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Tissue Culture of a Year-round Fruiting Variety of Artocarpus Heterophyllus L. in Bangladesh

F.M.S. Azam
Department of Biotechnology and Genetic Engineering University of Development Alternative Dhanmondi, Dhaka Bangladesh

M. Rahmatullah
Department of Biotechnology and Genetic Engineering University of Development Alternative Dhanmondi, Dhaka Bangladesh

Ather-uz-Zaman
Centre for Plant Tissue Culture PROSHIKA, Dhaka Bangladesh

Abstract/ Executive Summary:

Jackfruit (Artocarpus heterophyllus) is available in Bangladesh between June to August; however, a variety exists that fruits throughout the year. The edible portion is considered as a good source of carbohydrates, proteins, vitamins and minerals. In Bangladesh, malnutrition persists among 35-45% of the population and the child malnutrition rate is more than 50%. Under this circumstance, the wide cultivation of a jackfruit variety that bears fruit throughout the year can play a significant role in reducing malnutrition in Bangladesh. Since this particular variety is not widely available, we have developed a method for its clonal propagation. Healthy and juvenile shoot tips and nodal segments were collected from field-grown fruit-bearing trees and cultured in Murashige-Skoog (MS) media fortified with different concentrations and a combination of 6-benzyl amino purine and kinetin following the sterilization with 0.1% HgCl2. Sprouting and regeneration of micro shoots geared up when MS was enriched with 3.5 mg/L BAP. Proliferation frequency increased considerably and multiple shoots were regenerated as a clump of 2-3 shoots in pH 5.8 media within four weeks of inoculation through the synergistic effect of 6- benzyl amino purine (3.5 mg/L) and kinetin (1.5 mg/L). With the increase of subculture (up to the 10th maximum), the frequency of shoot proliferation was enhanced. The addition of 0.1 mg/L indole-3-acetic acid and 20% coconut water considerably increased shoot elongation and stimulated the growth of the shoots. About 80% rooting frequency was observed in ½ MS medium with 1.2 mg/L indole-3-butyric acids. Rooting percentage and their growth was much better in liquid media. After proper acclimatization, rooted plantlets were transferred to polythene bags containing garden soil, sand, and cowdung (1:1:1). After eight weeks of transplantation in an open field, more than 80% of the plants had survived. No morphological variants were observed during the passage of clonal propagation. This clonal propagation technique could play a significant role in improving productivity leading to reduced malnutrition.

Keywords: Jackfruit, Clonal propagation, Synergistic effect, Malnutrition improvement

Location:

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Fruit

Objectives:

This research study was undertaken to develop an efficient protocol using tissue culture technique for the large scale propagation of this year-round fruiting jackfruit variety by examining the effects of growth regulators and the Murasighe and Skoog (MS) medium (Murashige and Skoog, 1962).

Methodology:

This experiment was carried out at the Plant Tissue Culture Laboratory of PROSHIKA, Bangladesh in 2006 to 2007. Explant Collection and Sterilization Fifty to seventy explants were collected for each experiment. These were 10-15 day-old newly-sprouted healthy shoots from 10-12-year-old fruit-bearing trees of the year-round fruiting variety of jackfruit growing at the Bangladesh Agricultural Research Institute (BARI). The actual number of explants used in any particular experiment is detailed in the accompanying tables. After removal of expanded leaves, the shoot apices and nodes were washed thoroughly under running tap water for 30 min. then treated with Tween 80 plus Savlon for 10 min. and again rinsed with distilled water. The explants were surface disinfected with 0.1% HgCl2 (for 5-15 min.) followed by seven times of rinsing with sterile distilled water in a laminar airflow cabinet. The explants were finally trimmed, prepared, and cultured on the medium. Shoot Proliferation Stage Explants were implanted vertically on MS medium supplemented with different (0.2, 0.5, 0.8, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5 mg/L) concentrations of 6- benzyl amino purine (BAP). All media were supplemented with 3% sucrose and 0.8% Difco Bacto-agar. The pH was adjusted to 5.8 prior to autoclaving the media at 1 kg pressure per cm2 and 121°C for 20 min. All culture vessels were incubated in a growth room at 26±2°C with 16 hrs light provided by Philip white fluorescent tube lights with a photon flux density of 3,000 lux. Multiple Shooting In a second experiment, jackfruit explants were cultured on MS medium fortified with different concentrations and combinations of BAP (3.0, 3.5, 4.0 mg/L) and kinetin (Kn) from 0.2 to 3.0 mg/L to observe the synergistic effect on multiple shoot formation. Root Induction For root induction, micro-shoots were cultured on ½ MS medium enriched with different concentrations (0.2 to 2.0 mg/L) of indole-3-butyric acid (IBA). Acclimatization In vitro derived plantlets were gradually hardened through gradual exposure to optimum relative humidity and sunlight for seven days and then transferred to a net house. Plantlets were taken out from the culture vessels/tubes and then washed carefully under running tap water to remove any traces of agar. Each plantlet was transplanted to one of the small poly bags that contained different compositions and combinations of soil media to select the best formulation. The plantlets were immediately covered with a perforated polythene bag to prevent desiccation. Open Field Observation Acclimatized plants were observed for proper establishment in soil for fifteen days and field observation was done from January to February 2007 and data were recorded in one-week intervals. Data Assemblage Weekly growth observations were done up to four weeks after establishment, and experimental data were recorded. The parameters were the following: i. Shoot length in cm and number of leaves/shoot, ii. A number of multiple shoot/explant, iii. The average length of multiple shoots in cm, iv. A number of root/shoot, and v. Root length in cm. Recorded data were analyzed as mean ± SE according to Mian and Miyan (1984).

Source of Information: Proc. IS on Underutilized Plants Eds.: Jaenicke et al. Acta Hort. 806, ISHS 2009

Article Link (Online Journal):

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Achievement/Findings:

With this experiment, a protocol for the micropropagation of a unique variety of A. heterophyllus which fruits throughout the year was established from shoot tip and nodal explants. It may be noted that all previous studies were conducted with varieties of this species which fruits for only two to three months in any given year. To our knowledge, this is the first successful description of micropropagation of this unique variety. Successful micropropagation of this variety can lead to widespread cultivation and for obtaining this fruit throughout the year, which in turn can lead to a cheap source of nutritious food available to the general population and specifically to the nutritionally impoverished population of Bangladesh and other countries where this variety can be cultivated.

Knowledge Management: Report/Proceedings