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Abstract
Genes related to carbohydrate metabolism have evolved rapidly in eusocial bees, including honey bees. However, the characterisation of carbohydrate metabolism genes has not been reported in Apis andreniformis or Apis cerana indica. This study aimed to characterise phosphofructokinase (PFK) and pyruvate kinase (PK) genes in these honey bee species and to analyse the evolution of the genus Apis using these genes. This study found the first data regarding A. andreniformis PFK and PK-like nucleotide sequences. A BLAST-n algorithm-based search showed that A. andreniformis and A. c. indica PFK and PK genes were homologous with those of Apis florea and Apis cerana cerana from Korea, respectively. Multiple alignments of PFKs from five Apis species showed many exon gains and losses, but only one among the PKs. Thus, the exon–intron organisation of the PK genes may be more conserved compare with that of the PFKs. Another evolutionary pattern indicated that more nucleotide substitutions occurred in Apis’ PK than PFK genes. Deduced PFK amino acid sequences revealed a PFK consensus pattern of 19 amino acids, while the deduced PK amino acid sequences were predicted to have barrel and alpha/beta domains. Based on these two metabolism-related genes, The Neighbour-joining and Maximum likelihood phylogenetic trees are congruent and revealed that the A. andreniformis and A. florea group were in the basal position. Apis mellifera, A. cerana, and Apis dorsata formed a monophyletic clade, although the positions of A. mellifera and A. dorsata were different in the nucleotide- and amino acid-based phylogenetic trees.
Highlights
Honey bees use carbohydrates as the main fuel for flight and produce modified stored sugar to maintain the optimal hive temperature (Fischman et al 2011)
BLAST-n algorithm-based searches using the nucleotide collection database showed that A. andreniformis is closely related to A. florea, with 100% (GenBank XM_012485123.1) and 95% (GenBank XM_012487945.1) identities for PFK and Pyruvate kinase (PK)-like genes
Using a combination of PFK and PK-like nucleotide (Fig. 8) and amino acid (Fig. 9) sequences in the genus Apis and out group, this study found two topologies based on nucleotide sequence and amino acid phylogenetic tree
Summary
Honey bees use carbohydrates as the main fuel for flight and produce modified stored sugar (honey) to maintain the optimal hive temperature (Fischman et al 2011). Phosphofructokinase (PFK; EC 2.7.1.11) plays a key regulatory role in the glycolytic pathway It catalyses the reaction of fructose 6-phosphate using ATP to generate 1,6-diphosphate and ADP (Voet & Voet 1995). The first eukaryotic PFK sequence was characterised in cloned rabbit muscle and its 17-kb length was split into 22 exons, encoding 780 amino acids (Lee et al 1987). The exon–intron organisation was the same among human liver (Elson et al 1990), human muscle (Vaisanen et al 1992) and mouse liver (Rongnoparut et al 1991) PFKs. In insects, the PFK gene has been characterised in Drosophila melanogaster and spans 6.5 kb, which is split into 8 exons and encodes 787 amino acids. The amino acid sequence of D. melanogaster PFK showed a 50.9% identity with the human muscle PFK (Currie & Sullivan 1994)
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