Skip to main content
SpringerLink
Log in

Somatic embryogenesis and regeneration of Vigna radiata

  • Original Papers
  • Published:
Biologia Plantarum

Abstract

An efficient regeneration protocol via somatic embryogenesis was optimized for mung bean [Vigna radiata (L.) Wilczek; cv. Vamban 1]. Primary leaf explants were used for embryogenic callus induction in MMS medium (Murashige and Skoog salts with B5 vitamins) containing 2.0 mg dm−3 2,4-dichlorophenoxyacetic acid (2,4-D), 150 mg dm−3 glutamine and 3 % sucrose. Fast growing, highly embryogenic cell suspensions were established from 21-d-old calli in MMS medium supplemented with 0.5 mg dm−3 2,4-D and 50 mg dm−3 proline (Pro), and maximum recovery of globular (39.0 %), heart-shaped (26.3 %) and torpedo-stage (21.0 %) somatic embryos were observed in this medium. Mature cotyledonary-stage somatic embryos were cultured for 5 d in half strength B5 liquid medium containing 0.05 mg dm−3 2,4-D, 20 mg dm−3 Pro, 5 μM abscisic acid, 1000 mg dm−3 KNO3, 50 mg dm−3 polyethylene glycol (PEG 6000) and 30 g dm−3 D-mannitol. Mature somatic embryos were germinated after dessication for 3 d and complete development of plantlets accomplished in MMS medium containing 30 g dm−3 maltose, 0.5 mg dm−3 benzyladenine and 500 mg dm−3 KNO3. Profuse lateral roots, and regeneration frequency (up to 60 %) were observed in half-strength MMS medium containing 0.5 mg dm−3 indolebutyric acid (IBA). The regenerated plants were grown to fruiting and were morphologically normal and fertile.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

ABA:

abscisic acid

BA:

6-benzyladenine

B5:

Gamborg medium

CH:

casein hydrolysate

CIM:

embryogenic callus induction medium

2,4-D:

2,4-dichlorophenoxyacetic acid

GA3 :

gibberellic acid

Gln:

L-glutamic acid-5-amide

EIM:

somatic embryo induction medium

EMM:

somatic embryo maturation medium

KIN:

kinetin

ME:

malt extract

MMS:

MS salts with B5 vitamins

MS:

Murashige and Skoog medium

NAA:

α-naphthalene acetic acid

PEM:

proembryogenic mass

PEG:

polyethylene glycol

Pro:

L-proline

Put:

putrescine

RM:

rooting medium

TDZ:

thidiazuron

YE:

yeast extract

ZE:

zeatin

References

  • Ackerson, R.C.: Regulation of soybean embryogenesis by abscisic acid. — J. exp. Bot. 35: 403–413, 1984.

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Amutha, S., Ganapathi, A., Muruganantham, M.: Thidiazuron-induced high-frequency axillary and adventitious shoot regeneration in Vigna radiata (L.) Wilczek. — In Vitro cell. dev. Biol. Plant 42: 26–30, 2006.

    Article  CAS  Google Scholar 

  • Anbazhagan, V.R., Ganapathi, A.: Somatic embryogenesis in cell suspension cultures of pigeonpea (Cajanus cajan L.). — Plant Cell Tissue Organ Cult. 56: 179–184, 1999.

    Article  Google Scholar 

  • Betal, S., Sen Raychaudhuri, S.: Shoot tip multiplication in aromatic and non-aromatic varieties of Vigna radiata (L.) Wilczek. — Plant Cell Tissue Organ Cult. 11: 187–194, 2001.

    Google Scholar 

  • Chandra, M., Pal, A.: Differential response of the two cotyledons of Vigna radiata in vitro. — Plant Cell Rep. 15: 248–253, 1995.

    Article  CAS  Google Scholar 

  • Chen, H.K., Mok, M.C., Mok., D.W.S.: Somatic embryogenesis and shoot organogenesis from interspecific hybrid embryos of Vigna glabrescence and V. radiata. — Plant Cell Rep. 9: 77–79, 1990.

    Article  Google Scholar 

  • Devi, P., Radha, P., Sitamahalakshmi, L., Syamala, D., Manoj Kumar, S.: Plant regeneration via somatic embryogenesis in mung bean (Vigna radiata (L.) Wilczek). — Scientia Hort. 99: 1–8, 2004.

    Article  CAS  Google Scholar 

  • Duncan, D.B.: Multiple range and multiple F test. — Biometrics 11: 1–42, 1955.

    Article  Google Scholar 

  • Eapen, S., George, L.: Ontogeny of somatic embryos of Vigna aconitifolia, V. mungo and V. radiat. — Ann. Bot. 66: 219–226, 1990.

    Google Scholar 

  • Gamborg, O.L., Miller, R.A., Ojima, K.: Nutrient requirements of suspension cultures of soybean root cells. — Exp. Cell Res. 50: 151–158, 1968.

    Article  PubMed  CAS  Google Scholar 

  • Ganesh Kumari, K., Ganesan, M., Jayabalan, N: Somatic embryogenesis and plant regeneration in Ricinus communis. — Biol. Plant. 52: 429–436, 2008.

    Article  Google Scholar 

  • Girija, S., Ganapathi, A., Ananthakrishnan. G.: Somatic embryogenesis in Vigna radiata (L.) Wilczek. — Indian J. exp. Biol. 38: 1241–1244, 2000.

    PubMed  CAS  Google Scholar 

  • Gulati, A., Jaiwal, P.K.: Culture conditions effecting plant regeneration from cotyledons of Vigna radiata (L.) Wilczek. Plant Cell Tissue Organ Cult. 23: 1–7, 1990.

    Article  CAS  Google Scholar 

  • Gulati, A., Jaiwal, P.K.: In vitro induction of multiple shoots and plant regeneration from shoot tips of mung bean. — Plant Cell Tissue Organ Cult. 29: 199–205, 1992.

    Article  CAS  Google Scholar 

  • Jaiwal, P.K., Kumari, R., Ignacimuthu, S., Potrykus, I., Sautter, C.: Agrobacterium tumefaciens-mediated genetic transformation of mung bean (Vigna radiata L. Wilczek) — a recalcitrant grain legume. — Plant Sci. 161: 239–247, 2001.

    Article  PubMed  CAS  Google Scholar 

  • Kulothungan, S., Ganapathi, A., Shajahan, A., Kathiravan, K.: Somatic embryogenesis in cell suspension culture of cowpea (Vigna unguiculata (L.) Walp). — Israel J. Plant Sci. 43: 385–390, 1995.

    Google Scholar 

  • Mathews, H.: Morphogenetic responses for in vitro cultured seedling explants of mung bean (Vigna radiata (L.) Wilczek). — Plant Cell Tissue Organ Cult. 11: 233–240, 1987.

    Article  Google Scholar 

  • Mendoza, A.B., Kazumi, H., Nishimura, T., Futsuhara, Y.: Histological and scanning electron microscopic observations on plant regeneration in mung bean cotyledons cultured in vitro. — Plant Cell Tissue Organ Cult. 32: 37–143, 1993.

    Article  Google Scholar 

  • Mundhara, R., Rashid, A.: Recalcitrant grain legume Vigna radiata, mung bean, made to regenerate on change of hormonal and cultural conditions. — Plant Cell Tissue Organ Cult. 85: 265–270, 2006.

    Article  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Obendorf, R.L., Dickerman, A.M., Pflum, T.M., Kacalanos, M.A., Smith, M.E.: Drying rate alters soluble carbohydrates, desiccation tolerance and subsequent seedling growth of soybean (Glycine max L. Merrill) zygotic embryos during in vitro maturation. — Plant Sci. 132: 1–12, 1998.

    Article  CAS  Google Scholar 

  • Ozcan, S., Barghchi, M., Firek, S., Draper, J.: Efficient adventitious shoot regeneration and somatic embryogenesis in pea. — Plant Cell Tissue Organ Cult. 34: 271–277, 1993.

    Article  CAS  Google Scholar 

  • Patel, M.B., Bharadwaj, R., Joshi, A.: Organogenesis in Vigna radiata (L.) Wilczek. — Indian J. exp. Biol. 29: 619–622, 1991.

    Google Scholar 

  • Prem Anand, R., Ganapathi, A., Vengadesan, G., Anbazhagan, V.R., Kulothungan, S.: Plant regeneration from immature cotyledon derived callus of Vigna unguiculata (L.) Walp. — Curr. Sci. 80: 671–674, 2001.

    Google Scholar 

  • Prem Anand, R., Ganapathi, A., Anbazhagan, V.R., Vengadesan, G., Selvaraj, N.: High frequency plant regeneration via somatic embryogenesis in cell suspension cultures of cowpea, Vigna unguiculata (L.) Walp. — In Vitro cell. dev. Biol. Plant 36: 475–480, 2000.

    Article  Google Scholar 

  • Ramakrishnan, K., Gnanam, R., Sivakumar, P., Manickam, A.: In vitro somatic embryogenesis in cell suspension cultures of cowpea var. C152. — Plant Cell Rep. 24: 449–461, 2005.

    Article  PubMed  CAS  Google Scholar 

  • Rao, M.M., Lakshmi Sita, G.: Direct somatic embryogenesis from immature embryos of rosewood (Dalbergia latifolia Roxb.). — Plant Cell Rep. 15: 355–359, 1996.

    Article  CAS  Google Scholar 

  • Selvi, D.T., Ramaswamy, N.M., Sukumar, S., Sree Rangaswamy, S.R.: In vitro regeneration and protoplast culture studies in mung bean (Vigna radiata (L.) Wilczek). — In Vitro 31: 62, 1995.

    Google Scholar 

  • Sen, J., Kalia, S., Guha-Mukherjee, S.: Levels of endogenous free amino acids during various stages of culture of Vigna mungo (L.) Hepper. Somatic embryogenesis, organogenesis and plant regeneration. — Curr. Sci. 82: 429–433, 2002.

    CAS  Google Scholar 

  • Tazeen, S., Mirza, B.: Factors affecting Agrobacterium tumefaciens mediated genetic transformation of Vigna radiata (L.) Wilczek. — Pakistan J. Bot. 36: 887–896, 2004.

    Google Scholar 

  • Tivarekar, S., Eapen, S.: High frequency plant regeneration from immature cotyledons of mung bean. — Plant Cell Tissue Organ Cult. 66: 227–230, 2001.

    Article  CAS  Google Scholar 

  • Torres, A.C., Ze, N.M., Cantliffe, D.J.: Abscisic acid and somatic induction of synchronous somatic embryo development of sweet potato. — In Vitro cell. dev. Biol. Plant 37: 262–267, 2001.

    Article  CAS  Google Scholar 

  • Zale, J.M., Borchardt-Wier, H., Kidwell, K.K., Steber, C.M.: Callus induction and plant regeneration from mature embryos of a diverse set of wheat genotypes. — Plant Cell Tissue Organ Cult. 76: 277–281, 2004.

    Article  Google Scholar 

Download references

Author information

Author notes

  1. P. Sivakumar & K. Ramakrishnan

    Present address: Tamil Nadu Agricultural University, Tiruchirappalli, 620009, Tamil Nadu, India

Authors and Affiliations

  1. Centre for Plant Molecular Biology, Tamil Nadu Agricultural University, Coimbatore, 641003, India

    P. Sivakumar, R. Gnanam & A. Manickam

  2. Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad, 500007, India

    K. Ramakrishnan

Authors
  1. P. Sivakumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. R. Gnanam
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. K. Ramakrishnan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. A. Manickam
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to A. Manickam.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sivakumar, P., Gnanam, R., Ramakrishnan, K. et al. Somatic embryogenesis and regeneration of Vigna radiata . Biol Plant 54, 245–251 (2010). https://doi.org/10.1007/s10535-010-0043-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10535-010-0043-x

Additional key words

Search

Navigation