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Study on Genetic Diversity of BLRI Cattle Breed-1 Using Microsatellite

G. K. Deb
Biotechnology Division, BLRI, Savar, Dhaka-1341

S. R. Dey
Animal production research division, BLRI, Savar, Dhaka-1341

N. R. Sarker
Animal production research division, BLRI, Savar, Dhaka-1341

M. R. Mufti
Animal production research division, BLRI, Savar, Dhaka-1341

M. R. Rahman
Biotechnology Division, BLRI, Savar, Dhaka-1341

Abstract/ Executive Summary:

This study aimed to characterize BLRI Cattle Breed 1 (BCB-1) at genetic level and to find out their genetic distances with other native cattle (Red Chittagong cattle, RCC and Progenitor cattle of BCB-1 of Pabna and Sirajganj districts, Progenitor) of Bangladesh using three pairs of bovine microsatellite markers (ETH152,INRA32 and INRA63). Blood samples were collected randomly from fifteen animals of each group (BCB-1, RCC and Progenitor). Genomic DNA was extracted using BLRI developed DNA extraction kit. All the primer pairs were successfully amplified microsatellite loci. The PCR products were checked on 3% agarose gel for verification of quantity and size of amplicon. Data were analyzed using popGen32 (Version 1.31) computer program. Results showed the highest and lowest gene frequency values (0.850 and 0.150) were observed in allele A and allele B with ETH152 in RCC and RCB-1 cattle and INRA63 in RCB-1 cattle. All the three cattle populations showed significant deviation from Hardy-Weinberg equilibrium for at least one locus (RCC for ETH152 and INRA32, BCB-1 for ETH152 and INRA63 and progenitor for INRA63). The estimated mean observed (Ho) and expected heterozygosity (He) were 0.167±0.134, and 0.377±0.134 in RCC vs. 0.167±0.116 and 0.339±0.122 in BCB-1 vs. 0.800±0.200 and 0.393±0.097, respectively. However, mean heterozygosity of all three loci was same (0.351±0.111) in the studied cattle populations. All the studied microsatellite showed two polymorphic loci each. Average effective number of alleles was 1604±0.349, 1507±0.349 and 1621±0.261 in RCC, BCB-1 and progenitor cattle population respectively. The mean SI was 0.539±0.139 in RCC vs. 0.498±0.130 in BCB-1 vs. 0.558±0.100 in progenitor cattle. The mean heterozygote deficiency (FIS) values were 0.528 in RCC vs. 0.519 in BCB-1 vs. 0.514 in progenitor cattle. Among the three cattle populations genetic distances were low between BCB-1 and its progenitor. Dendrogram indicated segregation of the three cattle populations in to two main clusters. Cluster 1 consisted of RCC and that of cluster 2 consisted of BCB-1 and its progenitor. Our preliminary data inferred that BCB-1 is genetically separated from RCC and it has close relation with progenitor. This study is further conducting with numbers of news microsatellite markers for increasing accuracy of our results.

Keywords: BCB-1, Dendogram, genetic distances, heterozygosity, fixation index

Location: BLRI dairy farm, Shahzadpur Upazila of Sirajganj district, Bhangura, Faridpur and Sathia, Pabna.

Start Date: 01-07-2011

End Date: 30-06-2012

Program Area: Conservation and Biodiversity

Commodity: Cow

Objectives:

Methodology:

Collection of blood: Blood samples were collected from BCB-1, Red Chittagong and BCB-1 progenitor cattle populations in EDTA containing 10 ml collection vials using 21 G needle attached with a disposable 10 ml plastic syringe. gDNA extraction: gDNA was extracted using BLRI developed DNA extraction kit from frozen thawed blood samples. About 400 µL blood sample was taken in a 1.5 mL eppendorf tube and lysed with 1 mL lysis buffer through vortexing. DNA amplification and agarose gel electrophoresis: Amplification was performed in 20 µl reaction volume. The PCR reactions were started with an initial heating at 94⁰C for 5 minutes, followed by 35 cycles of 30s at 94⁰C, 40s at 56-60⁰C and 40s at 72⁰C and ended with a final extension at 72⁰C for 10 minutes. Data analysis: Only unambiguous, reproducible and scorable polymorphic fragments were taken into consideration for analysis. The data were used to estimate allelic frequency, genetic distances and dendogram using PopGen32 computer program.

Source of Information: BLRI. Proceedings of Annual Research Review Workshop, 2011-2012. Bangladesh Livestock Research Institute, Savar, Dhaka-1341

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Our preliminary data inferred that BCB-1 is genetically separated from RCC and it has close relation with its progenitor cattle.

Knowledge Management: Report/Proceedings