| Honeybees have long been the most important model organism for social biology,playing a key role in elucidating the mechanisms underlying many important social behaviors（such as waggle dance and labor division）.It is an important scientific problem in the field of social biology to study the behavior model of feeding larvae of honeybee and its paternal genetic influence.In-depth study of this scientific problem will help us further understanding the genetic basis and evolutionary trajectory of social behavior,and further enrich the theoretical system of social biology.In present study,Apis mellifera were used as the experimental materials,four experimental colonies were formed manually and marked their emerging workers,providing their own worker bee larvae（group B）and other colony worker bee larvae（group T）as different experimental treatment groups.The larval feeding behavior of the marked workers was observed and recorded using a self-designed and modified observation hive.Nursing behavior was divided into two different types:inspection behavior within 5s and feeding behavior5-300s.At the end of the behavioral observation experiment,samples of marker workers and larvae were collected.Genotype and patrilineal distribution of the marked workers and larvae using 11 SNP s.The observed behavioral data were analyzed with genotypic data and the results were showed that:1.One hundred and six,one hundred and three,seventy-nine and one hundred and eight nursing workers;12,10,17 and 11 successfully capped worker pupae were collected from observation experimental colonies B1,B2,T1 and T2,for a total of446 honey bee samples.The nursing workers belonged to 20,12,15,and 25patrilines.The results of the chi-square test of the patriline distribution of nursing workers in each colony showed that,the patriline distribution of nursing workers was non-random in both group B and group T,and that most of the bee workers（50%-70%）performing nursing tasks came from a few（3-6）patrilines in the colony.2.The random distribution as the H0,the inspection times and the feeding time of different patrilines worker bees in each experimental colony in group B and group T were tested by chi-square test.It was found that:inspection behavior showed a random distribution for group B2 and group T2(PB2=0.4805,PT2=0.1804)and a non-random distribution for group B1 and group T1(PB1=2.374*10-5,PT1=1.398*10-5);while feeding behavior all showed a non-random distribution(PB1=2.2*10-16,PB2=2.2*10-16,PT1=2.2*10-16,PT2=2.2*10-6).3.The random distribution as the H0,chi-square tests were performed on the inspection times and feeding time received by the larvae of different patrilines in group B and group T.It was found that:both inspection behavior and feeding behavior showed significant differences(inspection behavior:PB1=4.024*10-11,PB2=0.01173,PT1=2.2*10-16,PT2=2.2*10-16;feeding behavior:PB1=2.2*10-16,PB2=8.437*10-5,PT1=2.2*10-16,PT2=2.2*10-6).It shows that:the larvae were subjected to a non-random distribution of both inspection behavior and feeding behavior.4.We analyzed group B in comparison with group T,there was no significant difference in the mean inspection times of nursing bees,or in the mean feeding time,indicating that the colony origin of the larvae did not affect the nursing behaviors by workers.5.We used a chi-square test and make a heat map for the correlation between the nursing behaviors of nursing workers and larvae in both treatment groups.The test results showed that the different patrilines（genotype）nursing workers treated all target（including same patriline or same genotype）larvae equally when examining larvae(chi-square test PB1=1.00,PB2=1.00,PT1=0.999,PT2=1.00)in a random distribution model;the different patrilines（genotype）nursing workers fed larvae(PB1=0.00,PB2=0.00,PT1=0.00,PT2=0.00),showed a no kin-selection and non-random distribution model. |
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