Skip to main content
Log in

In vitro screening of rice genotypes for drought tolerance using polyethylene glycol

  • Original Paper
  • Published:
Acta Physiologiae Plantarum Aims and scope Submit manuscript

Abstract

The aim of the present investigation was to study in vitro somatic embryogenesis and to screen calli for drought tolerance using mature embryos as explants. Mature embryos of three aromatic (Pusa Basmati 1, Pant Sugandh Dhan 17, Taraori Basmati) and one non-aromatic (Narendra 359) indica rice (Oryza sativa L.) varieties were used for developing callus on Murashige and Skoog medium supplemented with 2, 4-dichlorophenoxy acetic acid (2, 4-D) (2.0 mg l−1 for Narendra 359 and 2.5 mg l−1 for Pusa Basmati 1, Taraori Basmati and Pant Sugandh Dhan 17). Screening of calli was done by sub-culturing calli for 15 days on Murashige and Skoog (MS) basal medium supplemented with different concentrations of polyethylene glycol (PEG)-6000 as chemical drought inducer. Callus volume decreased and total proline content was found to be increased significantly with increase in PEG concentration. Narendra 359 showed best response in terms of callus growth at 70 g l−1 of PEG. The highest percentage somatic embryogenesis among selected calli was observed in Pusa Basmati 1 and the lowest in Pant Sugandh Dhan 17. Excellent shooting and rooting (94%) was observed in MS + 0.1 mg l−1 naphthalene acetic acid (NAA) and MS + 2.0 mg l−1 2, 4-D. Regenerated plants were successfully acclimatized with 98% efficiency in greenhouse and grown under pot conditions up to maturity. It was observed that PEG treated somaclones accumulated more proline, chlorophyll content and developed more tiller and height than normal somaclones. Ten random amplified polymorphic DNA (RAPD) primers were used to amplify genomic DNA of somaclones of different varieties. Level of genetic polymorphism existing among these somaclones indicates that these markers can be used in breeding program for improving varieties through in vitro techniques.

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

Access this article

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

Instant access to the full article PDF.

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

Similar content being viewed by others

References

  • Adkins SW, Kunanuvatchaidach R, Godwin ID (1995) Somaclonal variation in rice: drought tolerance and other agronomic characters. Aust J Bot 43:201–209

    Article  Google Scholar 

  • Al-Bahrany AM (2002) Callus growth and proline accumulation in response to polyethylene glycol induced osmotic stress in rice, Oryza sativa L. Pak J Biol Sci 5:1294–1296

    Article  Google Scholar 

  • Al-Khayri JM, Al-Bahrany AM (2002) Callus growth and proline accumulation in response to sorbitol and sucrose-induced osmotic stress in rice (Oryza sativa L.). Biol Plant 45:609–611

    Article  CAS  Google Scholar 

  • Al-Khayri JM, Huang FH, Thompson LF, King JW (1992) Micropropagation of zoysiagrass (Zoysia japonica Steud). In: Bajaj YSP (ed) Biotechnology in agriculture and forestry, vol. 19, High-tech and micropropagation III. Springer, Berlin, pp 576–590

    Google Scholar 

  • Arnon DI (1949) Copper enzymes in isolated chloroplasts. Phenoloxidase in Beta vulgaris. Plant Physiol 24:1–15

    Article  PubMed  CAS  Google Scholar 

  • Baskaran P, Jayabalan N (2009) In vitro propagation of Psoralea corylifolia L. by somatic embryogenesis in cell suspension culture. Acta Physiol Plant 31:1119–1127

    Article  CAS  Google Scholar 

  • Bates LS, Waldren RP, Teare ID (1973) Rapid determination of free proline for water stress studies. Plant Soil 39:205–207

    Article  CAS  Google Scholar 

  • Brown PTH, Lange FD, Kranz E, Lörz H (1993) Analysis of single protoplast and regenerated plants by PCR and RAPD technology. Mol Gen Genet 237:311–317

    PubMed  CAS  Google Scholar 

  • Chuang SJ, Chen CL, Chen JJ, Chou WY, Sung JM (2009) Detection of somaclonal variation in micro-propagated Echinacea purpurea using AFLP marker. Sci Hort 120:121–126

    Article  CAS  Google Scholar 

  • Clark LJ, Cope RE, Whalley WR, Barraclough PB, Wade LJ (2002) Root penetration of strong soil in rainfed lowland rice: comparison of laboratory screens with field performance. Field Crops Res 76:189–198

    Article  Google Scholar 

  • Degenkolbe T, Do PT, Zuther E, Repsilber D, Walther D, Hincha DK, Köhl KI (2009) Expression profiling of rice cultivars differing in their tolerance to long-term drought stress. Plant Mol Biol 69:133–153

    Article  PubMed  CAS  Google Scholar 

  • Deo PC, Harding RM, Taylor M, Tyagi AP, Becker DK (2009) Somatic embryogenesis, organogenesis and plant regeneration in taro (Colocasia esculenta var. esculenta). Plant Cell Tiss Org Cult 99:61–71

    Article  Google Scholar 

  • Echenique V, Pli P, Mroginski L (1996) Plant regeneration in weeping love grass (Ergrostis curvlula) through inflorescence cultures. Plant Cell organ Cult 46:123–130

    Article  CAS  Google Scholar 

  • El-Tayeb MA, Hassanein AM (2000) Germination, seedling growth, some organic solutes and peroxidase expression of different Vicia faba lines as influenced by water stress. Acta Agron Hung 48:11–20

    Article  CAS  Google Scholar 

  • Endress R (1994) Plant cell biotechnology. Springer, Berlin, p 353

    Google Scholar 

  • Gomez KA, Gomez AA (1984) Statistical procedures for agricultural research, Second edition edn. John Wiley and Sons, New York

    Google Scholar 

  • Guo WL, Gong L, Ding ZF, Li YD, Li FX, Zhao SP, Liu B (2006) Genomic instability in phenotypically normal regenerants of medicinal plant Codonopsis lanceolata Benth. et Hook. F., as revealed by ISSR and RAPDs markers. Plant Cell Rep 25:896–906

    Article  PubMed  CAS  Google Scholar 

  • Hamilton EW, Heckathorn SA (2001) Mitochondrial adaptations to NaCl. Complex I is protected by antioxidants and small heat shock proteins, whereas complex II is protected by proline and betaine. Plant Physiol 126:1266–1274

    Article  PubMed  CAS  Google Scholar 

  • Handa AK, Bressan RA, Handa S, Hasegawa PM (1986) Proline accumulation and the adaptation of cultured plant cells to water stress. Plant Physiol 80:938–945

    Article  PubMed  CAS  Google Scholar 

  • Henry RJ (1998) Molecular and biochemical characterization of somaclonal variation. In: somaclonal variation and induced mutation in crop improvement. Kluwer Academic publishers, Dordrecht, pp 485–499

  • Heyser JW, Nabors MW (1981) Osmotic adjustment of cultures tobacco cells (Nicotiana tabacum var. Samsum) grown on sodium chloride. Plant Physiol 67:720–727

    Article  PubMed  CAS  Google Scholar 

  • Hiscox JD, Israelstam GF (1979) A method for the extraction of chlorophyll from leaf tissue without maceration. Can J Bot 57:1332–1334

    Article  CAS  Google Scholar 

  • Hossain A, Konisho M, Minami K, Nemoto K (2003) Somaclonal variation of regenerated plants in chili pepper (Capsicum annuum L.). Euphytica 130:233–239

    Article  Google Scholar 

  • Indra AP, Krishnaveni S (2009) Effect of hormones, explants and genotypes in in vitro culturing of sorghum. J Biochem Tech 1:96–103

    Google Scholar 

  • Jaccard P (1908) Nouvelles recherches sur la distribution lorale. Bull Soc Vaud Sci Nat 44:223–270

    Google Scholar 

  • Jiménez V (2000) Endogenous Hormone Levels in Wheat, Maize, Barley, Carrot, Grapevine and Citrus Tissues and relationship to their in vitro somatic embryogenesis. Verlag Grauder, Stuttgart, p 173

    Google Scholar 

  • Khan T, Singh AK, Pant RC (2006) Regeneration via somatic embryogenesis and organogenesis in different cultivars of cotton (gossypium spp). In Vitro Cell Dev Biol Plant 42:498–501

    Article  CAS  Google Scholar 

  • Lafitte HR, Li ZK, Vijayakumar CHM, Gao YM, Shi Y, Xu JL, Fu BY, Yu SB, Ali AJ, Domingo J, Maghirang R, Torres R, Mackill D (2006) Improvement of rice drought tolerance through backcross breeding: evaluation of donors and selection in drought nurseries. Field Crops Res 97:77–86

    Article  Google Scholar 

  • Lee KW, Choi GJ, Kim KY, Ji HC, Park HS, Yoon SH, Lee SH (2009) High frequency plant regeneration from mature seed derived callus of Italian ryegrass (Lolium multiflorum) cultivars. Afr J Biotech 8:6828–6833

    CAS  Google Scholar 

  • Liu LF, Lai KL (1991) Enhancement of regeneration in rice tissue cultures by water and salt stress. Biotech In Agric Forestry 14:47–57

    Google Scholar 

  • Mahmood T, Nazar N, Abbasi BH, Khan MA, Ahmad M, Zafar M (2010) Detection of somaclonal variations using RAPD fingerprinting in Silybum marianum (L.). J Med Plants Res 4:1822–1824

    CAS  Google Scholar 

  • Mallón R, Rodríguez-Oubiña J, González ML (2010) In vitro propagation of the endangered plant Centaurea ultreiae: assessment of genetic stability by cytological studies, flow cytometry and RAPD analysis. Plant Cell Tiss Organ Cult 101:31–39

    Article  Google Scholar 

  • Matsue Y, Uchimura Y, Sato H (2000) Comparison of productivity and growth habit between stored and newly harvested rice seeds and identification of the cultivars of stored rice seeds by RAPD method. Japan J Crop Sci 69:38–42

    Article  CAS  Google Scholar 

  • Matsumoto T, Wu JZ, Kanamori H et al (2005) The map-based sequence of the rice genome. Nature 436:793–800

    Article  Google Scholar 

  • Meneses A, Flores D, Muñoz M, Arrieta G, Espinoza AM (2005) Effect of 2, 4-D, hydric stress and light on indica rice (Oryza sativa) somatic embryogenesis. Rev Biol Trop 53:361–368

    PubMed  Google Scholar 

  • Mukherjee PK (1999) Rapid extraction of plant genomic DNA suitable for restriction analysis and PCR amplification. Asian Pacific J Mol Biol Biotechnol 7:95–96

    Google Scholar 

  • Munthali MT, Newbury HJ, Ford-Lloyd BV (1996) The detection of somaclonal variants of beet using RAPD. Plant Cell Rep 15:474–478

    Article  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Naghia PT, Malik JPS, Pandey MP, Singh NK (2002) Application of RAPD markers for genetic distance analysis of hybrid rice lines. Indian J Gen Plant Breed 62:1–4

    CAS  Google Scholar 

  • Nepomuceno AL, Oostrerhuis DM, Stewart JM (1998) Physiological responses of cotton leaves and roots to water deficit induced by polyethylene glycol. Environ Exp Bot 40:29–41

    Article  Google Scholar 

  • Oropeza M, Guevara P, García E, Ramírez JL (1995) Identification of sugarcane (Saccharum spp.) somaclonal variants resistant to sugarcane mosaic virus via RAPD markers. Plant Mol Biol Rep 13:182–191

    Article  CAS  Google Scholar 

  • Panjaitan SB, Abdullah SNA, Aziz MA, Meon S, Omar O (2009) Somatic embryogenesis from scutellar embryo of Oryza sativa L. var. MR219. Pertanika J Trop Agric Sci 32:185–194

    Google Scholar 

  • Parida AK, Das AB (2005) Salt tolerance and salinity effect on plants: a review. Ecotoxicol Environ Safety 60:324–349

    Article  PubMed  CAS  Google Scholar 

  • Raman R, Chahal GS, Dhaliwal HS (1994) Screening of genotype for callus induction and plant regeneration in rice. Crop Improv 21:1–2

    Google Scholar 

  • Rani TU, Reddy GM (1996) Factors affecting somatic embryogenesis and plant regeneration in five indica rice. Proc Indian Natn Sci Acad 62:43–50

    CAS  Google Scholar 

  • Rao KK (2003) Molecular tagging of a new bacterial blight resistance gene in rice using RAPD and SSR markers. Int Rice Res Newslet 28:16–17

    Google Scholar 

  • Reddy PC, Vajranabhaiah SN, Prakash AH (1994) Varietal responses of upland rice calli to polyethylene glycol (PEG-6000) stress. Adv Plant Sci 7:12–17

    Google Scholar 

  • Rensink WA, Buell CR (2005) Microarray expression profiling resources for plant genomics. Trends Plant Sci 10:603–609

    Article  PubMed  CAS  Google Scholar 

  • Rohlf FJ (1993) NT-SYS-pc: Numerical taxonomy and multivariate analysis system. Version 2.11W. Exteer Software, Setauket, NY

  • Roychoudhury A, Basu S, Sarkar SN, Sengupta DN (2008) Comparative physiological and molecular responses of a common aromatic indica rice cultivar to high salinity with non-aromatic indica rice cultivars. Plant Cell Rep 27:1395–1410

    Article  PubMed  CAS  Google Scholar 

  • Rueb S, Leneman M, Schilperoort RA, Hensgens LAM (1994) Efficient plant regeneration through somatic embryogenesis from callus induced on mature rice embryos (Oryza sativa L.). Plant Cell Tis Org Cult 36:259–264

    Article  Google Scholar 

  • Shankhadhar D, Shankhadhar SC, Mani SC (2000) In vitro selection for salt tolerance in rice. Biol Plant 3:477–480

    Article  Google Scholar 

  • Sharma VK, Hänsch R, Mendel RR, Schulze J (2005) Mature embryo axis-based high frequency somatic embryogenesis and plant regeneration from multiple cultivars of barley (Hordeum vulgare L.). J Exp Bot 56:1913–1922

    Article  PubMed  CAS  Google Scholar 

  • Singh N, Shukla A, Pant RC (2010) Rapid regeneration and efficient acclimatization of aromatic indica rice varieties. Agric Seg 1: AGS/1523

  • Taylor PWJ, Geijskes JR, Ko HL, Fraser TA, Henry RJ, Bircg RG (1995) Sensitivity of random amplified polymorphic DNA analysis to detect genetic change in sugarcane during tissue culture. Theor Appl Genet 90:1169–1173

    Article  CAS  Google Scholar 

  • Valladares S, Sánchez C, Martínez MT, Ballester A, Vieitez AM (2006) Plant regeneration through somatic embryogenesis from tissues of mature oak trees: true-to-type conformity of plantlets by RAPD analysis. Plant Cell Rep 25:879–886

    Article  PubMed  CAS  Google Scholar 

  • Verma P, Srivastava P, Singh SC, Mathur A (2010) Genetic fidelity of long-term micropropagated plantlets of Valeriana wallichii—an endangered medicinal plant. Cur Sci 99:436–438

    CAS  Google Scholar 

  • Xu K, Xu X, Fukao T, Canlas P, Maghirang-Rodriguez R, Heuer S, Ismail AM, Bailey-Serres J, Ronald PC, Mackill DJ (2006) Sub1A is an ethylene-response factor-like gene that confers submergence tolerance to rice. Nature 442:705–708

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Rohit Joshi.

Additional information

Communicated by R. Aroca.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Joshi, R., Shukla, A. & Sairam, R.K. In vitro screening of rice genotypes for drought tolerance using polyethylene glycol. Acta Physiol Plant 33, 2209–2217 (2011). https://doi.org/10.1007/s11738-011-0760-6

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11738-011-0760-6

Keywords

Navigation