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Effect of Plant Growth Regulators on Plant Regeneration of Hyacinth Bean (lablab Purpureus (l.) Sweet) Through Nodal Explants

M. T. Islam
Plant Genetic Resources Centre, Bangladesh Agricultural Research Institute (BARI), Joydebpur, Gazipur-1701, Bangladesh

M. S. Haque
Department of Biotechnology, Bangladesh Agricultural University (BAU), Mymensingh-2202, Bangladesh

S. Rahman
Plant Genetic Resources Centre, Bangladesh Agricultural Research Institute (BARI), Joydebpur, Gazipur-1701, Bangladesh

M. R. Molla
Plant Genetic Resources Centre, Bangladesh Agricultural Research Institute (BARI), Joydebpur, Gazipur-1701, Bangladesh

E. R. J. Keller
In Vitro Storage and Cryopreservation, Department of Genebank, Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), OT Gatersleben, Correns Strasse 3, D-06466 Stadt Seeland, Germany.

Abstract/ Executive Summary:

Nodal explants of three accessions namely BD-101, BD-122 and BD-8001 of hyacinth bean (Lablab purpureus (L.) Sweet) were cultured for four weeks on MS medium supplemented with different concentrations and combinations of NAA (0, 0.1 mgl^-1) and BAP (0.5, 1.0 and 2.5 mgl^-1) for plant regeneration. The highest shoot initiation was observed in 0.5 mgl^-1 BAP while the lowest shoot initiation was found in 2.5 mgl^-1 BAP with 0.1 mgl^-1 NAA. Earlier shoot initiation was exhibited in 0.5 mgl^-1 BAP with 0.1 mgl^-1 NAA. The highest number of leaflets and higher shoot lengths were observed in MS medium. Comparatively higher number of shoots was found in BAP (0.5-2.5 mgl^-1). The highest percentage of callus was initiated in medium supplemented with 1.0 mgl^{- 1} BAP. Earlier callus initiation and larger callus size were found in combination of BAP (0.5-2.5 mgl^-1) with 0.1 mgl^-1NAA. BD-122 cultured in MS medium was found superior for shoot regeneration through node culture. Four different concentrations of IBA (0.1-1.5 mgl^-1) were used for rooting. The highest percentage (86.67 %) of rooting was found in MS medium containing 0.1 mgl^-1 IBA at four weeks. Rooting frequency decreased with the increasing concentration of IBA. The accession BD-8001 had 99.60 % rooting in 0.5 mgl^-1IBA. The highest number of longest roots was exhibited in 0.1 mgl^-1 IBA. The regeneration protocol developed from nodal explants has applicability in improvement of hyacinth bean.

Keywords: Lablab purpureus, Nodal explant, Growth regulators, Regeneration

Location: In Vitro Conservation Laboratory of PGRC, BARI, Joydebpur Gazipur

Start Date:

End Date:

Program Area: Resource Development and Management

Commodity: Plant

Objectives:

The present investigation was undertaken to develop a reproducible protocol on plant regeneration using nodal explants of hyacinth bean.

Methodology:

Two experiments were conducted with three accessions viz. BD-101, BD-122 and BD-8001 of hyacinth bean at the In Vitro Conservation Laboratory of PGRC, BARI, Joydebpur Gazipur. The accessions are the landraces and are collected from different regions (districts) viz. Naogoan, Hobiganj and Rangamati in Bangladesh in the first experiment. The edible pod shape is elongate and green in BD-101, flat and light purple in BD-122, and flat and light green in BD-8001. The seed colors of the three accessions are yellow, mosaic and black. Plantlets developed through embryo culture were used as source materials. Four weeks aged plantlets were taken on sterilized Petri dishes. Stem segments with 1-2 nodes having axillary buds were aseptically excised from the apical portion without shoot tip under laminar flow and the nodal segments were used as explants. MS salts with vitamins (Murashige and Skoog, 1962) were supplemented with BAP (0.5, 1.0 and 2.5 mgl^-1) alone or in combination with NAA (0, 0.1 and 0.5 mgl^-1) in 7 treatments. The pH of the medium was adjusted to 5.8 before autoclaving and the media were solidified with agar 8 gl -1 . Ten ml media was poured in each culture tube (25 x 150 mm). The medium was autoclaved at 1.06 kg/cm-2 pressure and 121°C for 15 min. The nodal explants were placed vertically on the medium keeping the tips upside and gently pressed into the culture medium. The explants were incubated for four weeks in a culture room at 22 ± 1°C under about 2000 lux light provided by cool-white fluorescent tubes maintaining a 16/8 h alternate light/dark cycle and 60-70 % relative humidity. The culture tubes were checked daily to note the morphogenic development and the data were recorded on eight parameters (Table 1). The experimental set up was a 7 x 3 factorial in a completely randomized design with five replications each replication consisting of 10 explants. Data were subjected to ANOVA using the General Linear Model procedure. Significant interactions were analyzed. Mean separations were carried out by DMRT at 1 % using the MSTAT program. The percentage data were subjected to square root transformation method (Steel and Torrie, 1960; Gomez and Gomez, 1984). In the second experiment, in vitro raised shoots were transferred to rooting medium for plantlet formation. Shoots were carefully removed from the culture tubes. Each shoot was cut from the basal end and was transferred to MS medium supplemented with 0.1, 0.5, 1.0 or 1.5 mgl^-1 IBA One shoot was placed on the rooting medium and were incubated for four weeks. The experimental set up was 4 x 3 factorial CRD with three replications and each replication consisted of 10 explants. Similar methods were followed as of experiment-1 for data analysis in rooting and plantlets development.

Source of Information: Bangladesh J. Agril. Res. 44(1): 27-42, March 2019 ISSN 0258-7122 (Print), 2408-8293 (Online)

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Achievement/Findings:

It may be concluded that MS medium without any growth regulators is the superior for shoot proliferation through node culture. Percentages of callus initiation increased with the increase in the concentration of growth regulators. The high concentration of IBA suppressed root development. MS medium containing 0.1 mgl^-1 IBA emerged as the superior treatment for rooting of in vitro shoots in hyacinth bean. A protocol for in vitro regeneration from nodal explants of hyacinth bean has been developed in this study. This protocol is reproducible for in vitro culture and, short term preservation of hyacinth bean.

Knowledge Management: