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Virus Elimination in Potato Through Meristem Culture Followed by Thermotherapy

M. A. Ali
Department of Biotechnology, Faculty of Agriculture, Bangladesh Agricultural University, Mymensingh-2202

K. M. Nasiruddin
Department of Biotechnology, Faculty of Agriculture, Bangladesh Agricultural University, Mymensingh-2202

M. S. Haque
Department of Biotechnology, Faculty of Agriculture, Bangladesh Agricultural University, Mymensingh-2202

S. M. Faisal
Senior Scientific Officer,
Horticulture Division, RARS, BARI, Hathazari, Chittagong, Bangladesh

Abstract/ Executive Summary:

Virus elimination in potato through meristem culture followed by thermotherapy and virus indexing was studied. Three levels of thermotherapy, viz. 27±1^oC (control), 30±1^oC and 35±1^oC, sixteen combinations of BAP (Benzyl Amino Purine) plus GA₃(gibberellic Acid) concentrations viz. 0.0+0.0 (control), 0.0+0.2, 0.0+0.4, 0.0+0.6, 1.5+0.0, 1.5+0.2, 1.5+0.4, 1.5+0.6, 3.0+0.0, 3.0+0.2, 3.0+0.4, 3.0+0.6 , 4.5+0.0, 4.5+ 0.2, 4.5+0.4 and 4.5+0.6 were used in this study in three potato varieties viz. Diamant, Heera and Lalpakri. Among the thermo therapies, 27±1^oC showed the highest (24.55) survival response followed by 30±1^oC, 35±1^oC, respectively. The poorest (20.47) survival response of meristem derived plantlets was noticed in 35±1^oC which gave the highest percentage (43.79) of virus free plantlets followed by 30±1^oC. The best (25.85%) survival response was found in Lalpakri and the lowest (19.08%) survivality was recorded in Diamant. The highest (33.27) percentage of PVY (Potato Virus Y) free plantlets was observed in Heera. The combined treatment 3.0 mg L^-1 BAP and 0.2 mg L^-1 GA₃showed the highest (63.39) percentage of virus free plantlet production followed by 4.5 mg L^-1BAP and 0.2 mg L^-1 GA₃.

Keywords: Potato, Meristem, ELISA (Enzyme Linked Immuno-Sorbent Assay), BAP, GA3.

Location: Biotechnology Laboatory, Bangladesh Agricultural University, Mymensingh

Start Date: 00-01-2005

End Date: 00-12-2008

Program Area: Crop-Soil-Water Management

Commodity: Potato

Objectives:

1) To see the effect of thermotherapy on virus elimination in meristem cultured seed potato;

2) To establish a suitable and reproducible in vitro regeneration protocol of potato varieties for quality seed potato production.

Methodology:

The investigation was conducted at the USDA sponsored (United States Department of Agriculture) Biotechnology Laboratory at Bangladesh Agricultural University, Mymensingh during January 2005 to December 2008, as a part of Ph.D. research works. There were three factors in this experiment. Factor A consisted of three levels of thermotherapy, viz. 27±1⁰C (control), 30±1⁰C and 35±1⁰C, Factor B consisted of three potato varieties viz. Diamant, Heera and Lalpakri and Factor C consisted of sixteen combinations of cytokinin (BAP) plus gibberellin (GA3) concentrations viz. 0.0+0.0 (control), 0.0+0.2, 0.0+0.4, 0.0+0.6, 1.5+0.0, 1.5+0.2,1.5+0.4, 1.5+0.6, 3.0+0.0, 3.0+0.2, 3.0+0.4, 3.0+0.6 , 4.5+0.0, 4.5+ 0.2, 4.5+0.4 and 4.5+0.6. So, total number of treatments were 144 (3 × 3 × 16). Four explants were placed in a test tube and replicated 3 times. The experiment was done in Completely Randomized Design (CRD). In this study MS (Murashige and Skoog, 1962) medium was used as basal medium. Two sets, each containing plant materials of three potato varieties infected by PVY were selected. Each set consisted of 18 plants- affected by PVY, field symptoms and serological confirmation followed by ELISA test. After maturity the PVY infected potato tubers of three varieties were collected and stored carefully. PVY infected potato tubers of three varieties were kept under dark with 1 minute deeping in 1 ppm concentration of GA3 for sprouting. For meristem collection 0.2 - 0.5mm long size meritems were selected. After dissection the excised meristems were cultured in MS medium supplemented with the sixteen hormonal combinations. Calli derived from excised meristems were treated with three temperature regimes. All inoculations and manipulations were carried out aseptically under laminar airflow cabinet. The physical conditions for growth and development of cultures were maintained at the temperature of 25 ± 1⁰C and a light intensity of 2000-3000 lux provided by fluorescent tube. The photoperiod was maintained at 16 hours light and 8 hours dark (16L/8D) and the relative humidity was 60-70%.

Source of Information: SAARC JOURNAL OF AGRICULTURE, Vol. 11, Issue 1, 2013, pp. 71-80, ISSN 1682-8348, SAARC Agricultural Information Centre.

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The highest (33.27) percentage of PVY (Potato Virus Y) free plantlets was observed in Heera. The combined treatment 3.0 mg L^-1 BAP and 0.2 mg L^-1 GA₃ showed the highest (63.39) percentage of virus free plantlet production followed by 4.5 mg L^-1 BAP and 0.2 mg L^-1 GA₃.

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