Double Digested RAD Sequencing Technique for SNP Identification on Africanized Apis mellifera Honeybee Populations

The aim of this research was identified Single Nucleotide Polymorphisms (SNP´s) on Africanized honeybees populations with high and low honey production using ddRADseq (Double Digested RAD Sequencing) technique for genomic DNA sequencing. The experiment was conducted at Lageado Experimental Farm, UNESP, Botucatu, São Paulo, Brazil. Forty Africanized Apis mellifera L. beehives were used, managed for honey production and equal population, brood and food frames. During the blossom period, December 2013 to March 2014, supers were added to beehives as needed.  Honey production was evaluated individually and the ten most productive (High Honey Production, HHP), over 10 kg honey production for each beehive (12.21±1.5 kg), and less productive (Low Honey Production, LHP), less than 10 kg honey production for each beehive (4.8±3.9kg), were selected. Defense and hygienic behavior was evaluated. Genomic sequencing was done with larvae drone pool DNA from each group using ddRADseq technique and then sequenced by Illumina technology (DNA-Seq). SNP´s were identified by 1.12 STACKS program. The results were compared by ANOVA followed by Tukey test as to differences between averages. To correlate the data Pearson linear correlation was used. It was considered statistically different when p <0.05. HHP showed low defense behavior and after analysis by 1.12 STACKS were identified 187,500 and 529,172 SNPs in HHP and LHP groups, respectively, in Africanized Apis mellifera honeybees populations. It can be concluded that ddRADseq was a technique that can be used to SNP´s identification in different Africanized honeybees populations.