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High frequency conversion of non-embryogenic synseeds and assessment of genetic stability through ISSR markers in Gymnema sylvestre

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Abstract

The present study describes the first attempt of exploiting encapsulation technology for high plantlet recovery, short-term storage and conservation of Gymnema sylvestre—an antidiabetic liana. Indole-3-butyric acid (IBA) pretreated nodal segments (NS) were encapsulated in sodium alginate (Na2-alginate) matrix and the optimal culture conditions were evaluated in terms of maximum conversion capacity of synseeds into complete plantlets. Highest conversion frequency of synseeds was obtained on Murashige and Skoog’s (MS) medium supplemented with 5.0 µM 6-benzyladenine (BA). Augmentation of Na2-alginate matrix with plant growth regulators (PGRs) and additive further improved in vitro conversion rates and the synthetic endosperm composed of 3% Na2-alginate in MS + 2.5 µM BA + 2.5 µM gibberellic acid (GA3) + 50 µM adenine sulphate (AdS) stimulated maximum recovery (88.2 ± 0.48%) of complete plantlets from synseeds. Studies on short term cold storage of synseeds showed that nutrient encapsulation maintains the viability of NS for a storage period of 8 weeks. Ex-vitro conversion of synseeds was also carried out on soilrite and vermicompost (3:1) mixture under culture room conditions. Monomorphic DNA profiles produced through Inter-Simple Sequence Repeat (ISSR) markers confirmed the genetic uniformity between synseed derived and mother plantlets.

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Abbreviations

AdS:

Adenine-sulphate

BA:

6-Benzyladenine

GA3 :

Gibberellic acid

IBA:

Indole-3-butyric acid

ISSR:

Inter simple sequence repeats

MS:

Murashige and Skoog’s medium (1962)

PGRs:

Plant growth regulators

References

  • Anturlikar SD, Gopumadhavan S, Chauhan BL, Mitra SK (1995) Effect of D-400, a herbal formulation, on blood sugar of normal and alloxan-induced diabetic rats. Ind J Physiol Pharmacol 39:95–100

    CAS  Google Scholar 

  • Ara H, Jaiswal U, Jaiswal V (2000) Synthetic seed: prospects and limitation. Curr Sci 78:1438–1444

    Google Scholar 

  • Benelli C (2016) Encapsulation of shoot tips and nodal segments for in vitro storage of “Kober 5BB” Grapevine Rootstock. Horticulturae 2:1–8

    Article  Google Scholar 

  • Bhattacharya P, Kumar V, Van Staden J (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. https://doi.org/10.1007/s11240-018-1382-0

    Article  Google Scholar 

  • Bose S, Karmakar J, Fulzele DP, Basu U, Bandyopadhyay TK (2017) In vitro shoots from root explant, their encapsulation, storage, plant recovery and genetic fidelity assessment of Limonium hybrid ‘Misty Blue’—a florist plant. Plant Cell Tissue Organ Cult 129:313–324

    Article  CAS  Google Scholar 

  • Cangahuala-Inocente GC, Dal Vesco LL, Steinmacher D, Torres AC, Guerra MP (2007) Improvements in somatic embryogenesis protocol in Feijoa (Acacia sellowiana (Berg) Burret): induction, conversion and synthetic seeds. Sci Hortic 111:228–234

    Article  CAS  Google Scholar 

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

    Article  PubMed  CAS  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

    PubMed  CAS  Google Scholar 

  • Doyle J, Doyle J (1990) Isolation of DNA from small amounts of plant tissue. BRL Focus 12:13–15

    Google Scholar 

  • Germana` MA, Micheli M, Chiancone B, Macaluso L, Standardi A (2011) Organogenesis and encapsulation of in vitro-derived propagules of Carrizo citrange [Citrus sinensis (L.) Osb. × Poncirius trifoliate (L.) Raf]. Plant Cell Tissue Organ Cult 106:299–307

    Article  Google Scholar 

  • González-Benito ME, Kremer C, Ibáñez MA, Martín C (2016) Effect of antioxidants on the genetic stability of cryopreserved mint shoot tips by encapsulation-dehydration. Plant Cell Tissue Organ Cult 127:359–368

    Article  CAS  Google Scholar 

  • Jain AK, Patole SN (2001) Some threatened plants of Pachmari Biosphere reserve of Madhya Pradesh. J Ind Bot Soc 80:151–155

    Google Scholar 

  • Khilwani B, Kaur A, Ranjan R, Kumar A (2016) Direct somatic embryogenesis and encapsulation of somatic embryos for in vitro conservation of Bacopa monnieri (L.) Wettst. Plant Cell Tissue Organ Cult 127:433–442

    Article  CAS  Google Scholar 

  • Kumar AHG, Murthy HN, Paek KY (2002) Somatic embryogenesis and plant regeneration in Gymnema sylvestre. Plant Cell Tissue Organ Cult 71:85–88

    Article  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Onishi N, Sakamoto Y, Hirosawa T (1994) Synthetic seeds an application of mass production of somatic embryos. Plant Cell Tissue Organ Cult 39:137–145

    Article  Google Scholar 

  • Patel SS, Shah RS, Goyal RK (2009) Antihyperglycemic, antihyperlipidemic, antioxidant effect of Dihar, a polyherbal Ayurvedic formulation in Streptozotocin induced diabetes rats. Ind J Exp Biol 47:564–570

    Google Scholar 

  • Piccioni E (1997) Plantlets from encapsulated micropropagated buds of M.26 apple rootstock. Plant Cell Tissue Organ Cult 47:255–260

    Article  Google Scholar 

  • Reddy PS, Gopal GR, Sita GL (1998) In vitro multiplication of Gymnema sylvestre R. Br.—an important medicinal plant. Curr Sci 75:843–845

    Google Scholar 

  • Redenbaugh K, Slade D, Viss P, Fujii J (1987) Encapsulation of somatic embryos in synthetic seed coats. HortScience 22:803–809

    Google Scholar 

  • Saeed T, Shahzad A (2015) High frequency plant regeneration in Indian Siris via cyclic somatic embryogenesis with biochemical, histological and SEM investigations. Ind Crops Prod 76:623–637

    Article  CAS  Google Scholar 

  • 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 Tissue Organ Cult 120:519–530

    Article  CAS  Google Scholar 

  • Shaheen A, Shahzad A (2015) Nutrient encapsulation of nodal segments of an endangered white cedar for studies of regrowth, short term conservation and ethylene inhibitors influenced ex vitro rooting. Ind Crops Prod 69:204–211

    Article  CAS  Google Scholar 

  • Sharma S, Shahzad A, Teixeira da Silva JA (2013) Synseed technology-a complete synthesis. Biotechnol Adv 31:186–207

    Article  PubMed  CAS  Google Scholar 

  • Sharma S, Shahzad A, Mahmood S, Saeed T (2015) High-frequency clonal propagation, encapsulation of nodal segments for short-term storage and germplasm exchange of Ficus carica L. Trees 29:345–353

    Article  CAS  Google Scholar 

  • Standardi A, Piccioni E (1998) Recent perspectives on synthetic seed technology using nonembryogenic in vitro-derived explants. Int J Plant Sci 159:968–978

    Google Scholar 

  • Sugihara Y, Nojima H, Matsuda H, Murakami T, Yoshikawa M, Kimura I (2000) Antihyperglycemic effects of gymnemic acid IV, a compound derived from Gymnema sylvestre leaves in streptozotocin-diabetic mice. J Asian Nat Prod Res 2:321–327

    Article  PubMed  CAS  Google Scholar 

  • Suprasanna P, Anupama S, Ganapathi TR, Bapat VA (2001) In vitro growth and development of encapsulated shoot tips of different banana and plantain cultivars. J New Seeds 3:19–25

    Article  Google Scholar 

  • Thiyagarajan M, Venkatachalam P (2013) A reproducible and high frequency plant regeneration from mature axillary node explants of Gymnema sylvestre (Gurmur)—an important antidiabetic endangered medicinal plant. Ind Crops Prod 50:517–524

    Article  CAS  Google Scholar 

  • Tiwari P, Mishra BN, Sangwan NS (2014) Phytochemical and pharmacological properties of Gymnema sylvestre: an important medicinal plant. Biomed Res Int 2014:1–18

    Google Scholar 

  • Udayan PS, Tushar KV, George S, Balachandran I (2009) Notes on a few rare, endemic and red listed plants from Trumula forest of Tirupati Chittor district, Andhra Pradesh. J Econ Taxon Bot 33:277–284

    Google Scholar 

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Acknowledgements

Taiba Saeed gratefully acknowledges UGC for providing the financial assistance under the scheme of Maulana Azad National Fellowship (File No. MANF-MUS-UTT-2624). Authors are also thankful to DST- SERB, for financial support in the form of Young Scientist Projects (Vide Nos. SB/FT/LS-364/2012 and SB/YS/LS-156/2013).

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Authors and Affiliations

  1. Plant Biotechnology Laboratory, Department of Botany, Aligarh Muslim University, Aligarh, 202002, India

    Taiba Saeed, Anwar Shahzad, Naseem Ahmad & Shahina Parveen

Authors
  1. Taiba Saeed
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  2. Anwar Shahzad
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  3. Naseem Ahmad
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  4. Shahina Parveen
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Corresponding author

Correspondence to Anwar Shahzad.

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Communicated by Sergio J. Ochatt.

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Saeed, T., Shahzad, A., Ahmad, N. et al. High frequency conversion of non-embryogenic synseeds and assessment of genetic stability through ISSR markers in Gymnema sylvestre. Plant Cell Tiss Organ Cult 134, 163–168 (2018). https://doi.org/10.1007/s11240-018-1409-6

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