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Effect of Growth Regulators and Carbon Sources on Axillary Shoot Proliferation from Shoot-tip Explant and Successful Transplantation of Papaya (carica Papaya L.)

A. H. Kabir
Department of Botany, University of Rajshahi, Rajshshi 6205, Bangladesh.

M. A. Bari
Department of Botany, University of Rajshahi, Rajshshi 6205, Bangladesh.

A. K. M. N. Huda
Department of Genetics and Breeding, University of Rajshahi, Rajshshi 6205, Bangladesh.

M. A. Rezvy
Department of Botany, University of Rajshahi, Rajshshi 6205, Bangladesh.

I. Mahfuz
Department of Zoology, University of Rajshahi, Rajshshi 6205, Bangladesh.

Abstract/ Executive Summary:

The investigation was based on direct shoot formation from shoot tip explant, subsequent morphogenesis and rooting of the in vitro proliferated shoots and transplantation of regenerated plants under ex vitro condition. Through proper growth regulators, it was possible to differentiate multiple shoots from shoot tip of papaya (Shahi). In the present investigation, it was observed that shoot proliferation was best in MS medium containing BAP 1.0 mg L^-1+KIN 0.5 mg L^-1 and BAP 1.0 mg L^-1+NAA 0.5 mg L^-1. Maximum number of shoot per culture (28.2) and length of the largest shoot (1.7 cm) was found on MS medium supplemented with BAP 1.0 mg L^-1+KIN 0.5 mg L^-1. Shoot proliferation efficiency was found the best in 11 weeks old explant in MS medium containing BAP 1.0 mg L^-1+KIN 0.5 mg L^-1. Sucrose in 30 g L^-1 concentration as carbon source was proved to be best for shoot proliferation (82%). Maximum number of shoot per culture (41.2) and maximum length of shoot were obtained in the aforesaid concentration of carbon source. For adventitious rooting of regenerated shoot, MS medium containing IBA 1.0 mg L^-1+NAA 0.5 mg L^-1 was found to be the best. In this growth regulator, treatment 92% of rooting was observed. Longest length of the root (7.9 ± 0.121 cm) was found on MS medium supplemented with IBA 1.0 mg L^-1+IAA 0.5 mg L^-1. After transplanting the 20 days old rooted shoots into garden soil, compost and sand (2:1:1), 80% of survivability after 5 weeks was achieved.

Keywords: Micropropagation, Shoot-tip, Morphogenesis, Rooting, Transplantation

Location: Central Science Laboratory at the University of Rajshahi

Start Date: 00-03-2005

End Date: 00-07-2006

Program Area: Variety and Species

Commodity: Papaya

Objectives:

To establish a simple method of micro propagation of papaya through shoot tip culture for obtaining large-scale disease free seedlings.

Methodology:

Shahi, a local variety of Caria papaya L. (Papaya) was used as experimental material in this investigation. Shoot tips of field grown seedlings of Carica papaya L. (Shahi) were collected from Regional Horticultural Research Station, Rajshahi. Shoot tips collected from 10-12 weeks old field grown plants were disease free and healthy. This research was carried out in central science laboratory at the University of Rajshahi from the period of March, 2005 to July, 2006. The shoot tips of papaya were washed thoroughly under running tap water and then treated with 1% Savlon and 4-5 drops of Tween-80 for about 15-20 min with constant shaking. Then explants were washed 3-4 times with distilled water to make the material free from Savlon. Subsequently the explants were transferred to laminar airflow cabinet and transferred to 250/500 mL sterilized conical flask. After rinsing in 70% ethanol for 1-3 min the explants were immersed in 0.1% HgCl₂. The sterling with explants was constantly shaked during sterilization according to necessity of time to increase the release of free Cl₂. The surface sterilized shoot tip materials were taken in sterilized petridish. Shoot tips were trimmed to (1.0-1.5 cm) terminal parts of lightly furled leaves and 1.2 node of shoot tip explants. The photo period was maintained generally 14 h light 10 h dark. These explants were cultured on MS nutrient medium supplemented with BAP, KIN, NAA alone or in combination at different concentration. Days required to shoot initiation, percentage of explants showing shoot proliferation, number of total shoots, per explant, length of the longest shoots were considered as parameters for evaluating this experiment. At this stage, the proliferating cultures were subcultured again in the same initial medium in order to increase budding frequency. In this experiment, shoot tip explants from 10, 11 and 12 weeks old field grown seedlings were also used. Explants were cultured on MS nutrition medium supplemented with 1.0 mg L^-1 BAP and 0.5 mg L^-1 KIN for proliferating shoots. Local sugar, sucrose and glucose with three different concentration viz., 20, 30 and 40 g L^-1 for each were used for this experiment. The shoot tips were used as explants for this experiment and they were cultured on MS medium containing BAP 1.0 mg L^-1+KIN 0.5 mg L^-1 with three different concentration of each carbon source separately. The shoots proliferated from shoot tip explants in parent culture or subsequent subculture was induced for adventitious root formation to get full-fledged plantlets in vitro. The usable shoots (with length> 2 cm) were collected aseptically from shoot culture for rooting and cuttings were cultured individually in 150 x 25 mm tubes with 20 mL of rooting medium having different strength and combination of auxin. The observations on development pattern of roots were made throughout the entire culture period. Rooted plants were taken out from the culture tubes and washed carefully under running tap water for complete removal of remains of the medium. Pots (9x6 cm) were kept ready filled with garden soil, compost and sand in the proportion of 2:1:1, respectively. All the pots were covered with wet polythene bag for providing high humidity to the plants. After 45 days, the plants were transferred to the soil.

Source of Information: Biotechnology, 6(2): 268-272, 2007, ISSN 1682-296X

Article Link (Online Journal): DOI: 10.3923/biotech.2007.268.272, URL: http://scialert.net/abstract/?doi=biotech.2007.268.272

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

In the present investigation, it was observed that shoot proliferation was best in MS medium containing BAP 1.0 mg L^-1+KIN 0.5 mg L^-1 and BAP 1.0 mg L^-1+NAA 0.5 mg L^-1. Maximum number of shoot per culture (28.2) and length of the largest shoot (1.7 cm) was found on MS medium supplemented with BAP 1.0 mg L^-1+KIN 0.5 mg L^-1. Shoot proliferation efficiency was found the best in 11 weeks old explant in MS medium containing BAP 1.0 mg L^-1+KIN 0.5 mg L^-1. Sucrose in 30 g L^-1concentration as carbon source was proved to be best for shoot proliferation (82%). Maximum number of shoot per culture (41.2) and maximum length of shoot were obtained in the aforesaid concentration of carbon source. For adventitious rooting of regenerated shoot, MS medium containing IBA 1.0 mg L^-1+NAA 0.5 mg L^-1 was found to be the best.

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