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Germplasm conservation of economically important medicinal plant Nyctanthes arbor-tristis L. through encapsulation technique and maintenance under slow growth condition

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Abstract

An efficient encapsulation and slow growth conservation protocol was developed for Nyctanthes arbor-tristis L. an antiviral medicinal plant of the family Oleaceae. A gel matrix with 3% sodium alginate and 100 mM calcium chloride (CaCl2⋅2H2O) was found best for the encapsulation of nodal segments. The viability and shoot development potential of encapsulated nodal segments was optimized. Encapsulated nodal segments stored at 4 °C and 24 °C remained viable for up to 90 days and showed shoot development potential 42.89 ± 6.04% and 33.53 ± 7.15% respectively. Nodal segments maintained under slow growth conditions up to 180 days on one-eighth strength Murashige and Skoog (MS) medium supplemented with 0.5% sucrose was suitable for satisfactory viability (40.28 ± 2.04%), while further addition of 0.5 mg/l abscisic acid supported 40.36 ± 1.01% viability of the nodal segments. The best rooting response was achieved on half-strength MS medium supplemented with 4 mg/l indole-3-acetic acid. The field survival of rooted plants was 65%. The clonal fidelity of in-vitro derived plantlets was studied with start codon targeted primer profile, which showed the same banding mobility patterns as the source parent plant. The maximum banding profile was monomorphic and consistent, confirming the clonal stability of regenerated plants. The method developed will permit the in-vitro conservation of this species and facilitate an easy exchange of plant germplasm.

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Fig. 1
Fig. 2
Fig. 3

source parent mother plant of N. arbor-tristis L., and numeric number 1 to 5 denotes bands of tissue culture recovered acclimatized hardened plants

Fig. 4

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Abbreviations

ABA:

Abscisic acid

BAP:

6-Benzylaminopurine

IAA:

Indole-3-acetic acid

MS:

Murashige and Skoog

SA:

Sodium alginate

SCoT:

Start codon targeted

References

  • Agrawal J, Pal A (2013) Nyctanthes arbor-tristis Linn: a critical ethnopharmacological review. J Ethnopharmacol 146:645–658

    CAS  PubMed  Google Scholar 

  • Ahmad N, Anis M (2010) Direct plant regeneration from encapsulated nodal segments of Vitex negundo. Biol Plant 54:748–752

    Google Scholar 

  • Ahmad M, Anjum MA (2010) In-vitro storage of some pear genotype with the minimal growth technique. Turk J Agric for 34:25–32

    Google Scholar 

  • Ahmad N, Faisal M, Fatima N, Anis M (2012) Encapsulation of microcuttings for propagation and short-term preservation in Ruta graveolens: a plant with high medicinal value. Acta Physiol Plant 34:2303–2310

    CAS  Google Scholar 

  • Baskaran P, Kumari A, Staden JV (2018) In-vitro propagation via organogenesis and synthetic seeds of Urginea altissima (L.f.) Baker: a threatened medicinal plant. 3 Biotech 8:18. https://doi.org/10.1007/s13205-017-1028-7

    Article  PubMed  Google Scholar 

  • Bhattacharyya P, Kumar V, Staden JV (2018) In-vitro encapsulation based short term storage and assessment of genetic homogeneity in regenerated Ansellia africana (Leopard orchid) using gene targeted molecular markers. Plant Cell Tissue Organ Cult 133:299–310

    CAS  Google Scholar 

  • Chand S, Singh AK (2004) Plant regeneration from encapsulated nodal segments of Dalbergia sissoo Roxb.: a timber-yielding leguminous tree. J Plant Physiol 161:237–243

    CAS  PubMed  Google Scholar 

  • Cid B, Carvalho LPY, Luiz LCB (2008) Importance of abscisic acid (ABA) in the in-vitro conservation of cassava (Manihot esculenta Crantz). Chil J Agri Res 68:304–308

    Google Scholar 

  • Collard BCY, Mackill DJ (2009) Start codon targeted (SCoT) polymorphism: a simple noval DNA marker technique for generating gene –targeted marker in the plants. Plant Mol Biol Rep 27:86–93

    CAS  Google Scholar 

  • Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15

    Google Scholar 

  • El-Bahr MK, Abd El-Hamid A, Matter MA, Shaltout A, Bekheet SA, El-Ashry AA (2016) In- vitro conservation of embryogenic cultures of date palm using osmotic mediated growth agents. J Gen Eng Biotechnol 14:363–370

    CAS  Google Scholar 

  • Gupta P, Bajpai SK, Chandra K, Singh KL, Tandon JS (2005) Anti-viral profile of Nyctanthes arbor-tristis L. against encephalitis causing viruses. Indian J Exp Biol 43:1156–1160

    CAS  PubMed  Google Scholar 

  • Gupta AK, Harish RMK, Phulwaria M, Agarwal T, Shekhawat NS (2014) In-vitro propagation, encapsulation, and genetic fidelity analysis of Terminalia arjuna: a cardio-protective medicinal tree. Appl Biochem Biotechnol 173:1481–1494

    CAS  PubMed  Google Scholar 

  • Hatzilazarou S, Kostas S, Joachim M, Economou A (2020) Regeneration of Viburnum dentatum L from alginate-encapsulated shoot explants after short-term cold storage and assessment of genetic stability using ISSR analysis. Agronomy 10: 1660; doi:https://doi.org/10.3390/agronomy10111660.

  • Jahan AA, Anis M (2015) Retrieval of plantlets from cryopresesrved alginate buds of Nyctanthes arbor-tristis L.: an effectual approach for germplasm conservation. Int J Dev Res 5:4397–4402

    Google Scholar 

  • Jo EA, Tewari RK, Hahn EJ, Paek KY (2009) In-vitro sucrose concentration affects growth and acclimatization of Alocasia amazonica plantlets. Plant Cell Tissue Organ Cult 96:307–315

    CAS  Google Scholar 

  • Joshi K, Bahukhandi A, Parihar N, Rawat S, Bhatt ID (2019) Promoting conservation of threatened plant species in West Himalayan region through cultivation- a participatory approach. ENVIS Bull Himal Ecol 27:45–48

    Google Scholar 

  • Kalia RK, Rai MK, Kalia S, Singh R, Dhawan AK (2011) Microsatellite markers: an overview of the recent progress in plants. Euphytica 177:309–334

    CAS  Google Scholar 

  • Kaminska M, Skrzypek E, Wilmowicz E, Tretyn A, Trejgell A (2016) Effect of light conditions and ABA on cold storage and post-storage propagation of Taraxacum pieninicum. Plant Cell Tissue Organ Cult 127:25–34

    CAS  Google Scholar 

  • Kaminska M, Gołębiewski M, Tretyn A, Trejgell A (2018) Efficient long-term conservation of Taraxacum pieninicum synthetic seeds in slow growth conditions. Plant Cell Tissue Organ Cult 132:469–478

    CAS  Google Scholar 

  • Ma XM, Wu CF, Wang GR (2011) Application of artificial seeds in rapid multiplication of Pseudostellaria heterophylla. Afr J Biotechnol 10:15744–15748

    CAS  Google Scholar 

  • Micheli M, Hafiz IA, Standardi A (2007) Encapsulation of In-vitro derived explants of Olive (Olea europaea L.cv.Moraiolo) II. Effect of storage on capsule and derived shoot performance. Sci Horticult 113:286–292

    Google Scholar 

  • Mishra AK, Upadhyay R, Chaurasia JK, Tiwari KN (2016) Comparative antioxidant study in different flower extracts of Nyctanthes arbor-tristis L. (Oleaceae): an important medicinal plant. Braz J Bot 39:813–820

    Google Scholar 

  • Mishra AK, Upadhyay P, Dixit J, Kumar P, Tiwari KN, Mishra SK, More R, Singh J (2018) Ameliorative activity of ethanolic flower extract of Nyctanthes arbor-tristis L. against scopolamine-induced amnestic effect and profiling of active compounds using gas chromatography–Mass spectrometry and ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry. Pharmacog Mag 14:596–604

    CAS  Google Scholar 

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

    CAS  Google Scholar 

  • Nasiruddin M, Rafiul Islam AKM (2018) In-vitro slow-growth conservation for two genotypes of Solanum tuberosum l. Bangladesh J Bot 47:369–380

    Google Scholar 

  • Parveen S, Shahzad A (2014) Encapsulation of nodal segments of Cassia angustifolia Vahl. for short-term storage and germplasm exchange. Acta Physiol Plant 36:635–640

    CAS  Google Scholar 

  • Peng X, Ji Q, Wu H, Li Y (2015) Slow-growth conservation and clonal fidelity of Tetrastigma hemsleyanum microplants. In Vitro Cell Dev Biol Plant 51:463–470

    CAS  Google Scholar 

  • Prakash AV, Nair DS, Alex S, Soni KB, Viji MM, Regunath BR (2018) Calcium alginate encapsulated synthetic seed production in Plumbago rosea L. for germplasm exchange and distribution. Physiol Mol Biol Plants 24:963–971

    CAS  PubMed  PubMed Central  Google Scholar 

  • Rai MK, Asthana P, Singh SK, Jaiswal VS, Jaiswal U (2009) The encapsulation technology in fruit plants: a review. Biotechnol Adv 27:671–679

    PubMed  Google Scholar 

  • Raizada U, Nangia A (1989) Cyclic nocturnal opening and abcission of Nyctanthes flowers with a note on visiting thrips species. Curr Sci 58:93–96

    Google Scholar 

  • Rathore NS, Rai MK, Phulwaria M, Rathore N, Shekhawat NS (2014) Genetic stability in micropropagated Cleome gynandra revealed by SCoT analysis. Acta Physiol Plant 36:555–559

    CAS  Google Scholar 

  • Reddy MC, Murthy KSR, Pullaiah T (2012) Synthetic seeds: a review in agriculture and forestry. Afr J Biotechnol 11:14254–14275

    Google Scholar 

  • Rihan HZ, Kareem F, El-Mahrouk ME, Fuller MP (2017) Artificial seeds (principle, aspects and applications). Agronomy 7: 71; doi: https://doi.org/10.3390/agronomy7040071.

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

    Google Scholar 

  • Sagar R, Singh JS (2004) Local plant species depletion in a tropical dry deciduous forest of northern India. Environ Conserv 31:55–62

    Google Scholar 

  • Sah SK, Reddy KR, Li J (2016) Abscisic acid and abiotic stress tolerance in crop plants. Front Plant Sci 7:571. https://doi.org/10.3389/fpls.2016.00571

    Article  PubMed  PubMed Central  Google Scholar 

  • Seth S, Rath SC, Rout GR, Panigrahi J (2017) Somatic embryogenesis in Abutilon indicum (L.) Sweet and assessment of genetic homogeneity using SCoT markers. Plant Biosyst 151:704–714

    Google Scholar 

  • Shilpha J, Pandian S, Largia MJV, Sohn SI, Ramesh M (2021) Short-term storage of Solanum trilobatum L. synthetic seeds and evaluation of genetic homogeneity using SCoT markers. Plant Biotechnol Rep 15:651–661

    Google Scholar 

  • Singh AK, Varshney R, Sharma M, Agarwal SS, Bansal KC (2006) Regeneration of plants from alginate-encapsulated shoot tips of Withania somnifera (L.) Dunal, a medicinally important plant species. J Plant Physiol 163:220–223

    CAS  PubMed  Google Scholar 

  • Singh SK, Rai MK, Asthana P, Sahoo L (2010) Alginate-encapsulation of nodal segments for propagation, short-term conservation and germplasm exchange and distribution of Eclipta alba (L.). Acta Physiol Plant 32:607–610

    Google Scholar 

  • Subhashini DP, Satyanarayana B, Arundhati A, Raghava Rao T (2014) Plant regeneration from encapsulated nodal segments of Sterculia urens Roxb. an endangered gum-yielding tree. J Crop Sci Biotechnol 17:21–25

    Google Scholar 

  • Tavazza R, Rey NA, Papacchioli V, Pagnotta MA (2015) A validated slow-growth in-vitro conservation protocol for globe artichoke germplasm: a cost-effective tool to preserve from wild to elite genotypes. Sci Horticult 197:135–143

    Google Scholar 

  • Teixeira da Silva JA, Hossain MM, Sharma M, Dobranszki J, Cardoso JC, Songjun Z (2017) Acclimatization of in-vitro-derived Dendrobium. Hort Plant J 3:110–124

    Google Scholar 

  • Tyagi RK, Goswami R, Sanayaima R, Singh R, Tandon R, Agrawal A (2009) Micropropagation and slow growth conservation of cardamom (Elettaria cardamomum, Maton). In Vitro Cell Dev Biol-Plant 45:721–729

    Google Scholar 

  • Varshney A, Anis M (2014) Synseed conception for short-term storage, germplasm exchange and potentialities of regeneration genetically stable plantlets of desert date tree (Balanites aegyptiaca Del.). Agrofor Syst 88:321–329

    Google Scholar 

  • Vasudevan V, Subramanyam K, Elayaraja D, Karthik S, Vasudevan A, Manickavasagam M (2017) Assessment of the efficacy of amino acids and polyamines on regeneration of watermelon (Citrullus lanatus Thunb.) and analysis of genetic fidelity of regenerated plants by SCoT and RAPD markers. Plant Cell Tissue Organ Cult 130:681–687

    CAS  Google Scholar 

  • Vishwakarma K, Upadhyay N, Kumar N, Yadav G, Singh J, Mishra RK, Kumar V, Verma R, Upadhyay RG, Pandey M, Sharma S (2017) Abscisic acid signaling and abiotic stress tolerance in plants: a review on current knowledge and future prospects. Front Plant Sci 8:161. https://doi.org/10.3389/fpls.2017.00161

    Article  PubMed  PubMed Central  Google Scholar 

  • Yaseen M, Ahmad T, Sablok G, Standardi A, Hafiz IA (2013) Review: role of carbon sources for in vitro plant growth and development. Mol Biol Rep 40:2837–2849

    CAS  PubMed  Google Scholar 

  • Zayova E, Nedev T, Dimitrova L (2017) In vitro storage of Stevia rebaudiana Bertoni under slow growth conditions and mass multiplication after storage. Bio Bullet 3:30–38

    Google Scholar 

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Acknowledgements

The author Awadhesh Kumar Mishra (AKM) is thankful to University Grants Commission (UGC), New Delhi, India for fellowship support throughout the study of this work. The author Kavindra Nath Tiwari acknowledges Institute of Eminence (IoE), Banaras Hindu University, Varanasi, India (Scheme 6031) for supporting the research work.

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Authors and Affiliations

  1. Department of Botany, MMV, Banaras Hindu University, Varanasi, Uttar Pradesh, 221005, India

    Awadhesh Kumar Mishra, Kavindra Nath Tiwari & Pallavi Mishra

  2. Crop Improvement Division, Indian Institute of Vegetable Research, Shahanshapur, Varanasi, 221305, Uttar Pradesh, India

    Pallavi Mishra & Shailesh Kumar Tiwari

  3. Department of Pharmaceutical Engineering and Technology, Indian Institute of Technology, Banaras Hindu University, Varanasi, Uttar Pradesh, 221005, India

    Sunil Kumar Mishra

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Contributions

Author AKM, design, writing the manuscript and completed all experimental work. PM and SKT technically help in the analysis of data. KNT and SKM, is involve in proofreading, intellectual input and finally revise and agreed to the publication of manuscript.

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Correspondence to Kavindra Nath Tiwari.

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Mishra, A.K., Tiwari, K.N., Mishra, P. et al. Germplasm conservation of economically important medicinal plant Nyctanthes arbor-tristis L. through encapsulation technique and maintenance under slow growth condition. Plant Cell Tiss Organ Cult 149, 281–293 (2022). https://doi.org/10.1007/s11240-022-02244-1

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