松毛虫属（Dendrolimus) 部分种类亲缘关系与遗传分化的等位酶分析

采用等位酶聚丙烯酰胺凝胶电泳技术对松毛虫属5个种和亚种的野生种群进行了亲缘关系和遗传变异的研究.8种等位酶系统(乳酸脱氢酶LDH、苹果酸脱氢酶MDH、苹果酸酶ME、乙醇脱氢酶ADH、甲酸脱氢酶FDH、谷氨酸脱氢酶GDH、过氧化物酶POD、过氧化氢酶CAT)共检测到12个基因位点,其中6个位点为多态位点,检测到15个等位基因.松毛虫属5个种和亚种的总体水平多态位点比率P＝50%,平均有效基因数A = 1.917,平均期望杂合度He =0.267,平均遗传距离为0.0730～0.5701.遗传参数表明松毛虫属昆虫种间存在较高程度的遗传变异,聚类图和遗传距离数据表明赤松毛虫与马尾松毛虫亲缘关系最近,落叶松毛虫与思茅松毛虫亲缘关系最远.     

亚洲小车蝗不同地理种群遗传多样性的等位酶分析

采用等位酶电泳技术对亚洲小车蝗8个地理种群遗传多样性和遗传分化进行了研究,并对8种等位酶系统:苹果酸脱氢酶(MDH)、苹果酸酶(ME)、乙醇脱氢酶(ADH)、谷氨酸脱氢酶(GDH)、谷氨酸-草酰乙酸转氨酶(GOT)、异柠檬酸脱氢酶(IDH)、腺苷酸激酶(AK)和己糖激酶(HK)进行聚丙烯酰胺凝胶电泳分析。结果表明:在亚洲小车蝗8个地理种群中共检测到14个基因位点,其中7个位点为多态位点,检测到25个等位基因;种群总体水平多态位点比率P=42.86%,平均有效基因数A=1.786,平均期望杂合度He=0.072,种群平均遗传距离为0.069～0.235。聚类分析表明,遗传距离与地理距离存在一定相关性。亚洲小车蝗8个种群的遗传分化系数Fst=0.086,基因流Nm=3.142,表明亚洲小车蝗种群间有一定程度的遗传分化及基因交流。     

四纹豆象不同地理种群的遗传分化

【目的】通过对四纹豆象Callosobruchus maculatus Fabricius不同地理种群mtDNA-Cytb和COⅠ基因部分序列进行比较, 分析其不同地理种群间的遗传分化情况, 为揭示其与生物入侵的关系及入侵过程中种群系统发育地理格局与演变机制提供依据。【方法】用PCR产物直接测序法对分别来自中国海南、喀麦隆、韩国和泰国的四纹豆象4个地理种群的mtDNA-Cytb和COⅠ序列进行测序, 运用软件MEGA3.1对四纹豆象不同地理种群mtDNA-Cytb和COⅠ序列进行序列分析, 以绿豆象C. chinensis为外群构建了不同单倍型的分子系统树。【结果】34条420 bp Cytb序列中共检测到14个多态位点和5种单倍型, 33条822 bp COⅠ序列中检测到28个多态位点和9种单倍型, 其中4种单倍型为独享单倍型, 其余为全部或部分种群的共享单倍型。AMOVA分析结果显示, 四纹豆象4个地理种群间的遗传结构差异并不明显, 遗传差异主要发生在地理种群内。对4个地理种群进行了Fst值和基因流动统计, 结果表明4个地理种群间既存在着一定数量的基因交流, 也存在一定程度的遗传分化。【结论】根据单倍型分布格局初步推测, 中国不可能是四纹豆象的原产地, 而喀麦隆有可能是原产地之一, 并且喀麦隆种群与泰国种群之间的基因交流比较充分, 而中国种群与其他种群之间的遗传分化相对较大。     

三种松毛虫不同地理种群遗传多样性

运用13个ISSR引物对赤松毛虫(Dendrolimus spectabilis)、油松毛虫(Dendrolimus tabulaeformis)、落叶松毛虫(Dendrolimus superans)的种群遗传分化进行分析.13 个引物共产生195条带,其中184条具多态性,总的多态位点百分率为94.36%,扩增谱带具有明显的种间多态性.Shannon 信息指数和Nei信息指数均表明落叶松毛虫群体内的遗传多样性最高,油松毛虫与赤松毛虫则相差不多.各种松毛虫的不同地理居群出现了遗传分化,由Nei指数估计的群体间的遗传分化系数分别为15.50%、32.57%和41.92%.用UPGMA法对Nei′s遗传距离作聚类分析.聚类结果表明:不同地域的油松毛虫遗传距离与地理距离呈一定程度的相关趋势.     

基于微卫星标记的中国枣食芽象甲地理种群遗传多样性分析

【目的】枣食芽象甲Scythropus yasumatsui是我国北方枣树Zizyphus jujuba上的一种重要的灾害性害虫,在陕西、山西、河北、河南等枣树主产区普遍发生。本研究旨在揭示我国枣食芽象甲不同种群间的遗传分化和基因交流规律。【方法】利用枣食芽象甲转录组测序的SSR序列,使用荧光标记PCR及毛细管电泳分型方法,筛选出8个微卫星位点,对我国5个省份(山西、陕西、宁夏、河北和河南)10个地理种群共308头枣食芽象甲样本进行种群遗传多样性分析。【结果】8个SSR位点均存在无效等位基因且偏离哈迪 温伯格平衡。各位点的有效等位基因数(Ne)为2.113~8.016,多态信息含量(PIC)为0.561~0.908,期望杂合度(He)为0.476~0.865;种群间遗传分化系数(Fst)平均值为0.151;基因流(Nm)平均值为1.594。枣食芽象甲种群间遗传分化系数与地理距离之间显著正相关(r=0.596, P=0.0035),基于Nei’s遗传距离和Cavalli-Sforza & Edwards余弦遗传距离的系统进化树将10个地理种群均聚为3个相同的分支。【结论】结果说明,枣食芽象甲种群遗传多样性较高,不同地理种群间存在高度的遗传分化,且有一定的基因交流;地理隔离是影响枣食芽象甲地理种群遗传分化和基因交流的重要因素。     

蒙古沙葱萤叶甲种群遗传多样性的微卫星分析

【目的】沙葱萤叶甲 Galeruca daurica (Joannis)是一种近年来在内蒙古草原上猖獗成灾的新害虫,本研究旨在明确内蒙古沙葱萤叶甲不同地理种群间的遗传分化和基因交流程度。【方法】应用5对微卫星引物分析了沙葱萤叶甲8个地理种群的遗传多样性、基因流和遗传分化。【结果】5个位点等位基因数为10~18,有效等位基因数为9.2796~16.0388,多态信息含量值为0.6760~0.8985,期望杂合度为0.3430~0.5284,说明所选5个微卫星位点均为高度多态性位点。8个种群的期望杂合度为0.2216~0.3701,平均值为0.2680;种群间遗传分化系数为0.1244~0.4116,平均值为0.2521;种群间基因流为0.3574~1.7596,平均值为0.9622。8个地理种群根据遗传距离聚为3个分支,遗传距离与地理距离呈显著的正相关关系 (r=0.4854,P=0.0180)。【结论】沙葱萤叶甲种群遗传多样性低,不同地理种群之间基因流较小,遗传分化程度高;沙葱萤叶甲迁移能力弱和地理阻碍可能是限制其基因交流和导致遗传高度分化的主要原因。     

内蒙古沙葱萤叶甲种群遗传多样性的微卫星分析

【目的】沙葱萤叶甲Galeruca daurica(Joannis)是一种近年来在内蒙古草原上猖獗成灾的新害虫,本研究旨在明确内蒙古沙葱萤叶甲不同地理种群间的遗传分化和基因交流程度。【方法】应用5对微卫星引物分析了沙葱萤叶甲8个地理种群的遗传多样性、基因流和遗传分化。【结果】5个位点等位基因数为10~18,有效等位基因数为9.2796~16.0388,多态信息含量值为0.6760~0.8985,期望杂合度为0.3430~0.5284,说明所选5个微卫星位点均为高度多态性位点。8个种群的期望杂合度为0.2216~0.3701,平均值为0.2680;种群间遗传分化系数为0.1244~0.4116,平均值为0.2521;种群间基因流为0.3574~1.7596,平均值为0.9622。8个地理种群根据遗传距离聚为3个分支,遗传距离与地理距离呈显著的正相关关系(r=0.4854,P=0.0180)。【结论】沙葱萤叶甲种群遗传多样性低,不同地理种群之间基因流较小,遗传分化程度高;沙葱萤叶甲迁移能力弱和地理阻碍可能是限制其基因交流和导致遗传高度分化的主要原因。     

中国松毛虫属八个种和亚种亲缘关系的DNA指纹证据

利用DNA指纹谱方法探讨了中国松毛虫属8个种和亚种〔马尾松毛虫Dendrolimus punctatus punctatus (Walker),德昌松毛虫D. punctatus tehchangensis Tsai et Liu,文山松毛虫D. punctatus wenshanensis Tsai et Liu,思茅松毛虫D. kikuchii Matsumura,赤松毛虫D. spectabilis Butler,油松毛虫D. tabulaeformis Tsai et Liu,落叶松毛虫D. superans (Butler),云南松毛虫D. houi Lajonquiere之间的亲缘关系。13个随机引物在8种松毛虫中共检测到168个多态分子标记。分析表明,这8个种间的遗传距离的变化范围为0.3780～0.7360;马尾松毛虫与其亚种德昌松毛虫的遗传距离最近,为0.3780,与其另一亚种文山松毛虫以及油松毛虫的遗传距离次之,皆为0.5233;赤松毛虫、落叶松毛虫、云南松毛虫与马尾松毛虫的遗传距离则再次之,分别为0.6362,0.6770和0.6944;与马尾松毛虫遗传距离最远的是思茅松毛虫,为0.7360。8种松毛虫间具体的亲缘关系为: (D. superans (D. tabulaeformis (D. p. wenshanensis (D. p. tehchangensis, D. p. punctatus)(D. Kikuchii (D. spectabilis, D. houi))。     

华东地区中华蜜蜂六地理种群的遗传多样性及遗传分化

目的】利用23对微卫星标记对来自于南昌、黄山、桐庐、费县、宜兴、武夷山6个华东地区的中华蜜蜂Apis cerana cerana种群进行遗传多样性及遗传分化分析。【方法】通过计算多态信息含量、平均杂合度、等位基因数、遗传距离、基因流、F 统计量等参数, 评估各中蜂种群遗传多样性和各种群间遗传分化。【结果】各座位的等位基因数为5（A014）至30（AP043）。所有种群均显示较高水平的期望杂合度, 其中, 武夷山中蜂最低, 为0.4280; 南昌中蜂最高, 为0.6329。各中蜂种群间存在极显著的遗传分化, 平均分化系数(Fst)为0.344。基于Nei氏遗传距离运用NJ聚类法将6个中蜂种群划分为3类。【结论】华东6个中蜂种群的遗传多样性较高, 遗传分化显著; 分析遗传分化与地理距离的关系发现, 华东6个中蜂种群间的遗传分化与地理距离不存在显著相关。     

松毛虫质型多角体病毒的宿主域与交叉感染

自1956年从赤松毛虫Dendrolimus spectabilis上首次发现赤松毛虫质型多角体病毒1型（D. spectabilis cytovirus 1,DsCPV-1）以来,先后从马尾松毛虫D. punctatus、油松毛虫D. tabulaeformis、赤松毛虫、德昌松毛虫D. p. tehchangensis、文山松毛虫D. p. Wenshangensis和落叶松毛虫D. superans上发现了质型多角体病毒(cytoplasmic polyhedrosis virus,CPV)。病毒基因组dsRNA电泳图谱分析表明,这些松毛虫CPV的不同分离株均属于质型多角体病毒1型（cytovirus 1）。这些松毛虫CPV病毒可以感染鳞翅目10科35种昆虫,其中对多种昆虫具有很高的感染力和良好的杀虫效果,可以从中筛选替代宿主生产松毛虫CPV杀虫剂,用于害虫生物防治。松毛虫CPV接种某些昆虫后病毒的基因组dsRNA电泳图谱发生了改变,可能是异源病毒诱发了宿主自身潜伏型病毒的感染复制。     

基于Cyt b基因序列分析的松毛虫种群遗传结构研究

为了揭示松毛虫种群的遗传结构,采用DNA序列测定的方法测定了松毛虫不同种群的线粒体细胞色素b (Cyt b)基因的部分序列,并利用分子生物学软件分析其核苷酸组成、转换和颠换、氨基酸组成、遗传距离及亲缘关系。结果显示:在获得的Cyt b 基因387bp的序列中碱基A,T,C,G平均含量分别为40.1%、33.5%、9.5%、16.9%,A+T含量明显高于G+C含量表现出强烈的A、T偏向性,密码子第3位点的A+T含量高达86.5%,这种偏向性在种群间无明显差异。碱基替换主要发生在密码子第三位,转换大于颠换,且种群内替换高于种群间。该序列片段中共有39个核甘酸位点发生变异,遗传距离为0.000-0.100,显示出较小的遗传变异。蛋白质氨基酸由除谷氨酸以外的19种氨基酸组成。聚类分析结果表明马尾松毛虫和油松毛虫亚种遗传距离较近,种群间的遗传分化与生态环境有关。     

松毛虫雌性信息素的研究与应用

目前,在我国对松林危害最严重的害虫主要有马尾松毛虫Dendrolimnspunctatus(Walker)、落叶松毛虫D.superaus(Butler)、油松毛虫D.tabulaeformisTsaietLiu、赤松毛虫D.spectabilis(Butler)、思茅松毛虫D.kikuchiiMatsumura、云南松毛虫D.houiLajonqui埁re等。综述6种松毛虫的雌性信息素的成分鉴定、合成、EAG反应,人工合成雌性信息素的田间活性测定,性诱剂在林间的设置技术及种群监测等应用。     

Determination of Genetic Diversity of Vitis vinifera cv. Kabarcik Populations from the Coruh Valley Using SSR Markers

Northeastern Turkey is recognized as one of the most important germplasm centers for the grape in the world. In the present study, simple sequence repeat markers were used to investigate the genetic diversity between four Vitis vinifera cv. Kabarcik populations sampled from the Coruh Valley in Turkey, at altitudes of 800-1,150 m. The mean observed number of alleles per locus varied from 2 (loci VVMD7 and VVMD24) to 6 (VVS2) among populations. The population from the highest altitude showed the greatest average number of alleles, 4.5. With regard to the six loci examined in all populations, the mean observed heterozygosity was higher than the expected heterozygosity. Among the loci, VVS2 (probability of identity = 0.137) was found to be the most informative among populations. Genetic distances between populations ranged from 0.072 to 0.216. Genetic differentiation among populations was strongly related to geographic distances in all populations.     

Genetic structure of Pinus pinaster Ait. populations in Morocco revealed by nuclear microsatellites

Pinus pinaster is one of the most popular conifers used for reforestation in Morocco and represents an economically and ecologically important species for the region. In this study, nuclear microsatellites (ncSSRs) are used to compare genetic structure and diversity estimates of natural populations of Moroccan maritime pine. Samples were collected among 10 natural populations distributed in three biogeographically different regions, the Rif Mountain, the Middle and the High Atlas. Forty-five nuclear alleles at seven variable loci were found with a mean of 6.4 alleles per locus. A number of private alleles (17.1%) were shown in populations from Rif and Middle Atlas. Moreover, in Morocco, P. pinaster showed a lower genetic diversity than in other parts of its geographic range. Significant departures from Hardy–Weinberg equilibrium with excess homozygosity are observed indicating a high level of mating inside populations. Genetic diversity was structured with high variability among populations (Fst = 12%). Results show a correlation between genetic and geographic distances with an R-squared of 0.436. Two clusters were found using STRUCTURE, whereas three main clusters can be distinguished based on genetic distances of phylogenetic tree. Genetic relationships among maritime pine populations in Morocco appear to be related to historical, ecological as well as anthropogenic factors, suggesting the need for conservation strategies at the population level.     

基于微卫星的中国红火蚁种群遗传结构的研究

[背景]自入侵中国之后,红火蚁已给农林业、健康卫生、生态环境等造成了危害。红火蚁在中国的入侵、扩散路径及方式等仍然是待解决的问题。[方法]利用微卫星分子标记,对来自国内14个地区和国外1个地区共15个红火蚁地理种群的遗传多样性水平及种群遗传结构进行了研究。[结果]应用7对微卫星引物共检测到28个等位基因,15个红火蚁种群在各微卫星位点的基因型频率均符合Hardy-Weinberg平衡。各种群的平均表观杂合度HO、预期杂合度HE、Shannon信息指数Ⅰ、基因多样性指数Nei＇s和多态位点百分率P分别为0.2848、0.2708、0.3174、0.2629和43.63%,研究结果表明这15个红火蚁种群具有比较丰富的遗传多样性。种群间平均分化系数FST为0.4258,说明有42.58%的变异来源于种群间,表明红火蚁各种群之间有较高程度的分化,且遗传分化可能是由地理隔离和基因流障碍（Nem=0.7442）共同引起。遗传距离D显示,河源种群与其他种群间的遗传距离均相对高于其他各种群间的遗传距离,表明河源种群与其他地理种群之间存在较大的遗传差异,可能是较为原始的类型。[结论与意义]短距离的种群主要通过自然扩散方式传播,地理距离与亲缘关系有一定的相关性;长距离的种群主要依靠人为传播,因此地理距离与遗传距离不成正比。对于长距离的入侵事件,监控与检疫是关键的预防措施。     

Phylogenetic reconstruction and DNA barcoding for closely related pine moth species (Dendrolimus) in China with multiple gene markers

Unlike distinct species, closely related species offer a great challenge for phylogeny reconstruction and species identification with DNA barcoding due to their often overlapping genetic variation. We tested a sibling species group of pine moth pests in China with a standard cytochrome c oxidase subunit I (COI) gene and two alternative internal transcribed spacer (ITS) genes (ITS1 and ITS2). Five different phylogenetic/DNA barcoding analysis methods (Maximum likelihood (ML)/Neighbor-joining (NJ), "best close match" (BCM), Minimum distance (MD), and BP-based method (BP)), representing commonly used methodology (tree-based and non-tree based) in the field, were applied to both single-gene and multiple-gene analyses. Our results demonstrated clear reciprocal species monophyly for three relatively distant related species, Dendrolimus superans, D. houi, D. kikuchii, as recovered by both single and multiple genes while the phylogenetic relationship of three closely related species, D. punctatus, D. tabulaeformis, D. spectabilis, could not be resolved with the traditional tree-building methods. Additionally, we find the standard COI barcode outperforms two nuclear ITS genes, whatever the methods used. On average, the COI barcode achieved a success rate of 94.10-97.40%, while ITS1 and ITS2 obtained a success rate of 64.70-81.60%, indicating ITS genes are less suitable for species identification in this case. We propose the use of an overall success rate of species identification that takes both sequencing success and assignation success into account, since species identification success rates with multiple-gene barcoding system were generally overestimated, especially by tree-based methods, where only successfully sequenced DNA sequences were used to construct a phylogenetic tree. Non-tree based methods, such as MD, BCM, and BP approaches, presented advantages over tree-based methods by reporting the overall success rates with statistical significance. In addition, our results indicate that the most closely related species D. punctatus, D. tabulaeformis, and D. spectabilis, may be still in the process of incomplete lineage sorting, with occasional hybridizations occurring among them.     

Host range of Dendrolimus kikuchii nucleopolyhedrovirus

Dendrolimus kikuchii Matsumura nucleopolyhedrovirus (DekiNPV) has been considered as a biological control agent against D. kikuchii. We examined the infectivity and replication of DekiNPV in 11 other lepidopteran species, including Dendrolimus houi (Lajonquiere), Dendrolimus punctatus (Walker), D. punctatus wenshanensis (Tsai et Liu), Dendrolimus spectabilis (Butler), Hyphantria cunea (Drury), Lymantria dispar (L.), Ectropis grisescens (Warren), Helicoverpa armigera (Hübner), Spodoptera exigua (Hübner), Plutella xylostella (L.) and Spodoptera litura (Fabricius) larvae to define the host range of DekiNPV and identify suitable alternate hosts. Our study showed that DekiNPV is highly specific to D. kikuchii, but triggers covert infections in H. cunea, D. houi, D. punctatus, D. punctatus wenshanensis and D. spectabilis larvae into overt symptoms, indicating that DekiNPV is suitable as an ideal biocontrol agent for D. kikuchii, D. houi, D. punctatus, D. punctatus wenshanensis, D. spectabilis and H. cunea.     

马尾松毛虫及其寄主松属植物的地史初步考证

初步考证了我国重要的森林害虫———马尾松毛虫DendrolimuspunctatusWalker及其寄主松 (科 )属植物的化石起源 ,认为马尾松毛虫衍生为一个新的物种的时间 ,不应早于中生代的侏罗纪 ,很可能是侏罗纪与白垩纪早期的某个时期 ;并初步探讨了第三纪以来中国古地质、古气候变化对现今马尾松毛虫的分布、遗传多样性的空间格局及其历史成因的可能影响 ,文史还涉及许多重要的生物地质历史事件 ,对于理解马尾松毛虫在同样的地质时期的动态等具有重要的参考意义