摘要
【目的】中华蜜蜂Apis cerana cerana是一种兼有生态价值和经济价值的授粉昆虫。本研究拟揭示秦巴山区中华蜜蜂种群遗传多态性现状,探讨中华蜜蜂的种群分化机制及其影响因素。【方法】使用8个微卫星DNA标记评估秦巴山区17个样点共979个蜂群的中华蜜蜂遗传多态性,以长白山中华蜜蜂和阿坝中华蜜蜂作为外群进行种群遗传分化分析。【结果】秦巴山区中华蜜蜂95%的差异来源于样点内,样点间遗传分化系数(Fst)为0.002~0.037,基因流参数Nm为6.51~124.75,与外群的遗传分化分析中均显示秦巴山区中华蜜蜂不存在遗传分化。平均期望杂合度(He)为0.6877±0.1098,平均观察杂合度(Ho)为0.6364±0.1367,平均有效等位基因数(Ne)为3.7488±1.6201个,平均等位基因数(Na)为7.71±2.52个,多态信息含量(PIC)为0.6418±0.1152,香农指数(I)为1.5026±0.3754,这些参数均表明微卫星遗传多态性丰富。中蜂囊状幼虫病流行过的地区,其杂合度、多态信息含量以及等位基因数均显著低于未发病地区。【结论】秦巴山区中华蜜蜂种群数量大,分布均匀,基因流水平高,在650 km距离范围内没有发生种群分化;秦巴山区中华蜜蜂微卫星遗传多态性丰富,仅部分地区中华蜜蜂遗传多态性受中蜂囊状幼虫病的选择压而降低。
【Aim】The Chinese honeybee( Apis cerana cerana) plays vital roles in pollination for agroecosystem and honey bee products. This study aims to investigate the population differentiation and genetic polymorphism among Apis cerana cerana populations across the Qinling-Daba Mountain Areas,and to explore possible factors contributing to genetic differentiation and difference of genetic polymorphism. 【Methods】The genetic relationship was studied,and polymorphism was estimated among979 colonies of A. c. cerana from 17 localities combined with two outgroups,A. c. cerana colonies from Changbai Mountain and Aba,based on eight microsatellite loci. 【Results】The results showed that up to95% of the variation was observed within sampling localities while less variation between localities. The genetic differentiation coefficient( Fst) among 17 localities ranged from 0. 002 to 0. 037,and the gene flow( Nm) was from 6. 51 to 124. 75,indicating that no population differentiation was detected. The mean expected heterozygosity( He),observed heterozygosity( Ho),number of alleles( Na),number of effective alleles( Ne),polymorphism information content( PIC) and Shannon's information index( I)were 0. 6877 ± 0. 1098,0. 6364 ± 0. 1367,7. 71 ± 2. 52,3. 7488 ± 1. 6201,0. 6418 ± 0. 1152 and1. 5026 ± 0. 3754,respectively,suggesting the abundant genetic polymorphism. The values of the genetic polymorphism indices including the expected heterozygosity,number of alleles,number of effective alleles and polymorphism information content were significantly lower in localities which underwent Chinese sacbrood virus pandemic recently than those in the virus uninfected areas. 【Conclusion】In the context of the large population size and continuous distribution in Qinling-Daba Mountain Areas,there is no genetic differentiation among A. c. cerana populations across a distance of 650 km owing to a high level of gene flow. And the results also imply that the honeybee in Qinling-Daba Mountain Areas has rich genetic polymorphism. Significantly decreased polymorphism occurs in some localities that have undergone selective pressure from Chinese sacbrood virus prevalence.
引文
Bai HY,2014.The Response of Forest Vegetation in Qinling and Daba Mountain Ranges to Environmental Changes.Science Press,Beijing.1-5.[白红英,2014.秦巴山区森林植被对环境变化的响应.北京:科学出版社.1-5]
Barton NH,Briggs DEG,Eisen JA,Goldstein DB,Patal NH,2010.Evolution.Gold Spring Harbor Laboratory Press,Cold Spring Harbor,New York,USA.675-676.
Bohanak AJ,2002.IBD(isolation by distance):a program for analyses of isolation by distance.J.Hered.,93(2):153-154.
Cao LF,Su XL,Zhao DX,Hua QY,Hu FL,2013.Genetic diversity of microsatellite DNA for Apis cerana cerana in Zhejiang.Apiculture of China,64(2):10-11.[曹联飞,苏晓玲,赵东绪,华启云,胡福良,2013.浙江中华蜜蜂微卫星遗传多样性分析.中国蜂业,64(2):10-11]
Chahbar N,Munoz I,Dall’Olio R,De la Rúa P,Serrano J,Doumandji S,2013.Population structure of North African honey bees is influenced by both biological and anthropogenic factors.J.Insect Conserv.,17:385-392.
Chen J,Chen GH,Ji T,Yin L,Liu M,2008a.Study on microsatellite genetic polymorphism of Apis cerana cerana population from Yimeng mountain.Apiculture of China,59(4):11-14.[陈晶,陈国宏,吉挺,殷玲,刘敏,2008a.沂蒙山中华蜜蜂微卫星DNA遗传多样性分析.中国蜂业,59(4):11-14]
Chen J,Ji T,Yin L,Liu M,Chen GH,2008b.Analysis on genetic polymorphism of Apis cerana cerana population in Nanchang region with microsatellite marker.Journal of Anhui Agricultural Science,36(18):7600-7601.[陈晶,吉挺,殷玲,刘敏,陈国宏,2008b.利用微卫星标记分析南昌地区中华蜜蜂遗传多样性.安徽农业科学,36(18):7600-7601]
Chen LZ,Ma KP,2001.Biodiversity Science:Principles and Practice.Shanghai Science and Technology Press,Shanghai.93-122.[陈灵芝,马克平,2001.生物多样性科学:原理与实践.上海:上海科学技术出版社.93-122]
De la Rúa P,JafféR,Muoz I,Serrano J,Moritz RF,Kraus FB,2013.Conserving genetic diversity in the honeybee:comments on Harpur et al.(2012).Mol.Ecol.,22:3208-3210.
Gong YF,Zhang QK,2000.Classification and Evolution of Apis.Fujian Science and Technology Press,Fuzhou.21-26.[龚一飞,张其康,2000.蜜蜂分类与进化.福州:福建科学技术出版社.21-26]
Gu QH,Husemann M,Ding BQ,Luo Z,Xiong BX,2015.Population genetic structure of Bellamya aeruginosa(Mollusca:Gasropoda:Viviparidae)in China:weak divergence across large geographic distances.Ecol.Evol.,5(21):4906-4919.
Han HB,Zhou XR,Pang BP,Zhang MZ,Li HP,2013.Microsatellite marker analysis of the genetic diversity of Oedaleus asiaticus(Orthoptera:Acrididae)populations in Inner Mongolia,northern China.Acta Entomol.Sin.,56(1):79-87.[韩海斌,周晓榕,庞保平,张敏哲,李海平,2013.内蒙古亚洲小车蝗种群遗传多样性的微卫星分析.昆虫学报,56(1):79-87]
Harpur BA,Minaei S,Kent CF,Zayed A,2012.Management increases genetic diversity of honey bees via admixture.Mol.Ecol.,21:4414-4421.
Harpur BA,Minaei S,Kent CF,Zayed A,2013.Admixture increases diversity in managed honey bees:reply to De la Rúa et al.(2013).Mol.Ecol.,22:3211-3215.
Hurtrez-Boussès S,Hurtrez JE,Turpin H,Durand C,Durand P,De Meeus T,Meunier C,Renaud F,2010.Hydrographic network structure and population genetic differentiation in a vector of fasciolosis,Galba truncatula.Infect.Genet.Evol.,10(2):178-183.
Husemann M,Ray JW,King RS,Hooser EA,Danley PD,2012.Comparative biogeography reveals differences in population genetic structure of five species of stream fishes.Biol.J.Linn.Soc.,107:867-885.
Jara L,Muoz I,Cepero A,Martín-Hernández R,Serrano J,Higes M,De la Rúa P,2015.Stable genetic diversity despite parasite and pathogen spread in honey bee colonies.Sci.Nat.,102(9-10):1-8.
Kremen C,William NM,Aizen MA,Gemmill-Herren B,LeB uhn G,Minckley R,Packer L,Potts SG,Roulton T,Steffan-Dewenter I,Vazquez DP,Winfree R,Adams L,Crone EE,Greenleaf SS,Keitt TH,Klein AM,Regetz J,Ricketts TH,2007.Pollination and other ecosystem service produced by mobile organisms:a conceptual framework for the effects of land-use change.Ecol.Lett.,10(4):299-314.
Llewellyn KS,Loxdale HD,Harrington R,Brookes CP,Clark SJ,2003.Migration and genetic structure of the grain aphid(Sitobion avenae)in Britain related to climate and clonal fluctuation as revealed using microsatellites.Mol.Ecol.,12(1):21-34.
Loucifayad W,Achou M,Legout H,Alburaki M,Garnery L,2015.Genetic assessment of Algerian honeybee populations by microsatellite markers.Apidologie,46:392-402.
Michel S,Florence M,Vautrin D,Monnerot M,Cornuet JM,2007.Athird-generation microsatellite-based linkage map of the honey bee,Apis mellifera,and is comparison with the sequence-based physical map.Genome Biol.,8:66-77.
Nikolova SR,Bienkowska M,Gerula D,Ivanova EN,2015.Microsatellite DNA polymorphism in selectively controlled Apis mellifera carnica and Apis mellifera caucasica populations from Poland.Arch.Biol.Sci.,67(3):889-894.
Oleksa A,Chybicki I,Tofilski A,Burczyk J,2011.Nuclear and mitochondrial patterns of introgression into native dark bees(Apis mellifera mellifera)in Poland.J.Apicul.Res.,50:116-129.
Park SD,2001.Trypanotolerance in West African Cattle and the Population Genetic Effects of Selection.Ph D Dissertation,University of Dublin,Dublin,Ireland.
Peakall R,Smouse PE,2012.Gen AlE x 6.5:genetic analysis in Excel.Population genetic software for teaching and research-an update.Bioinformatics,28:2537-2539.
Pritchard JK,Stephens M,Donnelly P,2000.Inference of population structure using multilocus genotype data.Genetics,155:945-959.
Rousset F,2008.Genepop’007:a complete reimplementation of the Genepop software for Windows and Linux.Mol.Ecol.Resour.,8:103-106.
Slatkin M,Barton NH,1989.A comparison of three indirect methods for estimating average levels of gene flow.Evolution,43(43):1349-1368.
Takahashi JI,Shimizu S,Koyama S,Kimura K,Shimizu I,Yoshida T,2009.Variable microsatellite loci isolated from the Asian honeybee,Apis cerana(Hymenoptera;Apidae).Mol.Ecol.Resour.,9(3):819-821.
Takezaki N,Nei M,1996.Genetic distances and reconstruction of phylogenetic trees from microsatellite DNA.Genetics,144:389-399.
Wang CY,2007.Evolutionary Ecology.Higher Education Press,Beijing.126-152.[王崇云,2007.进化生态学.北京:高等教育出版社.126-152]
Wang P,Zhang M,Liu J,Xu L,Liu W,2015.Genetic diversity and structure of Libanotis buchtormensis(Fisch.)DC.in disjunct populations along the bilateral sides of deserts in northwestern China.Plant Sys.Evol.,301(9):2219-2230.
Wang SJ,2008.Breeding of queens resistant to Chinese Sacbrood Virus by five steps.Apiculture of China,59(10):48.[王双进,2008.五步法培育抗中囊病蜂王.中国蜂业,59(10):48]
Wright S,1978.Evolution and the Genetics of Population,Vol.4.Variability within and among Natural Population.University of Chicago Press,Chicago and London.
Xu XJ,Zhou SJ,Zhu XJ,Zhou BF,2013a.Microsatellite DNA genetic diversity of Apis cerana cerana in Hainan Island,southern China.Acta Entomol.Sin.,56(5):554-560.[徐新建,周姝婧,朱翔杰,周冰峰,2013a.海南岛中华蜜蜂遗传多样性的微卫星DNA分析.昆虫学报,56(5):554-560]
Xu XJ,Zhou SJ,Zhu XJ,Zhou BF,2013b.Microsatellite DNA genetic diversity of Apis cerana cerana from the Loess Plateau,Northwest China.Journal of Fujian Agriculture and Forestry University(Natural Science Edition),42(6):638-642.[徐新建,周姝婧,朱翔杰,周冰峰,2013b.黄土高原中华蜜蜂遗传多样性的微卫星DNA分析.福建农林大学学报(自然科学版),42(6):638-642]
Yin L,Ji T,Chen J,Chen GH,Liu M,2008.Study on genetic diversity of Apis cerana cerana populations from mountainous areas in South Anhui by microsatellite makers.Journal of Yangzhou University(Agricultural and Life Science Edition),29(4):19-22.[殷玲,吉挺,陈晶,陈国宏,刘敏,2008.利用微卫星标记分析皖南山区中华蜜蜂遗传多样性.扬州大学学报(农业与生命科学版),29(4):19-22]
Yu YL,Zhou SJ,Xu XJ,Zhu XJ,Zhou BF,2013.Analysis on genetic diversity of Aspis cerana cerana in Changbai Mountains.Journal of Fujian Agriculture and Forestry University(Natural Science Edition),42(6):643-647.[于瀛龙,周姝婧,徐新建,朱翔杰,周冰峰,2013.长白山中华蜜蜂(Apis cerana cerana)遗传多样性分析.福建农林大学学报(自然科学版),42(6):643-647]
Zayed A,2009.Bee genetics and conservation.Apidologie,40:237-262.
Zhang CX,Zhang XF,Chen TZ,Teng YZ,Liu WZ,Jiang YS,2011.Analysis of genetic diversity in twelve honeybee populations based on AFLP technique.Scientia Agricultura Sinica,44(18):3877-3885.[张春香,张学锋,陈廷珠,滕跃中,刘文忠,姜玉锁,2011.基于AFLP技术的12个蜜蜂种群的遗传多样性分析.中国农业科学,44(18):3877-3885]
Zhang PF,Zhou XR,Pang BP,Chang J,Shan YM,Zhang ZR,2015.Microsatellite marker analysis of the genetic diversity of Galeruca daurica(Coleoptera:Chrysomelidae)populations from Inner Mongolia.Acta Entomol.Sin.,58(9):1005-1011.[张鹏飞,周晓榕,庞保平,常静,单艳敏,张卓然,2015.内蒙古沙葱萤叶甲种群遗传多样性的微卫星分析.昆虫学报,58(9):1005-1011]
Zhang ZC,Luo GH,Zhang GF,Han GJ,Liu BS,Fang JC,2013.Genetic diversity of different geographical populations of Sesamia inferens as determined by AFLP.Chinese Journal of Applied Entomology,50(3):693-699.[张志春,罗光华,张谷丰,韩光杰,刘宝生,方继朝,2013.大螟不同地理种群的APFL遗传分析.应用昆虫学报,50(3):693-699]
Zhou BF,2002.The Bee Breeding Management.Xiamen University Press,Xiamen,Fujian.15-16.[周冰峰,2002.蜜蜂饲养管理学.福建厦门:厦门大学出版社.15-16]
Zhu XJ,Xu XJ,Zhou SJ,Wu XD,Zhou BF,2011a.Genetic analysis of Apis cerana cerana in Wuyi Mountain Nature Reserve based on microsatellite DNA.Fujian Journal of Agricultural Sciences,26(6):935-940.[朱翔杰,徐新建,周姝婧,吴显达,周冰峰,2011.武夷山自然保护区中华蜜蜂微卫星DNA遗传分析.福建农业学报,26(6):935-940]
Zhu XJ,Zhou BF,Wu XD,Xu XJ,Xia XC,2011b.Genetic diversity of Apis cerana cerana in Fujian based on microsatellite markers.Journal of Fujian Agriculture and Forestry University(Natural Science Edition),40(4):407-411.[朱翔杰,周冰峰,吴显达,徐新建,夏晓翠,2011.福建中华蜜蜂微卫星标记的遗传多样性分析.福建农林大学学报(自然科学版),40(4):407-411]