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Root Development Enhanced by Using Indole-3-butyric Acid and Naphthalene Acetic Acid and Associated Biochemical Changes of In Vitro Azalea Microshoots

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Abstract

Based on the importance of producing in vitro adventitious roots, this study was carried out to investigate the effects of indole-3-butyric acid (IBA) and naphthalene acetic acid (NAA) at a concentration of 2 mg L−1 on the formation of adventitious roots of azalea and their impact on biochemical changes and endogenous hormones. The rooting percentage, root number, and root length were increased in the microshoots of both studied cultivars (‘Mingchao’ and ‘Zihudie’) when the growth medium was supplemented with IBA. Additionally, peroxidase, indole acetic acid oxidase, hydrogen peroxide, and soluble protein contents were improved in both cultivars by auxin treatments especially during the first 7 days of the rooting period. However, application of IBA and NAA increased catalase and polyphenol oxidase in both cultivars during the first 14 and 28 days of culture. The increase in endogenous indole acetic acid (IAA) levels was accompanied by low activity of IAAO during most periods of root induction of microshoots in all treatments. Endogenous gibberellic acid levels were increased after 7 days of culture and then increased again after 28 days of culture. In contrast, the levels of endogenous zeatin riboside and isopentenyl adenosine were decreased with auxin treatments in the first period of the rooting process and then increased after 21 and 28 days of culture. The present study demonstrated that IBA at a concentration of 2 mg L−1 has a strong effect on azalea rooting. Moreover, the efficiency of IBA and NAA effects on biochemical changes during adventitious root induction was investigated, which may provide new horizons of in vitro rooting production and provide valuable information for the micropropagation of Rhododendron plants.

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References

  • Aina O, Quesenberry K, Gallo M (2015) Culture vessel and auxin treatments affect in vitro rooting and ex vitro survival of six Arachis paraguariensis genotypes. Sci Hortic 183:167–171

    Article  CAS  Google Scholar 

  • Anderson WC (1984) A revised tissue culture medium for shoot multiplication of rhododendron. J Am Soc Hortic Sci 109:343–347

    Google Scholar 

  • Benson EE (2000) Do free radicals have a role in plant tissue culture recalcitrance? In Vitro Cell Dev Biol Plant 36:163–170

    Article  CAS  Google Scholar 

  • Blaˇzková A, Sotta B, Tranvan H, Maldiney R, Bonnet M, Eihorn J, Kerhoas L, Miginiac E (1997) Auxin metabolism and rooting in young and mature clonesof Sequoia sempervirens. Physiol Plant 99:73–80

    Article  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Brownleader M, Hopkins J, Mobasheri A, Dey P, Jackson P, Trevan M (2000) Role of extensin peroxidase in tomato (Lycopersicon esculentum Mill.) seedling growth. Planta 210:668–676

    Article  PubMed  CAS  Google Scholar 

  • Carvalho LC, Vilela BJ, Vidigal P, Mullineaux PM, Amâncio S (2006) Activation of the ascorbate-glutathione cycle is an early response of micropropagated Vitis vinifera L. explants transferred to ex vitro. Int J Plant Sci 16:7759–7770

    Google Scholar 

  • Chao IL, Cho C-L, Chen L-M, Liu Z-H (2001) Effect of indole-3-butyric acid on the endogenous indole-3-acetic acid and lignin contents in soybean hypocotyl during adventitious root formation. J Plant Physiol 158:1257–1262

    Article  CAS  Google Scholar 

  • Cheniany M, Ebrahimzadeh H, Masoudi-nejad A, Vahdati K, Leslie C (2010) Effect of endogenous phenols and some antioxidant enzyme activities on rooting of Persian walnut (Juglans regia L.). Afr J Plant Sci 4:479–487

    CAS  Google Scholar 

  • Chu EP, Tavares AR, Kanashiro S, Giampaoli P, Yokota ES (2010) Effects of auxins on soluble carbohydrates, starch and soluble protein content in Aechmea blanchetiana (Bromeliaceae) cultured in vitro. Sci Hortic 125:451–455

    Article  CAS  Google Scholar 

  • Collet G (1985) Enracinement amélioré lors de la production in vitro de rosiers. Rev Suisse Vitic Arboric Hortic 17:259–263

    Google Scholar 

  • Da Costa Mello S, Angelotti-Mendonça J, Riboldi LB, Dall’Orto LTC, Suguino E (2016) Impact of indole-3-butyric acid on adventitious root development from cuttings of tea. HortTechnology 26:599–603

    Article  Google Scholar 

  • Da Costa CT, De Almeida MR, Ruedell CM, Schwambach J, Maraschin FDS, Fett-Neto AG (2013) When stress and development go hand in hand: main hormonal controls of adventitious rooting in cuttings. Front Plant Sci 4:133

    Article  PubMed  PubMed Central  Google Scholar 

  • Dabin P, Bouharmont J (1982) Application of in vitro cultures in azalea (Rhododendron simsii planch) (1). In Vitro Cult XXI IHC 131:89–94

    Google Scholar 

  • Dash GK, Senapati SK, Rout GR (2011) Effect of auxins on adventitious root development from nodal cuttings of Saraca asoka (Roxb.) de Wilde and associated biochemical changes. J Hortic For 3:320–326

    Google Scholar 

  • Eeckhaut T, Janssens K, De Keyser E, De Riek J (2010) Micropropagation of Rhododendron. In: Jain SM, Ochatt SJ (eds) Protocols for in vitro propagation of ornamentals plants, methods in molecular biology. Humana Press Inc, Totowa, pp 141–152

    Chapter  Google Scholar 

  • Feng D, Huang X, Liu Y, Willison JM (2016) Growth and changes of endogenous hormones of mulberry roots in a simulated rocky desertification area. Environ Sci Pollut R 23:11171–11180

    Article  CAS  Google Scholar 

  • Fu Z, Xu P, He S, da Silva JAT, Tanaka M (2011) Dynamic changes in enzyme activities and phenolic content during in vitro rooting of tree peony (Paeonia suffruticosa Andr.) plantlets. Maejo Int J Sci Technol 5:252–265

    CAS  Google Scholar 

  • Gaspar T, Kevers C, Hausman J, Berthon J, Ripetti V (1992) Practical uses of peroxidase activity as a predictive marker of rooting performance of micropropagated shoots. Agronomie 12:757–765

    Article  Google Scholar 

  • Gaspar T, Kevers C, Penel C, Greppin H, Reid DM, Thorpe TA (1996) Plant hormones and plant growth regulators in plant tissue culture. In Vitro Cell Dev Biol Plant 2:3277–3289

    Google Scholar 

  • González A, Tamés RS, Rodríguez R (1991) Ethylene in relation to protein, peroxidase and polyphenol xidase activities during rooting in hazelnut cotyledons. Physiol Plant 83:611–620

    Article  Google Scholar 

  • Góth L (1991) A simple method for determination of serum catalase activity and revision of reference range. Clin Chim Acta 196:143–151

    Article  PubMed  Google Scholar 

  • Guifeng L, Chuanping Y, Guanzheng Q, Xiangling Y (2001) Dynamic changes of four endogenous hormones in the larch hybrid during cuttings rooting. J NE Forestry Uni 6:000

  • GÜNEŞ T (2000) Peroxidase and IAA-oxidase activities during rooting in cuttings of three poplar species. Turk J Bot 24:97–102

    Google Scholar 

  • Guo X, Fu X, Zang D, Ma Y (2009) Effect of auxin treatments, cuttings’ collection date and initial characteristics on Paeonia ‘Yang Fei Chu Yu’cutting propagation. Sci Hortic 119:177–181

    Article  CAS  Google Scholar 

  • Gupta SD, Datta S (2003) Antioxidant enzyme activities during in vitro morphogenesis of gladiolus and the effect of application of antioxidants on plant regeneration. Biol Plant 47:179–183

    Article  CAS  Google Scholar 

  • Haissig BE (1986) Metabolic processes in adventitious rooting of cuttings. In: Jackson MB (ed) New root formation in plants and cuttings. Springer, Dordrecht, pp 141–189

    Chapter  Google Scholar 

  • Hartmann DE, Kester FT, Davies Jr, Geneve RL (2011) Plant propagation principles and practices, 8th edn. Prentice Hall, Englewood Cliffs

    Google Scholar 

  • Hatzilazarou SP, Syros TD, Yupsanis TA, Bosabalidis AM, Economou AS (2006) Peroxidases, lignin and anatomy during in vitro and ex vitro rooting of gardenia (Gardenia jasminoides Ellis) microshoots. J Plant Physiol 163:827–836

    Article  PubMed  CAS  Google Scholar 

  • Hisano H, Matsuura T, Mori IC, Yamane M, Sato K (2016) Endogenous hormone levels affect the regeneration ability of callus derived from different organs in barley. Plant Physiol Biochem 99:66–72

    Article  PubMed  CAS  Google Scholar 

  • Huang Y, Ji K-s, Zhai J-r (2007) Relationship between rooting ability and endogenous phytohormone changes in successive continuous generation cuttings of Buxus sinica var. parvifolia, an endangered woody species in China. For Stud China 9:189–197

    Article  CAS  Google Scholar 

  • Ilczuk A, Jacygrad E (2016) The effect of IBA on anatomical changes and antioxidant enzyme activity during the in vitro rooting of smoke tree (Cotinus coggygria Scop.). Sci Hortic 210:268–276

    Article  CAS  Google Scholar 

  • Khan S, Bi TB (2012) Direct shoot regeneration system for date palm (Phoenix dactylifera L.) cv. Dhakki as a means of micropropagation. Pak J Bot 44:1965–1671

    Google Scholar 

  • Kotis M, Yupsanis T, Syros T, Economou A (2009) Peroxidase, acid phosphatase, RNase and DNase activity and isoform patterns during in vitro rooting of Petunia × hybrida microshoots. Biol Plant 53:530–538

    Article  CAS  Google Scholar 

  • Lee TT, Starratt AN, Jevnikar JJ (1982) Regulation of enzymic oxidation of indole-3-acetic acid by phenols: structure-activity relationships. Phytochemistry 21:517–523

    Article  CAS  Google Scholar 

  • Li S, Xue L, Xu S, Feng H, An L (2007) Hydrogen peroxide involvement in formation and development of adventitious roots in cucumber. Plant Growth Regul 52:173–180

    Article  CAS  Google Scholar 

  • Liu Z-H, Hsiao I-C, Pan Y-W (1996) Effect of naphthalene acetic acid on endogenous indole-3-acetic acid, peroxidase and auxin oxidase in hypocotyl cuttings of soybean during root formation. Bot Bull Acad Sin 37:247–253

    CAS  Google Scholar 

  • Liu T, Hu Y, Li X (2008) Comparison of dynamic changes in endogenous hormones and sugars between abnormal and normal Castanea mollissima. Prog Nat Sci 18:685–690

    Article  CAS  Google Scholar 

  • Ludwig-Müller J (2000) Indole-3-butyric acid in plant growth and development. Plant Growth Regul 32:219–230

    Article  Google Scholar 

  • Macedo E, Vieira C, Carrizo D, Porfirio S, Hegewald H, Arnholdt-Schmitt B, Calado M, Peixe A (2013) Adventitious root formation in olive (Olea europaea L.) microshoots: anatomical evaluation and associated biochemical changes in peroxidase and polyphenol oxidase activities. J Hortic Sci Biotechnol 88:53–59

    Article  CAS  Google Scholar 

  • Mendes AF, Cidade L, Otoni W, Soares-Filho W, Costa MG (2011) Role of auxins, polyamines and ethylene in root formation and growth in sweet orange. Biol Plant 55:375–378

    Article  CAS  Google Scholar 

  • Mertens M, Werbrouck S, Samyn G, da Silva H, Debergh P (1996) Botelho dos Santos Moreira. In vitro regeneration of evergreen azalea from leaves. Plant Cell Tissue Org Cult 45:231–236. https://doi.org/10.1007/BF00043635

    Article  Google Scholar 

  • Metaxas DJ, Syros TD, Yupsanis T, Economou AS (2004) Peroxidases during adventitious rooting in cuttings of Arbutus unedo and Taxus baccata as affected by plant genotype and growth regulator treatment. Plant Growth Regul 44:257–266

    Article  CAS  Google Scholar 

  • Mishra BS, Singh M, Aggrawal P, Laxmi A (2009) Glucose and auxin signaling interaction in controlling Arabidopsis thaliana seedlings root growth and development. PLoS ONE 4:e4502

    Article  PubMed  PubMed Central  Google Scholar 

  • Molassiotis A, Dimassi K, Diamantidis G, Therios I (2004) Changes in peroxidases and catalase activity during in vitro rooting. Biol Plant 48:1–5

    Article  CAS  Google Scholar 

  • Nag S, Saha K, Choudhuri M (2001) Role of auxin and polyamines in adventitious root formation in relation to changes in compounds involved in rooting. J Plant Growth Regul 20:182–194

    Article  CAS  Google Scholar 

  • Naija S, Elloumi N, Jbir N, Ammar S, Kevers C (2008) Anatomical and biochemical changes during adventitious rooting of apple rootstocks MM 106 cultured in vitro. C R Biol 331:518–525

    Article  PubMed  CAS  Google Scholar 

  • Neves C, Sá MC, Amaˆncio S (1998) Histochemical detection of H2O2 by tissue printing as a precocious marker of rhizogenesis in grapevine. Plant Physiol Biochem 36:817–824

    Article  CAS  Google Scholar 

  • Osterc G, Štampar F (2011) Differences in endo/exogenous auxin profile in cuttings of different physiological ages. J Plant Physiol 168:2088–2092

    Article  PubMed  CAS  Google Scholar 

  • Osterc G, Petkovšek MM, Stampar F (2016) Quantification of IAA metabolites in the early stages of adventitious rooting might be predictive for subsequent differences in rooting response. J Plant Growth Regul 35:534–542

    Article  CAS  Google Scholar 

  • Palmer JO, Rankin SM, Yagi KJ, Tobe SS (1996) Juvenile hormone esterase activity during the reproductive cycle of the ring-legged earwig. Comp Biochem Physiol C Toxicol Pharmacol 114:145–151

    Google Scholar 

  • Perveen S, Anis M (2015) Physiological and biochemical parameters influencing ex vitro establishment of the in vitro regenerants of Albizia lebbeck. Agrofor Syst 89:721–733

    Article  Google Scholar 

  • Porfirio S, Calado ML, Noceda C, Cabrita MJ, da Silva MG, Azadi P, Peixe A (2016) Tracking biochemical changes during adventitious root formation in olive (Olea europaea L.). Sci Hortic 204:41–53

    Article  CAS  Google Scholar 

  • Preil W, Engelhardt M (1977) Meristem culture of azaleas (Rhododendron simsii). In: Symposium on tissue culture for horticultural purposes, vol 78, pp 203–208

  • Raymond J, Rakariyatham N, Azanza J (1993) Purification and some properties of polyphenoloxidase from sunflower seeds. Phytochemistry 34:927–931

    Article  CAS  Google Scholar 

  • Rout GR (2006) Effect of auxins on adventitious root development from single node cuttings of Camellia sinensis (L.) Kuntze and associated biochemical changes. Plant Growth Regul 48:111–117

    Article  CAS  Google Scholar 

  • Satisha J, Raveendran P, Rokade N (2008) Changes in polyphenol oxidase activity during rooting of hardwood cuttings in three grape rootstocks under Indian conditions. S Afr J Enol Vitic 29:94–97

    Google Scholar 

  • Scandalios JG (1993) Oxygen stress and superoxide dismutases. Plant Physiol 101:7–12

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Sheteiwy M, Shen H, Xu J, Guan Y, Song W, Hu J (2017) Seed polyamines metabolism induced by seed priming with spermidine and 5-aminolevulinic acid for chilling tolerance improvement in rice (Oryza sativa L.) seedlings. Environ Exp Bot 137:58–72

    Article  CAS  Google Scholar 

  • Sun YL, Hong SK (2010) Effects of plant growth regulators and L-glutamic acid on shoot organogenesis in the halophyte Leymus chinensis (Trin.). Plant Cell Tissue Org Cult 100:317–328

    Article  CAS  Google Scholar 

  • Syros T, Yupsanis T, Zafiriadis H, Economou A (2004) Activity and isoforms of peroxidases, lignin and anatomy, during adventitious rooting in cuttings of Ebenus cretica L. J Plant Physiol 16:69–77

    Article  Google Scholar 

  • Trifunović-Momčilov M, Motyka V, Dragićević I, Petrić M, Jevremović S, Malbeck J, Holík J, Dobrev PI, Subotić A (2016) Endogenous phytohormones in spontaneously regenerated Centaurium erythraea Rafn. plants grown in vitro. J Plant Growth Regul 35:543–552

    Article  CAS  Google Scholar 

  • Tsipouridis C, Thomidis T, Bladenopoulou S (2006) Rhizogenesis of GF677, Early Crest, May Crest and Arm King stem cuttings during the year in relation to carbohydrate and natural hormone content. Sci Hortic 108:200–204

    Article  CAS  Google Scholar 

  • Vatankhah E, Niknam V, Ebrahimzadeh H (2010) Activity of antioxidant enzyme during in vitro organogenesis in Crocus sativus. Biol Plant 54:509–514

    Article  CAS  Google Scholar 

  • Wiesman Z, Riov J, Epstein E (1988) Characterization and rooting ability of indole-3-butyric acid conjugates formed during rooting of mung bean cuttings. Plant Physiol 91:1080–1084

    Article  Google Scholar 

  • Wiszniewska A, Nowak B, Kołton A, Sitek E, Grabski K, Dziurka M, Długosz-Grochowska O, Dziurka K, Tukaj Z (2016) Rooting response of Prunus domestica L. microshoots in the presence of phytoactive medium supplements. Plant Cell Tissue Org Cult 125:163–176

    Article  CAS  Google Scholar 

  • Yan Y-H, Li J-L, Zhang X-Q, Yang W-Y, Wan Y, Ma Y-M, Zhu Y-Q, Peng Y, Huang L-K (2014) Effect of naphthalene acetic acid on adventitious root development and associated physiological changes in stem cutting of Hemarthria compressa. PLoS ONE 9:e90700

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Yan SP, Yang RH, Wang F, Sun LN, Song XS (2017) Effect of auxins and associated metabolic changes on cuttings of hybrid aspen. Forests 8:1–12

    Article  Google Scholar 

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Acknowledgements

This work was supported by the Science and Technology Major Project of Zhejiang Province, China (projects 2016C02056-12).

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Authors and Affiliations

  1. Physiology and Molecular Biology Laboratory of Ornamental Plants, Department of Horticulture, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, People’s Republic of China

    Mohamed S. Elmongy, Yan Cao, Hong Zhou & Yiping Xia

  2. Department of Vegetable and Floriculture, Faculty of Agriculture, Mansoura University, Mansoura, 35516, Egypt

    Mohamed S. Elmongy

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  1. Mohamed S. Elmongy
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  2. Yan Cao
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Correspondence to Yiping Xia.

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Elmongy, M.S., Cao, Y., Zhou, H. et al. Root Development Enhanced by Using Indole-3-butyric Acid and Naphthalene Acetic Acid and Associated Biochemical Changes of In Vitro Azalea Microshoots. J Plant Growth Regul 37, 813–825 (2018). https://doi.org/10.1007/s00344-017-9776-5

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