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Physiological and molecular characterization of water-stressed Chrysanthemum under robinin and chitosan treatment

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Severe water shortage limits horticultural crop growth and development, thereby compromising plant quality. Novel tools to enhance stress tolerance in medicinal horticultural crops are crucial to cope with growing environmental challenges to world crop performance. In this study, water solutions of robinin (25, 100, and 200 ppm) and/or foliar sprays of chitosan (0, 50, and 200 ppm) were applied to Chrysanthemum morifolium Ramat subjected to a 2 (2DWI) or 6 day (6DWI) irrigation intervals for 6 weeks. Morphological, physiological, and genetic markers associated with plant-response mechanisms to water stress were explored. Robinin + chitosan-treated plants showed increased morphological performance associated with enhanced chlorophyll, carbohydrates, proline, K+, Ca+2, phenols, leaf water potential, antioxidants, and leaf water content. Superoxide dismutase (SOD), peroxidase (POD), and ascorbate peroxidase (APX) enzymes were more active in robinin + chitosan-treated plants, while H2O2 accumulation was diminished. Higher expression levels of the Chrysanthemum antioxidant gene of zinc-finger transcription factor gene (Cm-BBX24), Chrysanthemum roots fu (DREB1A-1), Chrysanthemum heat shock protein CgHSP70, pyrroline-5-carboxylate synthetases (P5CS), pyrroline-5-carboxylate reductase (P5CR), and proline dehydrogenase (ProDH) were found in robinin- and chitosan-treated plants. Robinin + chitosan treatment stimulated the accumulation of carbohydrates, K+, Ca+2, proline, and chlorophylls to achieve osmotic adjustment and maintain turgor pressure. Accumulation of reactive oxygen species was controlled by enzymatic and non-enzymatic means, as well as the overexpression of stress-related genes (Cm-BBX24, DREB1A-1, CgHSP70, P5CS, P5CR, and ProDH) in robinin + chitosan-treated plants. Plant-response mechanisms for enhanced drought resistance interacted under robinin + chitosan treatment to improve plant performance under stress conditions.

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  1. AbdElgawad H, Zinta G, Hegab MM, Pandey R, Asard H, Abuelsoud W (2016) High salinity induces different oxidative stress and antioxidant responses in maize seedlings organs. Front Plant Sci 7:276

  2. Ahmad Z, Waraich EA, Akhtar S et al (2018) Acta Physiol Plant 40:80

  3. Ahmadi J, Pour-Aboughadareh A, Ourang SF, Mehrabi AA, Siddique KHM (2018) Acta Physiol Plant 40:90

  4. Akhmedzhanova V (1986) Robinin and kaempfereol from Vinca erecta. Chem Nat Compd 22:601

  5. Al-Ghamdi AA, Elansary HO (2018) Synergetic effects of 5-aminolevulinic acid and Ascophyllum nodosum seaweed extracts on Asparagus phenolics and stress related genes under saline irrigation. Plant Physiol Biochem 129:273–284

  6. Ali F, Bano A, Fazal A (2017) Recent methods of drought stress tolerance in plants. Plant Growth Regul 82:363–375

  7. Bistgani ZE, Siadat SA, Bakhshandh A, Pirbalouti AG, Hashemi M (2017) Interactive effects of drought stress and chitosan application on physiological characteristics and essential oil yield of Thymus daenensis Celak. Crop J 5:407–415

  8. Chen X-y, Yang Y, Ran L-p, Dong Z-d, Zhang E-j, Yu X-r, Xiong F (2017) Novel insights into miRNA regulation of storage protein biosynthesis during wheat caryopsis development under drought stress. Front Plant Sci 8:1707

  9. Cruz CM (2008) Drought stress and reactive oxygen species: production, scavenging and signaling. Plant Signal Behav 3:156–165

  10. Dubios M, Gilles K, Hamlton J, Rebers P, Smith F (1956) Colourimetric method for determination of sugars and related substances. Anal Chem 28:350–356

  11. Elansary HO (2017) Green roof Petunia, Ageratum, and Mentha responses to water stress, seaweeds, and trinexapac-ethyl treatments. Acta Physiol Plant 39:145

  12. Elansary HO, Mahmoud EA (2015) Basil cultivar chemotyping still favored over genotyping using core barcodes and possible resources of antioxidants. Essen Oil Res 27:82–87

  13. Elansary HO, Salem MZM (2015) Morphological and physiological responses and drought resistance enhancement of ornamental shrubs by trinexapac-ethyl application. Sci Hortic 189:1–11

  14. Elansary HO, Yessoufou K (2016) In vitro antioxidant, antifungal and antibacterial activities of five international Calibrachoa cultivars. Nat Prod Res 11:1339–1342

  15. Elansary HO, Zin El-Abedin TK (2019) Omeprazole alleviates water stress in peppermint and modulates the expression of menthol biosynthesis genes. Plant Physiol Biochem 139:578–586

  16. Elansary HO, Skalicka-Woźniak K, King IW (2016a) Enhancing stress growth traits as well as phytochemical and antioxidant contents of Spiraea and Pittosporum under seaweed extract treatments. Plant Physiol Biochem 105:310–320

  17. Elansary HO, Norrie J, Ali HM, Salem MZM, Mahmoud EA, Yessoufou K (2016b) Enhancement of Calibrachoa growth, secondary metabolites and bioactivity using seaweed extracts. BMC Complement Altern Med 16:341

  18. Elansary HO, Yessoufou K, Abdel-Hamid AME, El-Esawi MA, Ali HM, Elshikh MS (2017) Seaweed extracts enhance Salam turfgrass performance during prolonged irrigation intervals and saline shock. Front Plant Sci 8:830

  19. Elansary HO, Szopa A, Kubica P, Ekiert H, Ali HM, Elshikh MS, Abdel-Salam EM, El-Esawi M, El-Ansary DO (2018) Bioactivities of traditional medicinal plants in Alexandria. Evid Based Complement Altern Med 1463579:13

  20. Elansary HO, Szopa A, Kubica P, Ekiert H, Mattar MA, Al-Yafrasi MA, El-Ansary DO, El-Abedin TKZ, Yessoufou K (2019) Polyphenol profile and pharmaceutical potential of Quercus spp. Bark Extr Plants 8:486

  21. El-Esawi MA, Elansary HO, Elshanhory N, Abdel-Hamid AME, Ali HM, Elshikh MS (2017) Salicylic acid-regulated antioxidant mechanisms and gene expression enhance rosemary performance under saline conditions. Front Physiol 8:716

  22. Gu C, Chen S, Liu Z, Shan H, Luo H, Guan Z, Chen F (2011) Reference gene selection for quantitative real-time PCR in Chrysanthemum subjected to biotic and abiotic stress. Mol Biotechnol 49:192–197

  23. Gupta B, Huang B (2014) Mechanism of salinity tolerance in plants: physiological, biochemical, and molecular characterization. Int J Genomics 2014:701596

  24. He CF, Chen S, Lv G, Deng Y, Fang W, Liu Z, Guan Z, He C (2011) Chrysanthemum leaf epidermal surface morphology and antioxidant and defense enzyme activity in response to aphid infestation. J Plant Physiol 168:687–693

  25. Hoque MA, Banu MNA, Okuma E, Amako K, Nakamura Y, Shimoishi Y, Murata Y (2007) Exogenous proline and glycinebetaine increase NaCl-induced ascorbate-glutathione cycle enzyme activities, and proline improves salt tolerance more than glycinebetaine in tobacco Bright Yellow-2 suspension-cultured cells. J Plant Physiol 164:1457–1468

  26. Huang B, DaCosta M, Jiang Y (2014) Research advances in mechanisms of turfgrass tolerance to abiotic stresses: from physiology to molecular biology. Crit Rev Plant Sci 33:141–189

  27. Karuppusamy S (2009) A review on trends in production of secondary metabolites from higher plants by in vitro tissue, organ and cell cultures. J Med Plant Res 3:1222–1239

  28. Li Z, Li Y, Zhang Y, Cheng B, Peng Y, Zhang X, Ma X, Huang L, Yan Y (2018) Indole-3-acetic acid modulates phytohormones and polyamines metabolism associated with the tolerance to water stress in white clover. Plant Physiol Biochem 129:251–263

  29. Liu Q, Kasuga M, Sakuma Y, Abe H, Miura S, Yamaguchi-Shinozaki K, Shinozaki K (1988) Two transcription factors, DREB1 and DREB2, with an EREBP/AP2 DNA binding domain separate two cellular signal transduction pathways in drought and low-temperature-responsive gene expression, respectively, in Arabidopsis. Plant Cell 10:1391–1406

  30. Mehran A, Aghakouchak A, Nakhjiri N, Stewardson MJ, Peel MC, Phillips TJ, Wada Y, Ravalico JK (2017) Compounding impacts of human-induced water stress and climate change on water availability. Sci Rep 7:6282

  31. Nilsen ET, Orcutt DM (1996) Physiology of plants under stress: abiotic factors. Wiley, New York

  32. Pallas B, Clément-Vidal A, Rebolledo M-C, Soulié J-C, Luquet D (2013) Using plant growth modeling to analyze C source–sink relations under drought: inter- and intraspecific comparison. Front Plant Sci 4:437

  33. Pérez-López U, Robredo A, Lacuesta M, Sgherri C, Muñoz-Rueda A, Navari-Izzo F et al (2009) The oxidative stress caused by salinity in two barley cultivars is mitigated by elevated CO2. Physiol Plant 135:29–42

  34. Pichyangkura R, Chadchawan S (2015) Biostimulant activity of chitosan in horticulture. Sci Hortic 196:49–65

  35. Ramírez MÁ, Rodriguez AT, Alfonso L, Peniche C (2010) Chitin and its derivatives as biopolymers with potential agricultural applications. Biotecnol Appl 27:270–276

  36. Saxena R, Tomar RS, Kumar M (2016) Exploring nanobiotechnology to mitigate abiotic stress in crop plants. J Pharm Sci Res 8:974–980

  37. Seki M, Umezawa T, Urano K (2007) Shinozaki K (2007) Regulatory metabolic networks in drought stress responses. Curr Opin Plant Biol 10:296–302

  38. Sergiev I, Alexieva V, Ivanov S, Bankova V, Mapelli S (2004) Plant growth regulating activity of some flavonoids. CR Acad Bulg Sci 57:63

  39. Sharp RG (2013) A review of the applications of chitin and its derivatives in agriculture to modify plant-microbial interactions and improve crop yields. Agron 3:757–793

  40. Song A, Zhu X, Chen F, Gao H, Jiang J, Chen S (2014) A chrysanthemum heat shock protein confers tolerance to abiotic stress. Int J Mol Sci 15:5063–5078

  41. Tong Z, Hong B, Yang Y, Li Q, Ma N, Ma C, Gao J (2009) Overexpression of two chrysanthemum DgDREB1 group genes causing delayed flowering or dwarfism in Arabidopsis. Plant Mol Biol 71:115–129

  42. Yang F, Hu J, Li J, Wu X, Qian Y (2009) Chitosan enhances leaf membrane stability and antioxidant enzyme activities in apple seedlings under drought stress. Plant Growth Regul 58:131–136

  43. Yang Y, Ma C, Xu Y, Wei Q, Imtiaz M, Lan H, Gao S, Cheng L, Wang M, Fei Z (2014) A zinc finger protein regulates flowering time and abiotic stress tolerance in chrysanthemum by modulating gibberellin biosynthesis. Plant Cell 23:2038–2054

  44. Yildizli A, Çevik S, Ünyayar S (2018) Effects of exogenous myo-inositol on leaf water status and oxidative stress of Capsicum annuum under drought stress. Acta Physiol Plant 40:122

  45. Yin YG, Kobayashi Y, Sanuki A, Kondo S, Fukuda N, Ezura H, Sugaya S, Matsukura C (2010) Salinity induces carbohydrate accumulation and sugar-regulated starch biosynthetic genes in tomato (Solanum lycopersicum L. cv. ‘Micro-Tom’) fruits in an ABA- and osmotic stress-independent manner. J Exp Bot 61:563–574

  46. Yin H, Frette XC, Christensen LP, Grevsen K (2012) Chitosan oligosaccharides promote the content of polyphenols in Greek oregano (Origanum vulgare ssp. hirtum). J Agric Food Chem 60:136–143

  47. Yu S, Liao F, Wang F, Wen W, Li J, Mei H et al (2012) Identification of rice transcription factors associated with drought tolerance using the ecotilling method. PLoS ONE 7(2):e30765

  48. Zhang G, Chen M, Li L, Xu Z, Chen X, Guo J, Ma Y (2009) Overexpression of the soybean GmERF3 gene, an AP2/ERT type transcription factor for increased tolerance to salt, drought, and diseases in transgenic tobacco. J Exp Bot 60:3781–3796

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The work was supported by King Saud University, Researchers Supporting Project number (RSP-2019/118).

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Correspondence to Hosam O. Elansary.

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Elansary, H.O., Abdel-Hamid, A.M.E., Yessoufou, K. et al. Physiological and molecular characterization of water-stressed Chrysanthemum under robinin and chitosan treatment. Acta Physiol Plant 42, 31 (2020).

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  • Chrysanthemum morifolium
  • Irrigation intervals
  • Robinin
  • Gene expression
  • Chitosan