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In Vitro Plant Regeneration of Four Local Varieties of Chickpea (cicer Arietinum L.) Grown in Bangladesh

T. A. Banu
Plant Tissue Culture Section, Biological Research Division, Bangladesh Council of Scientific and Industrial Research, Dhanmondi, Dhaka-1205

R. H. Sarker
Department of Botany, University of Dhaka, Bangladesh.

M. I. Hoque
Department of Botany, University of Dhaka, Bangladesh.

Abstract/ Executive Summary:

In vitro regeneration system was developed through direct organogenesis from decapitated mature embryo explants of locally grown four chickpea varieties, namely, Barichhola-4, Hyprochhola, Binachhola-3 and Binachhola-4. Best response towards multiple shoot regeneration was obtained on MS medium supplemented with 0.5 mg/l BAP, 0.5 mg/l Kn, 0.2 mg/l NAA along with double concentrations of CaCl₂and NH₄NO₃. However good shoot health and expanded leaf was found on MS medium containing 1.0 mg/l kn. Apart from this, few experiments were conducted with decapitated embryo attached cotyledon. Using this explants highest number of multiple shoots were obtained on MSB medium containing 4× micronutrients of MS medium with 3.0 mg/l BAP and 0.04 mg/l NAA in all four varieties. Shoots regenerated on 1.0 mg/l kn supplemented medium showed good response towards rooting on MS medium supplemented with 0.2 mg/l IBA in all four varieties. It was observed that micro grafting is an alternative technique to in vitro rooting in chickpea.

Keywords: In vitro regeneration, Decapitated embryo, Chickpea

Location:

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Chickpea

Objectives:

To establish an efficient regeneration protocol in different chickpea varieties of Bangladesh

Methodology:

Four local varieties of chickpea (Cicer arietinum L.) were used in the present investigation namely Barichhola- 4, Hyprochhola, Binachhola-3, Binachhola-4. Among the seeds of these four varieties of chickpea (Cicer arietinum L.) Barichhola- 4 was collected from BARI, Joydebpur, Gazipur and other three varieties namely Hyprochhola, Binachhola-3 and Binachhlola-4 were collected from BINA, Mymensingh. Seeds were surface sterilized according to the protocol described by Sarker et al. (2003) and soaked over night in sterile distilled water or cultured in water- agar medium for germination. Decapitated mature embryo axes and decapitated mature embryo attached to the cotyledons were used as explants from overnight soaked seeds. The seeds were split open and mature embryo devoid of shoot and root meristems were taken as decapitated mature embryo axes explants. For shoot initiation and development all the explants were cultured on MS (Murashige and Skoog, 1962) or MSB (MS salts and B5 (Gamborg et al. 1968) vitamins) medium with various hormonal supplements namely BAP, Kn, NAA, In certain combinations, MS macrosalt namely CaCl₂ and NH₄HO₃ were used in higher concentrations than those innormal MS. For rooting, 3-5 cm long regenerated shoots were cultured on full or half strength of MS medium supplemented with various concentrations of IBA. All the cultures were maintained under 16 hours photoperiod at 25 ± 2oC. Sometimes healthy shoots, which were unable to induce roots, were grafted onto rootstocks taken from aseptically germinated 6 days old seedlings. The upper part of the rootstock was made a split cut with the help of a fine pointed scalpel and the cut of lower part of the scion (shoot) was made "V" shaped. After placing the scion onto the stock, thin cotton threads were used to tie the attachment region. The entire operation was done inside a laminar flow cabinet. Grafted shoots were cultured in test tubes containing ½ MS medium without any growth regulators. The plantlets with well-developed root system were transplanted in sterilized soil in small pots.

Source of Information: Bangladesh J. Sci. Ind. Res. 46(3), 379-384, 2011

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The results of the present investigation demonstrated the establishment of a reliable in vitro regeneration protocol from decapitated mature embryo axes and decapitated embryo attached with cotyledon explants for four selected Bangladeshi chickpea varieties. This protocol may be effectively used for the improvement of different chickpea varieties through Agrobacterium- mediated or any other suitable methods of genetic transformation.

Knowledge Management: Journal