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Standardization of Sucrose and 6-benzyl Aminopurine for in Vitro Micro Tuberization of Potato

Md. Afzal Hossain
Ministry of Environment & Forests, Bangladesh Secretariat, Bangladesh

Md. Abu Kawochar
Scientific Officer
Tuber crops research Centre (TCRC), Bangladesh Agricultural Research Institute, Joydebpur, Gazipur, Bangladesh

Abdullah-Al-Mahmud
International Potato Centre (CIP), SWCA, Banani, Dhaka, Bangladesh

Ebna Habib Md. Shofiur Rahaman
International Potato Centre (CIP), SWCA, Banani, Dhaka, Bangladesh

Md. Altaf Hossain
Tuber crops research Centre (TCRC), Bangladesh Agricultural Research Institute, Joydebpur, Gazipur, Bangladesh

Khondoker Md. Nasiruddin
Professor
Department of Biotechnology, Bangladesh Agricultural University, Bangladesh

Abstract/ Executive Summary:

In vitro micro tuberization from regenerated plantlets of potato varieties was observed in Biotechnology Laboratory, Department of Biotechnology, Bangladesh Agricultural University. Tubers of potato cultivars Diamant and Cardinal were used as initial experimental materials for meristem culture. The experiment was consisted of three factors; variety (Diamant, Cardinal), sucrose concentration (3%, 6%, 9%, 12%) and 6-benzyl aminopurine (BAP) levels (2.5, 5.0, 7.5 mg L^-1). As a whole, 24 treatments were laid out in complete randomized design with three replications. Among the varieties, Diamant required minimum days (6-17) for micro-tuber initiation, produced more number of micro-tubers (4.97) and produced more average weight of micro-tuber (120.39mg) but there had no significant difference. Among the sucrose levels, quickest (6-15 days) micro-tuber initiation, the highest number of micro-tubers vial^-1 (5.06) and the highest average weight of micro-tuber (137.31 mg) were found in 9% sucrose level. For different BAP levels, quickest (6-15 days) micro-tuber initiation, the highest number of micro-tubers vial^-1 (5.38) and the highest average weight of micro-tuber (126.31 mg) were found at 5.0 mg L^-1. The best combination for minimum duration (6-8 days) of micro-tuber initiation, the highest number of micro-tubers vial^-1 (6.00) and the highest average weight of micro-tuber (152.01 mg) was in Diamant with 9% sucrose at 5 mg L^-1. Concomitantly, the lowest number micro-tubers vial^-1 (2.00) and the lowest average weight of micro-tuber (89.98 mg) were found in Cardinal cultured with 3% sucrose media where at 7.5 mg L^-1 BAP and at 2.5 mg L^-1 BAP, respectively.

Keywords: 6-Benzyl aminopurine, Microtuberization, Plantlets, Potato

Location: Biotechnology Laboratory, Department of Biotechnology, Bangladesh Agricultural University, Mymensingh

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Commodity: Potato

Objectives:

To find out the appropriate doses of sucrose and growth regulators and their combination(s) with potato varieties for rapid microtuberization

Methodology:

Design and Treatments of the Experiment: The experiment was carried out in Biotechnology Laboratory, Department of Biotechnology, Bangladesh Agricultural University. The experiment was consisted of three factors; variety (Diamant, Cardinal), sucrose levels (3%, 6%, 9%, 12%) and BAP levels (2.5, 5.0, 7.5 mg L^-1). As a whole, 24 were laid out in complete randomized design with three replications. Explant, Culture Media and Regeneration of Plantlets: Meristem was used to regenerate for virus free plantlet production. From the regenerated virus free plantlets, plantlets again multiplied through rapid multiplication technique. Required amount of MS medium was prepared and supplemented with mentioned sucrose and BAP levels. Thus, prepared all groups of media were taken to separate test tubes and/or vials according to the number of treatments and replications. Then the test tubes/vials containing the media were sterilized by autoclaving. Stem segments having 1-5 nodes from plantlets of one month old which were collected from previous experiment were cultured for microtuberization. Single stem segments were placed to each test tube containing 10 ml medium and 4-5 stem segments were placed to each vial containing 40 ml medium. When placed in test tube, five test tubes were considered as one replication. Incubation: The cultures were incubated at a temperature of 25 ± 2^oC at 16 hrs photoperiod. Days to microtuber initiation were recorded. After two months of explantation, number of microtubers vial^-1 was calculated on the basis of total number of vials per treatment and average weight of microtuber was calculated on the basis of total number of microtubers produced. Data Collection and Statistical Analysis: After two months of explantation, data were recorded on i) Days to microtuber initiation, ii) Number of microtubers vial^-1, iii) Average weight of microtuber. Data were analyzed by statistical software MSTATC and means were adjudged by the Duncan’s Multiple Range Test.

Source of Information: American Journal of Agriculture and Forestry. Vol. 3, No. 2: pp. 25-30, (2015)

Article Link (Online Journal):

Funding Source:

1. United States Agency for International Development

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Microtuber production was affected by different levels of sucrose and 6-benzyl aminopurine (BAP). Genotypic variation was also found among the two genotypes regarding microtuber production. The best combination for rapid production of potato varieties was Diamant with 9% sucrose at 5 mg L-1 of 6-benzyl aminopurine. Hence the present protocol has the potential for the rapid multiplication of true clones without changing the genetic fidelity.

Knowledge Management: Journal