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In Vitro Regeneration of Papaya (carica Papaya L.) (2019--2020)

M.M. Khatun
Biotechnology Division, BARI, Joydebpur, Gazipur

A. Saha
Biotechnology Division, BARI, Joydebpur, Gazipur

K. Nahar
Biotechnology Division, BARI, Joydebpur, Gazipur

D. Khanam
Biotechnology Division, BARI, Joydebpur, Gazipur

M.A.Y. Akhond
Biotechnology Division, BARI, Joydebpur, Gazipur

Abstract/ Executive Summary:

The aim of this study was to develop an efficient regeneration protocol of papaya from immature seeds. Different concentrations of 2, 4-D were used for callus formation. The highest percentage of explant produced callus (39%) in 10 mg/l 2, 4-D concentration. In regeneration, the highest shoot number (32), highest shoot length (4.33 cm), highest leaf number (7.70), higher root number (2.40) and maximum root length (2.10 cm) was observed in 0.06 mg/l both BAP & NAA and 3.5 mg/l GA_3.In ex vitro condition all together 37% of plantlets were survived. They were well established in the field and produced fruit.

Keywords: Somatic embryogenesis, In vitro regeneration, Papaya

Location:

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Papaya

Objectives:

The study was aimed at finding out the embryogenic responses and plantlet regeneration from immature seeds of papaya.

Methodology:

Immature seeds of Shahi papaya were used for callus formation and regeneration in MS (Murashige and Skoog, 1962) medium. For callus formation, different concentrations of 2, 4-D were used in the medium. The pH of all media was adjusted to 5.8 before autoclaving. The media were autoclaved at 121⁰C and 1.06 kg cm^-2 pressure for 20 minutes. The seeds were washed with running tap water with jet trix for 30 min. After that, the seeds were sterilized by Sodium hypochloride solution with two drops of Tween-20 and washed 3-4 times with sterilized distilled water. Then, embryos were isolated and inoculated in a callus induction medium. All cultures were incubated at 25 ± 1⁰C with complete darkness. After callus formation, they were cultured on a somatic embryogenesis medium and kept in 16/8 hours light/dark photoperiod with a light intensity of 2000 lux under cool white fluorescent tubes. After 20 days they were again transferred to a regeneration medium.

Source of Information: Annual Research Report 2019-2020, Biotechnology Division, BARI, Joydebpur, Gazipur

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Immature seeds were cultured in four different concentrations of 2, 4-D for callus formation. The highest percentage of explants produced callus (39%) in T₂ concentration followed by T₁ (33%). On the other hand, a lower number of callus (28%) was observed in T₄ treatment. Calli were then transferred to a somatic embryogenesis medium. In this medium, some calli transformed to embryogenesis processes and some organogenesis processes. The produced embryos were cultured in MS medium without any growth regulators or with growth regulators that is the lower amount of weaker auxin-based medium. In both cases, some embryos were regenerated into intact plantlets and some regenerated shoots without roots. The shoots without roots were again cultured into a rooting medium for root induction. In the present investigation, high embryogenic responses were also observed in using auxin 2, 4-D in the medium. The result showed that the highest shoot number (32), the highest shoot length (3.77 cm), higher root number (4.33) and highest root length (2.47 cm) were obtained from T₂R₂ medium The well-developed plantlets with good root system were transferred to a greenhouse for acclimatization. After three to four days of hardening, the plantlets were removed from the culture tubes and all the adhering media were washed carefully so that, the root damage was the least. The plantlets were then planted into small plastic pots containing sand, soil and decomposed cowdung at the ratio of 1:1:1 and covered with plastic bags for 20 days. The bags were cut one side at 7 days to make a hole and at 15 days, the other side of the bags were cut. The next week, whole covers were removed to acclimatize the plantlets to normal greenhouse conditions. The plantlets were maintained in the greenhouse with proper care and watered whenever necessary. In our study, T₁ treated plantlets survived 25% and T₂treated plantlets survived 45%. In greenhouse conditions with proper management, the plantlets grew normally and found well established within three months. They were then ready for transplanting in the field. The plantlets developed from the present study were planted in the research field of Biotechnology Division, BARI, Gazipur. The plantlets were successfully established in the field and they were grown well. The plants were produced flower within three months from planting in the field and produced fruit within three months after flowering

Knowledge Management: Report/Proceedings