Cross-specific amplification of microsatellite DNA markers in Shorea platyclados

In the megadiverse forests of Southeast Asia,hundreds of timber species are economically important but the population genetics of only a few taxa are known.Cross-specific amplification of microsatellite loci among closely related taxa could enhance our ability to study and manage previously unstudied species. We successfully utilized STMS markers in Shorea platyclados, originally developed for Shorea curtisii. The six primer pairs we tried successfully produced PCR products of expected sizes. The number of alleles observed ranged from 10 to 14 and an average of 12 alleles were detected per locus. A high expected and observed heterozygosity was observed and it ranges from 0.718 to 0.827 among all populations across all six loci tested. Microsatellite DNA markers are highly polymorphic, co-dominant, reproducible, and amenable to high throughput genetic analyses. Overall, the crossspecific amplification of microsatellite loci appears to be complicated by numerous factors. While the approach may be effective for local management and conservation ofpoorly known species, the results must be carefully interpreted.     

Development of two microsatellite multiplex PCR systems for high throughput genotyping in Populus euphratica

Eighteen microsatellite primer pairs previously developed at Oak Ridge National Laboratory for Populus tremuloides Michx. and Populus trichocarpa Torr. & Gray were screened for amplification in Euphrates poplar, Populus euphratica Oliv. Thirteen loci were found to express polymorphisms ranging from two to 17 alleles. The eight most variable loci were selected to set up and optimize two multiplex polymerase chain reaction (PCR) assays. Three populations containing altogether 436 trees were used to characterize the selected loci and ascertain their applicability for parentage analysis and genotyping studies. Through cross-checking of clonal identity against sex of the genotyped trees we estimated the maximum error rate for merging genotypes to be less than 0.045. Foundation project: This study was supported by the Deutsche Forschungsgemeinschaft DFG (grant number SCHN 1080/1-1)     

笼养东北虎(Panthera tigiris altacia)的行为观察(英文)

从1998年3月至2001年3月,采用全事件取样法和焦点动物取样法对哈尔滨动物园的4只笼养东北虎(1雄3雌)进行了系统的行为观察。笼养东北虎的行为可以分成5种主要的方式:运动、卧息、睡眠、取食和其他行为(饮水、排尿、修饰、嬉戏和站立)。在所有的行为类型中,睡眠和运动所占的比值很高(占整个时间分配的75.18%);而卧息、取食和其他行为仅占24.82%。除了取食,一只雄性和其中两种雌性东北虎的其他4种主要行为具有相似的活动规律,在白昼和夜晚各有一个活动高峰;而另外一只雌性东北虎在夜晚则具有两个活动高峰期。表2参16。     

鹿类微卫星遗传标记的研究现状(英文)

微卫星DNA在染色体上随机即分布，是十分有效的遗传标记。目前，在鹿类发现约有200 多个微卫星位点，除了极少数位点是直接从DNA文库中筛选的以外，绝大多数都是将牛、羊 的微卫星位点“借用”到鹿类。这些位点被广泛应用到父权鉴定、遗传多样性和种群结构分 析、种群基因渗入的检测、孕期长短及越冬存活率等的遗传标记等方面的研究中。然而，微 卫星DNA的数量目前还远不能满足研究需要。将来要在以下方面开展工作：1）分离更多的鹿 类特异的微卫星位点，2）构建微卫星位点的遗传和物理图谱。这样，微卫星DNA才能在鹿类 研究中发挥更大的作用。     

Direct genotyping of the poplar leaf rust fungus,Melampsora medusae f. sp. deltoidae,using codominant PCR‐SSCP markers

Two anonymous DNA markers that are revealed by single‐strand conformational polymorphism (SSCP) analysis were developed for detection of polymorphisms in Melampsora medusae f. sp. deltoidae (Mmd). Mono‐uredinial isolates of Mmd were first obtained, DNA was extracted from urediniospores and random amplified polymorphic DNA (RAPD) products of eight mono‐uredinial isolates were separated on a SSCP gel to identify differences among them. Bands representing putative polymorphic loci among the eight isolates tested were excised from the SSCP gel and re‐amplified by polymerase chain reaction (PCR), and then cloned and sequenced. A primer pair was designed to amplify a DNA fragment of a size suitable for SSCP analysis (<600 bp) for two out of three DNA fragments sequenced. Each set of primers amplified a PCR product for all eight isolates that were initially used to generate them and the resulting PCR products were analysed by SSCP. Polymorphisms among isolates were identified for both putative loci. The two primer pairs amplified a PCR product of the expected size on an additional 32 mono‐uredinial isolates of Mmd tested. From the overall 40 mono‐uredinial isolates tested, 5 and 11 alleles were detected, and 12 and 34 isolates showed to be heterozygous, as indicated by the presence of more than two bands on the SSCP gel, at loci A and B, respectively. The primer pairs were tested for specificity against 106 fungal isolates belonging to various taxa, including other rusts, and against DNA extracted from greenhouse‐grown healthy poplar leaves. DNA amplification products of the expected size were obtained only when Mmd DNA was present. Optimization of PCR conditions with these two primer pairs allowed genotyping directly from single uredinia extracted from infected leaves, thus alleviating the need to culture the fungus to characterize individuals, hence making it possible to process large numbers of samples for population studies.     

18份樱属材料亲缘关系的ISSR分析

对福建山樱花、11份日本樱花品种和6份选优材料进行了标记,通过筛选出来的12条引物进行聚合酶链式反应(PCR)扩增。结果显示,共获得144个位点,多态性位点135个,多态性占93.75%。选取引物UBC808和UBC835构建指纹图谱,简单重复序列间扩增(ISSR)分子标记能有效地对18份樱属材料进行区分和鉴定。供试材料间的遗传相似系数在0.4532~0.8333之间,平均0.6337;遗传距离在0.1823~0.7651之间。通过非加权组平均法(UPGMA)进行聚类分析,在遗传相似性系数为0.5899时,可将18份樱属材料划分为4大类,其中选优材料与福建山樱花或日本樱花亲缘关系较近,但在花色、花期等方面有一定差异,且能区分和鉴定供试材料的亲缘关系。研究结果能为国内樱花新品种鉴定、优选和商品化等提供参考依据。     

半野化条件下华南虎繁殖行为研究

2010年1月—2011年12月期间，对韶关市华南虎(Panthera tigris amoyensis)繁育基地谱系号为387号（雌虎）和373号（雄虎）的两只华南虎采取视频监控和现场观察方法，进行繁殖行为研究。研究发现，华南虎在半野化模式饲养条件下，生长发育较快，雌雄华南虎分别在2.3岁和2.8岁开始发情；雌虎除怀孕期和哺乳期外，平均年发情4.5次，雄虎平均年发情10次；雌雄华南虎一年四季均有发情，每年7月和11月为发情高峰期。雌虎在2010年10月21日—2011年10月2日期间，共产崽3次，产前3 d 预产行为明显，怀孕期为100~102 d。     

香果树RAPD扩增条件的优化及遗传多样性初步分析

对浙江省天台山自然香果树种群的DNA进行了提取和RAPD扩增条件的优化,分别测试了镁离子、dNTP、模板DNA含量、引物浓度、DNA聚合酶量和牛血清白蛋白浓度对反应结果的影响,通过各因子的组合研究,可知香果树RAPD分析较适宜的扩增条件为:15μLPCR反应体积,1×Taq酶配套缓冲液,1 8mmol·L-1MgCl2,2UTaq酶;10ng模板DNA;20pmol引物;dATP、dCTP、dGTP、dTTP各0 1mmol·L-1。以此适宜条件对12个引物进行扩增,共扩增出51个DNA片段,多态位点百分率为45 1%,种群内遗传多样性较低,进一步研究不同种群香果树的遗传多样性具有一定的意义。     

Jackknife resampling for precision measurement of direct gene flow estimates

Gene flow from outside pollen sources reduces the expected genetic gain of seed orchards, and has been quantified by genotyping seed samples from crop years to determine the proportion of seed sired by outside sources. Random bulk seed sampling and microsatellite genotyping allows for fast, unbiased identification of orchard contamination rates. Bulk seed sampling has been employed in multiple studies of seed orchard gene flow, but a method of determining precision of gene flow estimates from this sampling method has yet to be described. In this study, we used jackknife resampling to generate random samples of migrant and within-population genotypes obtained from an empirical sample of Douglas-fir (Pseudotsuga menziesii [Mirb.] Franco) genotypes of five microsatellite loci. We generated samples of 50, 100, 150, 200, and 250 genotyped individuals from a sample of 400 seeds, with genotypes corresponding to low (6%), moderate (14%), or high (56%) pollen contamination levels. The mean gene flow estimate and confidence interval were calculated for each sample size and gene flow scenario to determine the precision of gene flow estimates obtained from each sample size in each contaminant gene flow scenario. Genotyping 150 individuals was sufficient to produce gene flow estimates with calculated confidence interval values below ± 1%, indicating that 150 genotyped individuals were sufficient for precise gene flow estimation in this population. This resampling method will enable precise and unbiased estimation of gene flow into seed orchards for future monitoring of gene flow, to allow better management of seed orchard genetic resources.     

笼养东北虎（Panthera tigiris altacia）的行为观察

从1998年3月至2001年3月，采用全事件取样法和焦点动物取样法对哈尔滨动物园的4只笼养东北虎（1雄3雌）进行了系统的行为观察。笼养东北虎的行为可以分成5种主要的方式：运动、卧息、睡眠、取食和其他行为（饮水、排尿、修饰、嬉戏和站立）。在所有的行为类型中，睡眠和运动所占的比值很高（占整个时间分配的75.18%）；而卧息、取食和其他行为仅占24.82%。除了取食，一只雄性和其中两种雌性东北虎的其他4种主要行为具有相似的活动规律，在白昼和夜晚各有一个活动高峰；而另外一只雌性东北虎在夜晚则具有两个活动高峰期。     

Using computer simulations to assess sampling effects on spatial genetic structure in forest tree species

Quantifying spatial genetic structure is key to inform forest management and restoration strategies. Reliable evaluations of genetic structure require sound sampling schemes because inappropriate sampling may over- and under-estimate spatial patterns of genetic structure. Sampling bias has been investigated through computer simulations mostly for animal species with continuous distributions. For tree species that have different life history traits, results from such studies may not apply. Here, I used spatially explicit landscape genetic simulations to assess the effects of spatial sampling scheme (random, systematic, and cluster), sampling intensity (35, 50, 65, and 80%), and the number of microsatellite loci (8, 14, and 20) on inferences of genetic structure under isolation by distance (IBD) in two forest tree species with varying dispersal distances and patchy distributions. Results showed that random sampling with 20 loci was the best performing sampling scheme, irrespective of sampling intensity and the strength of IBD. In contrast, the cluster and systematic sampling were sensitive to sample size. For the three sampling schemes, the number of loci had a large effect because with 8 loci there was an increasing chance of underestimating IBD. Increasing the number of samples over the number of loci, did not improve the performance of sampling schemes. Hence, researchers should put more effort on increasing the number of loci over increasing sample size. Results also showed that sampling error rates varied between species, and sampling bias appeared stronger for the species with a more aggregated spatial distribution.     

两种微卫星多样PCR系统应用于胡杨高效生产基因型

对已在橡树岭国家实验室建立起来的美洲山杨(Populus tremuloides Michx.)和毛果杨(Populus trichocarpa Torr.＆ Gray)18个微卫星引物进行筛选,用于胡杨相关基因的扩增.发现有13个基因位点可以表达基因多态性,范围是2～17个等位基因.选择了8个最易变的基因位点,来构建和优化2种多样PCR法.用来自3个种群的436株树木表征选择的位点,确定他们在亲缘关系分析、基因型研究的适用性.通过克隆基因型鉴定树木性别的交叉检验,我们估计并入基因型最大误差率小于O.045.     

浅谈野生虎监测

从野生虎监测的必要性出发,论述了野生虎监测目标;并根据我国野生虎的分布现状,以东北虎为例,提出了适宜于我国东北虎的监测尺度及在不同监测尺度下的监测要求和监测内容.     

美国山核桃群体遗传多样性的RAPD分析

运用随机扩增多态性DNA（RAPD）标记技术，采用Operon公司的10种随机引物，以采自美国东南部、北部、西部和我国江苏、浙江、湖北、湖南、云南等地的9个美国山核桃栽培群体45个单株的种子为材料，检测出多态位点42个。以这些多态位点为依据，计算了估测了多态性位点百分比、Shannon表型多样度指数、Virk群体内遗传距离和Nei氏群体间遗传距离，并用它们作为参数进行群体遗传多样性的RAPD分析。结果表明，参试的美国山核桃品种群体遗传变异明显，且群体间差异大于群体内差异，我国引种的群体遗传差异大于原产地美国的群体遗传差异。     

绥阳林区野生东北虎现状调查分析

对绥阳两个林场东北虎栖息地的生境概况、现状及保护措施进行了论述、分析和评价。     

枫香优良观赏种质的遗传多样性分析

利用ISSR标记技术对12份枫香观赏种质进行遗传多样性分析,通过正交实验优化枫香的ISSR-PCR扩增体系,即在20μL体系中加入PCR反应缓冲液2μL、镁离子1.5 mmol.L-1、Taq DNA聚合酶0.75 U、dNTP 0.15 mmol.L-1、引物0.4mmol.L-1、DNA模板80 ng。筛选出的19条ISSR引物共得到131条扩增带,其中多态性条带为94条,多态性条带比率为71.75%。12份种质的有效等位基因数是1.482,基因多样性0.3012,Shannon信息指数0.4674,表明遗传多样性水平相对较高。进一步的聚类将这些种质分成3类,其中4号样本和11号样本的遗传相似度最高。     

紫胶虫主要生产种的RAPD分子标记分析

用RAPD技术研究了紫胶虫7种主要生产种12个群体之间的亲缘关系,研究中每个群体使用了24个标本,共有288个个体用于研究.在试用的45种随机引物中,筛选出9种引物用于随机引物扩增,共得到121个清晰稳定的位点,其中118个为多态性位点,多态百分率为97.52%,每条引物检测到的位点数为11～16个,长度为150～3 000 bp.根据扩增结果,用UPGMA法对Nei's遗传距离进行聚类分析,构建分子系统树.7种紫胶虫聚类结果显示:紫胶虫各群体间的遗传距离为0.044 3～0.791 7,其中,种间的遗传平均距离为0.443 0,种内的遗传平均距离为0.170 7.根据聚类分析,讨论了紫胶虫主要生产种的亲缘关系.     

圈养白化孟加拉虎繁殖期行为时间分配的研究

于2012年10月至2013年9月在云南野生动物园，采用瞬时扫描法和全事件观察法对繁殖期白化孟加拉虎的行为进行了研究。结果表明：在发情交配期雌雄白化孟加拉虎的行为时间分配差异不显著（P〉0．05），均将较多的时间用于休息和运动；在妊娠期雌性白化孟加拉虎将较多的时间用于休息和运动，较少的时间用于摄食、睡眠和其它行为，对比妊娠期4只雌虎的行为，不同个体在同一种行为间差异性极显著（P〈0．01）；产仔期雌虎将更多的时间用于产仔和舔舐幼虎，其次是休息；在哺乳期雌虎将较多的时间用于休息，其次是哺乳和运动。通过单因素方差分析表明，在繁殖期不同阶段休息行为差异性极显著（P〈0．01），睡眠、运动和其它行为差异性不显著（P〉0．05）。     

Genetic diversity and differentiation of fall webworm (Hyphantria cunea Drury) populations

The molecular variation and genetic relationships among five populations of the fall webworm (Hyphantria cunea Drury) in China were assessed using microsatellite technology. Ten microsatellite primers, producing polymorphic bands, were used across 300 samples. The percentage of polymorphic loci (PPB) was 98.36%; the percentage of polymorphic loci in five populations ranged from high to low in the following order: Cangzhou population, Yantai population, Qinhuangdao population, Dandong population, and Shijiaz...     

Historical Change on Distribution and Quantity of the South-China Tiger (Panthera tigris amoyensis)

DistributionandQuantityofwildSouth-ChinatigerDistributionTheSouth-ChinatigersonlydistributeinChinaandtheskullstruCtureappeartobethemostsimilarwiththoseoftheirancestors.Asoneoffivesurvivedtigersubspecies,thistigersubspecieshasthelestquantityandisthemostendangeredintheworld.Therefore,the"InternationalTigerConservationStrategy"ScientificWorkshop,whichheldinMinnesotaoftheUnitedStatesofAmericaintheearlyof1986,pointedoutthattheSouth--Chinatigerwasoneofthemostendangeredanimalspecieswhichneede…