Mitochondrial D-loop sequence variation among Central Javanese Duck in Indonesia / Variação de sequências de D-Loop Mitocondrial entre Pato Central Java na Indonésia

This study was realized to determine the genetic variation of Central Javanese duck based on the D-Loop mtDNA gene. D-loop gene was amplified using PCR technique by specific primer and sequenced using dideoxy termination method with ABI automatic sequencer. ClustalW from MEGA-6.06 software program was employed for multiple alignments of nucleotide sequences. Nucleotide sequences of D-loop gene of mtDMA from the Central Javanese duck were aligned together with other Anas isolates from Genbank using ClustalW of MEGA-6.06 program. The estimation of genetic distance and phylogenetic tree construction were analyzed by Neighbor-Joining method, whereas the calculation of distance matrix was performed using Kimura 2-parameter. Multiple alignments obtained were 720 nucleotides at position 56 to 779 at the 5 "end. The results of the polymorphism analysis on D-loop sequences produced 23 haplotypes. However, this haplotype information does not represent the relationship among the geographical origins of duck with the certain duck species name. Moreover, a total number of 32 variable sites were identified. Insertions were detected in four sequences (126, 155, 771 and 779 nucleotide number). In the phylogenetic analysis, it is safe to conclude that the Central Javanese duck is closely related to Anas platyrhynchos and Anas zonorynchos.

Este estudo foi realizado para determinar a variação genética do pato de Java Central baseado no D-Loop mtDNA. A D-Loop foi amplificada utilizando a técnica de PCR com primer específico e sequenciado usando o método de terminação dideoxi com sequenciador ITB automático. ClustalW do programa software de MEGA-6.06 foi utilizado para alinhamentos de algumas sequências de nucleótidos. Sequências de nucleótidos de D-loop gene da mtDMA do pato de Java Central foram alinhados em conjunto com alguns isolados de Anas que foram obtidos de Genbank utilizando o programa ClustalW do programa MEGA-6.06. A estimativa da distância genética e a estrutura da árvore filogenética foram analisadas com o método Neighbor-Joining, enquanto que o cálculo da matriz de distância foi utilizado o parâmetro-2 Kimura. Os alinhamentos múltiplos obtidos foram 720 nucleótideos na posição 56 a 779 na extremidade 5’. As análises do polimorfismo sequencial do D-loop resultaram em 23 haplótipos. No entanto, esta informação haplótipo não representa a relação entre a origem geográfica de pato com o nome de determinadas espécies do pato. Além disso, num total de 32 sites favoráveis foram identificados. As inserções foram detectadas em quatro sequências (126, 155, 771 e 779 números de nucleotídeos). Na análise filogenética é seguro concluir que o pato de Java Central está estreitamente relacionada com a Anas platyrhynchos e Anas zonorynchos.

Mitochondrial D-loop sequence variation among Central Javanese Duck in Indonesia / Variação de sequências de D-Loop Mitocondrial entre Pato Central Java na Indonésia

 This study was realized to determine the genetic variation of Central Javanese duck based on the D-Loop mtDNA gene. D-loop gene was amplified using PCR technique by specific primer and sequenced using dideoxy termination method with ABI automatic sequencer. ClustalW from MEGA-6.06 software program was employed for multiple alignments of nucleotide sequences. Nucleotide sequences of D-loop gene of mtDMA from the Central Javanese duck were aligned together with other Anas isolates from Genbank using ClustalW of MEGA-6.06 program. The estimation of genetic distance and phylogenetic tree construction were analyzed by Neighbor-Joining method, whereas the calculation of distance matrix was performed using Kimura 2-parameter. Multiple alignments obtained were 720 nucleotides at position 56 to 779 at the 5 "end. The results of the polymorphism analysis on D-loop sequences produced 23 haplotypes. However, this haplotype information does not represent the relationship among the geographical origins of duck with the certain duck species name. Moreover, a total number of 32 variable sites were identified. Insertions were detected in four sequences (126, 155, 771 and 779 nucleotide number). In the phylogenetic analysis, it is safe to conclude that the Central Javanese duck is closely related to Anas platyrhynchos and Anas zonorynchos.(AU)

Este estudo foi realizado para determinar a variação genética do pato de Java Central baseado no D-Loop mtDNA. A D-Loop foi amplificada utilizando a técnica de PCR com primer específico e sequenciado usando o método de terminação dideoxi com sequenciador ITB automático. ClustalW do programa software de MEGA-6.06 foi utilizado para alinhamentos de algumas sequências de nucleótidos. Sequências de nucleótidos de D-loop gene da mtDMA do pato de Java Central foram alinhados em conjunto com alguns isolados de Anas que foram obtidos de Genbank utilizando o programa ClustalW do programa MEGA-6.06. A estimativa da distância genética e a estrutura da árvore filogenética foram analisadas com o método Neighbor-Joining, enquanto que o cálculo da matriz de distância foi utilizado o parâmetro-2 Kimura. Os alinhamentos múltiplos obtidos foram 720 nucleótideos na posição 56 a 779 na extremidade 5. As análises do polimorfismo sequencial do D-loop resultaram em 23 haplótipos. No entanto, esta informação haplótipo não representa a relação entre a origem geográfica de pato com o nome de determinadas espécies do pato. Além disso, num total de 32 sites favoráveis foram identificados. As inserções foram detectadas em quatro sequências (126, 155, 771 e 779 números de nucleotídeos). Na análise filogenética é seguro concluir que o pato de Java Central está estreitamente relacionada com a Anas platyrhynchos e Anas zonorynchos.(AU)

Mitochondrial D-loop sequence variation among Central Javanese Duck in Indonesia

 This study was realized to determine the genetic variation of Central Javanese duck based on the D-Loop mtDNA gene. D-loop gene was amplified using PCR technique by specific primer and sequenced using dideoxy termination method with ABI automatic sequencer. ClustalW from MEGA-6.06 software program was employed for multiple alignments of nucleotide sequences. Nucleotide sequences of D-loop gene of mtDMA from the Central Javanese duck were aligned together with other Anas isolates from Genbank using ClustalW of MEGA-6.06 program. The estimation of genetic distance and phylogenetic tree construction were analyzed by Neighbor-Joining method, whereas the calculation of distance matrix was performed using Kimura 2-parameter. Multiple alignments obtained were 720 nucleotides at position 56 to 779 at the 5 'end. The results of the polymorphism analysis on D-loop sequences produced 23 haplotypes. However, this haplotype information does not represent the relationship among the geographical origins of duck with the certain duck species name. Moreover, a total number of 32 variable sites were identified. Insertions were detected in four sequences (126, 155, 771 and 779 nucleotide number). In the phylogenetic analysis, it is safe to conclude that the Central Javanese duck is closely related to Anas platyrhynchos and Anas zonorynchos.