Agricultural Research Management Information System

Home
Research Summary
Search
About Us
- ARMIS
- Brochure
Contact Us
Report
User Request
Data Input
Help
- Operation Manual
  - PDF
  - Video
- Program Area & Commodity
We have reached 37600 number of research entries at this moment.

- Logout

Research Detail

Home
Research
Detail

In Vitro Morphogenesis in Chickpea (cicer Arietinum L.)

R. H. Sarker
Department of Botany, University of Dhaka, Dhaka-1000, Bangladesh

Shaikh Tahmina Awal
Department of Botany, University of Dhaka, Dhaka-1000, Bangladesh

Abstract/ Executive Summary:

Culture conditions were developed for in vitro plant regeneration with or without the intervening callus phase from two locally grown varieties of chickpea (Cicer arietinum L.) namely, Binasola-2 and Hyprosola. The best response regarding callus induction obtained when immature embryo was cultured on MS medium containing 3.0 mg/l BAP and 0.5 mg/l NAA in both Binasola-2 and Hyprosola. Maximum number of shoots obtained when immature embryo derived callus was cultured on MS medium containing 5.0 mg/l BAP + 0.5 mg/l Kn and only 5.0 mg/l BAP in Binasola-2 and Hyprosola, respectively. Highest frequency of direct shoot morphogenesis without the intervention of callus was achieved from the cotyledonary node and surrounding tissue on MS medium containing 0.25 mg/l BAP + 0.1 mg/l Kn for both the varieties. Root induction at the base of the isolated regenerated shoots was found to be the best when they were cultured on full strength of MS medium containing 0.05 mg/l IBA. Histological and histochemical studies through freezing microtome revealed the formation of several shoot buds from the peripheral region of callus tissue and development of shoot apex as well as the organization of vascular tissue within the developing shoots. Such studies also demonstrated the organizational features during the formation of roots.

Keywords: In vitro, Morphogenesis, Chickpea

Location: Bangladesh Institute of Nuclear Agriculture, Mymensingh

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Chickpea

Objectives:

To understand the structural features involve during the in vitro morphogenic development of this crop plant.

Methodology:

Seeds of the two varieties of chickpea (Cicer arietinum L.) namely, Binasola-2 and Hyprosola used in the present study were obtained from the Bangladesh Institute of Nuclear Agriculture (BINA), Mymensingh. Shoot tips, nodal segments and cotyledonary nodes of the above two varieties were collected from the aseptically grown seedlings. Medium containing 3% sucrose and 1.2% agar was used to germinate the seed and to grow seedlings aseptically. When required immature seeds were collected from field grown plants. Immature embryos obtained from 14-16 days old seeds were used during this investigation. These pods were washed with running tap water and then surface sterilized with 0.1% HgCl₂ solution for ten min. The pods were then washed four to five times with sterile distilled water. Following washing, the immature seeds were taken out from the pods and the seed coats were removed carefully. The cotyledons were then separated with a pair of forceps. The cotyledons with or without the immature embryo were cultured separately. Various explants were cultured on MS medium supplemented with different concentrations and combinations of a-napthalene acetic acid (NAA), 6-benzyl aminopurine (BAP) and kinetin (Kn) for callus induction and shoot formation. For inducing root development, 2-4 cm long regenerated shoots were excised and transferred to MS medium containing various concentrations of indole acetic acid (IAA), indole butyric acid (IBA) and NAA. Histological examinations during shoot formation were carried out using a freezing microtome (Coldtome, Sakura, Japan). Small segments (4 x 4 mm) of regenerating tissue were placed on a specimen holder with a small drop of distilled water at -20°C to change the tissue segment into a frozen block. The sample thus prepared was sliced into thin sections of 15-20 μ also under -20°C. Following slicing the sections were stained with either Coomassie brilliant blue R-250 or decolorised Aniline blue solution. The stained sections were observed under a Nikon photomicroscope fitted with eip-fluorescence illumination system, with a combination of UV-2A and V-2A filter cassette.

Source of Information: Plant Tissue Culture, Vol: 9, (2), pp: 141-150, Dec 1999

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

This study confirms the organogenic nature of callus tissue as well as formation of high frequency direct shoots from specific explant tissue. The histological studies support the evidences of root and shoot induction during in vitro morphogenesis in chickpea. The regeneration system developed in this study with or without the intervening callus phase may be exploited in the future genetic transformation work towards the development of improved chickpea varieties.

Knowledge Management: Journal