Introduction Mapping genes for natural variation in behavior 1991 2000 2004 1989 2005 2003 2003 2004 2005 2004 1998 2004 1998 1995 2004 2006 Behavioral genetics of foraging 1 1998 2004 Fig. 1 Arrows pln 1–4 Colored lines picture  pln-1 pln-2 pln-3 1995 2000 pln-1 pln-2 pln-3 Pln-1 pln2 2 Pln-2 pln-3 1995 2000 2 AmFor pln-4 2004 pln-1 1 2004 2006 Fig. 2 Solid bars Markers arrows Dashed lines underlined pln-4 AmFOR a Pln-1 Map on the left Map on the right b Pln-2 QTL map on the left map on the right c Pln-3 QTL map  2004 2004 2006 1 2006 2005 2004 2005 2005 2001 2005 2005 Behavioral genetics of defensive behavior 1987 2003 2004 1961 1985 1987 2002 1999 2003 1998 3 2003 1998 sting-1 2002 2003 Fig. 3 Arrows  Materials and methods QTL mapping and confirmation 1995 1998 2000 2004 2006 2001 2002 z z z z y μ δ y μ δ sting1 sting2 sting3 1999 1989 Cloning and sequencing marker fragments 2006 RACE and cDNA cloning Before expression analyses by quantitative real-time polymerase chain reaction (qRT-PCR), cDNA cloning was performed to confirm sequences and the gene prediction (location of introns and exons) and to provide information on sequence variation within some of the candidate genes. In the case of the serotonin receptor, this process provided complete sequence of the gene by making primers based on sequence of a putative G protein-coupled receptor (GPCR). Rapid amplification of cDNA ends (RACE) and cloning was performed using kits and manufacturers’ instructions. Total RNA was extracted from individual bees using the RNAqueous kit (Ambion, Austin TX). The cDNA was synthesized using the SMART PCR cDNA Synthesis kit (SMART, simple modular architecture research tool, BD Biosciences, Palo Alto CA). The cDNA clones were obtained using the TOPO-TA kit and the pCR4-TOPO vector (Invitrogen). Clones were sequenced from multiple worker bees. Several sequence reads were obtained from each clone. Comparative bioinformatics 2006 http://www.ncbi.nlm.nih.gov Quantitative real-time PCR 1998 free 2005 32 32 32 R 2 Results and discussion Bioinformatic analyses of putative gene functions and results of qRT-PCR allowed us to form hypotheses concerning gene networks involved in either foraging or defensive behaviors. Results of analyses and hypotheses concerning genes with potential to influence behavior are presented in the following two sections. Candidate genes for honeybee foraging behavior 2006 1 4 pln-1 Drosophila bazooka midway 2004 pln-1 1999 2002 Table 1 Annotation of peptides within 97% confidence intervals for foraging-behavior QTLs  Fig. 4 2005 2005 ILPs PI PIP IRS PI3K PIP5K PIG-P PDK1 PKB HR46 HR46 PTEN JH  pln-2 2 HR46 2004 HR46 2003 pln-2 AmTyr1 Drosophila hono 2004 pln-2 skittles 2 4 2003 Drosophila skittles 1998 1 pln-3 PI3K 68D 2004 2005 pln-3 2 3 pln-3 4 2001 Drosophila 2005 4 pln-3 bazooka pln-1 pln-2 bazooka 3 bazooka 2005 pln-3 pln-1 pln-2 2004 2006 AmFOR pln-4 IRS AmFOR Drosophila 2005 IRS Drosophila 2006 1 pln-3 pln-1 pln-2 pln-4 4 Drosophila Candidate genes for defensive behavior Sting-1 2002 2003 5 2 2003 Drosophila 2003 sting-1 Drosophila tango Tango 2003 tango 2003 2004 3 sting-1 Fig. 5 1 Markers Letters and numbers next to vertical bar underlined Dashed lines Drosophila a  Sting-1 b  Sting-2 c  Sting-3  Table 2 Annotation of peptides within 97% confidence intervals for defensive-behavior QTLs  Table 3 Expression of candidate genes in high-defensive bees relative to low-defensive worker bees  sting-2 Drosophila arr1 AmArr4 5 Drosophila arr1 2005 GABA-B-R1 2004 sting-3 sting-2 7 Am5HT 7 5 2005 1995 7 7 2001 2003 7 2005 sting-3 AmPKA-C1 2001 homer AmPKA-C1 2002 2004 14-3-3 Epsilon 3 GABA-B-R1 sting1 sting2 /discs lost AmArr4 /tango Am5HT 7 2005 The advantage of high recombination rates 4 Drosophila Drosophila 2006 2003 2005 4 2006 Drosophila sting2 2003 Table 4 Drosophila a b Genus Trait Number of QTL c Actual no. of genes  145 Apis Pollen foraging 4 320 222 113 107 Apis Defensive behavior 3 236 145 128 158 Drosophila Ovariole number 5 6,800 – 9,100 a Drosophila D. simulans D. sechellia 2006 Apis b c Conclusions 1 4 Am5HT 7