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Tissue culture methods using apices of nursery and orchard trees of Carica papaya cv. Sunrise Solo were evaluated. The explants were established in a modified Murashige and Tucker (1969) basal medium with half-strength inorganic salts, 0.5mgl-1 6-benzylaminopurine (BA) and 0.2mgl-1 naphthaleneacetic acid (NAA). Established explants were transferred to a proliferation medium consisting of Murashige and Tucker (1969) basal medium, 0.5mgl-1 BA and 0.1mgl-1 NAA, which caused extensive multiplication of shoots. Rooting was induced at a higher frequency by subculturing plantlets onto media with indole-3-butyric acid (IBA) than with NAA.
Key words tissue culture propagation - Carica papaya - in vitro establishment - proliferation - rooting
difference between in vivo and in vitro seed germination was observed to be the lowest in honeydew (6.3%) and highest in Disco (68%). In general, in vitro culture conditions increased the percentage and rate of seed germination in all cultivars. An average maximum germination of 95.5% was obtained after 78 days for naked embryos when cultured in the light at 30C on Murashige and Skoogs medium (MS) supplemented with TDZ (1.0 M/l). Author Keywords: Papaya; Seed; Germination; Carica papaya L.; In vitro germination; In vivo germination Abbreviations: BAP, 6-Benzyl amino purine; GA3, Gibberellic acid; NAA, Naphthalene acetic acid; MS, Murashige and Skoog medium; TDZ, Thidiazuron; 2,4-D, 2,4-Dichloro phenoxyacetic acid; 2,4,5-T, 2,4,5-Trichloro phenoxyacetic acid 2. Papaya (Carica papaya L.) is an important fruit species and is widely cultivated in many tropical and subtropical countries. The conventional methods of maintaining papaya germplasm under nursery and orchard conditions require extensive space and labor. It is very difficult to conserve papaya germplasm by conventional methods. In this study shoot tips from papaya were multiplied in vitro and cryopreserved by vitrification. Micropropagation was improved by testing growth regulator concentrations. Shoots grown on MS medium with 0.5 mg/L BA, 0.1 mg/L NAA and 1.0 mg/L GA3 were significantly better than on other treatments. Optimized vitrification procedures indicated that for papaya the best protocol was with shoot tips (2-3 mm) precultured for 3 days at 25CV on MS medium with 5% DMSO and 5% sucrose. Shoot tips (1.5~2.5 mm) were pretreated in 60% PVS2 for 50 min at room temperature, then they were completely dehydrated with 100% PVS2 for 30 min at 0C. After changing the solution with fresh PVS2 the materials were plunged directly into liquid nitrogen and stored for at least 24 hours. Shoot tips were rapidly warmed in a 40Cwater bath, and washed twice with 1.2 M sucrose solution and transferred to the optimized medium. Shoot tip regrowth was 52.6%. The shoots developed normally, rooted and survived transplantation.
and then transfer to liquid MS basal medium. Internal bacterial contamination was a serious obstacle.
Micropropagation of Papaya (Carica Papaya L.)
Micropropagation of papaya began three decade ago. Papaya is a polygamous fruit crop and both dioceous as well as gynodioceous varieties of papaya are being cultivated. Cloning of female papaya plants through in vitro shoot bud culture is an ideal approach. In vitro regeneration with shoot tip, excised from mature papaya plant, has been attempted (Ali & Hogan, 1976; Litz & Conover, 1978; Rajeevan & Pandey, 1986; Drew, 1988, 1992). However, most of the protocols are genotype dependent and could not be reproduced. Callus culture has also been reported in papaya (Arora & Singh, 1978; Litz et al., 1983). Papaya is recalcitrant to tissue culture. Slow rate of proliferation, poor establishment of axenic cultures and high mortality of plants during acclimatization are some of the main problems of papaya micropropagation. Most of the reports on papaya micropropagation using mature explants are therefore conflicting. Somatic embryogenesis system using hypocotyl (Fitch, 1993) or immature zygotic embryos (Fitch & Manshardt, 1990) has been successful in vitro regeneration. The micropropagation protocol described herein is based on direct somatic embryogenesis using immature zygotic embryo for somatic embryo induction and subsequent plant regeneration.