Plant Cell, Tissue and Organ Culture

, Volume 58, Issue 2, pp 99–110 | Cite as

Differences in shoot regeneration response from cotyledonary node explants in Asiatic Vigna species support genomic grouping within subgenus Ceratotropis (Piper) Verdc.

  • Renato A. Avenido
  • Kazumi Hattori


The efficiency of any plant regeneration system lies in part in its wide applicability to diverse genotypes. In Asiatic Vigna, cotyledon and cotyledonary node explants from 4-day-old seedlings of 27 genotypes were cultured in a medium consisting of MS salts, B5 vitamins, 3.0% sucrose and 1.0 mg l-1 BA. Direct and efficient multiple shoot regeneration (80–100%) from the cotyledonary nodes was obtained in all epigeal species namely radiata, mungo, aconitifolia, subspecies radiata var. sublobata, mungo var. silvestris and in the hypogeal but allotetraploid glabrescens. In contrast, two other hypogeal species V. angularis and V. umbellata failed to initiate shoots from the nodes. However, adventititious shoots developed at the basipetal cut (hypocotyl) in 35–67% of V. angularis explants. These results provide evidence in support of the existing genomic grouping within subgenus Ceratotropis, which designates AA, A1A1 and A1A1/- to epigeal, hypogeal and the allotetraploid species, respectively. Mean shoot production ranged from 3.3 to 10.4 shoots per explant during the first subculture and varied significantly among the responsive genotypes within 4 species. Additional shoots were obtained in all genotypes after subsequent subculture. However, cotyledons were not as regenerable as cotyledonary node explants. Although significant differences in rooting were observed among the shoots of the 15 genotypes, the response was generally higher in MS basal medium (MSO) than in MS with 1.0 mg l-1 IAA. Regenerated plants were successfully transferred to soil (50–100% survival rate) and all surviving plants were reproductively fertile.

cotyledonary node shoot regeneration species relationships subgenus Ceratotropis Vigna 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Avenido RA & Hautea DM (1990) In vitro organogenesis and flowering in mungbean (V.radiata L. Wilczek). Philipp. J. Crop. Sci. 15:169–173Google Scholar
  2. Avenido RA, Hautea DM, Mendoza CJ & Carandang SL (1991) Clonal propagation of F1 hybrids of mungbean (Vigna radiata L. Wilczek)_blackgram (V.mungo L. Hepper) by tissue culture. Philipp. J. Crop. Sci. 16:63–67Google Scholar
  3. Avenido RA, Power JB & Davey MR (1993) Differential plant regeneration responses from protoplasts of mungbean (Vigna radiata L.Wilczek) and mothbean (V.aconitifolia Jacq. Marechal). Philipp. J. Crop. Sci. 18:175–180Google Scholar
  4. Chandra M & Pal A (1995) Differential response of two cotyledons of Vigna radiata in vitro. Plant Cell Rep. 15:248–253CrossRefGoogle Scholar
  5. Chen HK, Mok MC & Mok DWS (1990) Somatic embryogenesis and shoot organogenesis from interspecific hybrid embryos of Vigna glabrescens and V.radiata. Plant Cell Rep. 9:77–79CrossRefGoogle Scholar
  6. Dana S (1980) Genomic relationship in the genus Vigna and its implications in the breeding programme. In: Gill KS (ed) Breeding Methods for the Improvement of Pulse Crops (pp. 357–367) Ludhiana, Punjab Agric Univ, IndiaGoogle Scholar
  7. Dana S & Karmakar PG (1990) Species relation in Vigna subgenus Ceratotropis and its implication in breeding. In: Janick J (ed) Plant Breed Rev. (pp. 19–42)Google Scholar
  8. Das DK, Shiva Prakash N & Bhalla-Sarin N (1998) An efficient regeneration system of blackgram (Vigna mungo L.) through organogenesis. Plant Sci. 134:199–206CrossRefGoogle Scholar
  9. Eapen S & George L (1990) Ontogeny of somatic embryos of Vigna aconitifolia, V.mungo and V.radiata. Ann. Bot. 66:219–226Google Scholar
  10. Endo Y & Ohashi H (1997) Cladistic analysis of phylogenetic relationships among tribes Cicereae, Trifolieae and Vicieae (Leguminosae). Am. J. Bot. 84:523–529CrossRefGoogle Scholar
  11. Fatokun CA, Danesh D, Young ND & Stewart EL (1993) Molecular and taxonomic relationships in the genus Vigna based on RFLP analysis. Theor. Appl. Genet. 86:97–104CrossRefGoogle Scholar
  12. Franklin CI, Trieu TN, Gonzales RA & Dixon RA (1991) Plant regeneration from seedling explants of green bean (Phaseolus vulgaris L.) via organogenesis. Plant Cell Tiss. Org. Cult. 24:199–206CrossRefGoogle Scholar
  13. Gamborg OL, Miller RA & Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cell. Exp. Cell Res. 50:151–158PubMedCrossRefGoogle Scholar
  14. Ge K, Wang Y, Yuan X, Huang P, Yang J, Nie Z, Testa D & Lee N (1989) Plantlet regeneration from protoplasts isolated from mesophyll cells of adzukibean (Phaseolus angularis Wight). Plant Sci. 63:209–216CrossRefGoogle Scholar
  15. Geetha N, Venkatachalam P & Rao GR (1997a) Plant regeneration and propagation of blackgram (V.mungo L. Hepper) through tissue culture. Trop. Agric. 74:73–76Google Scholar
  16. Geetha N, Venkatachalam P & Rao GR (1997b) In vitro plant regeneration from different seedling explants of blackgram (V.mungo L. Hepper) via organogenesis. Breed Sci. 47:311–315Google Scholar
  17. George L & Eapen S (1994) Organogenesis and embryogenesis from diverse explants in pigeonpea (Cajanus cajan L.). Plant Cell Rep. 13:417–420CrossRefGoogle Scholar
  18. Gill R, Eapen S & Rao PS (1987) Morphogenetic studies of cultured cotyledons of urd bean (V.mungo L. Hepper). J. Plant Physiol. 130:1–5Google Scholar
  19. Godbole DA, Kunachgi MN, Potdar UA, Krisnamurthy KV & Mascarenhas AF (1984) Studies on drought resistant legume: the mothbean, Vigna aconitifolia Jacq. Marechal II. Morphogenetic studies. Plant Cell Rep. 3:75–78CrossRefGoogle Scholar
  20. Grant JE, Cooper PA, McAra AE & Frew TJ (1995) Transformation of peas (Pisum sativum L.) using immature cotyledons. Plant Cell Rep. 15:254–258CrossRefGoogle Scholar
  21. Gulati A & Jaiwal PK (1990) Culture conditions effecting plant regeneration from cotyledons of Vigna radiata L. Wilczek. Plant Cell Tiss. Org. Cult. 23:1–7CrossRefGoogle Scholar
  22. Gulati A & Jaiwal PK (1992) In vitro induction of multiple shoots and plant regeneration from shoot tips of mungbean (Vigna radiata L. Wilczek). Plant Cell Tiss. Org. Cult. 29:199–205CrossRefGoogle Scholar
  23. Gulati A & Jaiwal PK (1993) In vitro selection of salt-resistant Vigna radiata (L.) Wilczek plants by adventitious shoot formation from cultured cotyledon explants. J. Plant. Physiol. 142:99–102Google Scholar
  24. Gulati A & Jaiwal PK (1994) Plant regeneration from cotyledonary nodes of mungbean (Vigna radiata L. Wilczek). Plant Cell Rep. 13:523–527CrossRefGoogle Scholar
  25. Ignamuthu S, Franklin G & Melchias G (1997) Multiple shoot formation and in vitro fruiting from cotyledonary nodes of Vigna mungo L. Hepper. Curr. Sci. 73:733–735Google Scholar
  26. Jaaska V & Jaaska V (1990) Isoenzyme variation in Asian beans. Bot. Acta. 103:281–290Google Scholar
  27. Jackson JA & Hobbs SLA (1990) Rapid multiple shoot production from cotyledonary node explants of pea (Pisum sativum L.). In Vitro Cell Dev. Biol. 26:835–838Google Scholar
  28. Jaiwal PK & Gulati A (1995) Current status and future strategies of in vitro culture techniques for genetic improvement of mungbean Vigna radiata L. Wilczek. Euphytica 86:167–181Google Scholar
  29. Kaga A, Tomooka N, Egawa Y, Hosaka K & Kamijima O (1996) Species relationships in the subgenus Ceratotropis (genus Vigna) as revealed by RAPD analysis. Euphytica88:17–24CrossRefGoogle Scholar
  30. Khatoon K & Ara N (1995) Somatic embryogenesis in the suspension cultures of Vigna radiata L. Pak. J. Bot. 27:105–109Google Scholar
  31. Kim JW & Minamikawa T (1996) Transformation and regeneration of French bean plants by the particle bombardment process. Plant Sci. 117:131–138CrossRefGoogle Scholar
  32. Kumar V & Davey MR (1991) Genetic improvement of legumes using somatic cell and molecular techniques. Euphytica 55:157–169CrossRefGoogle Scholar
  33. Kumar AS, Gamborg OL & Nabors MW (1988) Plant regeneration from cell suspension cultures of Vigna aconitifolia. Plant Cell Rep. 7:138–141CrossRefGoogle Scholar
  34. Maekawa F (1955) Topo-morphological and taxonomical studies in Phaseoleae, Leguminosae. Jap. J. Bot. 15:103–116Google Scholar
  35. Malik KA & Saxena PK (1992) Somatic embryogenesis and shoot regeneration from intact seedlings of Phaseolus acutifolius A., P. aureus L. Wilczek, P.coccineus L. and P.wrightii L. Plant Cell Rep. 11:163–168CrossRefGoogle Scholar
  36. Mathews VH (1987) Morphogenetic responses from in vitro cultured seedling explants of mungbean (Vigna radiata L.Wilczek). Plant Cell Tiss. Org. Cult. 11:233–240CrossRefGoogle Scholar
  37. McClean P & Grafton F (1989) Regeneration of dry bean (Phaseolus vulgaris L.) via organogenesis. Plant Sci. 60:117–122CrossRefGoogle Scholar
  38. Mendoza AB & Futsuhara Y (1990) Varietal differences on plant regeneration by tissue culture in mungbean (Vigna radiata L. Wilczek). Japan J. Breed. 40:457–467Google Scholar
  39. Mendoza AB, Hattori K, Nishimura T & Futsuhara Y (1993) Histological and scanning elecron microscopic observations on plant regeneration in mungbean cotyledon (Vigna radiata L. Wilczek) cultured in vitro. Plant Cell Tiss. Org. Cult. 32:137–143CrossRefGoogle Scholar
  40. Meurer CA, Dinkins RD & Collins GB (1998) Factors affecting soybean cotyledonary node transformation. Plant Cell Rep. 18:180–186CrossRefGoogle Scholar
  41. Murashige T & Skoog S (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol. Plant 15:473–497CrossRefGoogle Scholar
  42. Muthukumar B, Mariamma M, Veluthambi K & Gnanam A (1996) Genetic transformation of cotyledon explants of cowpea (Vigna unguiculata L. Wasp) using Agrobacterium tumefaciens. Plant Cell Rep. 15:980–985CrossRefGoogle Scholar
  43. Nagl W, Ignacinamuthu S & Becker J (1997) Genetic engineering and regeneration of Phaseolus and Vigna. State of the art and new attempts. J. Plant Physiol. 150:625–644Google Scholar
  44. Ozaki K (1986) Plantlet formation from the calli of primary leaf of adzukibean (Vigna angularis) Japan J. Breed. 36:416–419Google Scholar
  45. Pal M, Gosh M, Chandra M, Pal A & Biswas MM (1991) Transformation and regeneration of mungbean (Vigna radiata). Ind. J. Biochem. Biophys. 28:449–455Google Scholar
  46. Santalla M, Power JB & Davey MR (1998) Efficient in vitro shoot regeneration responses of Phaseolus vulgaris and P.coccineus. Euphytica 102:195–202CrossRefGoogle Scholar
  47. Sato T, Asaka D, Harada T & Matsukawa I (1993) Plant regeneration from protoplasts of adzukibean (Vigna angularis Ohwi & Ohashi). Japan J. Breed. 43:183–191Google Scholar
  48. Sato T (1995) Basic study of biotechnology in adzukibean (Vigna angularis Ohwi and Ohashi). Report of Hokkaido Pref. Agric. Exp. Station 87:1–68(Japanese)Google Scholar
  49. Shekhawat NS & Galston AW(1983) Isolation, culture and regeneration of mothbean Vigna aconitifolia protoplasts. Plant Sci. Lett. 32:43–51CrossRefGoogle Scholar
  50. Shiva Prakash N, Pental D & Bhalla-Sarin N (1994) Regeneration of pigeonpea (Cajanus cajan) from cotyledonary node via multiple shoot formation. Plant Cell Rep. 13:623–627CrossRefGoogle Scholar
  51. Smartt J (1981) Gene pools in Phaseolus and Vigna cultigens. Euphytica 30:445–449CrossRefGoogle Scholar
  52. Smartt J (1985) Evolution of grain legumes III. Pulses in the genus Vigna. Exp. Agric. 21:87–100CrossRefGoogle Scholar
  53. Takahashi W, Matsushita J, Kobayashi T, Tanaka O & Beppu T (1998) Plant regeneration from epicotyl segment and callus of Vigna angularis (cv Tanbadainagon) Japan J. Crop. Sci. 67:561–567(Japanese)Google Scholar

Copyright information

© Kluwer Academic Publishers 1999

Authors and Affiliations

  • Renato A. Avenido
    • 1
    • 2
  • Kazumi Hattori
    • 1
  1. 1.Laboratory of Plant Genetics and Breeding, Graduate School of Bioagricultural SciencesNagoya UniversityChikusa, NagoyaJapan
  2. 2.Institute of Plant Breeding, College of AgricultureUniversity of the Philippines Los Banos, CollegeLagunaThe Philippines

Personalised recommendations