摘要
结合已公布的大熊猫Ailuropoda melanoleuca基因组和本实验室所测6只大熊猫的转录组数据,筛选多态性微卫星位点并分析其组成及特征。结果显示:共获得326个多态性微卫星位点,其中二碱基多态性微卫星最多,共228个,占69.93%;三、四、五、六碱基所占比例分别为9.51%、14.11%、5.21%、1.22%。根据分析结果中缺失率与标准差2项指标以及位点序列长度,选取20个多态性二碱基微卫星位点,用于25只大熊猫个体血液DNA进行PCR验证并做后续分析。结果表明:不同位点的等位基因数为2~8,平均等位基因数为3.70,观测杂合度、期望杂合度分别为0~1.000、0.280~0.784,平均值分别为0.472和0.532。在Bonferroni校正后,证实4个位点显著偏离哈迪-温伯格平衡,所有位点未观察到显著连锁不平衡(P>0.01)。20个位点多态信息含量(PIC)在0.246~0.734,其中具有高度多态性的位点9个(PIC>0.50),11个位点呈中度多态性(0.25<PIC<0.50)。本研究筛选的微卫星位点有助于评估大熊猫的遗传多样性和种群结构,并制定有效的保护和管理策略。该方法也可为后续开发更多优良微卫星位点用于大熊猫种群遗传学研究提供资源。
Based on the published genomes of giant pandas(Ailuropoda melanoleuca),transcriptome of 6 giant pandas were sequenced in this study to screen the polymorphic microsatellite loci and analyze their composition and characteristics.The results showed that a total of 326 polymorphic microsatellite loci were obtained,of which the dinucleotide polymorphism microsatellites accounted for a maximum of 69.93%(228 in total).The proportions of tri-,tetra-,penta-,and hexanucleotide microsatellite loci were 9.51%,14.11%,5.21%,and 1.22%,respectively.According to the 2 indicators,deletion rate and standard deviation,and the length of the sequence,20 dinucleotide microsatellite loci were selected and verified in 25 giant panda individuals.Further analyses showed that the number of alleles at different loci ranged from 2 to 8,with an average value of 3.70.The observed heterozygosity and the expected heterozygosity ranged from 0 to 1.000 and 0.280 to 0.784 with the average values 0.472 and 0.532,respectively.After Bonferroni correction,it was confirmed that the 4 loci significantly deviated from the Hardy-Weinberg Equilibrium,and no significant linkage disequilibrium was observed at all loci(P>0.01).The polymorphic information content(PIC)of 20 loci ranged from 0.246 to 0.734,with 9 highly polymorphic loci(PIC>0.50),and 11 loci were moderately polymorphic(0.25<PIC<0.50).The microsatellite loci screened in this study can help to assess the genetic diversity and population structure of giant pandas,to develop effective conservation and management strategies,and provide resources for the subsequent development of more excellent microsatellite loci for the genetic study of giant panda population.
作者
涂洪梅
周闯
王冠楠
成美玲
岳碧松
孟杨
TU Hongmei;ZHOU Chuang;WANG Guannan;CHENG Meiling;YUE Bisong;MENG Yang(Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education,College of Life Sciences,Sichuan University,Chengdu 610065,China)
出处
《四川动物》
北大核心
2020年第1期15-22,共8页
Sichuan Journal of Zoology
基金
国家科技攻关项目(2016YFC0503200)
国家自然科学基金项目(31702032)
关键词
大熊猫
基因组
转录组
多态性
微卫星标记
Ailuropoda melanoleuca
genome
transcriptome
polymorphic
microsatellite
