In vitro micropropagation and genetic fidelity assesment in Flemingia macrophylla (Willd.) Merr: an ethnomedicinal plant

, , , , ,


Research Articles | Published:

Print ISSN : 0970-4078.
Online ISSN : 2229-4473.
Website:www.vegetosindia.org
Pub Email: contact@vegetosindia.org
Doi: 10.1007/s42535-020-00106-9
First Page: 286
Last Page: 295
Views: 602


Keywords: Flemingia macrophylla , Micro-propagation, ISSR, RAPD


Abstract


Flemingia macrophylla is an important ethnomedicinal plant of Fabaceae, with immense pharmaceutical properties. Now, it is becoming rare because of its poor seed dispersal mechanism and over-exploitation of the material. In this study we have successfully established an in vitro micro-propagation protocol employing explants like ex vivo leaf and cotyledonary node. Both the explants were cultured on MS medium supplemented with various combinations and concentrations of growth hormones. In vitro morphogenetic response of leaf explants are very poor when compared with cotyledonary nodes. Cotyledonary nodes cultured on MS medium supplemented with BAP (2.0 mg/l) in combination with IAA (0.2 mg/l) produced a high percentage of multiple shoots (13.76 ± 1.1). Elongated shoots (6.30 ± 0.01 cm) rooted on MS medium with IBA (0.8 mg/l) produced a high frequency of rooting (10.80 ± 1.23) with excellent root length (10.56 ± 1.70 cm). Plantlets with a fully developed root system were acclimatized using soil: manure (1:1) and shifted to the greenhouse. Investigations for the corroboration of genetic stability among the clones and mother plants were carried out by using ISSR and RAPD markers and the results were in clear evidence that all the plants are true-to-type. This scientific approach for in vitro propagation and conservation is prerequisite for better sustainability of the plant.



                Flemingia macrophylla
              , Micro-propagation, ISSR, RAPD


*Pdf Download

(*Only SPR Members can download pdf file; #Open Access;)

References


  1. Auer CA, Motyka V, Březinová A, Kamínek M (1999) Endogenous cytokinin accumulation and cytokinin oxidase activity during shoot organogenesis of Petunia hybrida. Physiol Plant 105:141–147

  2. Batista D, Sousa MJ, Pais MS (1996) Plant regeneration from stem and petiole-derived callus of Humulus lupulus L.(hop) clone Bragança and var. Brewer’s Gold. In Vitro Plant 32:37–41

  3. Benson EE (2000) Sepecial symposium: in vitro plant recalcitrance in vitro plant recalcitrance: an introduction. In Vitro Cell Dev Biol Plant 36:141–148

  4. Bhargavi C, Venkataramana B (2011) Anxiolytic and antiepileptic activity of aerial parts of Flemingia macrophylla in rodents. Asian J Pharm Res Health Care 3(1):2–8 

  5. Doyle J (1991) DNA Protocols for plants. In: Hewitt GM, Johnston AWB, Young JPW (eds) Molecular techniques in taxonomy. NATO ASI series (series H: cell biology), vol 57. Springer, Berlin, Heidelberg, pp 283–293

  6. Fang L (2008) Study on seed sterilization and bud proliferation in tissue culture of Flemingia philippinensis Merr. et Rolf. J Anhui Agric Sci 36(32):13981–13983

  7. Feng S-x, Ma X-j, Shi L-j, Ke F (2007) Primary study on germination character of Flemingia Philippinensis seeds. Seed 26(11):3–5

  8. Gaspar T, Kevers C, Penel C, Greppin H, Reid DM, Thorpe TAJIVC, Biology-Plant D (1996) Plant hormones and plant growth regulators in plant tissue culture. In Vitro Cell Dev Biol Plant 32:272–289

  9. Gurriaran M, Revilla M, Tames R (1999) Adventitious shoot regeneration in cultures of Humulus lupulus L.(hop) cvs. Brewers Gold and Nugget. Plant Cell Rep 18:1007–1011

  10. Heider B, Andersson MS, Schultze-Kraft R (2006) RAPD variation among North Vietnamese Flemingia macrophylla (Willd.) Kuntze ex Merr. accessions. In: Hawksworth DL, Bull AT (eds) Plant conservation and biodiversity. Springer, Dordrecht, pp 43–57

  11. Hilaire K, Daouda K, Michel Z, Justin K, Ochatt S (2009) In vitro plantlets regeneration in Bambara groundnut [Vigna subterranea (L.) Verdc. (Fabaceae)] through direct shoot bud differentiation on hypocotyl and epicotyl cuttings. Afr J Biotechnol 8(8):1466–1473

  12. Ho H-Y, Wu J-B, Lin W-C (2011) Flemingia macrophylla extract ameliorates experimental osteoporosis in ovariectomized rats. Evid Based Complement Alternat Med. https://doi.org/10.1093/ecam/nep179

  13. Hsieh P-C et al (2011) Hepatoprotective effect of the aqueous extract of Flemingia macrophylla on carbon tetrachloride-induced acute hepatotoxicity in rats through anti-oxidative activities. Am J Chin Med 39:349–365

  14. Jainul MA, Azam S, Chowdhury A, Mubarak R, Faruq K (2013) In-vitro thrombolytic and cytotoxic activity of methanolic extract of Flemingia macrophylla leaves. Asian J Pharm Clin Res 6(4):86–88

  15. Kalimuthu K, Paulsamy S, Senthilkumar R, Sathya M (2007) In vitro propagation of the biodiesel plant Jatropha curcas L. Plant Tissue Cult Biotechnol 17:137–147

  16. Kour B, Kour G, Kaul S, Dhar M (2014) In vitro mass multiplication and assessment of genetic stability of in vitro raised Artemisia absinthium L. plants using ISSR and SSAP molecular markers. Adv Botany 2014:727020

  17. Lai W-C et al (2013) Phyto-SERM constitutes from Flemingia macrophylla. Int J Mol Sci 14:15578–15594

  18. Lalnundanga L, Khanduri V (2010) Effect of Flemingia macrophylla on biological and physico-chemical characteristics of soil. American-Eurasian J Agric Environ Sci 8:206–211

  19. Madhusudhana Rao J, Sivaram-Babu S, Subrahmanyam K, Jagannadha-Rao K (1978) Homoflemingin and flemiwallichin C from the leaves of Flemingia wallichii W. & A. Curr Sci 47(16):584

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

  21. Prasad JS, Varma R (1977) 5, 7, 2', 4'-Tetrahydroxyisoflavone in Moghania [Maughania] macrophylla. Phytochemistry 16(7):1120

  22. Rana Y, Rana V, Jain VK, Jain B (2011) Acute toxicity study and analgesic activity of successive extracts of roots of Flemingia macrophylla (Willd.). Int J Phytopharm Res 2(2):62–67

  23. Rani V, Raina S (2000) Genetic fidelity of organized meristem-derived micropropagated plants: a critical reappraisal. In Vitro Cell Dev Biol Plant 36:319–330

  24. Rao KN, Srimannarayana G (1983) Fleminone, a flavanone from the stems of Flemingia macrophylla. Phytochemistry 22:2287–2290

  25. Rathore MS, Mastan SG, Yadav P, Bhatt V, Shekhawat N, Chikara J (2016) Shoot regeneration from leaf explants of Withania coagulans (Stocks) Dunal and genetic stability evaluation of regenerates with RAPD and ISSR markers. S Afr J Bot 102:12–17

  26. Rohela GK, Jogam P, Shabnam AA, Shukla P, Abbagani S, Ghosh MK (2018) In vitro regeneration and assessment of genetic fidelity of acclimated plantlets by using ISSR markers in PPR-1 (Morus sp.): an economically important plant. Sci Hort 241:313–321

  27. Rohela GK, Jogam P, Bylla P, Reuben C (2019) Indirect regeneration and rssessment of renetic fidelity of acclimated plantlets by SCoT, ISSR, and RAPD markers in Rauwolfia tetraphylla L.: an endangered medicinal plant. BioMed Res Int 2019:3698742

  28. Roy S (1995) Clonal propagation of an endangered tree of high economic value: Sterculia urens. Int J Mentel 12:32–33

  29. Sen J, Sharma A (1991) Micropropagation of Withania somnifera from germinating seeds and shoot tips. Plant Cell Tissue Organ Cult 26:71–73

  30. Shiao Y-J, Wang C-N, Wang W-Y, Lin Y-LJPm (2005) Neuroprotective flavonoids from Flemingia macrophylla. Planta Med 71:835–840

  31. Shinde S, Sebastian JK, Jain JR, Hanamanthagouda MS, Murthy HN (2016) Efficient in vitro propagation of Artemisia nilagirica var. nilagirica (Indian wormwood) and assessment of genetic fidelity of micropropagated plants. Physiol Mol Biol Plants 22:595–603

  32. Sita LG, Sreenatha K, Sujata S (1992) Plantlet production from shoot tip cultures of red sandalwood (Pterocarpus santalinus L.). Curr Sci 62:532–535

  33. Song JY, Sivanesan I, An CG, Jeong BR (2010) Adventitious shoot regeneration from leaf explants of miniature paprika (Capsicum annuum)‘Hivita Red’ and ‘Hivita Yellow.’ Afr J Biotechnol 9:2768–2773

  34. Souza RRd P, Silva PDdO, RRd R, MVd, Nery FC, Paiva RJCeA (2016) Optimization of jenipapo in vitro seed germination process. Ciência e Agrotecnologia 40:658–664

  35. Sriskandarajah S, Prinsen E, Motyka V, Dobrev PI, Serek M (2006) Regenerative capacity of cacti Schlumbergera and Rhipsalidopsis in relation to endogenous phytohormones, cytokinin oxidase/dehydrogenase, and peroxidase activities . J Plant Growth Regul 25:79–88

  36. Syiem D, Khup P (2007) Evaluation of Flemingia macrophylla L., a traditionally used plant of the north eastern region of India for hypoglycemic and anti-hyperglycemic effect on mice. Pharmacol Online 2:355–366


  37.  


Acknowledgements


Authors are greatly acknowledged to Prof. A. Sadanandam CSIR-Emeritus Scientist, Department of Biotechnology, Kakatiya University for his kind advice and encouragement. We are greatly acknowledged to Prof. V. S. Raju Department of Botany, Kakatiya University for authentication of plant and encouragement. Authors are also greatful to UGC, New Delhi for sanctioning SAP DRS programme to the department.


Author Information


Sirikonda Abhiteja
Department of Biotechnology, Kakatiya University, Warangal, India

Jogam Phanikanth
Department of Biotechnology, Kakatiya University, Warangal, India


Ellendula Raghu
Department of Biotechnology, Kakatiya University, Warangal, India


Kudikala Hemalatha
Department of Biotechnology, Kakatiya University, Warangal, India


Mood Kasim
Department of Biotechnology, Kakatiya University, Warangal, India

Allini Venkateswar Rao
Department of Biotechnology, Kakatiya University, Warangal, India
vrao.alleni@gmail.com