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Effects of thidiazuron on in vitro morphogenic response of Rhododendron sichotense Pojark. and Rhododendron catawbiense cv. Grandiflorum leaf explants

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Abstract

Thidiazuron (TDZ)-induced shoot organogenesis from in vitro-derived leaf explants of winter-hardy Rhododendron sichotense Pojark. and Rhododendron catawbiense cv. Grandiflorum was studied using two TDZ application methods: adding various concentrations to Anderson nutrient medium and a liquid pulse treatment. The highest frequency of shoot regeneration (93%) and maximum number of shoots per explant (24.6) from R. sichotense was obtained after a two-stage culture procedure that included exposure of leaf explants to 1.0 μМ TDZ followed by TDZ-free cultivation. TDZ at concentrations of 0.5 and 1.0 μМ was effective for leaf explants of R. catawbiense cv. Grandiflorum, inducing 60 and 85% regeneration frequencies, and 36.0 and 25.0 shoots per explant, respectively. The liquid-pulse treatment of leaf explants with 30 μM TDZ for 4 h was effective only for R. catawbiense cv. Grandiflorum. Histological analysis revealed that epidermal cells of the adaxial side at the leaf-blade base were involved in de novo shoot regeneration in both genotypes. However, the adventitious shoot buds of R. sichotense developed from protuberances, whereas shoot organogenesis of R. catawbiense cv. Grandiflorum occurred through the formation of embryo-like structures. Optimum root formation for both genotypes was achieved using indole-3-butyric acid liquid-pulse treatment of microshoots followed by ex vitro rooting. An efficient system was developed for multiplication of R. sichotense and R. catawbiense cv. Grandiflorum, which are cold-hardy genotypes promising for future genetic transformation.

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References

  • Aasim М, Hussain N, Umer EM, Zubair M, Hussain SB, Saeed S, Rafique TS, Sancak C (2010) In vitro shoot regeneration of fenugreek (Trigonella foenum-graecum L.) using different cytokinins. Afr J Biotechnol 9:7174–7179

    CAS  Google Scholar 

  • Ahmad N, Anis M (2012) An efficient and reproducible method for in vitro clonal multiplication of Rauvolfia tetraphylla L. and evaluation of genetic stability. Appl Biochem Biotechnol 168:1739–1752

    Article  PubMed  Google Scholar 

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

    Google Scholar 

  • Benmahioul B, Dorion N, Kaid-Harche M, Daguin F (2012) Micropropagation and ex vitro rooting of Pistachio (Pistacia vera). Plant Cell Tiss Organ Cult 108:353–358

    Article  Google Scholar 

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

  • Graner EM, Oberschelp GP, Brondani GE, Batagin-Piotto KD, de Almeida CV, de Almeida M (2013) TDZ pulsing evaluation on the in vitro morphogenesis of peach palm. Physiol Mol Biol Plants 19:283–288

    Article  PubMed Central  PubMed  Google Scholar 

  • Guo B, Abbasi BH, Zeb A, Xu LL, Wei YH (2011) Thidiazuron: a multi-dimentional plant growth regulator. Afr J Biotechnol 10:8984–9000

    CAS  Google Scholar 

  • Hebert CJ, Touchell DH, Ranney TG, LeBude AV (2010) In vitro shoot regeneration and polyploid induction of Rhododendron ‘Fragrantissimum Improved’. HortSci 45:801–804

    Google Scholar 

  • Huetteman CA, Preece JE (1993) Thidiazuron: a potent cytokinin for woody plant tissue culture. Plant Cell Tiss Organ Cult 33:105–119

    Article  CAS  Google Scholar 

  • Iapichino G, Chen THH, Fuchigami LH (1991) Adventitious shoot production from a vireya hybrid of Rhododendron. HortSci 26:594–596

    CAS  Google Scholar 

  • Iapichino G, McCulloch S, Chen THH (1992) Adventitious shoot formation from leaf explants of rhododendron. Plant Cell Tiss Organ Cult 30:237–241

    Article  CAS  Google Scholar 

  • Leva A (2012) Innovative protocol for “ex vitro rooting” on olive micropropagation. Cent Eur J Biol 6:352–358

    Google Scholar 

  • Li X, Krasnyanski SF, Korban SS (2002) Somatic embryogenesis, secondary somatic embryogenesis, and shoot organogenesis in Rosa. J. Plant Physiol 159:313–319

    Article  CAS  Google Scholar 

  • Lo KH, Giles KL, Sawhney VK (1997) Histological changes associated with acquisition of competence for shoot regeneration in leaf discs of Saintpaulia ionantha x confuse hybrid (African violet) cultured in vitro. Plant Cell Rep 16:421–425

    CAS  Google Scholar 

  • Madden JI, Jones CS, Auter CA (2005) Modes of regeneration in Pelargonium xhortorum (Geraniaceae) and three closely related species. In Vitro Cell Dev Biol – Plant 41:37–46

  • McCown BH, Lloyd GB (1983) A survey of the response of Rhododendron to in vitro culture. Plant Cell Tiss Organ Cult 2:77–85

    Article  Google Scholar 

  • Mertens M, Werbrouck S, Samyn G, Botelho H, Debergh P (1996) In vitro regeneration of evergreen azalea from leaves. Plant Cell Tiss Organ Cult 45:231–236

    Article  Google Scholar 

  • Mithila J, Hall JC, Victor JMR, Saxena PK (2003) Thidiazuron induces shoot organogenesis at low concentrations and somatic embryogenesis at high concentrations on leaf and petiole explants of African violet (Saintpaulia ionantha Wendl.). Plant Cell Rep 21:408–414

    CAS  PubMed  Google Scholar 

  • Murch SJ, Saxena PK (2001) Molecular fate of thidiazuron and its effects on auxin transport in hypocotyls tissues of Pelargonium x hortorum Bailey. Plant Growth Regul 35:269–275

    Article  CAS  Google Scholar 

  • Murthy BNS, Murch SJ, Saxena PK (1998) Thidiazuron: a potent regulator of in vitro plant morphogenesis. In Vitro Cell Dev Biol – Plant 34:267–275

  • Pascual L, Marin JA (2005) A liquid 2,4-D pulse increased shoot and root regeneration from leaf explants. Sci Hortic 106:582–592

    Article  CAS  Google Scholar 

  • Pausheva ZP (1988) Plant Cytology Practical Training. Agropromizdat, Moscow

    Google Scholar 

  • Pavingerova D (2009) The influence of thidiazuron on shoot regeneration from leaf explants of fifteen cultivars of Rhododendron. Biol Plant 54:797–799

    Article  Google Scholar 

  • Phulwaria M, Shekhawat NS (2013) An efficient in vitro shoot regeneration from immature inflorescence and ex vitro rooting of Arnebia hispidissima (Lehm). DC. - A red dye (Alkannin) yielding plant. Physiol Mol Biol Plants 19:435–441

    Article  PubMed Central  PubMed  Google Scholar 

  • Preece JE, Imel MR (1991) Plant regeneration from leaf explants of Rhododendron ‘P.J.M. Hybids’. Sci Hortic 48:159–170

    Article  CAS  Google Scholar 

  • Samyn G, De Schepper S, Van Bockstaele E (2002) Adventitious shoot regeneration and appearance of sports in several azalea cultivars. Plant Cell Tiss Organ Cult 70:223–227

    Article  CAS  Google Scholar 

  • Shaik NM, Arha M, Nookaraju A, Gupta SK, Srivastava S, Yadav AK, Kulkarni PS, Abhilash OU, Vishwakarma RK, Singh S, Tatkare R, Chinnathambi K, Rawal SK, Khan BM (2009) Improved method of in vitro regeneration in Leucaena leucocephala – a leguminous pulpwood tree species. Physiol Mol Biol Plants 15:311–318

    Article  PubMed Central  PubMed  Google Scholar 

  • Tomsone S, Gertnere D (2003) In vitro shoot regeneration from flower and leaf explants in Rhododendron. Biol Plant 46:463–465

    Article  Google Scholar 

  • Van Veen T (1969) Rhododendron in America. Sweeney Krist Dimm, Portland

    Google Scholar 

  • Vejsadová H, Petrova AS (2003) Somatic embryogenesis in Rhododendron catawbiense Grandiflorum. Acta Hortic 616:467–470

    Article  Google Scholar 

  • Vrishch DL, Varchenko LI, Urusov VM (2010) Rhododendron (Rhododendron L.) genus on Sikhote-Alin: geograrhy, ecology, genesis, economic prospects. Bulletin of KrasGAU 10:64–71

    Google Scholar 

  • Wamaitha MJ, Suwa K, Fukuda K, Mii M, Daimon H, Mishiba K (2010) Thidiazuron-induced rapid shoot regeneration via embryo-like structure formation from shoot tip-derived callus culture of sugarcane. Plant Biotechnol 27:365–368

    Article  CAS  Google Scholar 

  • Wojtania A, Gabryszewska E, Marasek A (2004) Regeneration of Pelargonium × hederaefolium ‘Bonete’ from petiole explants. Acta Physiol Plant 26:255–262

    Article  CAS  Google Scholar 

  • Woo SM, Wetzstein HY (2008) Morphological and histological evaluations of in vitro regeneration in Elliottia racemosa leaf explants induced on media with thidiazuron. J Amer Soc Hortic Sci 133:167–172

    Google Scholar 

Download references

Acknowledgments

This research was supported in part by Project no. 30.3 and no. 30.P of Program Presidium of the Russian Academy of Science “Living nature: state and the problem of development.” The authors wish to thank Geoffrey Harper for assistance in preparing the manuscript.

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Authors and Affiliations

  1. Biotechnology Laboratory, Central Siberian Botanical Garden, Siberian Branch of the Russian Academy of Sciences, 630090, Russian Federation, Novosibirsk, St. Zolotodolinskaya, 101

    Yulianna G. Zaytseva, Tatyana V. Poluboyarova & Tatyana I. Novikova

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  1. Yulianna G. Zaytseva
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  2. Tatyana V. Poluboyarova
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  3. Tatyana I. Novikova
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Correspondence to Yulianna G. Zaytseva.

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Editor: Zhezhi Wang

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Zaytseva, Y.G., Poluboyarova, T.V. & Novikova, T.I. Effects of thidiazuron on in vitro morphogenic response of Rhododendron sichotense Pojark. and Rhododendron catawbiense cv. Grandiflorum leaf explants. In Vitro Cell.Dev.Biol.-Plant 52, 56–63 (2016). https://doi.org/10.1007/s11627-015-9737-2

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