Abstract A procedure for the rapid tissue culture propagation of papaya is

being developed. 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

In vitro and in vivo germination of papaya (Carica

papaya L.) seeds

J. Bhattacharya and S. S. Khuspe

Plant Tissue Culture Division, National Chemical Laboratory, Pune 411 008, India

Accepted 19 December 2000

Available online 30 October 2001.

Abstract The effect of various pre-sowing treatments on seed germination in

soil of 10 widely cultivated cultivars of papaya were studied. Intact seeds, naked
embryos, halved seeds and halved embryos were germinated under in vitro
culture conditions and the effects of thidiazuron (TDZ), 6-benzyl amino purine
(BAP), naphthalene acetic acid (NAA), 2,4-dichloro phenoxyacetic acid (2,4-D)
and 2,4,5-trichloro phenoxyacetic acid (2,4,5-T) at different concentrations,
temperature and light were examined. Without pretreatment, the percentage
germination in soil varied between 3 and 71% (average 40.2%). Soaking for 24 h
in 200 ppm GA3 increased germination to 1279% (average 56.5%). The
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.

PROPAGATION OF PAPAYA THROUGH TISSUE CULTURE

Authors M.S. Rajeevan, R.M. Pandey
:
Abstract:
Tissue culture system using shoot tips from seedlings and lateral buds from
female plants was tried for the propagation of papaya variety Coorg Honey Dew.
Shoot tips and lateral buds could be established in Murashige and Skoog (MS)
medium containing Naphthalene acetic acid (NAA) 0.5 10 m and Kinetin (K)
2550 m. 6- Benzyl amino purine (BAP) 2 m was optimal for multiplication of
shoots. Indole butyric acid 10 m was effective for inducing profuse rooting and
normal plantlet formation.

Better survival of plantlets was obtained in the potting mix, sand: soil: cowdung
(1:1:1 v/v).

Explants from plantlets viz., stem segments (B5 medium with NAA 10 m and K
5 m), root (B5 medium containing NAA and K each at 10 m), petiole and leaf
segments (MS medium with NAA 0.5 m and BAP 2.5 m) could give rise to
callus in medium indicated in brackets.

TISSUE CULTURE PROPAGATION OF PAPAYA

Authors D. Shlesinger, O. Reuveni, U. Lavi
:
Abstract:
A rapid vegetative propagation of Papaya (Carica papaya L.) is needed for
establishing the selected tree of a breeding project.

We report here the procedure for tissue culture propagation of a dioecious

variety.

The explant were axillary buds of three years old field plants. Sterilization was
carried out by Rifampicin and NaOC1 solution. Initiation media was modified MS.
Multiplication media was MS supplemented with 0.5 ppm BA and 0.1 ppm NAA.
Rooting was carried out by immersing the explants in 1 ppm IBA for 1218 hours
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.