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Growth, lipid peroxidation, organic solutes, and anti-oxidative enzyme content in drought-stressed date palm embryogenic callus suspension induced by polyethylene glycol

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Abstract

Polyethylene glycol (PEG) can be used in somatic embryogenesis to enhance embryogenic development and improve the success of in vitro culture but PEG also causes osmotic stress in developing embryos. The effects of PEG on embryo growth and development in date palm cell suspension culture and associated antioxidant enzyme activities were evaluated. Callus maintained on MS basal media was transferred to regeneration liquid media supplemented with increasing levels (0–20%) of polyethylene glycol 6000 (PEG) to induce osmotic stress. The degree of embryogenic callus formation, its fresh weight, and the percentage of normal embryo callus shapes were increased with an increase in the level of PEG up to 10%. Total soluble protein (TSP), proline, glycine betaine (GB), total soluble phenol (TSPh), total sugars (TS), and total soluble organic acids (TOA) also increased whereas superoxide dismutase (SOD) activity decreased in response to PEG supplementation. Raising the PEG level increased malondialdehyde (MDA) concentration up to 10% PEG and thereafter decreased. Glutathione reductase (GR) and catalase (CAT) activities decreased at the highest levels of PEG. The proportion of normal embryo developmental shapes were about 50% compared with 20% abnormal shapes at optimum levels of PEG. Proliferation of somatic embryos was influenced by their developmental shapes. Cv. Samani accumulated more organic solutes compared with cv. Sewi in both control and stress inducing media. In contrast, lipid peroxidation, GR, SOD, and CAT activities were significantly higher in cv. Sewi than in cv. Samani indicating that the cv. Samani had the ability to tolerate a higher level of osmotic stress compared to cv. Sewi due to the enhanced osmotic re-balancing within its tissues.

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References

  • Acar O, Turkan I, Ozdemir F (2001) Superoxide dismutase and peroxidase activities in drought sensitive and resistant barley (Hordeum vulgare L.) varieties. Acta Physiol Plant 3:351–356

    Article  Google Scholar 

  • Ali SBGM, Kourosh V, Hassan BS, Siamak K, Charles L (2010) Enhancement of maturation and germination of somatic embryos in Persian walnut (Juglans regia L.) using osmolytes, hormones and cold treatments. Afr J Food Sci 4:735–743

    CAS  Google Scholar 

  • Al-Ka’aby HK and LH Abdul-Qadir (2011) Effect of water stress on callus induction from shoot tips of date palm (Phoenix dactylifera L.) cv. Bream cultured in vitro. Basrah J Date Palm Res 10:(2): 14 p

  • Al-Khateeb A (2008) Comparison effects of sucrose and date palm syrup on somatic embryogenesis of date palm (Phoenix dactylifera L.) Am J Biotechnol and Biochem 4(1):19–23

    Article  CAS  Google Scholar 

  • Al-Khayri JM (2013) Factors affecting somatic embryogenesis in date palm (Phoenix dactylifera L.) In: Aslam J, Srivastava PS, Sharma MP (eds) Somatic embryogenesis and genetic transformation in plants. Narosa Publishing House, New Delhi, pp 15–38

    Google Scholar 

  • Al-Khayri JM, Al-Bahrany AM (2004) Growth, water content and proline accumulation in drought-stressed callus of date palm. Biol Plant 48:105–108

    Article  CAS  Google Scholar 

  • Al-Khayri JM, Al-Bahrany AM (2012) Effect of abscisic acid and polyethylene glycol on the synchronization of somatic embryo development in date palm (Phoenix dactylifera L.) Biotechnology 11:318–325

    Article  CAS  Google Scholar 

  • Al-Mulla L, Bhat NR, Khalil M (2013) Salt tolerance of tissue-cultured date palm cultivars under controlled environment. Int J Biol Veter Agri Food Eng 7(8):476–479

    Google Scholar 

  • Alquarainy F (2007) Responses of bean and pea to vitamin C under salinity stress. Res J Ag Bio Sci 3(6):714–722

    Google Scholar 

  • Al-Zubaydi S, Jassim A, Zair H (2012) Effect of sodium chloride and proline on embryo formation and germination through in vitro micropropagation of date palm (Phoenix dactylifera L.) cv. Barhee J Agr Sci Tech 3:313–320

    Google Scholar 

  • Amirjani MR (2010) Effect of salinity stress on growth, mineral composition, proline content, antioxidant enzymes of soybean. Am J Pl Phys 5:350–360

    CAS  Google Scholar 

  • Bartels D, Sunkar R (2005) Drought and salt tolerance in plants. CRC Crit Rev Plant Sci 24:23–58

    Article  CAS  Google Scholar 

  • Beauchamp C, Fridovich I (1971) Superoxide dismutase: Improved assays and applicable to acrylamide gels. Anal Biochem 44:276–287

    Article  CAS  PubMed  Google Scholar 

  • Bekheet SA (2011) In vitro conservation of date palm germplasm. In: Jain SM, Al-Khayri JM, Johnson DV (eds) Date palm Biotechnology. Springer, Dordrecht, pp 337–360

    Chapter  Google Scholar 

  • Bergmeyer N (1970) In: Methoden der enzymatischen 1. Akademie Verlag, Berlin, pp 636–647

    Google Scholar 

  • Bohnert HJ, Jensen RG (1996) Strategies for engineering water stress tolerance in plants. Trends Biotechnol 14:89–97

    Article  CAS  Google Scholar 

  • Bor M, Özdemir F, Türkan I (2002) The effect of salt stress on lipid peroxidation and antioxidants in leaves of sugar beet (Beta vulgaris L.) and wild beet (Beta maritime L.) Plant Sci 164(1):77–84

    Article  Google Scholar 

  • Bourgeais-Chaillou P, Gurrier G (1992) Salt-responses in Lycopersicon esculentum calli and whole plants. L. Plant Physiol 140:494–501

    Article  CAS  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantitates of protein utilizing the principle of protein-duc binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Calic-Dragosavac D, Radojevic L (2010) Improvement of maturation and conversion of horse chestnut androgenic embryos. Biol Nyssana 1:49–55

    Google Scholar 

  • Costa M d SMN, Aloufa PM (2006) Organogeneses directa de Phoenix dactylifera L. Via peciolo cotiledonar. Pesquisa Agropecuária Tropical 36(3):195–198

    Google Scholar 

  • Cộte FX, Folliot M, Domergue R, Dubois C (2000) Field performance of embryogenic cell suspension-derived bananas plants (Musa AAA, cv. Granade naine). Euphutica 112:245–251

    Article  Google Scholar 

  • Dawood MG, Mervat SS, Hozayen M (2012) Physiological role of salicylic acid in improving performance, yield and some biochemical aspects of sunflower plant grown under newly reclaimed sandy soil. Aust J Basic & Appl Sci 6(4):82–89

    CAS  Google Scholar 

  • Dionisio Sese ML, Tobita S (1998) Antioxidant responses of rice seedlings to salinity stress. Plant Sci 135:1–9

    Article  CAS  Google Scholar 

  • Dubey RS, Rani M (1989a) Influence of NaCl salinity on growth and metabolic status of protein and amino acids in rice seedlings. J Agron Crop Sci 162:97–106

    Article  CAS  Google Scholar 

  • Dubey RS, Rani M (1989b) Salinity induces accumulation of free amino acids in germinating rice seeds differing in salt tolerance. J Agron Crop Sci 163:236–247

    Article  CAS  Google Scholar 

  • El Dawayati MM, Abd El Bar OH, Zaid ZE, Zein El Din AF (2012) In vitro morpho-histological studies of newly developed embryos from abnormal malformed embryos of date palm cv. Gundila under desiccation effect of polyethelyne glycol treatments. Ann Agric Sci 57(2):117–128

    Google Scholar 

  • El-Beltagi HS, Mohamed HI, Abdel Haleem MA, Zaki LM, Mogazy AM (2013) Physiological and biochemical effects of γ-irradiation on cowpea plants (Vigna sinensis) under salt stress. Not Bot Horti Agrobo 41(1):104–114

    CAS  Google Scholar 

  • El-Hadrami I, El Bellaj M, El Idrissi A, Aitil FJ, ElJaafari S, Saayf F (1998) Biotechnologies vegetales et amelioration du palmier dattier (Phoenix dactylifera L.) pivot de l’agriculture oasienne marocatne. Cahiers Agri 7:463–468

    Google Scholar 

  • El-Sharabasy SF, Wanas WH, Al-Kerdany AY (2008) Date palm cultivars in vitro screening to drought tolerance using isozymes. Arab J Biotech 11:263–272

    Google Scholar 

  • Farouk S (2011) Osmotic adjustment to wheat flag leaf in relation to flag leaf area and grain yield per plant. J Stress Phys & Biochem 7(2):117–138

    Google Scholar 

  • Filippou P, Bouchagier P, Skotti E, Fotopoulos V (2014) Proline and reactive oxygen/nitrogen species metabolism is involved in the tolerant response of the invasive plant species Ailanthus altissima to drought and salinity. Environ Exp Bot 97:1–10

    Article  CAS  Google Scholar 

  • Foyer CH, Halliwell B (1976) Presence of glutathione and glutathione reductase in chloroplasts: a proposed role in ascorbic acid metabolism. Planta 133:21–25

    Article  CAS  PubMed  Google Scholar 

  • Greive CM, Grattan SR (1983) Rapid assay for determination of water soluble quaternary amino compounds. Plant Soil 70:303–307

    Article  Google Scholar 

  • Helaly MNM, Hanan El-Hosieny AR (2011) In vitro selection and photosynthetic characterization of date palm regenerated by water stress. Am J Plant Phys 6(3):126–143

    Article  CAS  Google Scholar 

  • Herzog V, Fahimi H (1973) Determination of the activity of peroxidase. Anal Biochem 55:554–562

    Article  CAS  PubMed  Google Scholar 

  • Huang J, Redmann RE (1995) Solute adjustment to salinity and calcium supply in wild and cultivated barley. J Plant Nutr 18:1371–1380

    Article  CAS  Google Scholar 

  • Ishii K, Hosoi Y, Maruyama E, Kanetani SI (2008) Micropropagation of an endangered species Pinus armandii var. armamiana. Ann Forest Res 51:5–10

    Google Scholar 

  • Jain M, Mathur G, Koul S, Sarin NB (2001) Ameliorating effects of proline on salt stress lipid peroxidation in cell lines of groundnut (Archis hypogea L.) Plant Cell Report 20:463–468

    Article  CAS  Google Scholar 

  • Kamrun N, Mirza H, Mahabub AM, Masayuki F (2015) Exogenous glutathione confers high temperature stress tolerance in mung bean (Vigna radiata L.) by modulating antioxidant defense and methylglyoxal detoxification system. Environ Exp Bot 112:44–54

    Article  Google Scholar 

  • Kholova J, Sairam RK, Meana RC, Srivastava GC (2009) Response of maize genotypes to salinity stress in relation to osmolytes and metal ions contents, oxidative stress and antioxidant enzymes activity. Biol Plant 53(2):249–256

    Article  CAS  Google Scholar 

  • Lecouteux CG, Lzi FM, Bryan D, Mc Kresie BD (1993) Maturation of alfalfa (Medicago sativa L.) somatic embryos by abscisic acids, sucrose and chilling stress. Plant Sci 94:207–213

    Article  CAS  Google Scholar 

  • Ma QQ, Zou Q, Li YH, Li DQ, Wang W (2004) Amelioration of the water status and improvement of the anti-oxidant enzyme activities by exogenous glycine betaine in water-stressed wheat seedlings. Acta Agron Sinica 4:321–328

    Google Scholar 

  • Madhava Rao KV, Stresty TVS (2000) Antioxidative parameters in the seedlings of pigeonpea (Cajanus cajan L. Millspaugh) in response to Zn and Ni stresses. Plant Sci 157:113–128

    Article  CAS  PubMed  Google Scholar 

  • Magne C, Larher F (1992) High sugar content of extracts interferes with colorimetric determination of amino acids and free proline. Anal Biochem 200:358–362

    Article  Google Scholar 

  • Mirzaee M, Moieni A, Ghanati F (2013) Effects of drought stress on the lipid peroxidation and antioxidant enzyme activities in two canola (Brassica napus L.) cultivars. J Agr Sci Tech 15:593–602

    CAS  Google Scholar 

  • Mishra M, Shree Y, Pati R, Seal S, Shukla N (2010) Micropropagation of Mangifera indica L. cv. Kurakkan through somatic embryogenesis. Ind J Genetics Plant Breed 70:85–90

    Google Scholar 

  • Misra N, Gupta AK (2006) Interactive effects of sodium and calcium on proline metabolism in salt tolerant green gram cultivar. Amer J Plant Physiol 1(1):1–12

    Article  CAS  Google Scholar 

  • Misra N, Saxena P (2009) Effect of salicylic acid on proline metabolism in lentil grown under salinity stress. Plant Sci 177:181–189

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplast. Plant Cell Physiol 22:867–880

    CAS  Google Scholar 

  • Pottino BG (1981) Methods in plant tissue culture. Dept. of Hort. Agric. Collage, Maryland Univ., College, Park, Maryland, USA, pp 8-29

  • Rhodes D, Hanson AD (1993) Quaternary ammonium and tertiary sulfonium compounds in higher plants. Annu Rev Plant Physiol Plant Mol Biol 44:357–384

    Article  CAS  Google Scholar 

  • Sadasivam S, Manickam A (1996) Biochemical method, 2nd edn. New age International (P) Limited, New Delhi, pp 108–110

    Google Scholar 

  • Sakamoto A, Murata N (2002) The role of glycine betaine in the protection of plants from stress: clues from transgenic plants. Plant Cell Environment 25:163–1781

    Article  CAS  Google Scholar 

  • Sané D, Ould KM, Diouf D, Diouf D, Badiane FA, Sagna M, Borgel A (2005) Growth and development of date palm (Phoenix dactylifera L.) seedlings under drought and salinity stresses. Afr J Biotech 4(9):968–972

    Google Scholar 

  • Seckin B, Sekmen AH, Türkan I (2009) An enhancing effect of exogenous mannitol on the antioxidant enzyme activities in roots of wheat under salt stress. J Plant Growth Regul 28:12–20

    Article  CAS  Google Scholar 

  • Shalata A, Neumann PM (2001) Exogenous ascorbic acid (vitamin C) increases resistance to salt stress and reduces lipid peroxidation. J Exp Bot 52(364):2207–2211

    Article  CAS  PubMed  Google Scholar 

  • Singleton VL, Rossi JA (1965) Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. Am J Enol Vitic 16:144–158

    CAS  Google Scholar 

  • Soheilikhah Z, Karimi N, Ghasmpour HR, Zebarjadi AR (2013) Effects of saline and mannitol induced stress on some biochemical and physiological parameters of Carthamus tinctorius L. varieties callus cultures. AJCS 7(12):1866–1874

    CAS  Google Scholar 

  • Sperling O, Lazarovitch N, Schwartz A, Shapira O (2014) Effects of high salinity irrigation on growth, gas-exchange, and photoprotection in date palms Phoenix dactylifera L., cv. Medjool. Environ Exp Bot 99:100–109

    Article  CAS  Google Scholar 

  • Steel RGD, Torrie JH (1980) Principles and procedures of statistics: a biometrical approach, 2nd edn. McGraw Hill, New York ISBN-13: 978-0070609259

    Google Scholar 

  • Subbarao GV, Wheeler RM, Stutte GV, Levine LH (1999) How far can sodium substitute for potassium in red beet. J Plant Natr 22:1745–1761

    Article  CAS  Google Scholar 

  • Suganthi M, Arvinth S, Kumar RR (2012) Impact of osmotica and abscisic acid on direct somatic embryogenesis in tea. Int J Plant Res 2:22–27

    Article  Google Scholar 

  • Tajdoost S, Farboodnia T, Heidari R (2007) Salt pretreatment enhance salt tolerance in Zea mays L. seedling. Pakist J Biol Sci 10(12):2086–2090

    Article  CAS  Google Scholar 

  • Talbi S, Romero-Puertas MC, Hernández A, Terrón L, Ferchichi A, Sandalio LM (2015) Drought tolerance in a Saharian plant Oudneya africana: role of antioxidant defences. Environ Exp Bot 111:114–126

    Article  Google Scholar 

  • Tripathi SB, Gurumurthi K, Panigahi AK, Shaw BP (2007) Salinity induced changes in proline and betaine contents and synthesis in two aquatic macrophytes differing in salt tolerance. Biol Plant 51:110–115

    Article  CAS  Google Scholar 

  • Ünyayar S, Keles Y, Unal E (2004) Proline and ABA levels in two sunflower genotypes subjected to water stress. Bulg J Plant Physiol 30:34–47

    Google Scholar 

  • Velikova V, Yordanov I, Edreva A (2000) Oxidative stress and some antioxidant systems in acid rain-treated bean plants protective role of exogenous polyamines. Plant Sci 151:59–66

    Article  CAS  Google Scholar 

  • Verslues PE, Sharma S (2010) Proline metabolism and its implications for plant environment interaction. Arabidopsis Book, BioOne 8:1–23

    Google Scholar 

  • Viji M, Maheswari P, Karuppanapandian T, Manoharan K (2012) Effect of polyethylene glycol and mannitol on somatic embryogenesis of pigeonpea, Cajanus cajan (L.) Millsp. Afr J Biotechnol 11:10340–11034

    Article  Google Scholar 

  • Wang H, Xiao X, Meng-ying Yang M, Gao Z, Zang J, Fu X, Chen Y (2014) Effects of salt stress on antioxidant defense system in the root of Kandelia candel. Bot Stud 55:57

    Article  Google Scholar 

  • Xia W, Mason AS, Xiao Y, Liu Z, Yang Y (2014) Analysis of multiple transcriptomes of the African oil palm (Elaeis guineensis) to identify reference genes for RT-PCR. J Biotechnol 184:63–73

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

The authors would like to thank Prof. Dr. A. E. Hegazy at the Genetic Engineering Institute and Biotechnology Menofia, University Egypt, for his valuable help.

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Authors and Affiliations

  1. Agricultural Botany Department, Faculty of Agriculture, Mansoura University, Mansoura, Egypt

    Mohamed N. Helaly

  2. Horticultural Research Institute, Agricultural Research Centre, Giza, Egypt

    Hanan A.R. El-Hosieny

  3. Agricultural Botany Department, Faculty of Agriculture, Zagazig University, Zagazig, Egypt

    Naser M. El-Sarkassy

  4. School of Biological and Marine Sciences, Faculty of Science and Engineering, Plymouth University, Plymouth, UK

    Michael P. Fuller

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  1. Mohamed N. Helaly
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Correspondence to Michael P. Fuller.

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Helaly, M.N., El-Hosieny, H.A., El-Sarkassy, N.M. et al. Growth, lipid peroxidation, organic solutes, and anti-oxidative enzyme content in drought-stressed date palm embryogenic callus suspension induced by polyethylene glycol. In Vitro Cell.Dev.Biol.-Plant 53, 133–141 (2017). https://doi.org/10.1007/s11627-017-9815-8

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