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Enhanced proline accumulation and salt stress tolerance of transgenic indica rice by over-expressing P5CSF129A gene

  • Kumar, Vinay (Department of Botany, University of Pune) ;
  • Shriram, Varsha (Department of Botany, Annasaheb Magar College) ;
  • Kishor, P.B. Kavi (Department of Genetics, Osmania University) ;
  • Jawali, Narendra (Molecular Biology Division, Bhabha Atomic Research Centre) ;
  • Shitole, M.G. (Department of Botany, University of Pune)
  • Received : 2009.08.17
  • Accepted : 2009.11.09
  • Published : 2010.03.30

Abstract

[ ${\Delta}^1$ ]pyrroline-5-carboxylate synthetase (P5CS) is a proline biosynthetic pathway enzyme and is known for conferring enhanced salt and drought stress in transgenics carrying this gene in a variety of plant species; however, the wild-type P5CS is subjected to feedback control. Therefore, in the present study, we used a mutagenized version of this osmoregulatory gene-P5CSF129A, which is not subjected to feedback control, for producing transgenic indica rice plants of cultivar Karjat-3 via Agrobacterium tumefaciens. We have used two types of explants for this purpose, namely mature embryo-derived callus and shoot apices. Various parameters for transformation were optimized including antibiotic concentration for selection, duration of cocultivation, addition of phenolic compound, and bacterial culture density. The resultant primary transgenic plants showed more enhanced proline accumulation than their non-transformed counterparts. This proline level was particularly enhanced in the transgenic plants of next generation ($T_1$) under 150 mM NaCl stress. The higher proline level shown by transgenic plants was associated with better biomass production and growth performance under salt stress and lower extent of lipid peroxidation, indicating that overproduction of proline may have a role in counteracting the negative effect of salt stress and higher maintenance of cellular integrity and basic physiological processes under stress.

Keywords

References

  1. Ali G, Srivastava PS, Iqbal M (1999) Proline accumulation, protein pattern and photosynthesis in regenerants grown under NaCl stress. Biol Plant 42:89-95 https://doi.org/10.1023/A:1002127711432
  2. Anoop N, Gupta AK (2003) Transgenic indica rice cv IR-50 overexpressing Vigna aconitifolia $\delta$(1)-pyrroline-5-carboxylate synthetase cDNA shows tolerance to high salt. J Plant Biochem Biotechnol 12:109-116 https://doi.org/10.1007/BF03263170
  3. Bates LS, Waldren RP, Teare ID (1973) Rapid determination of free proline for water studies. Plant Soil 39:205-208 https://doi.org/10.1007/BF00018060
  4. Bhatnagar-Mathur P, Vincent V, Devi MJ, Lavanya M, Vani G, Sharma KK (2009) Genetic engineering of chickpea (Cicer arietinum L.) with the P5CSF129A gene for osmoregulation with implications on drought tolerance. Mol Breeding 23:591-606 https://doi.org/10.1007/s11032-009-9258-y
  5. Ge X, Chu Z, Lin Y, Wang S (2006) A tissue culture system for different germplasms of indica rice. Plant Cell Rep 25:392-402 https://doi.org/10.1007/s00299-005-0100-7
  6. Heath RL, Packer I (1968) Photoperoxidation in isolated chloroplast I. kinetics and stochiometry of fatty acid peroxidation. Arch Biochem Biophys 125:189-198 https://doi.org/10.1016/0003-9861(68)90654-1
  7. Hmida-Sayari A, Gargouri-Bouzid R, Bidani A, Jaoua L, Savoure A, Jaoua S (2005) Overexpression of $\Delta^{1}$-pyrroline-5-carboxylate synthetase increases proline production and confers salt tolerance in transgenic potato plants. Plant Sci 169:746-752 https://doi.org/10.1016/j.plantsci.2005.05.025
  8. Hong Z, Lakkineni K, Zhang Z, Verma DPS (2000) Removal of feedback inhibition of pyrroline-5-carboxylate synthetase results in increased proline accumulation and protection of plants from osmotic stress. Plant Physiol 122:1129-1136 https://doi.org/10.1104/pp.122.4.1129
  9. Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: $\beta$-Glucuronidase as sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901-3907
  10. Jogeswar G, Pallela R, Jakka NM, Reddy PS, Rao JV, Sreeniwasulu N, Kavi Kishor PB (2006) Antioxidative response in different sorghum species under short term salinity stress. Acta Physiol Plant 28:465-475 https://doi.org/10.1007/BF02706630
  11. Kant P, Kant S, Jain RK, Chaudhury VK (2007) Agrobacterium-mediated high frequency transformation in dwarf recalcitrant rice cultivars. Biol Plant 51:61-68 https://doi.org/10.1007/s10535-007-0012-1
  12. Kavi Kishor PB, Sangam S, Amrutha RN, Laxmi PS, Naidu NR, Rao KRSS, Rao S, Reddy KJ, Theriappan P, Sreenivasulu N (2005) Regulation of proline biosynthesis, degradation, uptake and transport in higher plants: its implications in plant growth and abiotic stress tolerance. Curr Sci 88:424-438
  13. Khanna HK, Raina SK (2002) Elite indica transgenic rice plants expressing modified Cry1Ac endotoxin of Bacillus thuringiensis show enhanced resistance to yellow stem borer (Scirpophaga incertulas). Transgenic Res 11:411-423 https://doi.org/10.1023/A:1016378606189
  14. Kumar SG, Reddy AM, Sudhakar C (2003) NaCl effects on proline metabolism in two high yielding genotypes of mulberry (Morus alba L.) with contrasting salt tolerance. Plant Sci 165:1245-1251 https://doi.org/10.1016/S0168-9452(03)00332-7
  15. Kumar KK, Muruthasalam S, Loganathan M, Sudhakar D, Balasubramanian P (2005) An improved Agrobacterium-mediated transformation protocol for recalcitrant elite indica rice cultivars. Plant Mol Biol Rep 23:67-73 https://doi.org/10.1007/BF02772648
  16. Kumar V, Shriram V, Nikam TD, Kavi Kishor PB, Jawali N, Shitole MG (2008) Assessment of tissue culture and antibiotic selection parameters useful for transformation of an important indica rice genotype Karjat-3. Asian Australas J Plant Sci Biotechnol 2:84-87
  17. Kumar V, Shriram V, Nikam TD, Jawali N, Shitole MG (2009) Antioxidant enzyme activities and protein profiling under salt stress in indica rice genotypes differing in salt tolerance. Arch Agron Soil Sci 55:379-394 https://doi.org/10.1080/03650340802595543
  18. Mansour MMF (2000) Nitrogen containing compounds and adaptation of plants to salinity stress. Biol Plant 43:491-500 https://doi.org/10.1023/A:1002873531707
  19. Matysik J, Bhalu AB, Mohanty P (2002) Molecular mechanisms of quenching of reactive oxygen species by proline under stress in plants. Curr Sci 82:525-532
  20. Misra N, Gupta AK (2005) Effect of salt stress on proline metabolism in two high yielding genotypes of green gram. Plant Sci 169:331-339 https://doi.org/10.1016/j.plantsci.2005.02.013
  21. Mohanty A, Kathuria H, Ferjani A, Sakamoto A, Mohanty P, Murata N, Tyagi AK (2002) Transgenic plants of an elite indica rice variety Pusa Basmati 1 harbouring the codA gene are highly tolerant to salt stress. Theor Appl Genet 106:51-57 https://doi.org/10.1007/s00122-002-1063-5
  22. Molinari HBC, Marur CJ, Filho JCB, Kobayashi AK, Pileggi M, Junior RPL, Pereira LFP, Vieira LGE (2004) Osmotic adjustment in transgenic citrus rootstock Carrizo citrange (Citrus sinensis Osb. X Poncirus trifoliata L. Raf.) overproducing proline. Plant Sci 167:1375-1381 https://doi.org/10.1016/j.plantsci.2004.07.007
  23. Munns R, Tester M (2008) Mechanisms of salinity tolerance. Annu Rev Plant Biol 59:651-681 https://doi.org/10.1146/annurev.arplant.59.032607.092911
  24. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Physiol Plant 15:473-497 https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  25. Nandakumar R, Babu S, Kalpana K, Raguchander T, Balasubramanian P, Samiyappan R (2007) Agrobacterium-mediated transformation of indica rice with chitinase gene for enhanced sheath blight resistance. Biol Plant 51:142-148 https://doi.org/10.1007/s10535-007-0027-7
  26. Parida AK, Dagaonkar VS, Phalak MS, Aurangabadkar LP (2008) Differential responses of the enzymes involved in proline biosynthesis and degradation in drought tolerant and sensitive cotton genotypes during drought stress and recovery. Acta Physiol Plant 30:619-627 https://doi.org/10.1007/s11738-008-0157-3
  27. Parvanova D, Ivanov S, Konstantinova T, Karanov E, Atanassov A, Tsvetkov T, Alexieva V, Djilianov D (2004) Transgenic tobacco plants accumulating osmolytes show reduced oxidative damage under freezing stress. Plant Physiol Biochem 42:57-63 https://doi.org/10.1016/j.plaphy.2003.10.007
  28. Pileggi M (2002) Genetic transformation of the lettuce cultivar Grand Rapids (Lectuca sativa L.) by Agrobacterium tumefaciens to improve osmotic stress tolerance. Gen Mol Res 1:176
  29. Pujni D, Chaudhary A, Rajam MV (2007) Increased tolerance to salinity and drought in transgenic indica rice by mannitol accumulation. J Plant Biochem Biotechnol 16:1-8 https://doi.org/10.1007/BF03321921
  30. Saharan V, Yadav RC, Yadav NR, Ram K (2004) Studies on improved Agrobacterium-mediated transformation in two indica rice (O. sativa L.). Afr J Biotechnol 3:572-575
  31. Sairam RK, Tyagi A (2004) Physiology and molecular biology of salinity stress tolerance in plants. Curr Sci 86:407-421
  32. Sairam RK, Srivastava GC, Agarwal S, Meena RC (2005) Differences in response to salinity stress in tolerant and susceptible wheat genotypes. Biol Plant 49:85-91 https://doi.org/10.1007/s10535-005-5091-2
  33. Sambrook J, Russell RW (2001) Molecular cloning: A laboratory manual. Cold Spring Harbour Laboratory Press, Cold Spring Harbour
  34. Sawahel WA, Hassan AH (2002) Generation of transgenic wheat plants producing high levels of the osmoprotectant proline. Biotech Lett 24:721-725 https://doi.org/10.1023/A:1015294319114
  35. Sridevi G, Dhandapani M, Veluthambi K (2005) Agrobacteriummediated transformation of White Ponni, a non-basmati variety of indica rice (Oryza sativa L.). Curr Sci 88:128-132
  36. Srivatanakul M, Park S, Sanders J, Salas M, Smith R (2000) Multiple shoot regeneration of kenaf (Hibicus cannabinus) from a shoot apex culture system. Plant Cell Rep 19:1165-1170 https://doi.org/10.1007/s002990000256
  37. Su J, Wu R (2004) Stress-inducible synthesis of proline in transgenic rice confers faster growth under stress conditions than that with constitutive synthesis. Plant Sci 166:941-948 https://doi.org/10.1016/j.plantsci.2003.12.004
  38. Task Force on Agricultural Biotechnology (2004) Agricultural biotechnology: safe and responsible use. Executive summary of the Task Force on Agricultural Biotechnology chaired by MS Swaminathan, submitted to the Ministry of Agriculture, Government of India. Curr Sci 87:425-426
  39. Vendruscolo ECG, Schuster I, Pileggi M, Scapim CA, Molinari HBC, Marur CJ, Vieira LGE (2007) Stress-induced synthesis of proline confers tolerance to water deficit in transgenic wheat. J Plant Physiol 164:1367-1376 https://doi.org/10.1016/j.jplph.2007.05.001
  40. Verbruggen N, Hermans C (2008) Proline accumulation in plants: a review. Amino Acids 35:753-759 https://doi.org/10.1007/s00726-008-0061-6
  41. Yamchi A, Jazii FR, Mousav A, Karkhane AA, Renu (2007) Proline accumulation in transgenic tobacco as a result of expression of arabidopsis $\Delta^{1}$-pyrroline-5-carboxylate synthetase (p5cs) during osmotic stress. J Plant Biochem Biotechnol 16:9-15 https://doi.org/10.1007/BF03321922
  42. Yookongkaew N, Srivatanakul M, Narangajavana J (2007) Development of genotype-independent regeneration system for transformation of rice (Oryza sativa ssp. indica). J Plant Res 120:237-245 https://doi.org/10.1007/s10265-006-0046-z
  43. Zhao F, Zhang H (2006) Expression of Suaeda salsa glutathione S-transferase in transgenic rice resulted in a different level of abiotic stress resistance. J Agric Sci 144:547-554 https://doi.org/10.1017/S0021859606006411

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