 were significantly upregulated in the R. solani-infected OE lines. We also found the antioxidant machineries to be upregulated during salinity as well as drought stress in the OE lines. Overall, the present study provides evidence that concurrent overexpression of G-protein subunits (RGG1 and RGB1) impart multiple (both biotic and abiotic) stress tolerance in rice which could be due to the enhanced expression of stress-marker genes and better management of reactive oxygen species (ROS)-scavenging/photosynthetic machinery. The current study suggests an improved approach for simultaneous improvement of biotic and abiotic stress tolerance in rice which remains a major challenge for its sustainable cultivation.


Antioxidants Biotic stress Defense marker genes Drought stress G-protein MAP kinase Overexpression ROS R. solani Salinity stress 



Ascorbate peroxidase




Empty vector control


Glutathiol reductase


Rice G-protein beta subunit1


Rice G-protein gamma subunit 1


Reactive oxygen species





DMS acknowledges post-doctoral fellowship from Department of Biotechnology (DBT), Govt. of India. Work on abiotic stress tolerance in NT laboratory is supported by the DBT, Government of India. SG acknowledges SPM fellowship from CSIR, India. RK acknowledges SRA fellowship from CSIR, India. Work in GJ lab was supported by core research grant from National Institute of Plant Genome Research, India and research funding from DBT, Govt. of India.

Compliance with ethical standards

Conflict of interest

The authors declare no competing financial interests. The seeds of OE lines can be freely obtained from the authors by following the regulatory guidelines associated with transgenic/genetically modified crops.

Supplementary material

425_2019_3241_MOESM1_ESM.doc (5.8 mb)
Supplementary material 1 (DOC 5904 kb)


  1. Acquaah G (2012) Principles of plant genetics and breeding, 2nd edn. Wiley, HobokenCrossRefGoogle Scholar
  2. Arnon DI (1949) Copper enzymes in isolated chloroplasts. polyphenoloxidase in Beta Vulgaris. Plant Physiol 24:1–15. CrossRefPubMedPubMedCentralGoogle Scholar
  3. Ayliffe M, Periyannan SK, Feechan A et al (2013) A simple method for comparing fungal biomass in infected plant tissues. Mol Plant-Microbe Interact 26:658–667. CrossRefPubMedGoogle Scholar
  4. Bai L, Liu Y, Mu Y et al (2018) Heterotrimeric G-protein γ subunit CsGG3.2 positively regulates the expression of cbf genes and chilling tolerance in cucumber. Front Plant Sci 9:488. CrossRefPubMedPubMedCentralGoogle Scholar
  5. Banu SA, Huda KMK, Tuteja N (2014) Isolation and functional characterization of the promoter of a DEAD-box helicase Psp68 using Agrobacterium-mediated transient assay. Plant Signal Behav 9:28992. CrossRefGoogle Scholar
  6. Bhardwaj D, Sheikh AH, Sinha AK, Tuteja N (2011) Stress induced β subunit of heterotrimeric G-proteins from Pisum sativum interacts with mitogen activated protein kinase. Plant Signal Behav 6(2):287–292. CrossRefPubMedPubMedCentralGoogle Scholar
  7. Bohnert HJ, Jensen RG (1996) Strategies for engineering water-stress tolerance in plants. Trends Biotechnol 14:89–97CrossRefGoogle Scholar
  8. Buysse J, Merckx R (1993) An improved colorimetric method to quantify sugar content of plant tissue. J Exp Bot 44:1627–1629. CrossRefGoogle Scholar
  9. Channamallikarjuna V, Sonah H, Prasad M et al (2010) Identification of major quantitative trait loci qSBR11-1 for sheath blight resistance in rice. Mol Breed 25:155–166. CrossRefGoogle Scholar
  10. Cha-Um S, Charoenpanich A, Roytrakul S, Kirdmanee C (2009) Sugar accumulation, photosynthesis and growth of two indica rice varieties in response to salt stress. Acta Physiol Plant 31:477–486. CrossRefGoogle Scholar
  11. Chen Y, Han Y, Meng Z et al (2016) Overexpression of the wheat expansin gene TaEXPA2 improved seed production and drought tolerance in transgenic tobacco plants. PLoS One 11:e0153494. CrossRefPubMedPubMedCentralGoogle Scholar
  12. Cheng Z, Li JF, Niu Y et al (2015) Pathogen-secreted proteases activate a novel plant immune pathway. Nature 521:213–216. CrossRefPubMedPubMedCentralGoogle Scholar
  13. Colaneri AC, Tunc-Ozdemir M, Huang JP, Jones AM (2014) Growth attenuation under saline stress is mediated by the heterotrimeric G protein complex. BMC Plant Biol 14:129. CrossRefPubMedPubMedCentralGoogle Scholar
  14. Ding L, Gao C, Li Y et al (2015) The enhanced drought tolerance of rice plants under ammonium is related to aquaporin (AQP). Plant Sci 234:14–21. CrossRefPubMedGoogle Scholar
  15. Dupre DJ, Robitaille M, Rebois RV, Hebert TE (2009) The role of Gßy subunits in the organization, assembly, and function of GPCR signaling complexes. Annu Rev Pharmacol Toxicol 49:31–56. CrossRefPubMedPubMedCentralGoogle Scholar
  16. Ferrero-Serrano Á, Su Z, Assmann SM (2018) Illuminating the role of the Gα heterotrimeric G protein subunit, RGA1, in regulating photoprotection and photoavoidance in rice. Plant Cell Environ 41:451–468. CrossRefPubMedGoogle Scholar
  17. Gampala SS, Kim T-W, He J-X et al (2007) An essential role for 14-3-3 proteins in brassinosteroid signal transduction in Arabidopsis. Dev Cell 13:177–189. CrossRefPubMedPubMedCentralGoogle Scholar
  18. Ghosh S, Gupta SK, Jha G (2014) Identification and functional analysis of AG1-IA specific genes of Rhizoctonia solani. Curr Genet 60:327–341. CrossRefPubMedGoogle Scholar
  19. Ghosh S, Kanwar P, Jha G (2017) Alterations in rice chloroplast integrity, photosynthesis and metabolome associated with pathogenesis of Rhizoctonia solani. Sci Rep 7:41610. CrossRefPubMedPubMedCentralGoogle Scholar
  20. Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Biochem 48:909–930CrossRefGoogle Scholar
  21. Guo C, Ge X, Ma H (2013) The rice OsDIL gene plays a role in drought tolerance at vegetative and reproductive stages. Plant Mol Biol 82:239–253. CrossRefPubMedGoogle Scholar
  22. Helliwell EE, Wang Q, Yang Y (2013) Transgenic rice with inducible ethylene production exhibits broad-spectrum disease resistance to the fungal pathogens Magnaporthe oryzae and Rhizoctonia solani. Plant Biotechnol J 11:33–42. CrossRefPubMedGoogle Scholar
  23. Jha G, Rajeshwari R, Sonti RV (2007) Functional interplay between two Xanthomonas oryzae pv, oryzae secretion systems in modulating virulence on rice. Mol Plant Microbe Interact 20:31–40. CrossRefPubMedGoogle Scholar
  24. Jones AM (2002) G-protein-coupled signaling in Arabidopsis. Curr Opin Plant Biol 5:402–407CrossRefGoogle Scholar
  25. Joo JH, Wang S, Chen JG et al (2005) Different signaling and cell death roles of heterotrimeric G protein α and β subunits in the Arabidopsis oxidative stress response to ozone. Plant Cell 17:957–970. CrossRefPubMedPubMedCentralGoogle Scholar
  26. Kalifa Y, Perlson E, Gilad A et al (2004) Over-expression of the water and salt stress-regulated Asr1 gene confers an increased salt tolerance. Plant Cell Environ 27:1459–1469. CrossRefGoogle Scholar
  27. Karasov TL, Chae E, Herman JJ, Bergelson J (2017) Mechanisms to mitigate the trade-off between growth and defense. Plant Cell 29:666–680. CrossRefPubMedPubMedCentralGoogle Scholar
  28. Kim H, Lee K, Hwang H et al (2014) Overexpression of PYL5 in rice enhances drought tolerance, inhibits growth, and modulates gene expression. J Exp Bot 65:453–464. CrossRefPubMedPubMedCentralGoogle Scholar
  29. Klopffleisch K, Phan N, Augustin K et al (2011) Arabidopsis G-protein interactome reveals connections to cell wall carbohydrates and morphogenesis. Mol Syst Biol 7:532. CrossRefPubMedPubMedCentralGoogle Scholar
  30. Li J, Li Y, Yin Z et al (2017) OsASR5 enhances drought tolerance through a stomatal closure pathway associated with ABA and H2O2 signalling in rice. Plant Biotechnol J 15:183–196. CrossRefPubMedGoogle Scholar
  31. Liu Q, Han R, Wu K et al (2018) G-protein βγ subunits determine grain size through interaction with MADS-domain transcription factors in rice. Nat Commun 9:852. CrossRefPubMedPubMedCentralGoogle Scholar
  32. 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. CrossRefPubMedPubMedCentralGoogle Scholar
  33. Maruta N, Trusov Y, Brenya E et al (2015) Membrane-localized extra-large g proteins and Gβγ of the heterotrimeric G proteins form functional complexes engaged in plant immunity in Arabidopsis. Plant Physiol 167:1004–1016. CrossRefPubMedPubMedCentralGoogle Scholar
  34. Meng X, Zhang S (2013) MAPK cascades in plant disease resistance signaling. Annu Rev Phytopathol 51:245–266. CrossRefGoogle Scholar
  35. Miao J, Yang Z, Zhang D et al (2018) Mutation of RGG2, which encodes a type B heterotrimeric G protein γ subunit, increases grain size and yield production in rice. Plant Biotechnol J 17:650–664. CrossRefPubMedPubMedCentralGoogle Scholar
  36. Misra S, Wu Y, Venkataraman G et al (2007) Heterotrimeric G-protein complex and G-protein-coupled receptor from a legume (Pisum sativum): role in salinity and heat stress and cross-talk with phospholipase C. Plant J 51:656–669. CrossRefPubMedGoogle Scholar
  37. Munns R, Tester M (2008) Mechanisms of salinity tolerance. Annu Rev Plant Biol 59:651–681. CrossRefPubMedGoogle Scholar
  38. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497. CrossRefGoogle Scholar
  39. Pattanagul W, Thitisaksakul M (2008) Effect of salinity stress on growth and carbohydrate metabolism in three rice (Oryza sativa L.) cultivars differing in salinity tolerance. Indian J Exp Biol 46:736–742PubMedGoogle Scholar
  40. Petrov V, Hille J, Mueller-Roeber B, Gechev TS (2015) ROS-mediated abiotic stress-induced programmed cell death in plants. Front Plant Sci 6:69. CrossRefPubMedPubMedCentralGoogle Scholar
  41. Sahoo RK, Tuteja N (2012) Development of Agrobacterium-mediated transformation technology for mature seed-derived callus tissues of indica rice cultivar IR64. GM Crops Food 3:123–128. CrossRefPubMedGoogle Scholar
  42. Sahoo RK, Ansari MW, Tuteja R, Tuteja N (2014) OsSUV3 transgenic rice maintains higher endogenous levels of plant hormones that mitigates adverse effects of salinity and sustains crop productivity. Rice 7:1–3. CrossRefGoogle Scholar
  43. Shrivastava P, Kumar R (2015) Soil salinity: a serious environmental issue and plant growth promoting bacteria as one of the tools for its alleviation. Saudi J Biol Sci 22:123–131. CrossRefPubMedGoogle Scholar
  44. Singh KB, Foley RC, Oñate-Sánchez L (2002) Transcription factors in plant defense and stress responses. Curr Opin Plant Biol 5:430–436CrossRefGoogle Scholar
  45. Subramaniam G, Trusov Y, Lopez-Encina C et al (2016) Type B heterotrimeric G protein γ -subunit regulates auxin and ABA signaling in tomato. Plant Physiol 170:1117–1134. CrossRefPubMedGoogle Scholar
  46. Suharsono U, Fujisawa Y, Kawasaki T et al (2002) The heterotrimeric G protein α subunit acts upstream of the small GTPase Rac in disease resistance of rice. Proc Natl Acad Sci USA 99:13307–13312. CrossRefPubMedGoogle Scholar
  47. Sun H, Qian Q, Wu K et al (2014) Heterotrimeric G proteins regulate nitrogen-use efficiency in rice. Nat Genet 46:652–656. CrossRefPubMedGoogle Scholar
  48. Swain DM, Sahoo RK, Srivastava VK et al (2017) Function of heterotrimeric G-protein γ subunit RGG1 in providing salinity stress tolerance in rice by elevating detoxification of ROS. Planta 245:367–383. CrossRefPubMedGoogle Scholar
  49. Taheri P, Tarighi S (2010) Riboflavin induces resistance in rice against Rhizoctonia solani via jasmonate-mediated priming of phenylpropanoid pathway. J Plant Physiol 167:201–208. CrossRefPubMedGoogle Scholar
  50. Takatsuji H (2014) Development of disease-resistant rice using regulatory components of induced disease resistance. Front Plant Sci 5:630. CrossRefPubMedPubMedCentralGoogle Scholar
  51. Tang N, Zhang H, Li X et al (2012) Constitutive activation of transcription factor OsbZIP46 improves drought tolerance in rice. Plant Physiol 158:1755–1768. CrossRefPubMedPubMedCentralGoogle Scholar
  52. Torres MA, Molina A, Dangl JL et al (2013) Functional interplay between Arabidopsis NADPH oxidases and heterotrimeric G protein. Mol Plant-Microbe Interact 26:686–694. CrossRefPubMedGoogle Scholar
  53. Tripathy BC, Oelmüller R (2012) Reactive oxygen species generation and signaling in plants. Plant Signal Behav 7:1621–1633CrossRefGoogle Scholar
  54. Trusov Y, Rookes JE, Chakravorty D et al (2006) Heterotrimeric G Proteins facilitate Arabidopsis resistance to necrotrophic pathogens and are involved in jasmonate signaling. Plant Physiol 140:210–220CrossRefGoogle Scholar
  55. Trusov Y, Sewelam N, Rookes JE et al (2009) Heterotrimeric G proteins-mediated resistance to necrotrophic pathogens includes mechanisms independent of salicylic acid-, jasmonic acid/ethylene- and abscisic acid-mediated defense signaling. Plant J 58:69–81. CrossRefPubMedGoogle Scholar
  56. Trusov Y, Chakravorty D, Botella JR (2012) Diversity of heterotrimeric G-protein subunits in plants. BMC Res Notes 5:608. CrossRefPubMedPubMedCentralGoogle Scholar
  57. Tuteja N (2009) Signaling through G protein coupled receptors. Plant Signal Behav 4:942–947. CrossRefPubMedPubMedCentralGoogle Scholar
  58. Tuteja N, Sahoo RK, Garg B, Tuteja R (2013) OsSUV3 dual helicase functions in salinity stress tolerance by maintaining photosynthesis and antioxidant machinery in rice (Oryza sativa L. cv. IR64). Plant J 76:115–127. CrossRefPubMedGoogle Scholar
  59. Urano D, Chen JG, Botella JR, Jones AM (2013) Heterotrimeric G protein signalling in the plant kingdom. Open Biol 3:120186. CrossRefPubMedPubMedCentralGoogle Scholar
  60. Urano D, Colaneri A, Jones AM (2014) Gα modulates salt-induced cellular senescence and cell division in rice and maize. J Exp Bot 65:6553–6561. CrossRefPubMedPubMedCentralGoogle Scholar
  61. Utsunomiya Y, Samejima C, Takayanagi Y et al (2011) Suppression of the rice heterotrimeric G protein β-subunit gene, RGB1, causes dwarfism and browning of internodes and lamina joint regions. Plant J 67:907–916. CrossRefPubMedGoogle Scholar
  62. Wang Z, Tan X, Zhang Z et al (2012) Defense to Sclerotinia sclerotiorum in oilseed rape is associated with the sequential activations of salicylic acid signaling and jasmonic acid signaling. Plant Sci 184:75–82. CrossRefPubMedGoogle Scholar
  63. Wang H, Meng J, Peng X et al (2015) Rice WRKY4 acts as a transcriptional activator mediating defense responses toward Rhizoctonia solani, the causing agent of rice sheath blight. Plant Mol Biol 89:157–171. CrossRefPubMedGoogle Scholar
  64. Wu H, Shabala L, Liu X et al (2015) Linking salinity stress tolerance with tissue-specific Na+ sequestration in wheat roots. Front Plant Sci 6:71. CrossRefPubMedPubMedCentralGoogle Scholar
  65. Xu J, Zhang S (2015) Mitogen-activated protein kinase cascades in signaling plant growth and development. Trends Plant Sci 20:56–64. CrossRefPubMedGoogle Scholar
  66. Yadav DK, Islam SMS, Tuteja N (2012) Rice heterotrimeric G-protein gamma subunits (RGG1 and RGG2) are differentially regulated under abiotic stress. Plant Signal Behav 7:733–740. CrossRefPubMedPubMedCentralGoogle Scholar
  67. Zhang L, Hu G, Cheng Y, Huang J (2008) Heterotrimeric G protein alpha and beta subunits antagonistically modulate stomatal density in Arabidopsis thaliana. Dev Biol 324:68–75. CrossRefPubMedGoogle Scholar
  68. Zhang H, Gao Z, Zheng X, Zhang Z (2012) The role of G-proteins in plant immunity. Plant Signal Behav 7:1284–1288. CrossRefPubMedPubMedCentralGoogle Scholar
  69. Zhang J, Zhang S, Cheng M et al (2018) Effect of drought on agronomic traits of rice and wheat: a meta-analysis. Int J Environ Res Public Health 15:839. CrossRefPubMedCentralGoogle Scholar
  70. Zhu Y, Chen H, Fan J et al (2000) Genetic diversity and disease control in rice. Nature 406:718–722. CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Durga Madhab Swain
    • 1
    • 2
    • 4
  • Ranjan Kumar Sahoo
    • 2
  • Ravindra Kumar Chandan
    • 1
    • 3
  • Srayan Ghosh
    • 1
  • Rahul Kumar
    • 1
  • Gopaljee Jha
    • 1
    Email author
  • Narendra Tuteja
    • 2
    Email author
  1. 1.Plant Microbe Interactions LaboratoryNational Institute of Plant Genome ResearchNew DelhiIndia
  2. 2.International Centre for Genetic Engineering and BiotechnologyNew DelhiIndia
  3. 3.School of Life SciencesCentral University of GujratGandhinagarIndia
  4. 4.Department of BiotechnologyRavenshaw UniversityCuttackIndia

Personalised recommendations