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.
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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.
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.
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.
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.
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.
Chandra, M., Pal, A.: Differential response of the two cotyledons of Vigna radiata in vitro. — Plant Cell Rep. 15: 248–253, 1995.
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.
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.
Duncan, D.B.: Multiple range and multiple F test. — Biometrics 11: 1–42, 1955.
Eapen, S., George, L.: Ontogeny of somatic embryos of Vigna aconitifolia, V. mungo and V. radiat. — Ann. Bot. 66: 219–226, 1990.
Gamborg, O.L., Miller, R.A., Ojima, K.: Nutrient requirements of suspension cultures of soybean root cells. — Exp. Cell Res. 50: 151–158, 1968.
Ganesh Kumari, K., Ganesan, M., Jayabalan, N: Somatic embryogenesis and plant regeneration in Ricinus communis. — Biol. Plant. 52: 429–436, 2008.
Girija, S., Ganapathi, A., Ananthakrishnan. G.: Somatic embryogenesis in Vigna radiata (L.) Wilczek. — Indian J. exp. Biol. 38: 1241–1244, 2000.
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.
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.
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.
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.
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.
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.
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.
Murashige, T., Skoog, F.: A revised medium for rapid growth and bioassays with tobacco tissue cultures. — Physiol. Plant. 15: 473–497, 1962.
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.
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.
Patel, M.B., Bharadwaj, R., Joshi, A.: Organogenesis in Vigna radiata (L.) Wilczek. — Indian J. exp. Biol. 29: 619–622, 1991.
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.
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.
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.
Rao, M.M., Lakshmi Sita, G.: Direct somatic embryogenesis from immature embryos of rosewood (Dalbergia latifolia Roxb.). — Plant Cell Rep. 15: 355–359, 1996.
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.
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.
Tazeen, S., Mirza, B.: Factors affecting Agrobacterium tumefaciens mediated genetic transformation of Vigna radiata (L.) Wilczek. — Pakistan J. Bot. 36: 887–896, 2004.
Tivarekar, S., Eapen, S.: High frequency plant regeneration from immature cotyledons of mung bean. — Plant Cell Tissue Organ Cult. 66: 227–230, 2001.
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.
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.
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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
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DOI: https://doi.org/10.1007/s10535-010-0043-x