Changes in Morphological Characteristics, Regeneration Ability, and Polysaccharide Content in Tetraploid Dendrobium officinale

in HortScience

Dendrobium officinale Kimura et Migo is a famous traditional Chinese medicinal plant. It produces various phytochemicals, particularly polysaccharides, which have nutraceutical and pharmaceutical values. To increase its biomass production and polysaccharide content, our breeding program has generated a series of polyploid cultivars through colchicine treatment of protocorm-like bodies (PLBs). The present study compared two tetraploid cultivars, 201-1-T1 and 201-1-T2, with their diploid parental cultivar, 201-1, in an established in vitro culture system. Tetraploid ‘201-1-T1’ and ‘201-1-T2’ had shorter leaves and shorter and thicker stems and roots, and they produced higher biomass compared with the diploid cultivar. The length and width of stomata significantly increased, but stomatal density decreased in tetraploid cultivars. The PLB induction rates from the stem node explants of the tetraploid cultivars were significantly higher than those of diploid. However, the PLB proliferation of tetraploids was lower than that of the diploid. The mean number of plantlets regenerated from tetraploid PLBs was also lower than that of the diploid after 4 months of culture. Polysaccharide contents in stems, leaves, and roots of 6-month-old tetraploid plantlets were significantly higher than those of diploids. The polysaccharide content in the stem of ‘201-1-T1’ was 12.70%, which was a 2-fold increase compared with the diploid cultivar. Our results showed that chromosome doubling could be a viable way of improving D. officinale in biomass and polysaccharide production.

Contributor Notes

This work was supported by the Science and Technology Planning Project of Guangdong Province, China (2015A020209118 and 2015A030302066) and the China Scholarship Council.L.X. and Z.-S.Z. are corresponding authors. E-mail: xieli@scau.edu.cn or zszhang@scau.edu.cn.
  • Alonso-SandeM.Teijeiro-OsorioD.Remuñán-LópezC.AlonsoM.J.2009Glucomannan, a promising polysaccharide for biopharmaceutical purposesEur. J. Pharm. Biopharm.72453462

    • Search Google Scholar
    • Export Citation
  • AtichartP.2013Polyploid induction by colchicine treatments and plant regeneration of Dendrobium chrysotoxumThai J. Agr. Sci.465963

  • BlancG.WolfeK.H.2004Functional divergence of duplicated genes formed by polyploidy during Arabidopsis evolutionPlant Cell1616791691

  • CakovaV.BonteF.LobsteinA.2017Dendrobium: Sources of active ingredients to treat age-related pathologiesAging Dis.8827849

  • ChaicharoenS.1995Studies on chromosome number, floral and characteristic fertility of diploid and allotetraploid Dendrobium superbiensKasetsart J. Natl. Sci.29150157

    • Search Google Scholar
    • Export Citation
  • ChaicharoenS.SaejewK.1981Autopolyploidy in Dendrobium phalaenopsisJ. Sci. Soc. Thailand72532

  • ChenJ.WeiX.2018Thidiazuron in micropropagation of aroid plants p. 95–113. In: N. Ahmad and M. Faisal (eds.). Thidiazuron: From urea derivative to plant growth regulator. Springer Nature Singapore Pte Ltd. Singapore

  • ChenX.TianL.ShanT.SunL.GuoS.2018Advances in germplasm resources and genetics and breeding of Dendrobium officinaleActa Pharm. Sinica5314931503(In Chinese with English abstract)

    • Search Google Scholar
    • Export Citation
  • ChenX.M.WangF.F.WangY.Q.LiX.L.WangA.R.WangC.L.GuoS.X.2012Discrimination of the rare medicinal plant Dendrobium officinale based on naringenin, bibenzyl, and polysaccharidesSci. China Life Sci.5510921099

    • Search Google Scholar
    • Export Citation
  • ChengJ.DangP.ZhaoZ.YuanL.ZhouZ.WolfD.LuoY.2019An assessment of the Chinese medicinal Dendrobium industry: Supply, demand and sustainabilityJ. Ethnopharmacol.2298188

    • Search Google Scholar
    • Export Citation
  • ChungH.H.ShiS.K.HuangB.ChenJ.T.2017Enhanced agronomic traits and medicinal constituents of autotetraploids in Anoectochilus formosanus Hayata, a top-grade medicinal orchidMolecules22113

    • Search Google Scholar
    • Export Citation
  • Colijn-HooymansC.M.HakkertJ.C.JansenJ.CustersJ.B.M.1994Competence for regeneration of cucumber cotyledons is restricted to specific developmental stagesPlant Cell Tissue Organ Cult.39211217

    • Search Google Scholar
    • Export Citation
  • ComaiL.2005The advantages and disadvantages of being polyploidyNat. Rev. Genet.6836846

  • Committee for the Pharmacopoeia of PR China2015Pharmacopoeia of PR China Part I. China Medical Science and Technology Press PR China (in Chinese)

  • CorneillieS.De StormeN.Van AckerR.FangelJ.U.De BruyneM.De RyckeR.M.GeelenD.WillatsW.G.T.VanholmeB.BoerjanW.A.2018Polyploidy affects plant growth and alters cell wall compositionPlant Physiol.1797487

    • Search Google Scholar
    • Export Citation
  • CorrêaJ.P.O.VitalC.E.PinheiroM.V.M.BatistaD.S.SaldanhaC.W.da CruzA.C.F.NotiniM.M.FreitasD.M.S.DaMattaF.M.OtoniW.C.2016Induced polyploidization increases 20-hydroxyecdysone content, in vitro photoautotrophic growth, and ex vitro biomass accumulation in Pfaffia glomerata (Spreng.) PedersenIn Vitro Cell. Dev. Biol. Plant.524555

    • Search Google Scholar
    • Export Citation
  • CuiJ.ChenJ.HennyR.J.2009Regeneration of Aeschynanthus radicans via direct somatic embryogenesis and analysis of regenerants with flow cytometryIn Vitro Cell. Dev. Biol. Plant453443

    • Search Google Scholar
    • Export Citation
  • CuiJ.LiuJ.DengM.ChenJ.HennyR.J.2008Plant regeneration through protocorm-like bodies induced from nodal explants of Syngonium podophyllum ‘White Butterfly’HortScience4321292133

    • Search Google Scholar
    • Export Citation
  • De OliveiraV.M.Forni-MartinsE.R.MagalhãesP.M.AlvesM.N.2004Chromosomal and morphological studies of diploid and polyploid cytotypes of Stevia rebaudiana (Bertoni) Bertoni (Eupatorieae, Asteraceae)Genet. Mol. Biol.27215222

    • Search Google Scholar
    • Export Citation
  • del PozoJ.C.Ramirez-ParraE.2014Deciphering the molecular bases for drought tolerance in Arabidopsis autotetraploidsPlant Cell Environ.3727222737

    • Search Google Scholar
    • Export Citation
  • DingG.ZhangD.DingX.ZhouQ.ZhangW.LiX.2008Genetic variation and conservation of the endangered Chinese endemic herb Dendrobium officinale based on SRAP analysisPlant Syst. Evol.276149156

    • Search Google Scholar
    • Export Citation
  • DouM.M.ZhangZ.H.LiZ.B.ZhangJ.ZhaoX.Y.2016Cardioprotective potential of Dendrobium officinale Kimura et Migo against myocardial ischemia in miceMol. Med. Rep.1444074414

    • Search Google Scholar
    • Export Citation
  • DuditsD.TörökK.CseriA.PaulK.NagyA.V.NagyB.SassL.FerencG.VankovaR.DobrevP.VassI.AyaydinF.2016Response of organ structure and physiology to autotetraploidization in early development of energy willow Salix viminalisPlant Physiol.17015041523

    • Search Google Scholar
    • Export Citation
  • GantaitS.MandalN.BhattacharyyaS.DasP.K.2011Induction and identification of tetraploids using in vitro colchicine treatment of Gerbera jamesonii Bolus cv. SciellaPlant Cell Tissue Organ Cult.106485493

    • Search Google Scholar
    • Export Citation
  • GaoS.L.ChenB.J.ZhuD.N.2002In vitro production and identification of autotetraploids of Scutellaria baicalensisPlant Cell Tissue Organ Cult.70289293

    • Search Google Scholar
    • Export Citation
  • GaoS.L.ZhuD.N.CaiZ.H.XuD.R.1996Autotetraploid plants from colchicine-treated bud culture of Salvia miltiorrhiza BgePlant Cell Tissue Organ Cult.477377

    • Search Google Scholar
    • Export Citation
  • GrossoV.FarinaA.GiorgiD.NardiL.DirettoG.LucrettiS.2018A high-throughput flow cytometry system for early screening of in vitro made polyploids in Dendrobium hybridsPlant Cell Tissue Organ Cult.1325770

    • Search Google Scholar
    • Export Citation
  • HannwegK.VisserG.SippelA.BertlingI.2016Micropropagation and in vitro polyploidisation of Siphonochilus aethiopicus (wild ginger) and its effect on selected horticultural characteristicsActa Hort.1113175181

    • Search Google Scholar
    • Export Citation
  • HeC.WuK.ZhangJ.LiuX.ZengS.YuZ.ZhangX.Teixeira da SilvaJ.A.DengR.TanJ.LuoJ.DuanJ.2017Cytochemical localization of polysaccharides in Dendrobium officinale and the involvement of DoCSLA6 in the synthesis of mannan polysaccharidesFront. Plant Sci.8173doi: 10.3389/fpls.2017.00173

    • Search Google Scholar
    • Export Citation
  • HeT.B.HuangY.P.YangL.LiuT.T.GongW.Y.WangX.J.ShengJ.HuJ.M.2016Structural characterization and immunomodulating activity of polysaccharide from Dendrobium officinaleIntl. J. Biol. Macromol.833441

    • Search Google Scholar
    • Export Citation
  • HegdeS.N.RameshsingC.N.VasundharaM.2015Characterization of Stevia rebaudiana Bertoni polyploids for growth and qualityMed. Plant7188195

    • Search Google Scholar
    • Export Citation
  • HossainM.M.KantR.VanP.T.WinartoB.ZengS.Teixera da SilvaJ.A.2013The application of biotechnology to orchidsCrit. Rev. Plant Sci.3269139

    • Search Google Scholar
    • Export Citation
  • JiangJ.L.YeW.LiY.Q.ZhouJ.J.LeiF.G.WeiD.Z.2014Growth and polysaccharides accumulation in autotetraploid Dendrobium officinalePlant Physiol. J.50519526(in Chinese with English abstract)

    • Search Google Scholar
    • Export Citation
  • JinQ.JiaoC.Y.SunS.W.SongC.CaiY.P.LinY.FanH.H.ZhuY.F.2016Metabolic analysis of medicinal Dendrobium officinale and Dendrobium huoshanense during different growth yearsPLoS One11117

    • Search Google Scholar
    • Export Citation
  • KetsaS.UthairatanakijA.PrayurawongA.2001Senescence of diploid and tetraploid cut inflorescences of Dendrobium ‘Caesar’Scientia Hort.91133141

    • Search Google Scholar
    • Export Citation
  • KnightC.A.BeaulieuJ.M.2008Genome size scaling through phenotype spaceAnn. Bot.101759766

  • KubalakovaM.DolezelJ.LebedaA.1996Ploidy instability of embryogenic cucumber (Cucumis sativus L.) callus cultureBiol. Plant.38475480

  • LamY.NgT.B.YaoR.M.ShiJ.XuK.SzeS.C.W.ZhangK.Y.2015Evaluation of chemical constituents and important mechanism of pharmacological biology in Dendrobium plantsEvid. Based Complement. Alternat. Med.2015841752doi: 10.1155/2015/841752

    • Search Google Scholar
    • Export Citation
  • LiH.ZhengX.S.LongC.L.2005Induction of polyploid of Dendrobium devonianumActa Bot. Yunnanica27552556(In Chinese with English abstract)

    • Search Google Scholar
    • Export Citation
  • LiJ.GaleS.W.KumarP.ZhangJ.FischerG.2018aPrioritizing the orchids of a biodiversity hotspot for conservation based on phylogenetic history and extinction riskBot. J. Linn. Soc.186473497

    • Search Google Scholar
    • Export Citation
  • LiM.DingB.HuangW.PanJ.DingZ.JiangF.2018bInduction and characterization of tetraploids from seeds of Bletilla striata (Thunb.) Reichb.fBioMed Res. Intl.Article ID 3246398doi: 10.1155/2018/3246398

    • Search Google Scholar
    • Export Citation
  • LinH.CarvalhoP.D.KhoD.TaiC.Y.PierreP.FinkG.R.PellmanD.2001Polyploids require Bik1 for kinetochore-microtubule attachmentJ. Cell Biol.15511731184

    • Search Google Scholar
    • Export Citation
  • LiuQ.ChenJ.CorlettR.T.FanX.YuD.YangH.GaoJ.2015Orchid conservation in the biodiversity hotspot of southwestern ChinaConserv. Biol.2915631572

    • Search Google Scholar
    • Export Citation
  • LuoD.QuC.LinG.S.ZhangZ.B.XieJ.H.ChenH.B.LiangJ.L.LiC.L.WangH.F.SuZ.R.2017Character and laxative activity of polysaccharides isolated from Dendrobium officinaleJ. Funct. Foods34106117

    • Search Google Scholar
    • Export Citation
  • MajdiM.KarimzadehG.MalboobiM.A.2014Spatial and developmental expression of key genes of terpene biosynthesis in feverfew (Tanacetum parthenium)Biol. Plant.58379384

    • Search Google Scholar
    • Export Citation
  • McConnellJ.KamemotoH.1993Morphology and meiotic behavior of three Dendrobium Amphidiploids and their diploid counterpartsHortscience28935937

    • Search Google Scholar
    • Export Citation
  • MishraM.K.1997Stomatal characteristics at different ploidy levels in Coffea LAnn. Bot.80689692

  • MorelG.M.1960Producing virus-free cymbidiumsAmer. Orchid Soc. Bull.29495497

  • MurashigeT.SkoogF.1962A revised medium for rapid growth and bioassays with tobacco tissue culturesPhysiol. Plant.15473497

  • ØvrebøJ.I.EdgarB.A.2018Polyploidy in tissue homeostasis and regenerationDevelopment145116

  • OsbornT.C.Chris-PiresJ.BirchlerJ.A.AugerD.L.ChenZ.F.LeeH.ComaiL.MadlungA.DoergeR.W.ColotV.MartienssenR.A.2003Understanding mechanisms of novel gene expression in polyploidsTrends Genet.19141147

    • Search Google Scholar
    • Export Citation
  • QianX.WangC.OuyangT.TianM.2014In vitro flowing and fruiting in culture of Dendrobium officinale Kimura et Migo. (Orchidaceae)Pak. J. Bot.4618771882

    • Search Google Scholar
    • Export Citation
  • SalmaU.KunduS.MandalN.2017Artificial polyploidy in medicinal plants: Advancement in the last two decades and impending prospectsJ. Crop Sci. Biotechnol.20919

    • Search Google Scholar
    • Export Citation
  • SattlerM.C.CarvalhoC.R.ClarindoW.R.2016The polyploidy and its key role in plant breedingPlanta243281296

  • SongT.H.ChenX.X.TangS.C.W.HoJ.C.M.LaoL.X.NgT.B.2016Dendrobium officinale polysaccharides ameliorated pulmonary function while inhibiting mucin-5AC and stimulating aquaporin-5 expressionJ. Funct. Foods21359371

    • Search Google Scholar
    • Export Citation
  • StoddardE.M.1965Identifying plants by epidermal characters. Conn. Agr. Exp. Stat. Circ. 227

  • SunQ.SunH.BellR.L.LiH.XinL.2011Variation of phenotype, ploidy level and organogenic potential of in vitro regenerated polyploids of Pyrus communisPlant Cell Tissue Organ Cult.107131140

    • Search Google Scholar
    • Export Citation
  • TalebiS.F.SaharkhizM.J.KermaniM.J.SharafiY.Raouf FardF.2017Effect of different antimitotic agents on polyploid induction of anise hyssop (Agastache foeniculum L.)Caryologia70184193

    • Search Google Scholar
    • Export Citation
  • TangH.X.ZhaoT.W.ShengY.J.ZhengT.FuL.Z.ZhangY.S.2017Dendrobium officinale Kimura et migo: A review on its ethnopharmacology, phytochemistry, pharmacology, and industrializationEvid. Based Complement. Altern. Med.Article ID 7436259doi: 10.1155/2017/7436259

    • Search Google Scholar
    • Export Citation
  • TantasawatP.KhairumA.ChaowisetW.WannajindapornW.2012Pronamide-induced polyploidy in rhynchostylis and DendrobiumActa Hort.937615620

  • Teixeira da SilvaJ.A.CardosoJ.C.Dobra’nszkiJ.ZengS.2015Dendrobium micropropagation: A reviewPlant Cell Rep.34671704

  • Teixeira da SilvaJ.A.GiangD.T.DobránszkiJ.ZengS.J.TanakaM.2014aPloidy analysis of Cymbidium, Phalaenopsis, Dendrobium and Paphiopedillum (Orchidaceae), and Spathiphyllum and Syngonium (Araceae)Biologia69750755

    • Search Google Scholar
    • Export Citation
  • Teixeira da SilvaJ.A.JinX.DobránszkiJ.LuJ.WangH.ZotzG.CardosoJ.C.ZengS.2016Advances in Dendrobium molecular research: Applications in genetic variation, identification and breedingMol. Phylogenet. Evol.95196216

    • Search Google Scholar
    • Export Citation
  • Teixeira da SilvaJ.A.ZengS.GaldianoR.F.JrDobra’nszkiJ.CardosoJ.C.VendrameW.A.2014bIn vitro conservation of Dendrobium germplasmPlant Cell Rep.3314131423

    • Search Google Scholar
    • Export Citation
  • TuH.Y.ZhangA.L.XiaoW.LinY.R.ShiJ.H.WuY.W.WuS.T.ZhongC.H.MoS.X.2018Induction and identification of tetraploid Hedychium coronarium through thin cell layer culturePlant Cell Tissue Organ Cult.135395406

    • Search Google Scholar
    • Export Citation
  • VichiatoM.R.M.VichiatoM.PasqualM.CastroD.M.DutraL.F.2007Tetraploidy induction and identification in Dendrobium nobile Lindl (Orchidaceae)Rev. Ciênc. Agron. Fortaleza38385390

    • Search Google Scholar
    • Export Citation
  • VichiatoM.R.M.VichiatoM.PasqualM.RodriguesF.A.CastroD.M.2014Morphological effects of induced polyploidy in Dendrobium nobile Lindl. (Orchidaceae)Crop Breed. Appl. Biotechnol.14154159

    • Search Google Scholar
    • Export Citation
  • WangA.H.WuQ.Q.YangL.XuH.J.ChenZ.L.2017Study on polyploid of Dendrobium ochreatum induced by colchicine. J. Southwest Univ. (Natural Science Ed.). 39:55–60 (In Chinese with English abstract)

  • WangK.P.WangH.X.LiuY.G.ShuiW.Z.WangJ.F.CaoP.WangH.J.YouR.X.ZhangY.2018Dendrobium officinale polysaccharide attenuates type 2 diabetes mellitus via the regulation of PI3K/Akt-mediated glycogen synthesis and glucose metabolismJ. Funct. Foods40261271

    • Search Google Scholar
    • Export Citation
  • XieL.ZhouS.S.WangM.G.ZengR.Z.GuoH.R.ZhangZ.S.2017Creation and micropropagation of polyploids in Cymbidium hybridumActa Hort.16107113

    • Search Google Scholar
    • Export Citation
  • XieS.Z.LiuB.ZhangD.D.ZhaX.Q.PanL.H.LuoJ.P.2016Intestinal immunomodulating activity and structural characterization of a new polysaccharide from stems of Dendrobium officinaleFood Funct.727892799

    • Search Google Scholar
    • Export Citation
  • XuJ.HanQ.B.LiS.L.ChenX.J.WangX.N.ZhaoZ.Z.ChenH.B.2013Chemistry, bioactivity and quality control of Dendrobium, a commonly used tonic herb in traditional Chinese medicinePhytochem. Rev.12341367

    • Search Google Scholar
    • Export Citation
  • YangL.ShiS.XiangZ.X.2013In vitro induction, identification and physiological characteristics of autotetraploid Dendrobium officinaleActa Bot Boreal.-Occident.3321892193(In Chinese with English abstract)

    • Search Google Scholar
    • Export Citation
  • YuanZ.CongG.ZhangJ.2014Effects of exogenous salicylic acid on polysaccharides production of Dendrobium officinaleS. Afr. J. Bot.957884

    • Search Google Scholar
    • Export Citation
  • YuanZ.Q.ZhangJ.Y.LiuT.2017Enhancement of polysaccharide accumulation in Dendrobium officinale by exogenously applied methyl jasmonateBiol. Plant.61438444

    • Search Google Scholar
    • Export Citation
  • Yun-SooK.Eun-JooH.HosakatteN.M.Kee-YoeupP.2004Effect of polyploidy induction on biomass and ginsenoside accumulations in adventitious roots of ginsengJ. Plant Biol.47356360

    • Search Google Scholar
    • Export Citation
  • ZhanZ.G.ChengX.U.2011Study on colchiploid of Dendrobium officinale induced by colchicinesJ. Zhejiang Univ. (Sci. Ed.).38321325(In Chinese with English abstract)

    • Search Google Scholar
    • Export Citation
All Time Past Year Past 30 Days
Abstract Views 284 284 154
Full Text Views 23 23 9
PDF Downloads 17 17 7