Summary
Protoplasts were isolated from hypocotyl of V. mungo (L.) Hepper or hypocotyl-derived callus of V. sublobata (Phaseolus sublobata Roxb.) and V. unguiculata (L.) Walp (syn. V. sinensis (L.) Saviex Hassk) using an enzyme solution comprising Cellulase 2.5%, Macerozyme, Hemicellulase and Driselase each at a 0.5% level in 0.5 M sorbitol. Isolated protoplasts were cultured in Murashige and Skoog's (1962) basal liquid medium supplemented with BA, NAA, 2,4-D (1 mg/l each) and sucrose (14%). After four weeks, protoplast colonies were transferred to the same medium with a reduced level of sucrose (7%). Colonies proliferated into actively growing calli. Further attempts to regenerate plants from such calli were not successful. However, protoclones of V. unguiculata differentiated roots on auxin/cytokinin supplemented media. Alternative methods for shoot differentiation from protoplastderived cultures were tried by the use of Agrobacterium tumefaciens “shooter” strains pGV 2215 or pGV 2298 or wild type strain B6S3.
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References
Arcioni S, Davey MR, Dos Santos AVP, Cocking EC (1982) Somatic embryogenesis in tissues from mesophyll and cell suspension protoplasts of Medicare coerulea and M. glutinosa. Z Pflanzenphysiol 106:105–110
Bharal S, Rashid A (1980) Isolation of protoplasts from stem and hypocotyl of the legume Vigna sinensis and some factors affecting their regeneration. Protoplasma 102:307–313
Davey MR, Bush E, Power JB (1974) Cultural studies of a dividing legume leaf protoplast system. Plant Sci Lett 3:127–133
Davey MR (1983) Recent developments in the culture and regeneration of plant protoplasts. In: Potrykus I, Harms CT, Hinnen A, Hutter R, King PJ, Shillito RD (eds) Protoplasts. Birkhauser, Basel, pp 19–29
Dos Santos AVP, Otuka DE, Cocking EC, Davey MR (1980) Organogenesis and somatic embryogenesis in tissues derived from leaf protoplasts and leaf expiants of Medicago sativa. Z Pflanzenphysiol 99:261–270
Gill R, Eapen S (1986) Plant regeneration from hypocotyl protoplasts of mothbean (Vigna aconitifolia). Curr Sci 55:100–102
Gresshoff PM (1980) In vitro culture of white clover:callus, suspension, protoplast culture and plant regeneration. Bot Gaz 141:157–164
Grosser JW, Collins GB (1984) Isolation and culture of Trifolium rubens protoplasts with whole plant regeneration. Plant Sci Lett 37:165–170
Johnson LB, Stuteville DL, Higgins RK, Skinner DZ (1981) Regeneration of alfalfa plants from protoplasts of selected Regen S clones. Plant Sci Lett 20:297–304
Kao KN, Michayluk MR (1980) Plant regeneration from mesophyll protoplasts of alfalfa. Z Pflanzenphysiol 96:135–141
Krishnamurthy KV, Godbole DA, Mascarenhas AF (1984) Studies on a drought resistant legume:the mothbean, Vigna aconitifolia (Jacq) Marechal. 1. Protoplast culture and organogenesis. Plant Cell Rep 3:30–32
Lu DY, Pental D, Cocking EC (1982) Plant regeneration from seedling cotyledon protoplasts. Z Pflanzenphysiol 107:59–63
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:471–497
Shekhawat NS, Galston AW (1983) Isolation, culture, and regeneration of Moth bean Vigna aconitifolia leaf protoplasts. Plant Sci Lett 32:43–51
Shekhawat NS, Galston AW (1983) Mesophyll protoplasts of fenugreek (Trigonella foenum-graecutn):isolation, culture, and shoot regeneration. Plant Cell Rep 2:119–121
Steffen A, Eriksson T, Schieder O (1986) Shoot regeneration of mesophyll protoplasts transformed by A grobacterium tumefaciens generally not achievable with untransformed protoplasts. Theor Appl Genet 72:135–140
Takebe I, Labib G, Melchers G (1971) Regeneration of whole plants from isolated mesophyll protoplasts of tobacco. Naturwissenschaften 58:318–320
Wilson VM, Haq N, Evans PK (1985) Protoplast isolation, culture and plant regeneration in the winged bean (Psophocarpus tetragonolobus L.) DC. Plant Sci 41:61–68
Xu ZH, Davey MR, Cocking EC (1982b) Organogenesis from root protoplasts of the forage legume Medicago sativa and Trigonella foenum-graecum. Z Pflanzenphysiol 107:231–235
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Communicated by G.S.Khush
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Gill, R., Eapen, S. & Rao, P.S. Callus induction from protoplasts of V. unguiculata, V. sublobata and V. mungo . Theoret. Appl. Genetics 74, 100–103 (1987). https://doi.org/10.1007/BF00290091
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DOI: https://doi.org/10.1007/BF00290091