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Association of Gdf9 Gene Polymorphisms with Litter Size in Indigenous Sheep of Bangladesh

Faruk Hossain
Department of Animal Breeding and Genetics, Faculty of Animal Husbandry, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Sharmin Akter Suma
Department of Animal Breeding and Genetics, Faculty of Animal Husbandry, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Mohammad Shamsul Alam Bhuiyan
Department of Animal Breeding and Genetics, Faculty of Animal Husbandry, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Abstract/ Executive Summary:

Sheep in Bangladesh are well known as efficient users of low quality roughages, well adapted to hot and humid agro-climatic conditions, capable of bi-annual lambing with multiple births and resistant to common prevalent diseases. They play an important role in the supply of animal protein. Present study aimed to investigate possible association of GDF9 gene polymorphisms with litter size. A total of 126 DNA samples extracted from the blood of indigenous sheep from 5 locations namely Tangail, Noakhali, Naogaon, Gaibandha and Satkhira with known litter size were used to study the association of GDF9 polymorphism by PCR based RFLP method. Two polymorphic regions of GDF9(FecG1 and FecG8) were amplified by PCR, digested with respective restriction enzymes and 126 sheep were genotyped. Current study revealed that genotype and allele frequency for FecG1 varied among the sheep from different locations. The genotype (GG,AG, AA) frequency were 51.59%, 45.24% and 3.17% and the allele (G and A) frequencies in the overall population were 74.21% and 25.79%, respectively. There was a significant association of FecG1 of GDF9 gene polymorphism with litter size. The homozygous GGgenotype had the lowest litter size (1.59±0.09; n=65) and homozygous AA genotype had the highest litter size (2.00±0.41; n=4). No genotypic variations were found for FecG8.Findings of this study specially the polymorphism of FecG1 together with genotyping of some sheep could be utilized in the selection program to increase the lamb production potentiality of indigenous sheep of Bangladesh.

Keywords: Sheep, GDF9, Gene, Polymorphism, Litter size

Location: Animal Genomics and Breeding Laboratory, Department of Animal Breeding and Genetics, Bangladesh Agricultural University, Mymensingh

Start Date:

End Date:

Program Area: Animal Health and Management

Commodity: Sheep

Objectives:

The present study was undertaken on native sheep from different agro-ecological zones of Bangladesh to identify the polymorphisms of FecG1 and FecG8 fragments of GDF9 gene in native sheep and to reveal the association of GDF9 gene polymorphisms with litter size.

Methodology:

Phenotypic data and sample collection- Data were collected from five distinctly located sheep populations from five regions and a total of 310 farmers were interviewed from the selected locations for collecting data on the litter size of individual sheep. A total of 126 blood samples from ewes with known litter size were collected from a foresaid five locations and analyzed for genotyping. Blood was taken from the jugular vein using venoject tubes (Addbio manufacturing company, South Korea) coated with Di Sodium Ethylene Di Amine Tetra Acetate (EDTA) as anticoagulant. The collected blood sample in the venoject tube was kept at 4-6°C in icebox and carried as soon as possible to Animal Genomics and Breeding Laboratory, Department of Animal Breeding and Genetics, Bangladesh Agricultural University, Mymensingh and preserved at 4-6°C until DNA extraction. DNA extraction- DNA was extracted following the standard protocol (Hanrahan et al., 2004) from the whole blood using PrimePrep^TM genomic DNA extraction kit, (GeNet Bio, South Korea) according to the manufacturer’s instruction manual. In this method, RBC lysis buffer was used prior to the extraction in order to breakdown of RBC and concentrating WBC. Extracted DNA was stored in -20°C until use. DNA quantification and detection- After extraction, the concentration of DNA was measured by Nanodrop 1000 spectrophotometer (Model ND1000) according to the instruction of (Hanrahan et al., 2004). Agarose gel electrophoresis for visualizing/detecting the isolated DNA was performed. The Purity of extracted DNA was assessed by calculating the OD260/OD280 nm ratio using NanoDrop (Model ND1000) spectrophotometer. The range of concentration and the purity determined by OD 260/280 ratio from Nanodrop 1000. PCR amplification- DNA samples were amplified in Biometra gradient thermocycler using standard PCR protocol (Polley et al., 2010). All samples were subjected to polymerase chain reaction. PCR was carried out in 16 µl reaction mixture containing 8.00 µl PCR master mix, 0.7µl primer forward, 0.7µl primer reverse, 4.2 µl deionized water and with 2.4 µl of DNA sample. A no-template negative control was used every time. Statistical analysis- The data were analyzed using Generalized Linear Model (GLM) procedure of Statistical Analysis System (SAS) (SAS Institute Inc., 2009) computer package, version 9.1.3

Source of Information: Res. Agric., Livest. Fish.7(2): 283-292, August 2020

Article Link (Online Journal): DOI: https://doi.org/10.3329/ralf.v7i2.48871

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

PCR-RFLP based genotyping analysis revealed that the genotype and allele frequency for FecG1 polymorphism of GDF9 gene in the sheep populations varies between locations. Current study showed that there was an association of FecG1 polymorphism of GDF9 gene with litter size in indigenous sheep of Bangladesh. In addition, it was observed that the litter size had significant (P<0.02) effect on genotype. The homozygous GG genotype showed lowest litter size (1.59±0.09) and homozygous AA genotype represented the highest litter size (2.00±0.41). On the other hand, there were no genotypic differences for the polymorphism of FecG8 fragment of GDF9 gene in any of the sheep population in this study. Therefore, in can be concluded that there is an association of polymorphism of FecG1 fragment of GDF9 gene with litter size in indigenous sheep of Bangladesh and the sheep with AA genotype which might be the suitable candidate as breeding animal in a selection program for improving prolificacy of indigenous sheep of Bangladesh.

Knowledge Management: Journal