目的 通过研究云南省盈江县那邦镇微小按蚊在不同年份的群体遗传特征,对比微小按蚊在遗传多样性方面的变化.方法 分别于2014年5月和2021年5月在云南省盈江县那邦镇用诱蚊灯采集蚊虫,经形态学分类鉴定后,单管单只保存待用.使用试剂盒提取微小按蚊的DNA.用8对荧光引物扩增模板DNA中的微卫星序列后,产物送测序公司进行毛细管电泳检测.使用PopGen32分别计算单个微卫星位点和群体的等位基因数(number of alleles,Na)、有效等位基因数(effective number of alleles,Ne)、观测杂合度(observed heterozygosity,Ho)、期望杂合度(expected heterozy-gosity,He)、香农指数(Shannon's information index,Ⅰ).PIC-CALC 软件计算多态信息含量(polymorphism information con-tent,PIC).结果 2014年捕获158只共6种按蚊,2021年捕获529只共5种按蚊.2014年和2021年的微小按蚊在按蚊中的构成比差异有统计学意义(x2=70.48,P＜0.01).8个微卫星位点共检测到85个等位基因.Na为6～20个,平均值为10.625.Ne为1.717～7.797个,平均值为4.011;PIC最高的是am4位点,最低的是am25位点.群体方面,在2014年中发现77个等位基因;在2021年中发现62个等位基因.2014年Ne为1.630～8.658,平均为4.147;2021年Ne为1.760～6.744,平均为 3.698.2014年平均 Ho为 0.641,2021年为 0.650;2014年平均 He 为 0.699,2021年为 0.691.2014年I 为 0.774～2.493,平均为1.579;2021年I为0.938～2.224,平均为1.464.2014年的4个指标Na、Ne、I和PIC均高于2021年,分别是Na:9.625＞7.750,Ne:4.147＞3.698,I:1.579＞1.464,PIC:0.655＞0.640.结论 盈江县那邦镇微小按蚊 2014年和 2021年的群体均呈高度多态性.但2021年的群体遗传多样性低于2014年.
Comparison of genetic diversity of Anopheles minimus in Nabang Town,Yingjiang County,Yunnan Province between 2014 and 2021
Objective To compare the changes in the genetic diversity of Anopheles minimus through the research on the population genetic characteristics of Anopheles minimus between different years in Nabang Town,Yingjiang County,Yunnan Province.Methods Anopheles mosquitoes were collected by light traps in Nabang Town,Yingjiang County,Yunnan Province in May 2014 and May 2021.After morphological identification,each mosquito was individually stored in separate tubes for fur-ther analysis.DNA of Anopheles minimus was extracted using kits.Microsatellite sequences in the template DNA were ampli-fied using eight pairs of fluorescent primers,and the resulting products were subjected to capillary electrophoresis by a sequenc-ing company.PopGen32 software was used to calculate the observed number of alleles(Na),effective number of alleles(Ne),ob-served heterozygosity(Ho),expected heterozygosity(He),and Shannon's information index(Ⅰ)for individual microsatellite loci and population groups.PIC-CALC software was used to calculate the polymorphic information content(PIC).Results A total of 158 mosquitoes belonging to 6 Anopheles species were captured in 2014,while 529 mosquitoes belonging to 5 Anopheles spe-cies were captured in 2021.The composition ratio of Anopheles minimus among the mosquito species differed significantly be-tween 2014 and 2021(x2=70.48,P＜0.01).For 8 microsatellite loci,a total of 85 alleles were detected,a range of 6-20 alleles per locus and an average of 10.625 alleles.Ne ranged from 1.717 to 7.797,with an average of 4.011.The highest PIC was found in the am4 locus,and the lowest in the am25 locus.For the population groups,77 alleles were found in 2014,and 62 alleles were found in 2021.Ne ranged from 1.630 to 8.658,with an average of 4.147 in 2014.Ne ranged from 1.760 to 6.744,with an average of 3.698 in 2021.The average Ho was 0.641 in 2014 and 0.650 in 2021,while the average He was 0.699 in 2014 and 0.691 in 2021.The Shannon's index ranged from 0.774 to 2.493 in 2014,with an average of 1.579,and from 0.938 to 2.224 in 2021,with an average of 1.464.Na,Ne,I,and PIC were all higher in 2014 compared to 2021,with Na:9.625＞7.750,Ne:4.147＞3.698,I:1.579＞1.464,and PIC:0.655＞0.640,respectively.Conclusions The populations of Anopheles minimus in Nabang Town,Yingjiang County,exhibited high levels of polymorphism in both 2014 and 2021.However,the genetic diversity of the population in 2021 was lower than that in 2014.
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