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Adventitious Shoot Regeneration from Immature Embryo Explant Obtained from Female × Female Momordica Dioica

A. Hoque
Department of Botany, University of Rajshahi, Rajshahi

M. Hossain
Department of Botany, University of Rajshahi, Rajshahi

S. Alam
Department of Botany, University of Rajshahi, Rajshahi

S. Arima
Faculty of Agriculture, Saga University, Japan.

R. Islam
Department of Botany, University of Rajshahi, Rajshahi

Abstract/ Executive Summary:

Higher percentage of adventitious shoots regenerated from the immature embryo explant than that of immature cotyledon of the crosses made between female and female (pollen was collected from induced bisexual flower) of tetraploid kakrol (Momordica dioica) on MS induction medium supplemented with different concentrations and combinations of BAP, NAA and GA₃. The best response of shoot proliferation was obtained in embryo explants grown in supplemented with 10.8 mg/l BA, 1.08 mg/l NAA and 0.54 mg/l GA₃; whereas, shoot regeneration on cotyledon was achieved on a MS supplemented with 16.2 mg/l BA, 2.7 mg/l NAA and 0.54 mg/l GA₃. In both cases shoot primordia emerged continuously. This system exhibited a potential for repeated harvesting of the shoots from the original explant as the latter continued to expand and regenerate new shoots, upon repeated periodical subculturing on to fresh medium. Regenerated shoots were excised from both sources and rooted in MS supplemented with different concentrations of IAA. There was no significant difference of root proliferation rate than their sources. Rooted plantlets were acclimatized successfully and later established in the field for the production of female plant and also evaluation of somaclonal variation.

Keywords: Immature cotyledon, Immature embryo, Momordica dioica, Organogenesis

Location:

Start Date:

End Date:

Program Area: Variety and Species

Commodity:

Objectives:

The present paper describes a rapid, simple and comparatively efficient shoot regeneration system from immature embryo- and cotyledon explants from the seeds of female × female of kakrol, thus establishing a technique that can be commercially exploited for mass production of female kakrol plants

Methodology:

Seeds from unripe fruits (18 days after anthesis) of the cross between female × female tetraploid kakrol [Momordica dioica (Roxb.) Willd.] were collected from the experimental field. Seeds were washed with distilled water (200 ml) containing three drops of Tween 80 for five min to remove the outer mucilaginous sheath. Tween 80 was removed by washing five - six times with distilled water. Surface sterilization of seeds was carried out with 0.1% HgCl₂ (w/v) for ten min and subsequently the seeds were rinsed four times with sterile distilled water. The seed coat was removed and explants (6) consisting of embryo and fully grown cotyledons were cultured in Petri dishes on to a solid (30 ml, agar 0.6%) MS containing sucrose 3%, and without growth regulators led to the swelling of the explants, facilitated dissection of the embryos on the next day. After swelling the explants, embryo (2 mm) and cotyledon were dissected carefully and cultured separately on to the MS containing different concentrations and combinations of BA (5.4 - 16.2 mg/l), NAA (0.54 - 2.7 mg/l) and GA₃ (0.54 mg/l). For shoot multiplication, a part of nodular callus (6 - 7 mm³) which originated from both explants was cultured on to the selected regeneration media (30 ml) in conical flasks of 250 ml.Each experiment was randomized completely, with three replications per treatment and 18 explants per replication. Observations were recorded every week for shoot proliferation and rooting. The comparison of means was analyzed using DMRT (Duncan 1955) at p < 0.05.

Source of Information: Plant Tissue Cult. & Biotech. 17(1): 29-36, 2007 (June)

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

In conclusion, to our knowledge this is the first report for in vitro adventitious shoot regeneration from embryo axes of seeds from female × female of tetraploid kakrol. Our method allows plant regeneration through adventitious shoot bud formation in kakrol within four weeks, and the young plants can be transferred to soil seven - eight weeks thereafter. The protocol described here provides a rapid and prolific shoot regeneration system that has opened possibilities for commercial production of female kakrol plants and at the same time would provide somaclonal variants to plant breeders to come up with improved varieties through their selection.

Knowledge Management: Journal