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In vitro shoot tip multiplication of bambara groundnut [Vigna subterranea (L.) Verdc.]

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Abstract

A rapid, simple and efficient protocol for direct in vitro multiple shoot induction and plantlet recovery was achieved from shoot tip explants of Bambara groundnut [Vigna subterranea (L.) Verdc.]. Shoot tips, were isolated from in vitro-grown seedlings and cultured on Murashige and Skoog basal medium (MS) containing Gamborg’ vitamins (B5) and supplemented with different concentrations of the plant growth regulators Thidiazuron (TDZ), N6-benzylaminopurine (BAP), Kinetin (KIN) and Adenine sulfate (ADS). TDZ 0.45 μM was found to be best for shoot multiplication with a mean of 11shoots per explant. Among the carbon sources tested, the best response in terms of mean number of shoots per explant was obtained with sucrose (11). The mean number of shoots per explant induced among the studied landraces of Bambara groundnut varied from 9 to 13. Individual shoots, aseptically excised, produced normal roots within 2 weeks on the basal MS medium supplemented with Indole Butyric Acid (IBA) or Naphthalene acetic acid (NAA). The highest number of roots per shoot and the longest roots were obtained on MS medium with 2 mg/L IBA. Rooted plantlets were successfully hardened under greenhouse conditions and subsequently established in the field conditions, with a recorded survival rate of 70 and 80 %, respectively. The transferred plants in the field were morphologically normal and fertile. This protocol can be efficiently used for mass propagation, germplasm preservation and probably also for gene transfer of Vigna subterranea (L.).

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References

  • Aasim M, Khawar KM, Ozcan S (2009) In vitro micropropagation from plumular apices of Turkish cowpea (Vigna unguiculata L.) cultivar Akkiz. Scientia Hortic 122:468–471

    Article  CAS  Google Scholar 

  • Aasim M, Hussain N, Umer EM, Zubair M, Hussain SB, Saeed S, Rafique TS, Sancak C (2010) In vitro shoot regeneration of fenugreek (Trigonella foenum-graecum L.) using different cytokinins. Afr J Biotechnol 9(42):7174–7179

    CAS  Google Scholar 

  • Abdelwahd R, Hakam N, Labhilili M, Udupa SM (2008) Use of an adsorbent and antioxidants to reduce the effects of leached phenolics in in vitro plantlet regeneration of faba bean. Afr J Biotechnol 7:997–1002

    CAS  Google Scholar 

  • Adu-Dapaah HK, Sangwan RS (2003) Agronomic and Biotechnical Approaches to Bambara Groundnut Improvement. In: Proceedings of the International Bambara Groundnut Symposium, Botswana College of Agriculture, Botswana, 8–12 August, pp 245–254

  • 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(3):385–392

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Akpalu MM, Atubilla IA, Oppong-Sekyere D (2013) Assessing the level of cultivation and utilization of Bambara groundnut (Vigna subterranea (L.) Verdc.) in the sumbrungu community of bolgatanga, upper east region, Ghana. Int J Pl An Env Sci 3(3):68–75

    Google Scholar 

  • Amutha S, Ganapathi A, Muruganantham M (2003) In vitro organogenesis and plant formation in Vigna radiata (L.) Wilczek. Plant Cell Tiss Organ Cult 72:203–207

    Article  CAS  Google Scholar 

  • Anchirinah VM, Yridoe EK, Bennet-Lattey SO (2001) Enhancing sustainable production and genetic resource conservation of Bambara groundnut: a survey of indigenous agricultural knowledge systems. Outlook Agric 30:281–286

    Article  Google Scholar 

  • Anwar F, Sharmila P, Pardha SP (2008) An ideal protocol for in vitro regeneration, efficient rooting and stable transplantation of Chickpea. Physiol Mol Biol Plant 14:329–335

    Article  CAS  Google Scholar 

  • Anwar F, Sharmila P, Pardha SP (2010) No more recalcitrant: Chickpea regeneration and genetic transformation. Afr J Biotechnol 9(6):782–797

    Article  CAS  Google Scholar 

  • Anwar F, Alghamdi SS, Ammar MH, Siddique KHM (2011) An efficient in vitro regeneration protocol for faba bean (Vicia faba L.). J Med Plants Res 5(28):6460–6467

    CAS  Google Scholar 

  • Aragao FJL, Ribeiro SG, Barros LMG, Brasileiro ACM, Maxwell DP, Rech EL, Faria JC (1998) Transgenic beans (Phaseolus vulgaris L.) engineered to express viral antisense RNAs show delayed and attenuated symptoms of bean golden mosaic geminivirus. Mol Breed 4:491–499

    Article  CAS  Google Scholar 

  • Atif RM, Patat-Ochatt EM, Svabova L, Ondrej V, Klenoticova H, Jacas L, Griga M, Ochatt SJ (2013) Gene transfer in legumes. Progr Bot 74:37–100

    Article  CAS  Google Scholar 

  • Barpete S, Khawar KM, Özcan S (2014) Differential competence for in vitro adventitous rooting of grass pea (Lathyrus sativus L.). Plant Cell Tissue Organ Cult 119(1):39–50

    Article  CAS  Google Scholar 

  • Basalma D, Uranbey S, GürleK D, Özcan S (2008) TDZ-induced plant regeneration in Astragalus cicer L. Afr J Biotechnol 7(8):955–959

    CAS  Google Scholar 

  • Brar MS, Al-Khayri JM, Shamblin CE, Mc New RW, Morelock TE, Anderson EJ (1997) In vitro shoot tip multiplication of Cowpea [Vigna unguiculata (L.) Walp.] In Vitro Cell Dev Biol-Plant 33:114–118

    Article  CAS  Google Scholar 

  • Brunning JL, Kintz JL (1977) Computational handbook of statistics. 2nd ed Scott Foresman, Glenview

    Google Scholar 

  • Chaudhury D, Madanpotra S, Jaiwal R, Saini R, Kumar PA, Jaiwal PK (2007) Agrobacterim tumefaciens-mediated high frequency genetic transformation of an Indian cowpea (Vigna unguiculata L. Walp) cultivar and transmission of transgenes into progeny. Plant Sci 172:692–700

    Article  CAS  Google Scholar 

  • Fleissner KWE (2006) The improvement of Bambara groundnut production in Northern Namibia by means of breeding strategies and agronomic investigations. Die Dissertation wurde am 22. Juni 2006 bei der Technischen Universität München eingereicht und durch die Fakultät Wissenschaftszentrum Weihenstephan für Ernährung, Landnutzung und Umwelt am 10. angenommen 151pp.

  • Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158

    Article  CAS  PubMed  Google Scholar 

  • Gaspar T, Kevers C, Faivre-Rampant O, Crèvecoeur M, Penel C, Greppin H, Dommes J (2003) Changing concepts in plant hormone action. In Vitro Cell Dev Biol-Plant 39:85–106

    Article  CAS  Google Scholar 

  • Gulati A, Jaiwal PK (1990) Culture conditions effecting plant regeneration from cotyledon of mungbean [Vigna radiata (L.) Wilczek]. Plant Cell Rep 13:523–527

    Article  Google Scholar 

  • Guo B, Abbasi BH, Amir Zeb, Xu LL, Wei YH (2011) Thidiazuron: A multi-dimensional plant growth regulator. Afr J Biotechnol 10(45):8984–9000

    Article  CAS  Google Scholar 

  • Hazarika BN (2006) Morpho-physiological disorders in in vitro culture of plants. Sci Hort 108:105–120

    Article  CAS  Google Scholar 

  • Huetteman CA, Preece JE (1993) Thidiazuron: a potent cytokinin for woody plant tissue culture. Plant Cell Tissue Organ Cult 33:105–119

    Article  CAS  Google Scholar 

  • Jahan I, Alam N, Roy PK (2015) Micropropagation of yardlong bean (Vigna unguiculata (L.) Walsp. Ssp. Sesquipedalis L. Verdc.) through in vitro culture. Bangladesh J Bot 43(2):345–350

  • Jitsopakula N, Thammasirib K, Ishikawac K (2013) Efficient adventitious shoot regeneration from shoot tip culture of Vanda coerulea, a Thai orchid. Scias 39:449–455

    Google Scholar 

  • Kavango Farming Systems And Extension Project TEAM (1995) Survey of an Inland Community of Kavango, Mbora Village, Kavango Farming Systems Research and Extension Project, Rundu, Namibia

    Google Scholar 

  • Khatun MK, Haque MS, Islam S, Nasiruddin KM (2008) In vitro regeneration of mungbean (Vigna radiata L.) from different explants. Progress Agric 19:13–19

  • Koné M, Patat-Ochatt EM, Conreux C, Sangwan RS, Ochatt SJ (2007) In vitro morphogenesis from cotyledon and epicotyl explants and flow cytometry distinction between landraces of Bambara groundnut [Vigna subterranea (L.) Verdc], an under-utilised grain legume. Plant Cell Tiss Organ Cult 88:61–75

    Article  Google Scholar 

  • Koné M, Kouakou TH, Koné D, Kouadio YJ, Zouzou M, Ochatt SJ (2009a) Factors affecting regeneration of bambara groundnut [Vigna subterranea (L.)Verdc.] from mature embryo axes. In Vitro Cell Dev Biol-Plant 45:769–775

  • Koné M, Kouakou TH, Koné D, Kouadio YJ, Zouzou M, Ochatt SJ (2009b) 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

  • Koné M, Koné T, Kouakou TH, Konaté S, Ochatt SJ (2013) Plant regeneration via direct shoot organogenesis from cotyledon explants of Bambara groundnut, Vigna subterranea (L.) Verdc. Biotechnol Agron Soc Environ 17(4):584–592

    Google Scholar 

  • Koné M, Koné T, Silué N, Soumahoro AB, Kouakou TH (2015) In Vitro Seeds Germination and Seedling Growth of Bambara Groundnut (Vigna subterranea (L.) Verdc. (Fabaceae)). Sci World J. doi:10.1155/2015/595073

    Google Scholar 

  • Lacroix B, Assoumou N’Dong Y, Sangwan RS (2003) Efficient in vitro direct shoot regeneration systems in Bambara groundnut [Vigna subterranea (L.) Verdc.] Plant Cell Rep 21:1153–1158

    Article  CAS  PubMed  Google Scholar 

  • Liu THA, Lin JJ, Wu RY (2006) The effects of using trehalose as a carbon source on the proliferation of Phalaenopsis and Doritaenopsis protocorm-like-bodies. Plant Cell Tiss Organ Cult 86(1):125–129

    Article  Google Scholar 

  • Malik KA, Saxena PK (1992) Thidiazuron induces high-frequency shoot regeneration in intact seedlings of pea (Pisum sativum (L.)), chickpea (Cicer arietinum (L)) and lentil (Lens culinaris). Austr. J Plant Physiol 19:731–740

    CAS  Google Scholar 

  • Mao JQ, Zaidi MA, Altosaar I (2006) In vitro regeneration of Vigna unguculata (L.) Walp.cv. Blackeye cowpea via shoot organogenesis. Plant Cell Tiss Organ Cult 87:121–125

    Article  CAS  Google Scholar 

  • Massawe FJ, Schenkel W, Basu S, Temba EM (2003) Artificial Hybridisation in Bambara Groundnut. In: Proceedings of the International Bambara Groundnut Symposium, Botswana College of Agriculture, Botswana, 8–12, p 193–210

  • McDonagh JF (1998) A Review of Soil Fertility in Northern Namibia: an Aid for assessing Research Needs, University of East Anglia Norwich, UK

    Google Scholar 

  • Mello MO, Dias CTS, Amaral AFC, Melo M (2001) Growth of Bauhinia forficata Link, Curcuma zedoaria Roscoe and Phaseolus vulgaris L. cell suspension cultures with carbon sources. Sci Agric 58(3):481–485

    Article  CAS  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 

  • Nagori R, Purohit SD (2004) In vitro plantlet regeneration in Annona squamosa through direct shoots bud differentiation on hypocotyls segments. Sci Hortic 99:89–98

    Article  CAS  Google Scholar 

  • Ncube B, Twomlow SJ, van Wijk MT, Dimes JP, Giller KE (2007) Productivity and residual benefits of grain legumes to sorghum under semi-arid conditions in southwestern Zimbabwe. Plant Soil 299:327–336

    Article  Google Scholar 

  • Ochatt SJ, Abirached-Darmency M, Marget P, Aubert G (2007) The Lathyrus Paradox “Poor men’s diet” or a remarkable genetic resource for protein legume breeding? In: Ochatt SJ, Jain SM (eds) Breeding of neglected and under-utilized crops, spices and herbs. Science Press, Plymouth, pp 41–60

    Google Scholar 

  • Owonubi SJ, Ojuederie OB, Olayede MN (2011) Organogenesis of Bambara groundnut (Vigna subterranea (L.) Verdc.) through in vitro culture from nodal segments. Int J Plant Dev Biol 1(5):53–57

    Google Scholar 

  • Park SY, Kim YW, Moon HK, Murthy HN, Choi YH, Cho HM (2008) Micropropagation of Salix pseudolasiogne from nodal explants. Plant Cell Tiss Organ Cult 93:341–346

    Article  Google Scholar 

  • Parveen S, Venkateshwarlu M, Srinivas D, Reddy KJM, Ugandhar T (2012) Direct in vitro shoots proliferation of Chick pea (Cicer arietinum L.) from shoot tip explants induced by thidiazuron. Biosci Discover 3(1):01–05

    Google Scholar 

  • Perez-Tornero O, Tallon CI, Porras I (2009) An efficient protocol for micropropagation of lemon (Citrus limon) from mature nodal segments. Plant Cell Tiss Organ Cult 100(3):263–271

    Article  Google Scholar 

  • Petersen KK, Hansen J, Krogstrup P (1999) Significance of different carbon sources and sterilization methods on callus induction and plant regeneration of Miscanthus_ogiformis Honda ‘Giganteus. Plant Cell Tiss Organ Cult 58:189–197

    Article  Google Scholar 

  • Raghavendra T, Sudhakar P (2014) Pattern of plant regeneration from shoot tip explants of pigeonpea (Cajanus cajan L. milllsp) var lrg-41. Int J Appl Biol Pharm Technol 1(5):261–266

    Google Scholar 

  • Rajeswari V, Paliwal K (2008) In vitro adventitious shoot organogenesis and plant regeneration from seedling explants of Albizia odoratissima L. f. (Benth.). In vitro Cell Dev Biol-Plant 44 (2):78–83

    Article  CAS  Google Scholar 

  • Saini R, Jaiwal PK (2002) Age, position in mother seedling, orientation, and polarity of the epicotyl segments of black gram (Vigna mungo L. Hepper) determines its morphogenic response. Plant Sci 163:101–109

    Article  CAS  Google Scholar 

  • Saini R, Jaiwal P (2005) Transformation of a recalcitrant grain legume, Vigna mungo L. Hepper, using Agrobacterium tumefaciens-mediated gene transfer to shoot apical meristem cultures. Plant Cell Rep 24:164–171

    Article  CAS  PubMed  Google Scholar 

  • Sambe MAN, Sagna M, Sy MO (2010) Seed germination and in vitro regeneration of Parkia biglobosa (Jacq.) Benth. Afr J Biotechnol 9(21):3099–3108

    CAS  Google Scholar 

  • Smith C (1995) “Carbohydrate chemistry” In Plant Biochemistry and Molecular Biology, Lea PJ, Leegood RC, (eds) John Wiley & Sons, Chichester, pp 73–111

    Google Scholar 

  • Snedecor GW, Cochran WG (1967) Statistical Methods 6th. Ed., Ames: The Iowa State University Press, p 593

    Google Scholar 

  • Sopalun K, Thammasiri K, Ishikawa K (2010) Micropropagation of the Thai orchid Grammatophyllum speciosum Blume. Plant Cell Tiss Organ Cult 101:143–150

    Article  Google Scholar 

  • Stanton WR, Orraca-Tetteh R, Doughty J, Steele W (1966) (Voandzeia subterranea (L.) Thours). In: Grain Legumes in Africa. Food and Agriculture Organization (UN), Rome, Italy. pp. 127–132

    Google Scholar 

  • Tang Y, Chen L, Li XM, Li J, Luo Q, Lai J, Li HX (2012) Effect of culture conditions on the plant regeneration via organogenesis from cotyledonary node of cowpea (Vigna unguiculata L. Walp). Afr J Biotechnol 11(14):3270–3275

    Article  CAS  Google Scholar 

  • Thomas JC, Katterman FR (1986) Cytokinin activity induced by thidiazuron. Plant Physiol 81:681–683

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Thorpe TA (1982) Carbohydrate Utilization and Metabolism. In: Bonga JM, Durzan DJ (eds) Tissue culture in forestry. Martinus Nijhoff/Dr. W. Junk Publishers, The Hague, pp 325–368.doi:10.1007/978-94-017-3538-4-11

    Chapter  Google Scholar 

  • Tripathi L, Singh AK, Singh S, Singh R, Chaudhary S, Sanyal I, AmlaDV (2013) Optimization of regeneration and Agrobacterium-mediated transformation of immature cotyledons of chickpea (Cicer arietinum L.) Plant Cell Tiss Organ Cult 113(3):513–527

  • Vijayan S, Beena MR, Kirti PB (2006) Effective and simple regeneration of mungbean (Vigna radiata (L). Wilzcek) using cotyledonary node explants. J Plant Bioch Biotechnol 15:131–134

    Article  CAS  Google Scholar 

  • Yadav SK, Sreenu P, Maheswari M, Vanaja M, Venkateswarlu B (2010) Efficient shoot regeneration from double cotyledonary node explants of green gram(Vigna radiata L Wilczek). Indian J Biotechnol 9:403–407

    CAS  Google Scholar 

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Acknowledgments

The authors wish to acknowledge to the Department of Natural Sciences of University Nangui Abrogoua for providing necessary facilities for this investigation.

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  1. Laboratoire de Physiologie Végétale, UFR Biosciences, Université Félix Houphouët Boigny, 22 BP 582, Abidjan 22, Côte d’Ivoire

    Nakpalo Silué

  2. Laboratoire de Biologie et Amélioration des Productions Végétales, UFR Sciences de la Nature, Université Nangui Abrogoua, 02 BP 801, Abidjan 02, Côte d’Ivoire

    Tchoa Koné, André Brahima Soumahoro & Mongomaké Koné

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  1. Nakpalo Silué
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Silué, N., Koné, T., Soumahoro, A.B. et al. In vitro shoot tip multiplication of bambara groundnut [Vigna subterranea (L.) Verdc.]. Plant Cell Tiss Organ Cult 127, 603–611 (2016). https://doi.org/10.1007/s11240-016-1096-0

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