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Genetic Relationship in Ten Sugarcane Varieties of Bangladesh and Thailand Based on Ssr Markers

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

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

M. K. Begum
Bangladesh Sugarcane Research Institute, Ishurdi-6620, Pabna, Bangladesh.

N. Jahan
Department of Biotechnology and Genetic Engineering,Islamic University, Kushtia, Bangladesh

F. Waresi
Department of Biotechnology and Genetic Engineering,Islamic University, Kushtia, Bangladesh.

Abstract/ Executive Summary:

The experiment was carried out at the Biotechnology Laboratory, Bangladesh Sugarcane Research Institute (BSRI), Ishurdi, Pabna, Bangladesh. Five BSRI released sugarcane varieties (Isd 24, Isd 28, Isd 29, Isd 30 and Isd 31), five Thai varieties (K84200, K764, K8865, K8887 and K8892) and nine microsatellite primers (SMC687 CG, SMC384BS, SMC119CG, SMC336BS, MSSCIR8, SMC703BS, MSSCIR74, SMC 766BS and SMC18SA) were used for the investigation. Polymorphic information content of microsatellite markers ranged from 0.44 to 0.90. The most polymorphic marker was MSSCIR74 with polymorphic information content of 0.90. The highest linkage distance (28.0) was recorded between the varieties Isd 31 and K8887 from other varieties investigated. The lowest linkage distance (9.0) was observed between variety K8892 and K887. Genetic relationships among the varieties showed that at the average linkage distance of 23.5 varieties were grouped into two major clusters. One variety of Bangladesh included in the one cluster while others included in other cluster. Variety Isd 24 of Bangladesh separated from the other varieties of both countries. The variety Isd 24 is one of the varieties having chewing quality with superior performances against the abiotic (drought, flood and water-logging) stress than the other varieties of Bangladesh. Results of this investigation indicate the wide genetic distances between the sugarcane varieties of Bangladesh and Thailand.

Keywords: Sugarcane, Genetic relationship, SSR marker, Varieties.

Location: Biotechnology Laboratory, Bangladesh Sugarcane Research Institute (BSRI), Ishurdi, Pabna, Bangladesh

Start Date: 00-00-2006

End Date: 00-00-2007

Program Area: Variety and Species

Commodity: Sugarcane

Objectives:

To analyze the genetic relationship in ten sugarcane varieties of Bangladesh and Thailand based on SSR markers.

Methodology:

The experiment was carried out at the Biotechnology Laboratory, Bangladesh Sugarcane Research Institute (BSRI), Ishurdi, Pabna, Bangladesh in the cropping season 2006-2007. Five BSRI released sugarcane varieties (Isd 24, Isd 28, Isd 29, Isd 30 and Isd 31), five Thai varieties (K84200, K764, K8865, K8887 and K8892) were used. The fresh tops from the 8 month old field grown sugarcane were collected and outer leaf sheaths were removed leaving inner spindle to get meristem cylinder. Then the meristem cylinder (spindle base) was cut down into small pieces (about 5mm long with 3mm dia) with sterile scissors and weighed out the required amount (0.2g) using a fine balance. Care was taken during sampling from injury of meristem cylinder after removing of outer leaf sheaths. A modified method of were used to isolate total genomic DNA. Cut pieces of meristem cylinder (0.2g) was taken in small mortar (dia 6.5cm and 3.5cm depth) and homogenized with pestle in 800 µl of extraction buffer (200 mM Tris HCI, pH 8.0; 50 mM EDTA.H₂O, pH 8.0; 2.2 M NaCI; 2% CTAB; 0.06% Sodium Sulfite) until finely shredded within 40-50 second. Total grinded sample was taken into 2 ml eppendorf tube, then added 150 µl of each 5% SDS, 10% PVP, 20% CTAB were mixed well quickly and incubated at 65°C at water bath for 40 minutes. During incubation 3-4 times inversion was done. After incubation the samples were cooled to room temperature and equal volume of Phenol t Chloroformt Isoamyl Alcohol (25 t 24 t 1) was added, mixed well by inversion and centrifuged at 10,000 rpm at room temperature for 30 minutes. Then aqueous phase (about 800 IJI) was recovered and transferred to a fresh ice-cold 2ml eppendorf tube and an equal volume of ice-cold isopropanol was added followed by 120 µl of 6M NaCI. The samples were incubated at -20°C fer at least 1 hr and centrifuged at 10,000 rpm at room temperature for 20 minutes. The upper layer of solution was discarded carefully by using adjustable micropipette and 70% ice-cold ethanol of about 2.5 times of the solution was added. The same was again centrifuged at 10,000 rpm at room temperature for 10 minutes. After pellet formation the solution was discarded from the tube carefully so that the DNA pellet remains constant and undisturbed. Then 70% ethanol was added up to the slant of the tube. The ethanol was discarded from the tube carefully so that the DNA pellet remains constant and undisturbed. The DNA pellet was dried for at least 30 minutes by putting the tubes upside down on a filter paper. Then the DNA pellet was re-suspended in 50µl of TE buffer (10 mM Tris HCI, pH 8.0; 1 mM EDTA.H20, pH 8.0) and stored at -20°C for subsequent use.

Source of Information: Bangladesh Journal of Sugarcane, Vol: 31, pp:45-51, 2009, ISSN 1991-8925

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Funding Source:

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Total Budget (Tk.) :

Achievement/Findings:

Genetic relationships among the varieties showed that at the average linkage distance of 23.5 varieties were grouped into two major clusters. One variety of Bangladesh included in the one cluster while others included in other cluster. Variety Isd 24 of Bangladesh separated from the other varieties of both countries. The variety Isd 24 is one of the varieties having chewing quality with superior performances against the abiotic (drought, flood and water-logging) stress than the other varieties of Bangladesh. Results of this investigation indicate the wide genetic distances between the sugarcane varieties of Bangladesh and Thailand.

Knowledge Management: Journal