湖南龙山不同海拔中华蜜蜂种群遗传多样性的微卫星DNA分析

Microsatellite DNA analysis of the genetic diversity of Apis cerana cerana(Hymenoptera:Apidae)populations at different altitudes in Longshan,Hunan,central China

【目的】利用11对微卫星标记对湖南龙山中华蜜蜂Apis cerana cerana种群遗传多样性进行分析,评估种群内遗传变异和种群间的遗传分化。【方法】从湖南省龙山县采集不同海拔(1080和665 m)中华蜜蜂各30群,共60群。从每群10~20头成年工蜂中随机挑选1头提取DNA作为模板,利用11对微卫星引物进行PCR。基于PCR扩增产物,通过Microsatellite-Toolkit软件计算高海拔种群(G)和低海拔种群(D)各基因位点的优势等位基因频率(Pi)、期望杂合度(He)、多态信息含量(PIC)。根据FSTAT程序计算种群内近交系数(Fis)。用SPSS 25.0软件分析两个种群(G和D)间Pi,He,PIC和Fis的差异显著性。【结果】湖南省龙山县中华蜜蜂高海拔种群(G)和低海拔种群(D)的Pi分析表明,两个种群均具有较高的遗传多样性;高海拔种群的He,PIC和Fis平均值分别为0.593,0.556和0.121,均略低于低海拔种群的0.631,0.587和0.187。两个种群间Pi,He,PIC和Fis无显著性差异(PPi=0.721>0.05,PHe=0.759>0.05,PPIC=0.802>0.05,PFis=0.767>0.05)。【结论】位于武陵山区龙山县地域高海拔与低海拔中华蜜蜂种群具有较高遗传多样性,但遗传分化程度不高,提示海拔因素可能不直接影响中华蜜蜂种群遗传多样性。

【Aim】To analyze the genetic diversity of Apis cerana cerana populations in Longshan,Hunan,central China by using 11 pairs of microsatellite markers,so as to evaluate the genetic variation within the population and genetic differentiation between populations.【Methods】A total of 60 colonies of A.c.cerana were collected from Longshan County,Hunan Province at different altitudes(1080 and 665 m).Ten to twenty adult worker bees were randomly collected from each colony,of which one individual was randomly selected to extract DNA as a template,and PCR was performed using 11 pairs of microsatellite primers.Based on PCR amplification products,the dominant allele frequency(Pi),expected heterozygosity(He)and polymorphism information content(PIC)of each locus in the high altitude population(G)and the low altitude population(D)were calculated by Microsatellite-Toolkit software.The inbreeding coefficient(Fis)within population was calculated according to the FSTAT program.The significance of differences in Pi,He,PIC and Fis between the two populations(G and D)was analyzed by using SPSS 25.0 software.【Results】The Pi analysis indicated that both the high altitude(G)and low altitude(D)populations of A.c.cerana in Longshan County,Hunan Province had high genetic diversity.The average values of He,PIC and Fis in the high altitude population(G)were 0.593,0.556 and 0.121,respectively,which were slightly lower than those of the low altitude population(D)(0.631,0.587 and 0.187,respectively).There was no significant difference in Pi,He,PIC and Fis between the high and low altitude populations(PPi=0.721>0.05,PHe=0.759>0.05,PPIC=0.802>0.05,PFis=0.767>0.05).【Conclusion】A.c.cerana populations from high and low altitudes in Longshan County in Wuling Mountain area have high genetic diversity,but the degree of genetic differentiation is not high,suggesting that the altitude factor may not directly affect the population genetic diversity of A.c.cerana.