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Comparative Study on Meristem Culture of Three Potato Cultivars Diamant, Cardinal and Granula and Their Shoot Formation

Marufa Khatun
Department of Genetic Engineering and Biotechnology, University of Rajshahi, Rajshahi-6205, Bangladesh.

Md. Shahadat Hossain
Department of Genetic Engineering and Biotechnology, University of Rajshahi, Rajshahi-6205, Bangladesh.

Md. Khalekuzzan
Department of Genetic Engineering and Biotechnology, University of Rajshahi, Rajshahi-6205, Bangladesh.

Amina Rownaq
Department of Genetic Engineering and Biotechnology, University of Rajshahi, Rajshahi-6205, Bangladesh.

Abstract/ Executive Summary:

The main aim of this study was to analyze the comparative study on meristem culture of three important potato cultivars, diamant, cardinal and granula and their shoot formation using meristem tips. Apical meristems were isolated from shoot tips of 25-35 days old field grown plants. After surface sterilization the meristems (0.3 mm) were isolated from the shoot tips. After isolation the apical meristems were placed quickly on “M” shaped filter paper bridge in culture tubes containing liquid MS medium supplemented with various concentrations and combinations of different plant growth regulators. Meristems showed their first growth response by increase in size and became greenish white in colour. They continued their growth and developed shoots with roots. In the present investigation, among the three cultivars of potato, cardinal were the best responsive cultivars for the resuming new growth of cultured meristems on MS medium supplemented with KIN (0.4 mg l^-1) + GA3 (0.5 mg l^-1). Cultivar diamant showed comparatively better growth response in the primary culture and granula showed better results in shoot length formation in MS0 medium. Cultivar cardinal proved to be best potato cultivar in case of meristem culture than diamant and granula cultivars.

Keywords: Meristem culture, Shoot formation, Potato cultivars

Location: Biotechnology Laboratory, Institute of Biological Sciences, University of Rajshahi, Rajshahi

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Commodity: Potato

Objectives:

To see the comparative study on meristem culture of three potato cultivars diamant, cardinal and granula and their shoot formation using meristem tips in order to sort out the steady protocol.

Methodology:

The present experiment was carried out in the Biotechnology Laboratory, Institute of Biological Sciences, University of Rajshahi, Rajshahi-6205, Bangladesh. The meristem of three potato cultivars viz., diamant, cardinal and granula were used as explants. Apical meristems were isolated from shoot tips of 25-35 days old field grown plants. The explants were taken in a conical flask and thoroughly washed under running tap water for 30 min to reduce the level of surface microorganisms. Then the explants were taken in reagent bottle containing distilled water with few drops of tween-80 (wetting agent) and 2-3 drops of savlon for about 10-12 min and subsequently rinsed with constant shaking. This was followed by a second washing with distilled water to remove all traces of treated chemicals. Surface sterilization was carried out by 0.1% HgCl₂ with gentle shaking for 4 min. The sterilized explants were then washed 5-7 times with sterile distilled water immediately to remove all traces of HgCl₂. This procedure was carried out in aseptic condition of laminar airflow cabinet. After surface sterilization the meristems (0.3 mm) were isolated from the shoot tips. After isolation the apical meristems were placed quickly on “M” shaped filter paper bridge in culture tubes containing liquid MS medium supplemented with various concentrations of KIN, GA₃. The meristem responed after 4 weeks of inoculation and then subcultured in MS medium without growth regulators for shoot and root induction. Data were recorded on number of root developments/ explants, shoot length and root formation. Before shoot formation, a serological identification was done in the cultured plants and to detected virus. In this detection the double antibody sandwitch enzyme-linked immunsorbent assay (DAS- ELISA) methods were followed. Virus free plantlets were used for shoot formation in MS semi-solid medium. After 4-5 weeks it was subcultured on MS₀ semisolid medium or MS semisolid medium having NAA, IBA singly or in combinations for root and shoot developments. The explants were cultured on MS medium with 3% (W/V) sucrose which was solidified with 0.7% (W/V) agar. The pH of the media was adjusted to 5.7 prior to autoclaving at 121⁰C for 20 min. The cultures were incubated in a culture room at 25±2⁰C with a photoperiod of 16 hour at 3000 lux light intensity provided by cool white fluorescent tubes. For each treatment, 10 replications were used and all experiments were repeated thrice. Well developed plantlets were removed carefully from the culture vessels, washed gently under running tap water and planted in plastic pots containing a potting mixture of sand, soil and farmyard manure in the ratio of 1:1:1. The potted plantlets were covered by polythene sheet to maintain suitable humidity and then transferred to the fields.

Source of Information: Asian J. Med. Biol. Res. 2015, 1 (3), 537-544 (ISSN 2411-4472)

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The main aim of our study to minimize this limitation associated with meristem culture and plantlet elongation for commercial production of disease free potato seeds. The potato crop has been emerged as an important commercial crop for the farmers particularly in the lean period of the country. Protocol for rapid production of meristem culture and disease free potato seed would be a mile stone for our farmers in their socio-economic development.

Knowledge Management: Journal