【作者】 余林生；

【导师】 邹运鼎； 李增智；

【作者基本信息】 安徽农业大学 ， 森林保护学， 2007， 博士

【摘要】 本项目在总结国内外东方蜜蜂种群动态及群体多样性研究进展的基础上,结合皖南山区生态环境,采用昆虫种群生态学及现代分子生物学研究方法,系统研究了皖南山区中华蜜蜂种群数量动态特征及群体多样性,研究结果如下:（1）对皖南山区对中华蜜蜂（Apis cerana cerana Fab.）种群数量动态与结构特征研究结果为:皖南中蜂蜂王产卵力强,分蜂性弱,同一蜂种的不同蜂群的分蜂性也有差异,维持群势大,有效采集时间长,越夏期蜂王停产期短,群势下降不明显,越冬期短,越冬饲料消耗量少,越冬稳定。皖南中蜂蜂王产卵量和种群内蜜蜂数量明显受气候和蜜粉源植物影响,周年分为春季蜂群繁殖期、自然分蜂时期、蜂群越夏期、蜂群秋季更新期和蜂群越冬期,呈现“两高”（春、秋）和“两低”（夏、冬）的变化模式曲线。皖南中蜂性比值为278.5:1～381.2:1,雄蜂呈季节性出现,大多是在春末夏初的分蜂季节里培育雄蜂,种群内年龄组配在不断变化。秋季培育适龄越冬蜂的蜂群,工蜂腺体处于发育初期,越冬期蜂群群势下降率低,越冬安静,饲料消耗率低,越冬稳定性强,幽王断子提高了适龄越冬蜂数量和质量。秋季适龄越冬蜂的培育与幽王断子的技术措施影响越冬期中蜂种群的年龄组配,年龄组配是蜜蜂种群的结构特征,而蜜蜂种群的结构特征影响蜂群越冬稳定性与越冬效果。在新法饲养中蜂的季节性管理中,必须采取培育适龄越冬蜂并适时幽王断子的技术措施。（2）中华蜜蜂卵在温度低于30℃条件下不能孵化,在32℃、34℃和36℃发育历期差异显著（P＜0.05）,受精卵在32～38℃下的孵化率差异不显著（P＞0.05）,而未受精卵在32℃条件下的孵化率与其它温度下的孵化率的差异显著（P＜0.05）。温度对中华蜜蜂蜂王和雄蜂封盖子期影响,导致蜂王初生重、性成熟时间差异显著（P＜0.05）。蜂王和雄蜂认巢飞行次数分别为1.23～1.31、1.08～1.13,持续时间分别为0.12～0.13h、0.16～0.20h;蜂王和雄蜂交配飞行次数分别为1.10～1.12、1.01～1.05,持续时间分别为0.22～0.23h、0.18～0.23h;蜂王与雄蜂交配最适宜温度为20～28℃。蜂王交配飞行一次侧输卵管的精子数为3.37～4.15×10⁶,自然交配产卵蜂王受精囊精子数为3.55～3.62×10⁶。蜂王初生重与产卵量之间呈正相关γ=0.715（p＜0.05）,周年蜂王产卵量受气候和蜜粉源影响明显,早春中蜂群势越强,蜂团中心温度越高,蜂团体积越大,蜂王开始产卵越早。通过对新法饲养培育过不同代幼虫的中蜂巢脾研究表明,由于茧衣和粪便积聚,使巢房的容积和形状发生变化,从而影响了后代的培育。中蜂咬毁旧脾,给管理带来麻烦,堆积的蜡屑又助长了巢虫的滋生。防止的根本措施是适应它喜新脾习性,勤换新脾。在安徽皖南山区,中华蜜蜂发育历期:蜂王平均为14.3d（332～353h）,工蜂平均为19.1d（446～468h）,雄蜂平均为24.1d（556～576h）,中华蜜蜂初生重:蜂王平均为167.7mg,工蜂平均为94.9mg,雄蜂平均为146.4mg,季节（春、夏、秋）温度的差异,使得中蜂发育历期相应缩短或延长,发育历期的变化引起初生重的差异极显著（p＜0.01）。幼虫的体重增重速度在卵孵化后90h以内较慢,90h以后增重速度很快,在种用蜂王和雄蜂培育过程中,确保蜂王和雄蜂幼虫发育处在稳定的蜂巢环境中,提高种用蜂王和雄蜂的质量。（3）对新法饲养和旧法饲养的皖南山区中华蜜蜂群体温度变化及调控研究结果为:中蜂蜂群巢脾中心点巢温受气温影响不明显,基本稳定,而巢脾边缘巢温与气温呈正相关,随气温升降而升降。新法和旧法饲养的中华蜜蜂蜂团中心温度分别为春季（32.57±0.29）℃、（32.78±0.60）℃,夏季（34.03±0.33）℃、（34.26±0.31）℃,秋季（32.19±0.26）℃、（32.39±0.26）℃,冬季（26.14±0.39）℃、（27.47±0.31）℃;边缘温度分别为春季（22.24±1.77）℃、（22.49±1.78）℃,夏季（33.71±0.64）℃、（34.08±0.66）℃,秋季（19.27±0.54）℃、（19.68±0.52）℃,冬季（14.20±0.09）℃、（14.81±0.07）℃。新法和旧法饲养的蜂团中心温度与环境温度之间差异均极显著（P＜0.01）;夏季和冬季新法和旧法饲养的蜂团边缘温度与环境温度之间差异均显著（P＜0.05）,春、秋季差异不显著（p＞0.05）;冬季旧法饲养蜂团中心与边缘温度均高于新法。在环境温度剧变条件下,通过群体调节作用,冷库中繁殖群蜂团中心温度保持30.5～32.6℃;温室中断子群蜂团中心温度保持27.8～31.8℃。中蜂越冬蜂团中心温度与外界环境温度日变化同步,伴随着越冬蜂群群势下降,越冬蜂团中心温度和相对湿度波动范围增大,蜂群越冬稳定性下降。旧法饲养比新法饲养中蜂蜂群越冬稳定性强。（4）中蜂与意蜂食物（蜜源植物）资源生态位宽度分别是0.923、0.765,中蜂对蜜源植物采集喜好性差异小,而意蜂差异大,中蜂对意蜂生态位重迭为0.142,中蜂对意蜂生态位相似性为0.755;油菜花期,中蜂与意蜂时间资源生态位宽度分别是0.879、0.801,中蜂对意蜂生态位重迭为0.146,中蜂对意蜂生态位相似性为0.875;枇杷花期,中蜂与意蜂时间资源生态位宽度分别是0.760、0.677,中蜂对意蜂生态位重迭为0.139,中蜂对意蜂生态位相似性为0.901;中蜂与意蜂空间资源生态位宽度分别是0.797、0.670,中蜂对意蜂生态位重迭为0.179,中蜂对意蜂生态位相似性为0.903;冬季温度低中蜂耐寒性强于意蜂,海拔高、温度低中蜂比意蜂种群分布广。西方蜜蜂与中蜂在生态位上虽然有许多重叠,但其个体特性却存在许多差异,中蜂对本地植物授粉资源的生态位宽度大于西方蜜蜂。中蜂的灭绝,就会降低当地植物授粉总量,使多种植物授粉受到影响,逐渐减少一些种类的数量,直至最终绝灭,结果导致山林中植物多样性减少。中蜂是我国自然生态体系中不可缺少的重要环节,具有重要的生态价值,保护中蜂也就是保护蜜蜂遗传基因的多样性,对于恢复植被、保护物种多样性、改善生态环境具有重要作用,具有显著的生态效益。在植被结构和生态条件不同的皖南山区和皖西大别山区、江淮地区及淮北平原,栖息环境的改变和种间竞争的生存竞争等因素是影响中蜂群体分布的主要因素。皖南山区和皖西大别山区自然植被完整,生态条件好,蜜粉源植物丰富,中蜂群体数量多,分布区域广,分布密度高,中蜂蜂王自然交配极少受到干扰,种间竞争处于优势,是中蜂栖息与繁衍的理想场所。江淮地区和淮北平原自然植被少,生态系统受到不同程度破坏,蜜粉源植物种类少,花期短而集中,中蜂蜂王自然交配受到意蜂雄蜂干扰,种间竞争处于劣势,中蜂群体数量锐减,分布区域缩小,群体分布密度大幅度下降,淮北平原比江淮地区更显著。（5）通过测定皖南山区5个不同采样点的15群中华蜜蜂,每群15只工蜂,每只蜜蜂样共8个形态特征,结合测定的性状特征数据进行差异显著性检验及主成分分析、辨别分析和聚类分析,发现形态特征存在差异,其中喙长、右前翅长、第三节背板宽、第三节背板颜色、第四节背板宽、第四节背板颜色差异显著;皖南中蜂种群内遗传变异丰富,种群中许多形态性状特征表现出较明显的地理变异性。皖南中蜂种群形态性状的多态性、多型性及其生态地理变异式样,均具有生态适应意义。对来自皖南山区以及浙江和江西的共17群中华蜜蜂线粒体DNA多态性的检测,并将皖南中蜂线粒体DNA的PCR扩增序列,同江西、浙江中蜂的扩增序列进行比较,共发现四种线粒体单体（haplotype）:A₁A₂A₃和B₃为第一种,C₁C₂ C₃为第二种,B₂为第三种,其余为第四种,皖南中蜂线粒体DNA序列同江西、浙江中蜂无明显差异。通过对皖南中蜂mtDNA COⅡ基因的440个碱基对进行测序分析发现,440个碱基对中,G+C含量从15.1%～15.7%不等,A+T含量从84.3%～84.9%不等;440个碱基对里共有8个位点变异,T-C和A-T转换分别为3个和2个。简略位点为4个。分析17群中华蜜蜂的440个碱基序列里,位点变异从0个到6个不等,变异率从0%到1.36%。同一地区的蜂群之间,如安徽省黄山市徽州区、屯溪区、黄山区,其碱基序列位点变异分别为2个、3个和1个。个别不同地区的蜂群之间如歙县和祁门县的碱基序列差异为0。皖南中蜂（A～E）的遗传相似系数均在0.967以上,皖南中蜂与浙江中蜂的遗传相似系数均在0.971以上,皖南中蜂与江西中蜂的遗传相似系数均在0.973以上,说明皖南中蜂与浙江、江西中蜂亲缘关系非常近。通过Cluster V构建的系统进化树分析,皖南中蜂A与B₃为一类,C为一类,B₂为一类,E₁为一类,D、B₁、E₂、E₃和江西、浙江中蜂为一类。更多还原

【Abstract】 On the basis of a review on the research of oriental honey bees,population dynamics and colony diversity of the Chinese honey bee,Apis cerana cerana,in montanic area of southern Anhui were studied systematically,by use of methods of population dynamics ahd molecular biology.The following results were achieved.1.Study on population dynamics and structure characterization of Apis cerana cerana Fab.led to the following results:The queens of the Chinese honey bee in this area had high fecundity and poor swarming capacity,but the capacity of different colonies were different.It kept strong colony potential,with long effective collecting period.Queens had short oversummering oviposition-off,without clear potential decline,and short and overwinterring period,with little and stable food consumption. The fecundity of a queen,high in spring and autumn,and low in summer and winter, and population size,big in spring and fall,and small in summer and winter,were impacted obviously by climate and honey and nectar plant.Five different periods appeared in bee colony:spring reproduction period,natural warming period, oversummering period,autumn refreshment period,and overwintering period.Sex ratio ranged within 278.5:1～381.2:1.Drones emergence varied seasonally and were bred mostly in swarming seasons.Age composition varied continually.The colony potential of the swarm which bred spados,with early developed glands,in autumn for overwintering declinedat a low rate and kept in quite status,with low forage consumption,indicating an overwintering stability.Cutting off offspring by imprisonment of queens improved both quantity and quality of adaptive overwintering bees.Age composition of a overwintering swarm,which impacting stability and result of overwintering,was influenced by breeding adaptive overwintering bees and cutting off offspring through imprisonment of queens.The techniques of breeding adaptive overwintering bees and cutting offspring by imprisonment of queens should be taken in the seasonal management of new feeding methods.2.The eggs of the Chinese honey bee did not hatch,and development duration differed significantly at 32,34 and 36℃（P＜0.05）,sterilized eggs hatched at a non significantly different rate（P＞0.05）,while unsterilized eggs at a significantly different rate（P＜0.05）.The queens and the cell-covered stage drones were influenced by temperature,resulting in significantly different newborn weight and mature time（P＜0.05）.Frequency for hiving discerning flying for queens and drones were 1.23～1.31 and 1.08～1.13,respectively,and flying duration was 0.12～0.13 and 0.16～0.20h,respectively.Frequency for coupling flight was 1.10～1.12 and 1.01～1.05, respectively and flying duration was 0.22～0.23 and 0.18～0.23h,respectively.The most suitable temperature for coupling was 20～28℃.Semen number was 3.37×10⁶～4.15×10⁶ in lateral oviduct in one coupling flight and 3.55×10⁶～3.62×10⁶ in spermathecal sac in natural coupulation.The newborn weight of a queen was positively correlated to oviposition amount,with r=0.715（P＜0.05）.Yearly oviposition of queen was impacted obviously by climate and origin of honey and nectar.The greater the colony potential was and the higher the temperature at the center of swarm was,the bigger the swarm size was,the earlier started oviposition the queens.Honeycomb of the honey bee of larvae of different generations bred by new breeding method was studied.The result indicated that the volume and shape were changed due to accumulated cocoon and feces,which affected breeding offspring.The bees tended to destroy old comb by chewing,causing much trouble to management, while the cumulated wax debris promoted wax moth infestation.The fundamental management strategy was to adapt the preference behavior to a new comb by frequent comb change.In this area,development duration was 14.3 d（332～353h）for queens, 19.1 d（446～468h）for spados,and 24.1 d（556～576h）,and weight of a queen was 167.7 mg for queens,94.4 for spados,and 146.4 mg for drones.The changing seasonal temperature made the development duration of the bee shortened or elongated,which caused extreme significant difference of newborn weight（P＜0.01）. The rate of weight gain was slow in 90 h after hatching,sped up after 90h. Accordingly,the larvae for queens and drones should be kept in stable hive environment in order to raise the quality of breeding queens and drones.Colony temperature and its adjustment were investigated in the honeybees bred by both new and old breeding methods in the montanic area.The result showed that the temperature of the honeybee swarm bred at comb center by new breeding method was 32.57±0.29℃34.0±0.33℃32.19±0.26℃,and 26.14±0.39℃,respectively in spring,summer,autumn,and winter,while the temperature at comb edge was 22.24±1.77℃,33.71±0.64℃19.27±0.54℃and 14.2±0.09℃respectively in the seasons.The temperature of the honeybee swarm bred at comb center by old breeding method was 32.78±0.60℃34.26±0.31℃,32.39±0.26℃and 27.47±0.31℃, respectively,while the temperature at comb edge was 22.49±1.77℃34.08±0.66℃, 19.68±0.52℃,and 14.81±0.07℃,respectively.The temperature at comb center of the swarms bred by both new and old breeding methods was extremely significantly different to environment temperature（P<0.01）,and that at comb edge was significantly different to environment temperature in summer and winter（P<0.05）, but not significantly different in spring and autumn（P>0.05）.In winter,the temperatures at both center and edge of the swarm bred by old breeding methods were higher than those by new method.The center temperature was kept at 30.5～32.6℃through group regulation in swarms bred in a cooler,while kept at 27.8～31.8℃in an incubator.The center temperature of an overwintering swarm changed in accord with environment temperature.As the colony potential declined,the center and edge temperatures changed with more fluctuation,and overwintering stability declined.4.Food resource niche breadths of the Chinese honeybee and the Italian honeybee were 0.923 and 0.765,respectively,and niche overlap index and niche similarity of the 2 bee species were 0.142 and 0.755,respectively.During flowering phase of oil rape,time resource niche breadths were 0.879 and 0.901,respectively, and the index and the similarity were 0.146 and 0.875,respectively;during flowering phase of loquat,the breadth 0.760 and 0.677,respectively,and index and similarity 0.139 and 0.901,respectively.Spatial resource niche breadths of the 2 bee species were 0.797 and 0.670,niche overlap index 0.179,and niche similarity 0.903.There were many niche overlaps between the 2 honeybees,but there were many differences between their respective characteristics,and the niche breadth in local pollination resource for the Chinese honeybee was wider than that for the Italian honeybee.If the former gets extinct,total local pollination will be decreased,resulting in that pollination of many plants is influenced,some species will decrease gradually until extinction,and as a result,plant diversity will decline in the montanic area. Accordingly,the Chinese honeybee is a necessary link of important ecological value in the natural ecosystems in China.Its protection is the protection of honeybee genetic diversity,which plays an important role on vegetation recovery,species diversity preservation,and ecological environment improvement.In montanic area of south Anhui and Baibieshan montanic area of west Anhui, area of Yangtze-Huaihe Rivers,as well as Huaibei Plains,change of habitat and interspecific competition are the main limiting factors for colony distribution.The montanic area of south Anhui and Baibieshan montanic area of west Anhui,with wide distribution area and high distribution density in favorable ecological environment including intact vegetation,abundant honey and nectar plant,queen couple with little interference,are excellent site for the Chinese honeybee,where the its colonies are large,and play a superior role in interspecific competition.Comparatively,in areas of Yangtze-Huaihe Rivers,and Huaibei Plains,where ecosystems have been somewhat ruined,with simple vegetation,monotonous honey and nectar plant,short and concentrated flowering phase,the natural mating of the Chinese honeybees,inferior in the interspesific competition,is interfered by the Italian honeybee,Apis mellifera ligustica,and colonies have decreased sharply,with its distribution shrunk and density declined to a great extent.It happened more seriously in Huaibei Plains than in the area of Yangtze-Huaihe Rivers.5.Eight morphorlogical features of each spado in 15 feral colonies,15 bees each colony,were tested for difference significance test and principal component analysis, discriminant test,and cluster analysis.The results showed that they were morphologically different,with significant differences in lengths of beak length and right fore-wing,breadth and color of the 3^rdand 4^thdorsal plate,which displayed great polymorphism and polytypism and clear ecogeological variation types,suggesting highly ecological adaptation.Seventeen colonies of the Chinese honeybee from montanic areas of southern Anhui,Zhejiang and Jiangxi were tested for mtDNA diversity.Their mtDNA’s amplified were sequenced and compared.Four mt haplotypies were discovered:①A₁A₂A₃ and B₃,②C₁C₂C₃,③B₂,and④the rest.Their mtDNA sequences were not significantly different.The sequencing result of a 440bp long fragment of mtDNA Cytochrome c-oxidase gene（COⅡ）revealed that among the fragment,G+C content varied from 15.1～15.7%,and A+T from 84.3～84.9;there were 8 variation sites totally in the 440bp’s,with 3 T-C transitions,2 A-T transversions,and 4 parsimony sites,but there were 0-6 sites（0～1.36%）in the 440bp’s of each colony.Among the colonies of Huizhou District,Tunxi District,and Huangshan District in the same area,there were 2,3,and 1 variation sites,respectively.However,between some colonies from Qimen County and Sexian County in different areas,there was no variation site in their COⅡbene nucleotide sequences.The genetic similarities were all over 0.967 between the Chinese honeybee colonies from southern Anhui,over 0.971 between those from southern Anhui and Zhejiang,over 0.973 between those from southern Anhui and Jiangxi,suggesting that the honeybee of southern Anhui is closely related to those of Zhejiang and Jiangxi.Based on a cladogram analysis by Cluster V,there were 5 clustered groups:A and B₃;C;B₂;E₁;D,B₁,E₂,E₃ and those from Jiangxi and Zhejiang.更多还原