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Multivariate screening approach indicated adaptive tolerance to salt stress in the seedlings of an agroforestry tree, Eucalyptus tereticornis Sm.

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Abstract

Eucalyptus tereticornis is one of the fast-growing agroforestry trees providing raw material for paper and plywood industries. Its plantations are hampered by rising soil salinity and identification of salt-tolerant (ST) seedlings could be beneficial to undertake plantation in such soils so to meet the growing demand of the industries. Three different in vitro screening approaches (direct, indirect, and gradual exposure to salt stress) were employed in the present study for the identification of ST seedlings. The screening via gradual exposure to salt stress (0–400 mM NaCl) was highly efficient and promoted tolerance level up to 400 mM NaCl. Shoot growth of seedlings demonstrated a 91.93% increase due to physiological adaptation to salt stress. The indirect screening approach was also effective but only a 12.8% rise in shoot length was recorded when cultured on a medium fortified with 400 mM NaCl. On the other hand, the direct screening approach could only identify seedlings with a tolerance level of 200 mM NaCl. Further, when salt-sensitive (SS) and ST seedlings were exposed to salt stress (0, 200, and 400 mM NaCl) for 14 days, a significant increase in chlorophyll, osmolyte accumulation, and antioxidant enzyme activity was only observed in ST seedlings. The tolerance of ST seedlings was also associated with significantly higher transcript levels of genes encoding superoxide dismutase, peroxidase, and catalase. Overall, these findings will be useful for faster screening of tree species and help in understanding the physio-biochemical basis of salt tolerance in the seedlings of E. tereticornis.

Key message

In the present study, the potential of in vitro screening for the identification of ST seedlings in Eucalyptus tereticornis was investigated. Gradual increase in salt concentration during the exposure period of seedlings resulted in acquiring tolerance up to 400 mM NaCl, which could be due to osmolyte accumulation and overexpression of genes encoding antioxidant enzymes.

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References

  • Abdelaziz M, Xuan T, Mekawy A, Wang H, Khanh T (2018) Relationship of salinity tolerance to Na+ exclusion, Proline accumulation, and antioxidant enzyme activity in rice seedlings. Agriculture 8:166

    CAS  Google Scholar 

  • Acosta-Motos JR, Ortuño MF, Bernal-Vicente A, Diaz-Vivancos P, Sanchez-Blanco MJ, Hernandez JA (2017) Plant responses to salt stress: adaptive mechanisms. Agronomy 7:18

    Google Scholar 

  • Adams MA, Richter A, Hill AK, Colmer TD (2005) Salt tolerance in Eucalyptus spp.: identity and response of putative osmolytes. Plant Cell Environ 28:772–787

    CAS  Google Scholar 

  • Aebi H (1984) Catalase in vitro. Methods in enzymology, vol 105. Elsevier, Amsterdam, pp 121–126

    Google Scholar 

  • Aggarwal D, Kumar A, Reddy MS (2015) Genetic transformation of endo-1,4-β-glucanase (Korrigan) for cellulose enhancement in Eucalyptus tereticornis. Plant Cell Tissue Organ Cult 122:363–371

    CAS  Google Scholar 

  • Ahanger MA, Tomar NS, Tittal M, Argal S, Agarwal R (2017) Plant growth under water/salt stress: ROS production; antioxidants and significance of added potassium under such conditions. Physiol Mol Biol Plants 23:731–744

    PubMed  PubMed Central  Google Scholar 

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

    CAS  PubMed  PubMed Central  Google Scholar 

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

    CAS  Google Scholar 

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

    CAS  PubMed  Google Scholar 

  • Boland DJ, Brooker MIH, Chippendale GM, Hall N, Hyland BPM, Johnston RD, Kleinig DA, McDonald MW, Turner JD (1984) Forest trees of Australia. Nelson and CSIRO, Melbourne

    Google Scholar 

  • Bussotti F, Pollastrini M, Gessler A, Luo Z-B (2018) Experiments with trees: from seedlings to ecosystems. Environ Exper Bot 152:1–6

    Google Scholar 

  • Chang S, Puryear J, Cairney J (1993) A simple and efficient method for isolating RNA from pine trees. Plant Mol Biol Rep 11:113–116

    CAS  Google Scholar 

  • Cha-um S, Somsueb S, Samphumphuang T, Kirdmanee C (2013) Salt tolerant screening in eucalypt genotypes (Eucalyptus spp.) using photosynthetic abilities, proline accumulation, and growth characteristics as effective indices. Vitro Cell Dev Biol Plant 49:611–619

    CAS  Google Scholar 

  • Chen D, Keiper F, De Filippis L (1998) Physiological changes accompanying the induction of salt tolerance in Eucalyptus microcorys shoots in tissue culture. J Plant Physiol 152:555–563

    CAS  Google Scholar 

  • Colomba EL, Tommasino E, Luna C, Griffa S, Carloni E, Ribotta A, Quiroga M, Grunberg K (2013) Differential salt-stress response during germination and vegetative growth in in vitro selected somaclonal mutants of Cenchrus ciliaris L. S Afr J Bot 87:157–163

    Google Scholar 

  • Demidchik V, Straltsova D, Medvedev SS, Pozhvanov GA, Sokolik A, Yurin V (2014) Stress-induced electrolyte leakage: the role of K+-permeable channels and involvement in programmed cell death and metabolic adjustment. J Exp Bot 65:1259–1270

    CAS  PubMed  Google Scholar 

  • Dubois M, Gilles KA, Hamilton JK, Pt R, Smith F (1956) Colorimetric method for determination of sugars and related substances. Anal Chem 28:350–356

    CAS  Google Scholar 

  • Fuller MP, Hamza JH, Rihan HZ, Al-Issawi M (2012) Germination of primed seed under NaCl stress in wheat. International Scholarly Research Notices

  • Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Biochem 48:909–930

    CAS  PubMed  Google Scholar 

  • Gong M, Chen B, Li Z-G, Guo L-H (2001) Heat-shock-induced cross adaptation to heat, chilling, drought and salt stress in maize seedlings and involvement of H2O2. J Plant Physiol 158:1125–1130

    CAS  Google Scholar 

  • Grattapaglia D, Vaillancourt RE, Shepherd M, Thumma BR, Foley W, Külheim C, Potts BM, Myburg AA (2012) Progress in Myrtaceae genetics and genomics: Eucalyptus as the pivotal genus. Tree Genet Genomes 8:463–508

    Google Scholar 

  • Gupta B, Huang B (2014) Mechanism of salinity tolerance in plants: physiological, biochemical, and molecular characterization. Int J Genomics. https://doi.org/10.1155/2014/701596

    Article  PubMed  PubMed Central  Google Scholar 

  • Hayat S, Hayat Q, Alyemeni MN, Wani AS, Pichtel J, Ahmad A (2012) Role of proline under changing environments: a review. Plant Signal Behav 7:1456–1466

    CAS  PubMed  PubMed Central  Google Scholar 

  • Heath RL, Packer L (1968) Photoperoxidation in isolated chloroplasts: I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys 125:189–198

    CAS  PubMed  Google Scholar 

  • Hniličková H, Hnilička F, Orsák M, Hejnák V (2019) Effect of salt stress on growth, electrolyte leakage, Na+ and K+ content in selected plant species. Plant Soil Environ 65:90–96

    Google Scholar 

  • Hohmann M, Stahl A, Rudloff J, Wittkop B, Snowdon R (2016) Not a load of rubbish: simulated field trials in large-scale containers. Plant Cell Environ 39:2064–2073

    CAS  PubMed  Google Scholar 

  • Kader M (2005) A comparison of seed germination calculation formulae and the associated interpretation of resulting data. J Proc R Soc NSW 138:65–75

    Google Scholar 

  • Kaur A, Singh D, Gupta N, Kumar A (2018) In vitro propagation of important rootstocks of apple for rapid cloning and improvement. Biotechnologies of crop improvement, vol 1. Springer, New York, pp 215–241

    Google Scholar 

  • Koźmińska A, Wiszniewska A, Hanus-Fajerska E, Boscaiu M, Al Hassan M, Halecki W, Vicente O (2019) Identification of salt and drought biochemical stress markers in several Silene vulgaris populations. Sustainability 11:800

    Google Scholar 

  • Kumar A, Palni L, Nandi S (2003) The effect of light source and gelling agent on micropropagation of Rosa damascena Mill. and Rhynchostylis retusa (L.) Bl. J Hortic Sci Biotech 78:786–792

    Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    CAS  PubMed  Google Scholar 

  • McEwen CM Jr (1971) Monoamine oxidase [rabbit serum]. Methods in enzymology, vol 17. Elsevier, Amsterdam, pp 686–692

    Google Scholar 

  • Munns R, James RA (2003) Screening methods for salinity tolerance: a case study with tetraploid wheat. Plant Soil 253:201–218

    CAS  Google Scholar 

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

    CAS  Google Scholar 

  • Negrão S, Schmöckel S, Tester M (2017) Evaluating physiological responses of plants to salinity stress. Ann Bot 119:1–11

    PubMed  Google Scholar 

  • Niknam S, McComb J (2000) Salt tolerance screening of selected Australian woody species—a review. For Ecol Manag 139:1–19

    Google Scholar 

  • Nounjan N, Charoensawan V, Chansongkrow P, Siangliw JL, Toojinda T, Chadchawan S, Theerakulpisut P (2018) High performance of photosynthesis and osmotic adjustment are associated with salt tolerance ability in rice carrying drought tolerance QTL: physiological and co-expression network analysis. Front Plant Sci 9:1135

    PubMed  PubMed Central  Google Scholar 

  • Pandolfi C, Azzarello E, Mancuso S, Shabala S (2016) Acclimation improves salt stress tolerance in Zea mays plants. J Plant Physiol 201:1–8

    CAS  PubMed  Google Scholar 

  • Pandolfi C, Bazihizina N, Giordano C, Mancuso S, Azzarello E (2017) Salt acclimation process: a comparison between a sensitive and a tolerant Olea europaea cultivar. Tree Physiol 37:380–388

    CAS  PubMed  Google Scholar 

  • Panuccio M, Jacobsen S-E, Akhtar SS, Muscolo A (2014) Effect of saline water on seed germination and early seedling growth of the halophyte quinoa. AoB plants. https://doi.org/10.1093/aobpla/plu047

    Article  PubMed  PubMed Central  Google Scholar 

  • Polle A, Rennenberg H (2019) Physiological responses to abiotic and biotic stress in forest trees. Multidiscip Digit Publ Inst. https://doi.org/10.3390/books978-3-03921-515-7

    Article  Google Scholar 

  • Purohit M, Srivastava S, Srivastava P (1998) Stress tolerant plants through tissue culture. Plant tissue culture and molecular biology: application and prospects. Narosa Publishing House, New Delhi, pp 554–578

    Google Scholar 

  • Rahneshan Z, Nasibi F, Moghadam AA (2018) Effects of salinity stress on some growth, physiological, biochemical parameters and nutrients in two pistachio (Pistacia vera L.) rootstocks. J Plant Interact 13:73–82

    CAS  Google Scholar 

  • Rai MK, Kalia RK, Singh R, Gangola MP, Dhawan A (2011) Developing stress tolerant plants through in vitro selection—an overview of the recent progress. Environ Exper Bot 71:89–98

    Google Scholar 

  • Rajabi Dehnavi A, Zahedi M, Ludwiczak A, Cardenas Perez S, Piernik A (2020) Effect of salinity on seed germination and seedling development of sorghum (Sorghum bicolor (L.) Moench) genotypes. Agronomy 10:859

    Google Scholar 

  • Regni L, Del Pino AM, Mousavi S, Palmerini CA, Baldoni L, Mariotti R, Mairech H, Gardi T, D’Amato R, Proietti P (2019) Behaviour of four olive cultivars during salt stress. Front Plant Sci 10:867

    PubMed  PubMed Central  Google Scholar 

  • Sabo VA, Knezevic P (2019) Antimicrobial activity of Eucalyptus camaldulensis Dehn. plant extracts and essential oils: a review. Ind Crops Prod 132:413–429

    Google Scholar 

  • Sairam R, Srivastava G (2002) Changes in antioxidant activity in sub-cellular fractions of tolerant and susceptible wheat genotypes in response to long term salt stress. Plant Sci 162:897–904

    CAS  Google Scholar 

  • Santangeli M, Capo C, Beninati S, Pietrini F, Forni C (2019) Gradual exposure to salinity improves tolerance to salt stress in rapeseed (Brassica napus L.). Water 11:1667

    CAS  Google Scholar 

  • 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

    CAS  PubMed  Google Scholar 

  • Singh D, Kumar A (2020) In vitro screening and characterization of selected elite clones of Eucalyptus tereticornis Sm. for salt stress. J Plant Growth Regul. https://doi.org/10.1007/s00344-020-10138-9

    Article  Google Scholar 

  • Singh D, Kaur S, Kumar A (2020) In vitro drought tolerance in selected elite clones of Eucalyptus tereticornis Sm. Acta Physiol Plant 42:17

    CAS  Google Scholar 

  • Smart RE, Bingham GE (1974) Rapid estimates of relative water content. Plant Physiol 53:258–260

    CAS  PubMed  PubMed Central  Google Scholar 

  • Sumner JB, Howell SF (1935) A method for determination of saccharase activity. J Biol Chem 108:51–54

    CAS  Google Scholar 

  • Sundari BKR, Dasgupta MG (2012) Selection and validation of reference genes for real-time qRT-PCR normalization in different tissues of Eucalyptus tereticornis. Silvae Genet 61:280–286

    Google Scholar 

  • Sychta K, Słomka A, Sliwinska E, Migdałek G, Kuta E (2020) From cells highly tolerant to Zn and Pb to fully fertile plants—selection of tolerant lines with in vitro culture. Plant Physiol Biochem 146:231–237

    CAS  PubMed  Google Scholar 

  • Tahjib-UI-Arif M, Sohag AAM, Afrin S, Bashar KK, Afrin T, Mahamud A, Polash MAS, Hossain M, Sohel M, Taher A (2019) Differential response of sugar beet to long-term mild to severe salinity in a soil-pot culture. Agriculture 9:223

    CAS  Google Scholar 

  • Tavakkoli E, Rengasamy P, McDonald GK (2010) High concentrations of Na+ and Cl ions in soil solution have simultaneous detrimental effects on growth of faba bean under salinity stress. J Exp Bot 61:4449–4459

    CAS  PubMed  PubMed Central  Google Scholar 

  • Tavakkoli E, Fatehi F, Coventry S, Rengasamy P, McDonald GK (2011) Additive effects of Na+ and Cl ions on barley growth under salinity stress. J Exp Bot 62:2189–2203

    CAS  PubMed  PubMed Central  Google Scholar 

  • Thiam M, Champion A, Diouf D, Ourèye SYM (2013) NaCl effects on in vitro germination and growth of some senegalese cowpea (Vigna unguiculata (L.) Walp.) cultivars. ISRN biotechnol. https://doi.org/10.5402/2013/382417

    Article  PubMed  PubMed Central  Google Scholar 

  • Trueman SJ, Hung CD, Wendling I (2018) Tissue culture of Corymbia and Eucalyptus. Forests 9:84

    Google Scholar 

  • Vangelisti A, Zambrano LS, Caruso G, Macheda D, Bernardi R, Usai G, Mascagni F, Giordani T, Gucci R, Cavallini A (2019) How an ancient, salt-tolerant fruit crop, Ficus carica L., copes with salinity: a transcriptome analysis. Sci Rep 9:2561

    PubMed  PubMed Central  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

    CAS  Google Scholar 

  • Woodward AJ, Bennett IJ (2005) The effect of salt stress and abscisic acid on proline production, chlorophyll content and growth of in vitro propagated shoots of Eucalyptus camaldulensis. Plant Cell Tissue Organ Cult 82:189–200

    CAS  Google Scholar 

  • Wu W, Zhang Q, Ervin E, Yang Z, Zhang X (2017) Physiological mechanism of enhancing salt stress tolerance of perennial ryegrass by 24-epibrassinolide. Front Plant Sci 8:1017

    PubMed  PubMed Central  Google Scholar 

  • Xie X, He Z, Chen N, Tang Z, Wang Q, Cai Y (2019) The roles of environmental factors in regulation of oxidative stress in plant. Biomed Res Int. https://doi.org/10.1155/2019/9732325

    Article  PubMed  PubMed Central  Google Scholar 

  • Yaish MW, Kumar PP (2015) Salt tolerance research in date palm tree (Phoenix dactylifera L.), past, present, and future perspectives. Front Plant Sci 6:348

    PubMed  PubMed Central  Google Scholar 

  • Zhang M, Fang Y, Ji Y, Jiang Z, Wang L (2013) Effects of salt stress on ion content, antioxidant enzymes and protein profile in different tissues of Broussonetia papyrifera. S Afr J Bot 85:1–9

    Google Scholar 

  • Zhang X, Yin H, Chen S, He J, Guo S (2014) Changes in antioxidant enzyme activity and transcript levels of related genes in Limonium sinense kuntze seedlings under NaCl stress. J Chem. https://doi.org/10.1155/2014/749047

    Article  Google Scholar 

  • Zhu J, Fan Y, Shabala S, Li C, Lv C, Guo B, Xu R, Zhou M (2020) Understanding mechanisms of salinity tolerance in barley by proteomic and biochemical analysis of near-isogenic lines. Int J Mol Sci 21:1516

    CAS  PubMed Central  Google Scholar 

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Acknowledgements

The authors would like to acknowledge TIFAC-CORE for creating the infrastructure and providing necessary facilities to carry out the research work.

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DS: experimental design, data compilation, analysis and interpretation, writing the manuscript. AK: data interpretation, commenting and finalizing the manuscript.

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Correspondence to Anil Kumar.

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Singh, D., Kumar, A. Multivariate screening approach indicated adaptive tolerance to salt stress in the seedlings of an agroforestry tree, Eucalyptus tereticornis Sm.. Plant Cell Tiss Organ Cult 145, 545–560 (2021). https://doi.org/10.1007/s11240-021-02025-2

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