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Tissue Culture Variability on Callus Induction and Plant Regeneration of Five Sugarcane Varieties

N. Islam
Bangladesh Sugarcane Research Institute Ishurdi-6620, Pabna

M. A. Hossain
Bangladesh Sugarcane Research Institute Ishurdi-6620, Pabna

M. A. S. Miah
Bangladesh Sugarcane Research Institute Ishurdi-6620, Pabna

Abstract/ Executive Summary:

The investigation was conducted at Biotechnology Laboratory of Bangladesh Sugarcane Research Institute (SSRI), Ishurdi , Pabna, Bangladesh to investigate the tissue culture variability on callus induction and plant regeneration of five sugarcane varieties viz., Isd 18, Isd 28, Isd 32, Isd 33 and Isd 34 from unexpanded leaf sheath explant. Explants were inoculated on MS medium containing 2.0-4.0 mg/I of 2, 4-0 for callus induction. Best results were obtained from all tested varieties at 3.0 mg/I of 2, 4-0. Combination of cytokinin (SA) with auxins (NAA or ISA) showed the best performance for shoot proliferation to that of concentration of cytokinin (SA) alone. Among different media, 1.0 mg/I SA + 0.5 mg/I NAA showed best result for multiplication of shoots. The highest number of shoot (11.8) was obtained from the variety Isd 33 followed by Isd 18 and Isd 32 whereas, the variety Isd 34 showed the lowest (9.7) performance among the five varieties. In vitro proliferated shoots were rooted on medium supplemented with different auxins. Best results of rooting were observed on MS medium supplemented with 5.0 mg/I NAA for all varieties. The highest number of healthy roots (14.7) was produced in Isd 34 followed by Isd 28, Isd 33 and Isd 18 but the lowest (7.9) number in Isd 32 when 5.0mg/1 NAA supplemented in medium. Interactions of varieties and media exerted remarkable effects on shoot and root formation capabilities. The plantlets were successfully transferred to soil after root formation and ninety percent were survived. Results indicate varietals variability of sugarcane for callus induction and plant regeneration through tissue culture.

Keywords: Sugarcane, Variability, Tissue culture, Callus and Regeneration.

Location: Biotechnology Laboratory, BSRI, Ishurdi, Pabna

Start Date: 01-01-2008

End Date: 31-12-2008

Program Area: Variety and Species

Commodity: Sugarcane

Objectives:

To evaluate the callus induction, shoot multiplication and root forming potentials of five varieties and to compare different media and select suitable media for specific variety.

Methodology:

The study was conducted at Biotechnology Laboratory of Bangladesh Sugarcane Research Institute (BSRI), Ishurdi, Pabna, Bangladesh. The young leaf-sheath of five sugarcane varieties viz., Isd 18, Isd 28, Isd 32, Isd 33 and Isd 34 were used as experimental material. Disease-free and actively growing top shoot of sugarcane varieties viz, Isd 18, Isd 28, Isd 32, Isd 33 and Isd 34 were collected from 3-4 months old plants as explants from experimental field of BSRI. Then the plant materials were washed thoroughly under running tap water in the laboratory to reduce dust surface contaminants. Outer whorl of leaves was removed and surface sterilization induced treatment of the materials with 1% savlon for 10 minutes with constant shaking. Then the explants were washed 3-4 times with sterilized distilled water and taken under running laminar airflow cabinet and immersed in 0.1 % HgCI₂ into a 250 ml sterilized conical flask and shaking for 10-12 minutes. To remove traces of the sterilant, the material was then washed 4-5 times with sterile distilled water. Then the explants were prepared from the surface sterilized materials for establishing aseptic culture for callus induction and plant regeneration. For callus induction 0.5 cm of sugarcane leaf sheath explants were aseptically cultured on MS medium with varying concentrations (2, 3 and 4 mg/I) of 2,4-0 after sterilizing the plant material. The cultures were kept under dark for one week at a temperature of 25±1⁰C. For evaluating the optimum growth requirements for the shoot induction, the calli were cultured on different shoot induction media viz., 1.0 mg/I BA, 1.0 mg/I BA + 0.5 mg/I NAA, 1.0 mg/I SA + 0.5 mg/I Kn. After initiation of auxiliary shoots, the whole explant was rescued aseptically on a sterile petridish and was cut into convenient sizes by a sterile scalpel and then transferred on the appropriate medium for proliferating of shoots. Then each of the microcuttings of shoot was cultured on freshly prepared MS medium supplemented with IBA, NAA and IAA at the concentration of 5.0 mg/1. Unless mentioned specially, all cultures were grown in an air-conditioned culture room illuminated by 40W white florescent tube light. The temperature of the culture room was maintained at 25±1⁰C and light intensity varied from 2000-3000 lux provided by cool-white florescent tubular lamp.

Source of Information: Bangladesh Journal of Sugarcane, Vol. 32, pp. 13-21, April 2011, ISSN- 1991-8925

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The results were obtained from all tested varieties at 3.0 mg/I of 2, 4-0. Combination of cytokinin (SA) with auxins (NAA or ISA) showed the best performance for shoot proliferation to that of concentration of cytokinin (SA) alone. Among different media, 1.0 mg/I SA + 0.5 mg/I NAA showed best result for multiplication of shoots. The highest number of shoot (11.8) was obtained from the variety Isd 33 followed by Isd 18 and Isd 32 whereas, the variety Isd 34 showed the lowest (9.7) performance among the five varieties. In vitro proliferated shoots were rooted on medium supplemented with different auxins. Best results of rooting were observed on MS medium supplemented with 5.0 mg/I NAA for all varieties. The highest number of healthy roots (14.7) was produced in Isd 34 followed by Isd 28, Isd 33 and Isd 18 but the lowest (7.9) number in Isd 32 when 5.0mg/1 NAA supplemented in medium. Interactions of varieties and media exerted remarkable effects on shoot and root formation capabilities. The plantlets were successfully transferred to soil after root formation and ninety percent were survived. Results indicate varietals variability of sugarcane for callus induction and plant regeneration through tissue culture.

Knowledge Management: Journal