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Shoot Regeneration and Root Induction in Brinjal by Growth Regulators

Md. Abdul Alim
Assistant Production Manager
ACI (Seed) Ltd.

Bhabendra Kumar Biswas
Professor
Department of Genetics and Plant Breeding, Hajee Mohammad Danesh Science and Technology University (HSTU), Dinajpur, Bangladesh;

Md. Hasanuzzaman
Professor
Department of Genetics and Plant Breeding, Hajee Mohammad Danesh Science and Technology University (HSTU), Dinajpur, Bangladesh;

Pronay Bala
PhD Fellow
Department of Crop Physiology & Ecology, HSTU, Dinajpur

Santanu Roy
Program officer
Natural Resources Management Project, Caritas Fisheries Program, Mirpur, Dkaka, Bangladesh

Abstract/ Executive Summary:

The experiment was carried out to study the effect of genotypes and growth regulators on root induction and shoot regeneration of brinjal ( Solanum melongena) genotypes. Leaf segments of three varieties of Solanum were cultured on MS medium with different concentrations and combinations of plant growth regulators. Among the tested genotypes, Protab showed highest percentage of shoot regeneration (65.67%). Early and maximum rate of regeneration was found in MS + 1 mg/L NAA (naphthalene acetic acid) + 0.1 mg/LBAP (6-benzylamino purine) for all the genotypes. The highest number of roots per shoot was counted in Protab (73.33%) on 1 /2 MS + 2 mg/L IBA (indole butyric acid) + 0.4 mg/L BAP. Based on the overall performance, the variety Protab appeared as the best for shoot and root formation and ultimately successful regeneration of plants.

Keywords: Regeneration, Induction, Root, Brinjal, Growth regulators

Location: Biotechnology Laboratory at the Department of Biotechnology of Hajee Mohammad Danesh Science and Technology University, Dinajpur.

Start Date: 00-09-2011

End Date: 00-01-2012

Program Area: Variety and Species

Commodity: Brinjal

Objectives:

To develop a suitable regeneration protocol for the tested three brinjal genotypes grown in Bangladesh.

Methodology:

The experiment was carried out during the period from September, 2011 to January, 2012 in the Biotechnology Laboratory at the Department of Biotechnology of Hajee Mohammad Danesh Science and Technology University, Dinajpur. The varieties of Brinjal used in the present investigation namely-Protab, Green Ball and Ghemma Begun (Local) were collected from A.R. Mallik Seeds Co. Metal Agro Ltd. and locally from Dinajpur. The surface sterilization of seeds was carried out under a Laminar Air Flow Cabinet. The floated seeds were discarded and others were rinsed in 70% ethyl alcohol for one minute, and then thoroughly washed with sterilized water. The alcohol treated seeds were immersed into 0.1% HgCl₂ solution for 8-10 minutes, few drops Tween-20 per 100 ml were also added at that time. The seeds were then washed 5-6 times with sterilized distilled water. Sterilized seeds were placed into seed germination medium in vials. Six seeds were placed in each vial. The culture was then incubated in dark till the germination of seeds. These vials were then transferred to 16 hours light for normal seedling growth . Twenty one days old seedlings were used as source of contamination free explants. The seedlings raised in axenic culture were used as the source of different kinds of explants. The leaf segments were used as explants. Leaf segment from each germinated seedling were cut into small pieces using sterilized scalpel under a Laminar Air Flow cabinet. Four pieces of leaf segments were arranged on each vials and gently pressed into the surface of the sterilized culture medium with various combinations and concentrations of growth regulators viz.,T₁ = MS medium containing with 1 mg/L NAA (Napthaline acetic acid) + 0.1 mg/L BAP (6-benzylamino purine). T₂ = MS medium containing with 1.5 mg/L NAA + 0.5 mg/l BAP,T₃ = MS medium containing with 1.5 mg/L NAA + 1 mg/l BAP,T₄ = MS medium containing with 1 mg/L NAA + 1.5 mg/l BAP and T₅ = MS medium containing with 0.5 mg/L NAA + 2 mg/L BAP. MS media with different concentrations and combinations of BAP and NAA (e.g., T₁ =1/2 MS medium containing 2 mg/LIBA + 0.4 mg/L BAP, T₂ = 1/2 MS medium containing with 1 mg/L IBA + 0.3 mg/l BAP and T₃ = 1/2 MS medium containing with 0.5 mg/L IBA + 0.2 mg/L BAP) were used for root formation. The subculture vials were again incubated at 25+2 °C with moderate light intensity. All cultures were examined regularly and the vials showing symptoms of contamination were discarded. The plantlets with sufficient root system were separat ed from the vials. Agar was gently washed out with running tap water from root. The plantlets transplanted to small pots containing garden soil, sands and cowdung at the ratio of 1: 2: 1. Immediately after transplantation, the plants along with pots were covered with moist polythene bag to prevent desiccation. To reduce sudden shock, the pots were kept in the controlled environment in a growth room. The interior of the polythene bags were sprayed with water at every 24 hours to maintain high humidity around the plantlets. At the time plantlets were also nourished with Hoagland’s solution. After 2-3 days, the polythene bags were gradually perforated to expose the plants to natural environment. The polythene bags were completely removed after 7-10 days. The plantlets at this stage were placed in natural environment for 3-10 hours daily. The established plants were calculated based on the number of plantlets placed in the pot and number of plants finally established or survived. Percent plant establishment = (Number of established plantlets/Total number of plantlets) x 100. The experiments were arranged in Completely Randomized Design (CRD). The analysis of variance for different characters was analyzed using MSTAT-C and means were compared by the Duncan’s Multiple Range Test (DMRT).

Source of Information: Agric. Livest. Fish. Vol. 2, No. 1, April 2015: 43-51

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

By considering the overall investigation and comparing the performance of three brinjal genotypes, it was found that Protab was the best cultivar in case of shoot regeneration and root induction. The findings from the present investigation of the effect of genotypes and growth regulators on shoot regeneration and root induction of brinjal ( Solanummelongena) could be efficiently utilized for the advanced biotechnological research, as for example gene transfer and crop improvement.

Knowledge Management: Journal