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Regeneration of Blackgram (vigna Mungo L.) on Changes of Hormonal Condition

Shariful Alam Mony
Bangladesh Agricultural University, Department of Biotechnology, Mymensingh-2202, Bangladesh

Mirza Hassanuzzaman
Sher-e-Bangla Agricultural University, Department of Agronomy, Dhaka-1207, Bangladesh

Kamrun Nahar
Sher-e-Bangla Agricultural University, Department of Agricultural Botany, Dhaka-1207, Bangladesh

Md. Shahidul Haque
Bangladesh Agricultural University, Department of Biotechnology, Mymensingh-2202, Bangladesh

Abstract/ Executive Summary:

The study comprised of experiments for shoot regeneration and plantlet formation from cotyledonary node of Vigna mungo by culturing them on low concentration of BAP followed by transfer to hormone free MS medium. Cotyledonary node explants were cultured on different concentrations of BAP (0, 1, 2.5 and 50 mg l^-1). Shoot regeneration occurred from cotyledonary nodes irrespective of the presence or absence of BAP in the medium. However, culture of cotyledonary node explants for 10 days on medium containing 1.0 mg l^-1 BAP followed by transfer to hormone free medium gave higher number of shoots (9.33/explant) compared to culture of the explants on hormone free medium for 15 days followed by transfer to medium containing 1.0 mg l-1 BAP (8.33/explants). The regenerated shoots were transferred to rooting medium supplemented with different concentrations of IBA and NAA. The high frequency (100.0%) of rooting was observed with MS medium supplemented with 0.5 mg l^-1 IBA. The rooted plants were transferred to pots for hardening.

Keywords: blackgram regeneration, Cotyledonary node hormone free medium

Location: Biotechnology Lab, Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202

Start Date: 00-00-2008

End Date: 00-00-2009

Program Area: Variety and Species

Commodity: Black gram

Objectives:

To develop protocols for shoot regeneration and plantlet formation from cotyledonary node of Vigna mungo by culturing them on low concentration of BAP followed by transfer to hormone free MS medium

Methodology:

The seeds of Vigna mungo (2n=22) were collected from Bangladesh Institute of Nuclear Agriculture. Cotyledonary nodes were used as explants in the present study. Surface sterilization of mature seeds was carried out under laminar air flow cabinet. Seeds were washed by sterile distilled water for 3 to 5 minutes. Later, they were rinsed in 70% ethyl alcohol followed by washing with sterile distilled water for 3 times. Finally, surface disinfection was done with 0.1% HgCl₂ for five minutes with continuous agitation. The seeds were then washed 5 times with sterile distilled water to remove the sterilant. The mature seeds were placed on a solidified agar medium containing no growth regulators in sterile vials. Seven to ten days after germination, cotyledonary nodes of seedlings were dis-sected and cultured on MS medium supplemented with different concentrations of BAP (0, 1, 2.5, 5 and 10 mg l^-1) for direct shoot regeneration. Shoots regenerated on medium without growth regulator were transferred to medium containing different concentration of BAP (1, 2.5 and 5 mg l^-1). Again, shoots regenerated on medium containing different concentration of BAP were transferred to medium without BAP. For rooting, 5-6 cm shoots excised from multiple shoots were implanted to rooting medium consisting of MS medium supplemented with IBA (0, 0.25, 0.5, 1, 2, 3, 4 or 5 mg l^-1) and NAA (0.2 or 0.5 mg l^-1). MS medium supplemented with different concentrations and combinations of growth regulators as per treatments was used for shoot induction, shoot multiplication and rooting of shoots. Hormones were added separately to different media according to their requirements. pH ofthe media was adjusted to 5.8 and then agar 9.0 g l^-1 was added to solidify the medium. All instruments, glass wares and culture media were sterilized by autoclaving at 1.16 kg cm^-2 pressure at 121°C for 30 min. The temperature of the growth room was maintained at 25±2°C. A 16 h light period was maintained with light intensity of 2000 lux. At least 20 explants were cultured in each treatment and all treatments were repeated thrice. One month old adequately rooted shoots when reached a height of 7-8 cm were brought out from culture room and kept in room temperature for 7 days. Plantlets were then taken out from the vials and agar was washed out gently from the roots. These plantlets were transplanted in small pots containing a mixture of compost, soil and sand (1:2:1) and were covered with transparent sheets. They were nurtured under room temperature in presence of sufficient light. In that conditions the plantlets were watered every alternate day. The data for the parameters recorded in the present study were statistically analyzed by the programmed MSTATC. The experiment was conducted following Completely Randomized Design (CRD). The one way analysis of variances for different parameters was performed and the means were compared by Duncan’s multiple range test (DMRT) at 5% level of probability for interpretation of results using MSTAT-C Computer programmed.

Source of Information: Not. Bot. Hort. Agrobot. Cluj 38 (3) , 140-145, 2010

Article Link (Online Journal):

Funding Source:

1. Government

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The high frequency (100.0%) of rooting was observed with MS medium supplemented with 0.5 mg l^-1 IBA. The rooted plants were transferred to pots for hardening.

Knowledge Management: Journal