Abstract
Black Gram (Vigna mungo L.), widely known as Mashkalai in Bangladesh is an important protein source used as human food as well as fodder. BARI Mash 2 is a popular black gram variety released by Bangladesh Agriculture Research Institute (BARI) which is cultivated throughout the country and very popular especially in the char areas. Establishment of a reliable regeneration system for BARI Mash 2 has been tried for further genetic improvement. A rapid, reproducible and efficient in vitro regeneration method was developed using hypocotyl and young leaf explants through callus formation. The frequency of callus formation was highest (75%) on Murashige and Skoog (MS) medium supplemented with a high concentration (31.66 ?M) of 2,4-Dichlorophenoxyacetic Acid (2,4-D) using the young leaf as explants source. Callus induction rate was less in hypocotyls in the same medium. No further progress was observed from those calluses. MS medium containing 16.11?M of ?- Naphthalene acetic acid (NAA) showed the 70% calli induction from hypocotyls segment. These calli were amenable to produce multiple shoots (5-6 shoot) in the medium containing 17.75 ?M of 6 Benzyl aminopurine (BAP) alone and the combination of BAP (17.75 ?M ) and NAA (2.68 ?M). Shoots were rooted most effectively (55%) in half strength MS basal medium containing 7.38 ?M of Indole-butyric Acid (IBA). Well rooted plantlets were successfully acclimatized, transferred to the soil and found to produce flowers and fruits. The efficient and reproducible regeneration protocol described here allows for successful in vitro regeneration of BARI Mash 2 that is vital for future genetic manipulation.Jahangirnagar University J. Biol. Sci. 6(1): 23-33, 2017 (June)
Highlights
Callus induction: In this experiment, hypocotyl and leaf segments were chosen as explants for callus initiation
Explants were cultured on Murashige and Skoog (MS) medium containing different concentrations of Auxin (2,4-D and Naphthalene Acetic Acid (NAA)) to identify and select the optimum amount of growth regulators for maximum callus initiation and shoots regeneration
It was observed that callus formation was highly affected by 2,4-D concentration in different explants and increase the level of 2,4-D accelerated the callus initiation in short period of time
Summary
The present investigation was attempted to develop an efficient and reproducible protocol for successful in vitro regeneration and multiple shoot formation through callus using different explants such as hypocotyls and leaf segments of BARI Mash 2. For in vitro germination the surface of BARI Mash 2 seeds were sterilized using different concentration and incubation time on commercial Clorox (1% Sodium Hypochlorite) and Tween 20. Explants were cultured on MS medium containing different concentrations of Auxin (2,4-D and NAA) to identify and select the optimum amount of growth regulators for maximum callus initiation and shoots regeneration.
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