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Callus Induction and High-frequency Plant Regeneration of Pineapple (ananas Comosus (l.) Merr.)

M. Atique Akbar
Department of Botany, Jahangirnagar University, Savar, Dhaka-1342, Bangladesh

Biplab K. Karmakar
Department of Botany, Jahangirnagar University, Savar, Dhaka-1342, Bangladesh

Shyamal K. Roy
Department of Botany, Jahangirnagar University, Savar, Dhaka-1342, Bangladesh

Abstract/ Executive Summary:

Meristem tips of the crown of Ananas comosus, cultured on MS supplemented with 1.5 mg/l NAA and 1.0 mg/l Kn, callused within three weeks. This callus, when subcultured on MS with 1.5 mg/l Kn + 0.5 mg/l NAA produced a large number of shoots. After four weeks, shoots were excised and implanted in shoot elongation medium consisting of MS basal salts fortified with 15% coconut water only. After harvesting shoots, the old callus was transferred to the fresh medium of the same constituents, where the old callus expanded and many new buds also emerged. Shoots rooted well within two weeks, when they were excised individually and implanted in half strength MS with 2.0 mg/l IBA. Eighty per cent plantlets survived when transferred to open field. Regenerated plants were morphologically uniform with normal leaf shape and growth pattern.

Keywords: Pineapple, Callus, Regeneration, Acclimatization

Location: Madhupur pineapple fields, Tangail

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Pineapple

Objectives:

To identify the Callus Induction and High-frequency Plant Regeneration of Pineapple (Ananas comosus (L.) Merr.)

Methodology:

The samples were collected in April - May from Madhupur pineapple fields of Tangail district in Bangladesh. Explants were prepared for further experiments according to the procedure described. After surface sterilization apices were excised at the base and divided into pieces of explants each containing 2 - 3 nodes. The basal medium used for all experiments was MS mineral formulation containing standard salts and vitamins, 30 g/l sucrose and 8 g/l agar (BDH, England). The media were variously supplemented with 2,4-D, BA, Kn alone or in combinations with NAA or IBA and coconut water (5 - 15%). The pH was adjusted to 5.8 ± 0.05 before adding agar, and the media were autoclaved at 1.1 kg cm^_-2 for 20 min at 120⁰C. Cultures were incubated at 25 ± 1⁰C with a photoperiod of 16 h/day at 30 μmol s^_-1,m^_-2 light intensity by cool white fluorescent light. Cultures were initiated in 150 ∇ 25 mm glass tubes containing 12.5 ml of medium. The cultures were regularly subcultured at four weeks intervals on fresh medium in 100 ml conical flasks or juice bottles containing 40 ml of medium. The optimum hormone levels and other supplements in the culture medium essential to maximize production of uniform plantlets were selected. Shoots about 3 - 6 cm in length obtained from the elongation media were separated aseptically from the culture vessels and transferred to rooting media, containing different combinations of auxins. Following generation of sufficient root systems the regenerated plantlets were considered ready for transplantation. For hardening, the vessels containing rooted shoots were kept at normal room temperature and light for 14 days. Thereafter, the rooted shoots were removed from the culture, washed thoroughly to remove remnants of agar from roots and transplanted to small pots containing garden soil, compost and sand (1 : 1: 1 v/v). Plants were well covered with polythene sheet for three weeks to ensure high humidity while irrigating regularly. After three months the plantlets were transferred to open field.

Source of Information: Plant Tissue Cult. 13 (2) : 109-116, 2003 (December)

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The results show that the highest number of shoots was found to regenerate in the medium fortified with 1.5 mg/l Kn + 0.5 mg/l NAA. BA and Kn alone. BANAA combination was not so efficient to initiate shoot regeneration from callus as Kn-NAA combination. The percentage of calli forming shoots, and the number of regenerated shoots per callus varied according to the growth regulator treatment. MS containing Kn alone resulted in shoot differentiation, more efficiently than that observed in BA, but at a lower frequency compared to NAA supplemented media. The technique described here appears to be a promising method of propagation of pineapple. As the potentiality of shoot multiplication from callus continued for a long time, regenerants may be characterized by somaclonal variation. Several species of Dubosia, Cuphea, Amaranthus and Salvia Produced regenerants through callus-mediated adventitious shoot differentiation. Such regenerants may prove to be a potential source of somaclonal variants, giving birth to traits of agronomic importance. The regenerated plants of pineapple are currently being screened for agronomically useful genetic variants. Shoots rooted well within two weeks, when they were excised individually and implanted in half strength MS with 2.0 mg/l IBA. Eighty per cent plantlets survived when transferred to open field. Regenerated plants were morphologically uniform with normal leaf shape and growth pattern.

Knowledge Management: Journal