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Micropropagation, encapsulation, and conservation of Decalepis salicifolia, a vanillin isomer containing medicinal and aromatic plant

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Abstract

An efficient in vitro propagation and synthetic seed production protocol was established for the conservation of Decalepis salicifolia (Bedd. ex Hook.f.) Venter, an endemic and critically endangered ethnomedicinal species. Murashige and Skoog (MS) media supplemented with 2.2 μM BAP and 5.7 μM IAA produced shoot tip proliferation with an average shoot length of 5.81 ± 0.03 cm in 3 wk. Axillary bud proliferation was found to be best in 11.1 μM BAP with an average of 3.5 ± 0.06 shoots per explant. Shoot tip and nodal explants were encapsulated in sodium alginate and their regeneration was achieved following storage at 4°C up to 12 wk. Successful rooting was obtained in modified MS medium with low nitrate and high sucrose concentration. Quarter strength MS media containing 2.5 mM each of NH4NO3 and KNO3 along with 8% sucrose produced an average number of 12.4 ± 1.18 roots with an average length of 4.3 ± 0.08 cm. The in vitro derived rooted plants were successfully hardened (92.8%) and established in field with 100.0% survival rate. Inter-simple sequence repeat (ISSR) markers proved the genetic fidelity among the in vitro grown plant showing 99.5% similarity with the mother plant. Micropropagation derived acclimatized plants produced 2-hydroxy-4-methoxybenzaldehyde in amounts similar to seed-derived field-grown plants of the same age. The in vitro propagation protocol developed for D. salicifolia can be utilized to reduce exploitation pressure from their natural habitats and augment ecorestoration, conservation, and cultivation of this critically endangered and industrially valuable plant. The synthetic seeds technique will serve as propagules in ex situ gene banks, as well as supports cost-effective germplasm conservation.

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References

  • Ahmad N, Khan MI, Ahmed S, Javed SB, Faisal M, Anis M, Rehman S, Umair SM (2013) Change in total phenolic content and antibacterial activity in regenerants of Vitex negundo L. Acta Physiol Plant 35:791–800

    CAS  Google Scholar 

  • Ahmad Z, Shahzad A, Sharma S (2018a) Enhanced multiplication and improved ex vitro acclimatization of Decalepis arayalpathra. Biol Plant 62:1–10

    CAS  Google Scholar 

  • Ahmad Z, Shahzad A, Sharma S, Parveen S (2018b) Ex vitro rescue, physiochemical evaluation, secondary metabolite production and assessment of genetic stability using DNA based molecular markers in regenerated plants of Decalepis salicifolia (Bedd.Ex Hook.F.) Venter. Plant Cell Tiss Org Cult 132:497–510

  • Ahmed R, Anis M (2014) Rapid in vitro propagation system through shoot tip cultures of Vitex trifolia L.—an important multipurpose plant of the Pacific traditional medicine. Physiol Mol Biol Plants 20:385–392

    CAS  PubMed  PubMed Central  Google Scholar 

  • Al-Khalifah NS, Hadi S, Khan F (2005) Influence of sucrose concentration on in vitro growth of five rose (Rosa hybrida L.) cultivars. Plant Tiss Cult 15:43–49

  • Anitha S, Pullaiah T (2002) In vitro propagation of Decalepis hamiltonii. J Trop Med Plants 3:227–232

    Google Scholar 

  • Borner A (2006) Preservation of plant genetic resources in the biotechnology era. Biotechnol J 1:1393–1404

    PubMed  Google Scholar 

  • Calamar A, De Klerk GJ (2002) Effect of sucrose on adventitious root regeneration in apple. Plant Cell Tiss Org Cult 70:207–212

  • Carpentier SC, Vertommen A, Swennen R, Witters E, Fortes C, Souza MT Jr, Panis B (2010) Sugar-mediated acclimation: the importance of sucrose metabolism in meristems. J Proteome Res 9:5038–5046

    CAS  PubMed  Google Scholar 

  • Chen X, Qu Y, Sheng L, Liu J, Huang H, Xu L (2014) A simple method suitable to study de novo root organogenesis. Front Plant Sci 5:208

    PubMed  PubMed Central  Google Scholar 

  • Danso KE, Ford-Lloyd BV (2003) Encapsulation of nodal cuttings and shoot tips for storage and exchange of cassava germplasm. Plant Cell Rep 21:718–725

    CAS  PubMed  Google Scholar 

  • Eliasson L, Bertell G, Bolander E (1989) Inhibitory action of auxin on root elongation not mediated by ethylene. Plant Physiol 91:310–314

    CAS  PubMed  PubMed Central  Google Scholar 

  • Faisal M, Anis M (2007) Regeneration of plants from alginate encapsulated shoots of Tylophora indica (Burm.F.) Merrill, an endangered medicinal plant. J Hortic Sci Biotechnol 82:351–354

    CAS  Google Scholar 

  • Gabryszewska EA (2015) Effect of different sucrose and nitrogen salt levels in the medium and temperature on in vitro propagation of Helleborus niger L. Acta Agrobot 68:161–171

    Google Scholar 

  • Gangaprasad A, Decruse SW, Seeni S, Nair GM (2005) Micropropagation and ecorestoration of Decalepis arayalpathra (Joseph & Chandra.) Venter—an endemic and endangered ethnomedicinal plant of Western Ghats. Indian J Biotechnol 4:265–270

    CAS  Google Scholar 

  • Gangaprasad A, Nair LG, Radhakrishnan K, Seeni S, Nair GM, Pushpangadan P (2003) Micropropagation of Utleria salicifolia endemic and endangered ethnomedicinal plant of Western Ghats. J Med Aromatic Plant Sci 25:19–24

    Google Scholar 

  • George J, Ravishankar GA, Pereira J, Divakar S (1999) Bioinsecticide from swallowroot (Decalepis hamiltonii) Wight & Arn protects food grains against insect infestation. Curr Sci 774:501–502

    Google Scholar 

  • George S, Geetha SP, Udayan PS, Tushar KV, Augustine A, Balachandran I (2007) Micropropagation and in vitro conservation of Utleria salicifolia Bedd. Ex Hook.F. (Periplocaceae)—a little known, monotypic, endemic and critically endangered medicinal plant of Western Ghats. In: XIXth Kerala Science Congress, Kannur University, Kerala

  • George S, Sebastian DP, Kavitha CV, Raghavan SC, Augustine A, Kusumanchi P (2016) In vitro antiproliferative effect of the root extracts from Decalepis hamiltonii, Hemidesmus indicus and Utleria salicifolia. Devagiri J Sci 2:52–57

    Google Scholar 

  • George S, Sulaiman CT, Balachandran I, Augustine A (2011) Decalepis salicifolia (Bedd. Ex Hook. F.) Venter (Apocynaceae)—a new source for 2-hydroxy-4-methoxybenzaldehyde. Med Plants Int J Phytomed Rel Ind 3:259–260

    Google Scholar 

  • Giridhar P, Rajasekaran T, Nagarajana S, Ravishankar GA (2004) Production of 2-hydroxy-4-methoxybenzaldehyde in roots of tissue culture raised and acclimatized plants of Decalepis hamiltonii Wight & Arn., an endangered shrub endemic to southern India and evaluation of its performance vis-à-vis plants from natural habitat. Indian J Exp Biol 42:106–110

  • Gollagunta V, Adelberg JW, Rieck J, Rajapakse N (2005) Sucrose in storage media and cultivar affects post-storage regrowth of in vitro Hosta propagules. Plant Cell Tiss Org Cult 80:191–199

  • Hung CD, Trueman SJ (2012) Alginate encapsulation of shoot tips and nodal segments for short-term storage and distribution of the eucalyptus Corymbia torelliana × C. citriodora. Acta Physiol Plant 34:117–128

    CAS  Google Scholar 

  • Hyndman SE, Hasegawa PM, Bressan RA (1981) The role of sucrose and nitrogen in adventitious root formation on cultured rose shoots. Plant Cell Tiss Org Cult 1:229–238

  • Kanchanapoom K, Sunheem S, Kanchanapoom K (2010) In vitro propagation of Adenium obesum (Forssk.) Roem. and Schult. Not Bot Hort Agrobot Cluj 38:209–213

    CAS  Google Scholar 

  • Kiran K, Sonbarse PP, Veeresh L, Shetty NP, Parvatam G (2018) Expression profile of phenylpropanoid pathway genes in Decalepis hamiltonii tuberous roots during flavour development. 3 Biotech 8:365

    PubMed  PubMed Central  Google Scholar 

  • Kolachevskaya OO, Lomin SN, Arkhipov DV, Romanov GA (2019) Auxins in potato: molecular aspects and emerging roles in tuber formation and stress resistance. Plant Cell Rep 9:1–18

    Google Scholar 

  • Kozai T (1991) Photoautotrophic micropropagation. In Vitro Cell Dev Biol Plant 27:47–51

    Google Scholar 

  • Kozlowski TT (1992) Carbohydrate sources and sinks in woody plants. Bot Rev 58:107–222

    Google Scholar 

  • Kubo I, Kinst-Hori I (1999) 2-Hydroxy-4-methoxybenzaldehyde: a potent tyrosinase inhibitor from African medicinal plants. Planta Med 65:19–22

    CAS  PubMed  Google Scholar 

  • Kulus D (2019) Managing plant genetic resources using low and ultra-low temperature storage: a case study of tomato. Biodivers Conserv 28:1003–1027

    Google Scholar 

  • Kumar GP, Sivakumar S, Siva G, Vigneswaran M, Kumar TS, Jayabalan N (2016) Silver nitrate promotes high-frequency multiple shoot regeneration in cotton (Gossypium hirsutum L.) by inhibiting ethylene production and phenolic secretion. In Vitro Cell Dev Biol - Plant 52:408–418

  • Lata H, Chandra S, Khan IA, ElSohly MA (2009) Propagation through alginate encapsulation of axillary buds of Cannabis sativa L.—an important medicinal plant. Physiol Mol Biol Plants 15:79–86

    CAS  PubMed  PubMed Central  Google Scholar 

  • Ling WX, Zhong Z (2012) Seasonal variation in rooting of the cutting from tetraploid locust in relation to nutrients and endogenous plant hormones of the shoot. Turk J Agric For 36:257–266

    Google Scholar 

  • MacGregor DR, Deak KI, Ingram PA, Malamy JE (2008) Root system architecture in Arabidopsis grown in culture is regulated by sucrose uptake in the aerial tissues. Plant Cell 20:2643–2660

    CAS  PubMed  PubMed Central  Google Scholar 

  • Mahanga GM, Akenga TO, Lwande W, Ndiege IO (2005) 2-Hydroxy-4-methoxybenzaldehyde: larvicidal structure–activity studies. Bull Chem Soc Ethiop 19:61–68

    CAS  Google Scholar 

  • Makowczyńska J, Sliwinska E, Kalemba D, Piątczak E, Wysokińska H (2016) In vitro propagation, DNA content and essential oil composition of Teucrium scorodonia L. ssp. scorodonia. Plant Cell Tiss Org Cult 127:1–3

  • Manzanera JA, Pardos JA (1990) Micropropagation of juvenile and adult Quercus suber L. Plant Cell Tiss Org Cult 21:1–8

  • Matysiak N, Gabryszewska E (2016) The effect of in vitro culture conditions on the pattern of maximum photochemical efficiency of photosystem II during acclimatisation of Helleborus niger plantlets to ex vitro conditions. Plant Cell Tiss Org Cult 125:585–593

  • Maya C, Kumar NSP (2016) Enhancement of phytochemicals in accessions of Decalepis hamiltonii Wight & Arn. Int J Eng Sci 5:32–36

    Google Scholar 

  • Mazri MA, Meziani R, El Fadile J, Ezzinbi AE (2016) Optimization of medium composition for in vitro shoot proliferation and growth of date palm cv. Mejhoul. 3. Biotech 6:111–121

    Google Scholar 

  • Mishra J, Singh M, Palni LM, Nandi SK (2011) Assessment of genetic fidelity of encapsulated microshoots of Picrorhiza kurrooa. Plant Cell Tiss Org Cult 104:181–186

  • Mishra P, Kumar A, Sivaraman G, Shukla AK, Kaliamoorthy R, Slater A, Velusamy S (2017) Character-based DNA barcoding for authentication and conservation of IUCN red listed threatened species of genus Decalepis (Apocynaceae). Sci Rep 7:14910

    PubMed  PubMed Central  Google Scholar 

  • Mohana DC, Raveesha KA (2010) Antimycotic, antibiodeteriorative and antiaflatoxigenic potency of 2-hydroxy-4-methoxybenzaldehyde isolated from Decalepis hamiltonii on fungi causing biodeterioration of maize and sorghum grains. J Mycol Plant Pathol 40:197–206

    CAS  Google Scholar 

  • Molsaghi M, Moieni A, Kahrizi D (2014) Efficient protocol for rapid Aloe vera micropropagation. Pharm Biol 52:735–739

    CAS  PubMed  Google Scholar 

  • Mukonyi KW, Ndiege IO (2001) 2-Hydroxy-4-methoxybenzaldehyde: aromatic taste modifying compound from Mondia whytei Skeels. Bull Chem Soc Ethiop 15:137–142

    CAS  Google Scholar 

  • Muller D, Leyser O (2011) Auxin, cytokinin and the control of shoot branching. Ann Bot 107:1203–1212

    PubMed  PubMed Central  Google Scholar 

  • Murashige T (1974) Plant propagation through tissue cultures. Annu Rev Plant Physiol 25:135–166

    CAS  Google Scholar 

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

    CAS  Google Scholar 

  • Ngomuo M, Mneney E, Ndakidemi P (2013) The effects of auxins and cytokinin on growth and development of (Musa sp.) var.“Yangambi” explants in tissue culture. Am J Plant Sci 4:2174

    Google Scholar 

  • Otiende M, Nyabundi JO, Ngamau K, Opala P (2017) Effects of cutting position of rose rootstock cultivars on rooting and its relationship with mineral content and endogenous carbohydrates. Sci Hortic 225:204–212

    CAS  Google Scholar 

  • Park SY, Kim YW, Moon HK, Murthy HN, Choi YH, Cho HM (2008) Micropropagation of Salix pseudolasiogyne from nodal explants. Plant Cell Tiss Org Cult 93:341

  • Pawlicki N, Welander M (1995) Influence of carbohydrate source, auxin concentration and time of exposure on adventitious rooting of the apple rootstock Jork 9. Plant Sci 106:167–176

    CAS  Google Scholar 

  • Pullman GS, Bai K, Hane M, Ruland D, Cruse-Sanders JM, Boyd RS, Johnson S (2019) Seed cryopreservation and micropropagation of the federally threatened species, Price’s potato-bean (Apios priceana BL Robins.). In Vitro Cell Dev Biol - Plant 55:558–568

  • Radhakrishnan K, Pandurangan AG, Pushpangadan P (1998) Utleria salicifolia—a new ethnobotanical record from Kerala, India. Fitoterapia 69:403–405

    Google Scholar 

  • Rai MK, Asthana P, Jaiswal VS, Jaiswal U (2010) Biotechnological advances in guava (Psidium guajava L.): recent developments and prospects for further research. Trees 24:1–12

    CAS  Google Scholar 

  • Rao CV, Ojha SK, Radhakrishnan K, Govindarajan R, Rastogi S, Mehrotra S, Pushpangadan P (2004) Antiulcer activity of Utleria salicifolia rhizome extract. J Ethnopharmacol 91:243–249

    PubMed  Google Scholar 

  • Ravikumar K, Sankar RV, Prabhakaran V (2001) Utleria salicifolia Bedd. Ex Hook.F. (Periplocaceae) a monotypic, endemic and critically endangered, red listed medicinal plant relocated from its type locality. My Forest 37:495–498

    Google Scholar 

  • Ray A, Bhattacharyaa S (2008) Storage and plant regeneration from encapsulated shoot tips of Rauvolfia serpentina—an effective way of conservation and mass propagation. S Afr J Bot 74:776–779

    CAS  Google Scholar 

  • Remya B, Latha PG, Caroline ELM, Suja SR, Shyamal S, Shine VJ, Sini S, Anuja GI, Shikha P, Rajasekharan S (2010) Hepatoprotective studies on Utleria salicifolia Bedd. Medicinal Plants- Int J Phytomed Rel Ind 2:110–119

    Google Scholar 

  • Romano A, Noronha C, Martins-Loucao MA (1995) Role of carbohydrates in micropropagation of cork oak. Plant Cell Tiss Org Cult 40:159–167

  • Roycewicz P, Malamy JE (2012) Dissecting the effects of nitrate, sucrose and osmotic potential on Arabidopsis root and shoot system growth in laboratory assays. Philos Trans R Soc B 367:1489–1500

    CAS  Google Scholar 

  • Saeed T, Shahzad A, Ahmad N, Parveen S (2018) High frequency conversion of non-embryogenic synseeds and assessment of genetic stability through ISSR markers in Gymnema sylvestre. Plant Cell Tiss Org Cult 134:163–168

  • Saha S, Sengupta C, Ghosh P (2015) Encapsulation, short-term storage, conservation and molecular analysis to assess genetic stability in alginate-encapsulated microshoots of Ocimum kilimandscharicum Guerke. Plant Cell Tiss Org Cult 120:519–530

  • Sharma S, Shahzad A (2012) Encapsulation technology for short-term storage and conservation of a woody climber, Decalepis hamiltonii Wight and Arn. Plant Cell Tiss Org Cult 111:191–198

  • Sharma S, Shahzad A, Ahmad A, Anjum L (2014) In vitro propagation and the acclimatization effect on the synthesis of 2-hydroxy-4-methoxy benzaldehyde in Decalepis hamiltonii Wight and Arn. Acta Physiol Plant 36:2331–2344

    CAS  Google Scholar 

  • Silue N, Kone T, Soumahoro AB, Kone M (2016) In vitro shoot tip multiplication of bambara groundnut [Vigna subterranea (L.) Verdc.]. Plant Cell Tiss Org Cult 127:603–611

  • Sima BD, Desjardins Y (2001) Sucrose enhances phosphoenolpyruvate carboxylase activity of in vitro Solanum tuberosum L. under non-limiting nitrogen conditions. In Vitro Cell Dev Biol - Plant 37:480–489

  • Singh AK, Sharma M, Varshney R, Agarwal SS, Bansal KC (2006) Plant regeneration from alginate-encapsulated shoot tips of Phyllanthus amarus Schum and Thonn, a medicinally important plant species. In Vitro Cell Dev Biol - Plant 42:109–113

  • Singh S, Purohit VK, Prasad P, Nautiyal AR (2015) Micropropagation of Valeriana wallichii DC. (Indian valerian) through nodes. Indian J Biotechnol 14:127–130

    Google Scholar 

  • Siva G, Sivakumar S, Kumar GP, Vigneswaran M, Vinoth S, Selvan AM, Ahamed AP, Manivannan K, Kumar RR, Thajuddin N, Kumar TS (2015) Optimization of elicitation condition with jasmonic acid, characterization and antimicrobial activity of psoralen from direct regenerated plants of Psoralea corylifolia L. Biocatal Agric Biotechnol 4:624–631

    Google Scholar 

  • Sudha CG, Krishnan PN, Pushpangadan P, Seeni S (2005) In vitro propagation of Decalepis arayalpathra, a critically endangered ethnomedicinal plant. In Vitro Cell Dev Biol - Plant 41:648–654

  • Thorpe T (1982) Carbohydrate utilization and metabolism. In: Bonga JM, Durzan DJ (eds) Tissue culture in forestry. Martinus Nijhoff Publishers, London, pp 325–368

    Google Scholar 

  • Van Le QU, Samson GU, Desjardins Y (2001) Opposite effects of exogenous sucrose on growth, photosynthesis and carbon metabolism of in vitro plantlets of tomato (L. esculentum Mill.) grown under two levels of irradiances and CO2 concentration. J Plant Physiol 158:599–605

    CAS  Google Scholar 

  • Vargas VML, Flota FV (2006) Plant cell culture protocols. In: Methods in molecular biology, 2nd edn. Springer Science & Business Media, Berlin, p 133

    Google Scholar 

  • Ved D, Saha D, Ravikumar K, Haridasan K (2015) Utleria salicifolia. The IUCN red list of threatened species 2015: e.T50130765A50131450 https://doi.org/10.2305/IUCN.UK.2015-4.RLTS.T50130765A50131450.en

  • Venkatachalam P, Jinu U, Sangeetha P, Geetha N, Sahi SV (2018) High frequency plant regeneration from cotyledonary node explants of Cucumis sativus L. cultivar ‘green long’ via adventitious shoot organogenesis and assessment of genetic fidelity by RAPD-PCR technology. 3. Biotech 8:60

    Google Scholar 

  • Wojtania A, Skrzypek E, Marasek-Ciolakowska A (2019) Soluble sugar, starch and phenolic status during rooting of easy- and difficult-to-root magnolia cultivars. Plant Cell Tiss Org Cult 136:499–510

  • Woodward AW, Bartel B (2005) Auxin: regulation, action, and interaction. Ann Bot 95:707–735

    CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang H, Forde BG (2000) Regulation of Arabidopsis root development by nitrate availability. J Exp Bot 1:51–59

    Google Scholar 

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Acknowledgments

Director, CSIR-CIMAP, Lucknow, India is gratefully acknowledged for providing the laboratory facilities (CIMAP Institutional Communication No. CIMAP/PUB/2018/9). Financial assistance to the author V.R. through INSPIRE fellowship (IF150398) from Department of Science and Technology (DST), New Delhi, India is gratefully acknowledged. Council of Scientific and Industrial Research (CSIR), New Delhi is acknowledged for funding the work through Phytopharmaceutical Mission Project (HCP-010).

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  1. Plant Biology and Systematics, CSIR—Central Institute of Medicinal and Aromatic Plants (CIMAP), Research Center, Bengaluru, India

    Vereena Rodrigues, Amit Kumar, Sivaraman Gokul & Velusamy Sundaresan

  2. Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, India

    Vereena Rodrigues, Amit Kumar & Velusamy Sundaresan

  3. Chemical Sciences Division, CSIR—Central Institute of Medicinal and Aromatic Plants (CIMAP), Lucknow, India

    Ram S. Verma

  4. Plant Biotechnology Division, CSIR—Central Institute of Medicinal and Aromatic Plants (CIMAP), Lucknow, India

    Laiq ur Rahman

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  1. Vereena Rodrigues
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Rodrigues, V., Kumar, A., Gokul, S. et al. Micropropagation, encapsulation, and conservation of Decalepis salicifolia, a vanillin isomer containing medicinal and aromatic plant. In Vitro Cell.Dev.Biol.-Plant 56, 526–537 (2020). https://doi.org/10.1007/s11627-020-10066-z

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