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The Significance of Non-controlled Natural Light, Temperature and Humidity in the Commercial Micropropagation of Solanum Tuberosum L. Cultivar Diamant.

F. J. Zapata?Arias
BRAC Agriculture and Biotechnology Laboratory, Gazipur, Bangladesh

Shahana Akter
BRAC Agriculture and Biotechnology Laboratory, Gazipur, Bangladesh

S. M. Asadul Haque
BRAC Agriculture and Biotechnology Laboratory, Gazipur, Bangladesh

Shilpi Akther
BRAC Agriculture and Biotechnology Laboratory, Gazipur, Bangladesh

Firoza Khatun
BRAC Agriculture and Biotechnology Laboratory, Gazipur, Bangladesh

Md. A. Mazid
BRAC Agriculture and Food Security Program, Bangladesh

Mahabub Hossain
BRAC International, Dhaka, Bangladesh.

Abstract/ Executive Summary:

The aim of our study was to reduce the unit cost of in vitro micropropagation of the Solanum tuberosum cultivar Diamant widely cultivated in Bangladesh with the guarantee that the quality and quantity of the in vitro plants produced was not jeopardized. This was done by entirely replacing the conventional micropropagation conditions of maintaining the in vitro plants in a controlled room whose temperature varies between 25 and 30⁰C, its humidity between 60 to 70% and its light intensity of 20,238 to 20,409 for 19 hours; with a room whose roof was made of corrugated plastic sheets that allow a partial passage of natural light. Under this conditions the amount of light, temperature and humidity were not controlled. During the time work the temperatures in this non?conventional room fluctuate between 14 and 400C, the light intensities were between 20,017 to 20,687 Lux and the humidity between 40 to 90%. Experiments were initiated in May, 2009 through March, 2014 covering summer, rainy and winter seasons. After two years of laboratory research and two years of field studies, we have not found differences between yield production of the micropropagated plants grown under control and non-controlled conditions, very often the latter plants were robust and adapted faster when transfer to field conditions. All plants used in the field experiments were no more than seven in vitro passages. A RBCD yield trail of the plants was done during two seasons and it was no found any difference in yield between them. Moreover, a yield trail of the minitubers to produce breeder seed (second generation) was done during the season 2013-2014 and no differences were found between the controls and the tubers derived from the low cost. This paper shows a new approach of in vitro micropropagation of plants by reducing the total cost of electricity in the culture room by using non-controlled natural light, ambient temperature and humidity regimes. More importantly, this significant cost reduction (savings) is achieved without compromising the quality and quantity of the in vitro plants produced. Furthermore, the simplicity of this technology will allow application by small laboratories and progressive farmers with a minimal capital outlay.

Keywords: Potato, Micropropagation, Natural light, Natural temperature, Natural humidity, Cost reduction

Location: BRAC Agriculture and Biotechnology Laboratory, Gazipur, Bangladesh

Start Date: 00-05-2009

End Date: 00-03-2014

Program Area: Variety and Species

Commodity: Potato

Objectives:

To reduce the total cost per in vitro plant of the potato cultivar Diamant

Methodology:

Tubers of high yielding cultivar Diamant were obtained from the Bangladesh Agricultural Development Corporation (BADC) and grown under dark conditions for 30 to 35 days at 20 to 22ºC. Under these conditions the tubers sprout and the buds placed on a test tube containing 15 ml of MS and 30 g/l of sugar semi?solidified with 6 grams of agar with a pH adjusted to 5.8. Healthy growing plants were placed in fresh medium and heat shocked for 30 days at 38⁰C. Shoot tips were isolated and grown into adult plants under controlled conditions of 16 hrs of light and constant temperature and humidity. Samples of in vitro plants were taken to the Bangladesh Agriculture Research Institute (BARI) and checked for the presence or absence of the following viruses: Potato Virus X (PVX), Potato Virus Y (PVY), Potato Leaf Roll Virus (LRV), Potato Virus M (PVM) and Potato Virus S (PVS) using the Enzyme Linkage Immunosorbent Assay (ELISA). Four nodal stem cuttings with around 32 nodes were cultured in a magenta box containing MS major and minor salts, 30 g/l of commercial sugar, solidified with 6 g/l of agar and the pH adjusted to 5.8 The in vitro plants were maintained in the controlled room under the following conditions: the room temperature was kept between 26 to 30ºC; the humidity was maintained between 60 to 70% and the illumination was for 16 hrs. The illumination was provided by cool- white fluorescent tubes from Hitachi 36 watt pre?heat, F40T8/Day. The light intensities measured in Lux with a light meter 840006 from Super Scientific, Taiwan. The humidity was measured in percentage with a Hygrometer Super Scientific 736920, Germany. In vitro plants growing under non?controlled conditions were maintained in a room (4.9 × 5.3 × 3 meters) with a roof made of corrugated plastic sheets that allow a partial passage of natural light with four windows two of 1.2 × 1.3 m and two of 1.27 × 1.50 m protected by a metal net of 1 mm of mesh. The bottles were placed in a heavy duty 5?tier chrome plated wire shelving 60” x 24” x 76” each shelf. In each level we could place up to 150 flasks. A separation between the levels was of 30 cm and the total height was of 1.5 After this procedure were planted in a field in single hill plots with 64 plants per replication with a total of 256 plants per treatment using an RCBD design. The line to line distance was of 60 cm and the plant to plant distance of 15 cm. The field for the yield studies was prepared as follows: 150 of urea, 110 of TSP, 150 of MP, 50 gypsum, 4 of zinc sulfate, 50 of magnesium and 6 of boric acid (all in Kilograms per acre). Mini?tubers were harvested after approximately 100 days when the yield data were calculated. meters.

Source of Information: Plant Tissue Cult. & Biotech. 24(2): 131?139, 2014 (December)

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

We observed that in the lower rack the levels of the natural light experiment the in vitro plants were elongated while at the top levels they were shorter but their color was always deep green. As against this, the plants under controlled conditions at all rack levels were elongated and pale. Water drops formed inside the bottles under both conditions. We overcame this problem of high humidity inside the culture bottles by using cotton plugs instead of plastic cups. Usually the plants grown under controlled conditions needed hardening, while the ones grown under natural conditions did not. During the time work the temperatures in this non-conventional room fluctuate between 14 and 40⁰C, the light intensities were between 20,017 to 20,687 Lux and the humidity between 40 to 90%. Experiments were initiated in May, 2009 through March, 2014 covering summer, rainy and winter seasons. After two years of laboratory research and two years of field studies, we have not found differences between yield production of the micropropagated plants grown under control and non?controlled conditions, very often the latter plants were robust and adapted faster when transfer to field conditions. All plants used in the field experiments were no more than seven in vitro passages. A RBCD yield trail of the plants was done during two seasons and it was no found any difference in yield between them. Moreover, a yield trail of the minitubers to produce breeder seed (second generation) was done during the season 2013-2014 and no differences were found between the controls and the tubers derived from the low cost.

Knowledge Management: Journal