Advertisement

Physiology and Molecular Biology of Plants

, Volume 19, Issue 2, pp 261–268 | Cite as

In vitro selection and characterization of polyethylene glycol (PEG) tolerant callus lines and regeneration of plantlets from the selected callus lines in sugarcane (Saccharum officinarum L.)

  • Srinath Rao
  • Jabeen FTZ
Research Article

Abstract

A system for in vitro selection of drought tolerant callus lines in sugarcane was developed. High molecular weight PEG was used as selective agent. Selected callus line grew better than non-selected callus when grown on different concentrations of PEG. The activity of antioxidant enzymes like CAT, POX, APX and SOD were high in selected callus than in non-selected callus. Osmolytes like proline and ascorbic acid were at higher levels in selected callus than in non-selected callus, however at higher concentrations (20–30 %) of PEG, levels of proline and ascorbic acid decreased. The frequency of organogenesis and number of plantlets decreased in selected callus than in non-selected callus. The results can be used for in vitro screening and manipulations of sugarcane for improvement of drought tolerance

Keywords

Antioxidant enzymes Callus PEG Selection Proline Sugarcane 

Abbreviations

2,4-D

2,4-dichloro phenoxy acitic acid

BAP

6-Benzyl aminopurin

NAA

α-Napthalene acitic acid

Kn

Kinetin

CAT

Catalase

POX

Peroxidase

APX

Ascorbate peroxidase

SOD

Superoxide dismutase

MS

Murashige and Skoog

ROS

Reactive Oxygen Species

Notes

Acknowledgements

The authors thank head department of Botany Gulbarga University Gulbarga for the facilities provided during the course of the investigation,

References

  1. Aazami MA, Torabi M, Jalili E (2010) In vitro response of promising tomato genotypes for tolerance to osmotic stress. Afr J Biotechnol 9(26):4014–4017Google Scholar
  2. Abdel-Raheem AT, Ragab AR, Kasem ZA, Omar FD, Samera AM (2007) In vitro selection for tomato plants for drought tolerance via callus culture under polyethylene glycol (PEG) and mannitol treatments. Afr Crop Sci Conf Proc 8:2027–2032Google Scholar
  3. Aqeel-Ahmad MS, Javed F, Ashraf M (2007) Iso-osmotic effect of NaCl and PEG on growth, cations and free proline accumulation in callus tissue of two indica rice (Oryza sativa L.) genotypes. Plant Growth Regul 53:53–63CrossRefGoogle Scholar
  4. Bates LS, Waldren RP, Teare ID (1973) Rapid determination of free proline for water stress studies. Plant Soil 39:205–207CrossRefGoogle Scholar
  5. Begum MK, Islam MO, Miah MAS, Hossain MA, Islam N (2011) Production of Somaclone In vitro for Drought Stress Tolerant Plantlet Selection in Sugarcane (Saccharum officinarum L.). Agriculturists 9(1&2):18–28Google Scholar
  6. Beyer WF, Fridovich I (1987) Assaying for superoxide dismutase activity: some large consequences of minor changes in condition. Annal Biochem 161:559–566CrossRefGoogle Scholar
  7. Bhaskaran S, Smith RH, Newton RJ (1985) Physiological changes in cultured Sorgham cells in response to induced water stress 1. Free proline. Plant Physiol 79:239–248CrossRefGoogle Scholar
  8. Bradford MM (1976) A rapid and sensitive method for the quantitation of micro gram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  9. Bueno P, Piqueras A, Kurepa J, Savoure A, Veerbruggen N, Montegu MV, Inze D (1998) Expression of antioxidant enzymes in response to abscisic acid and high osmoticum in BY-2 cell cultures. Plant Sci 138(1):27–34CrossRefGoogle Scholar
  10. Chandlee JM, Scandalios JG (1984) Analysis of variants affecting the catalase development program in maiz scutellum Theor. Appl Genetics 69:71–77Google Scholar
  11. Davies KJA (1987) Protein damage and degradation by oxygen radicals. J Biol Chem 262:9895–9901PubMedGoogle Scholar
  12. El-Haris MK, Barakat MN (1998) Evaluation of the in vitro selected drought tolerant wheat lines under drought stress conditions. Alex J Agric Res 43:293–302Google Scholar
  13. Errabii T, Gandonou CB, Essalmani H, Abrini J, Idaomar M, Senhaji NS (2006) Effects of NaCl and mannitol induced stress on sugarcane (Saccharum sp.) callus cultures. Acta Physiol Plant 29:95–102CrossRefGoogle Scholar
  14. Errabii T, Gandonou CB, Essalmani H, Abrini J, Idaomar M, Senhaji NS (2008) Growth, proline and ion accumulation in sugarcane callus cultures under drought-induced osmotic stress and its subsequent relief. Afri J Biotechnol 5:1488–1493Google Scholar
  15. Fridovich I (1986) Biological effects of superoxide radical. Arch Biochem Biophys 247:1–11PubMedCrossRefGoogle Scholar
  16. Gangopadhyay G, Basu S, Mukherjee BB, Gupta S (1997) Effect of salt and osmotic shocks on unadapted and adapted callus lines of tobacco. Plant Cell Tiss Org Cult 49:45–52CrossRefGoogle Scholar
  17. Handa S, Handa AK, Hasegawa PM, Bressan RA (1986) Proline accumulation and the adaptation of cultured plant cells to water stress. Plant Physiol 80:938–945PubMedCrossRefGoogle Scholar
  18. Hassan NS, Shaaban LD, El-Sayed AH, Seleem EE (2004) In Vitro Selection for Water Stress Tolerant Callus Line of Helianthus annus L Cv Myak. Int J Agric Biol http://www.ijab.org
  19. Heath RL, Packer (1968) Photooxidation in isolated chloroplasts. Arch Biochem Biophys 125:189–198PubMedCrossRefGoogle Scholar
  20. Jabeen FTZ (2007) Selection of drought tolerant cell lines in sugsrcane (Saccharum offcinarum Linn.) through cell and tissue culture technique. PhD. Thesis submitted to Gulbarga University Gulbarga Karnataka IndiaGoogle Scholar
  21. Jadhav YR, Garkar RM, Najan BR, Khade KK, Choudhary AN (1998) Nira (Co-86032) A new variety of sugarcane of Maharastra. J Maharastra agricuniv 23(1):10–12Google Scholar
  22. Kumar KB, Khan PA (1982) Peroxidase and Polyphenol oxidase in excised ragi (Elaucine coracona cv. 2020 leaves during senescence Indian. Expt Bot 20:412–416Google Scholar
  23. Kumar RR, Karjol K, Naik GR (2011) Effect of polyethylene glycol induced water stress on physiological and biochemical responses in Pigeon pea (Cajanus cajan L. Millsp.). RRST-Plant Physiol 3:148–152Google Scholar
  24. Larson RA (1988) The antioxidants of higher plants. Phytochemistry 27:969–978CrossRefGoogle Scholar
  25. Li L, van Stadan J (1998) Effects of plant growth regulators on the antioxidant systems in callus of two maiz cultivars subjected to water stress. Plant Growth Regul 24:55–66CrossRefGoogle Scholar
  26. Ming SY, ZhiDong Z, YaDong L, Wu L, Liu HG (2009) Effect of PEG stress on resistance of low bush blueberry callus. J Jilan Agric Univ 31(5):538–542Google Scholar
  27. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiol Plant 15:473–497CrossRefGoogle Scholar
  28. Musa Y (2011) The use of Polyethylene glycol (PEG) as selection agent of callus and plantlets of some sugarcane varieties for drought tolerance. J Agrivigor 10(2):130–140Google Scholar
  29. Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbatespecific peroxidase in spinach chloroplast. Plant Cell Physiol 22:867–880Google Scholar
  30. Nath AK, Kumari S, Sharma DR (2005) In Vitro selection and chaterization of water stress tolerant cultures of bell pepper. Indian J Plant Physiol 10(1):14–19Google Scholar
  31. Patade VY, Bhargava S, Suprasanna P (2011) Salt and drought tolerance of sugarcane under iso-osmotic salt and water stress: growth osmolyte accumulation and antioxidant defense. J plant interact 6(4):275–282Google Scholar
  32. Patade VY, Bhargava S, Suprasanna P (2012) Effects of NaCl and iso-osmotic PEG stress on growth, Osmolytes accumulation and antioxidant defense in cultured sugarcane cells. Plant Cell Tissue Organ Cult 108:279–286CrossRefGoogle Scholar
  33. Prabha C, Bharti S (1980) Effect of ascorbic acid on proline accumulation in cowpea leaves under water stress conditions. Indian J Plant Physiol 23:317–318Google Scholar
  34. Sabbah S, Tal M (1990) Development of callus and suspension cultures of potato resistant to NaCl and mannitol and their response to stress. Plant Cell Tissue Organ Cult 21:119–124CrossRefGoogle Scholar
  35. Sadasivam S, Manickam A (1992) Biochemical methods for agricultural sciences, vol 80. Wiley Eastern Limited, New Delhi, pp 179–180Google Scholar
  36. Santos-Diaz N, Ochoa-Alejo N (1994) Effect of water stress on growth, osmotic potential and solute accumulation in cell cultures from chilli pepper (a mesophyte) and Creosote bush (a xerophytes). Plant Sci 96(1–2):21–29CrossRefGoogle Scholar
  37. Shah AH, Shah SH, Ahmad H, Swati ZA (2012) Adaptation to polyethylene stress maintains totipotency of cell lines of Oryza sativa L. CV Swat-1 for a longer period. Pak J Bot 44(1):313–316Google Scholar
  38. Siddeswar G, Kavi Kishor PB (1989) Plant regeneration from polyethylene glycol adapted callus of rice. Curr Sci 58:926–928Google Scholar
  39. Singh AK, Sharma DR (2008) In vitro screening and regeneration of water stress tolerant culture of tomato. Indian J Plant Physiol 13(1):33–38Google Scholar
  40. Srivastava DK, Gupta VK, Sharma DR (1995) In vitro selection and characterization of water stress tolerant callus cultures of tomato (Lycopersicon esculentum L. Mill cv. Ktl.). Indian J Plant Physiol 38:99–104Google Scholar
  41. Yaser F, Uzal O, Ozpay T (2010) Changes of lipidperoxidation andchlorophyll amount of greenbean genotypes underdrought stress. Afr J Agric Res 5(19):2705–2709Google Scholar

Copyright information

© Prof. H.S. Srivastava Foundation for Science and Society 2013

Authors and Affiliations

  1. 1.Department of BotanyGulbarga UniversityGulbargaIndia

Personalised recommendations