黑龙江省完达山东部林区东北虎猎物生物量

研究一个地区猎物种群生物量能否满足捕食动物种群数量的需求,这对于了解濒危大型食肉动物是否受到来自于食物缺乏的威胁和制定相应的保护措施极其重要。为了掌握黑龙江省完达山东部林区东北虎食物需求与猎物生物量之间的关系,于2008年冬季至2009早春积雪覆盖期采用随机布设样线,通过收集有蹄类动物在雪地上留下的足迹等活动的方法,在东方红林业局和迎春林业局管辖境内3 692.06 km²的区域布设大样方48个,并在大样方里共布设样线240条开展有蹄类动物种群数量调查,确定东北虎猎物生物量。调查结果表明:研究地区野猪(成体502 606只,亚成体209 210只)、马鹿(成体331 357只,亚成体67 72只)和狍子(成体810 815只,亚成体202 203只)的生物量分别为74 767.50 87 825.00 kg、79 744.50 85 984.50 kg 和 31 337.00 31 525.50 kg,3种有蹄类动物生物量共计1 85 849.00 205 335.00 kg。研究地区猎物总生物量为209 619.89 231 598.24 kg。如果按8%的生物提供给东北虎,3种主要猎物生物量可满足5.22 6.92只东北虎个体的食物需求,研究地区猎物总生物量则可满足5.89 7.81只东北虎个体的食物需求。此外,对足迹遇见率与抽样强度、抽样强度与足迹遇见率的均值标准误差之间关系的分析表明,在完达山东部林区布设120条样线(抽样距离600 km)、150条样线(抽样距离750 km)和115条样线(抽样距离675 km)能满足野猪、马鹿、狍子种群数量调查准确性的最低需求。     

Comparative landscape genetic analyses show a Belgian motorway to be a gene flow barrier for red deer (Cervus elaphus), but not wild boars (Sus scrofa)

While motorways are often assumed to influence the movement behaviour of large mammals, there are surprisingly few studies that show an influence of these linear structures on the genetic make-up of wild ungulate populations. Here, we analyse the spatial genetic structure of red deer (Cervus elaphus) and wild boars (Sus scrofa) along a stretch of motorway in the Walloon part of Belgium. Altogether, 876 red deer were genotyped at 13 microsatellite loci, and 325 wild boars at 14 loci. In the case of the red deer, different genetic clustering tools identified two genetic subpopulations whose borders matched the motorway well. Conversely, no genetic structure was identified in the case of the wild boar. Analysis of isolation-by-distance patterns of pairs of individuals on the same side and on different sides of the motorway also suggested that the road was a barrier to red deer, but not to wild boar movement. While telemetry studies seem to confirm that red deer are more affected by motorways than wild boar, the red deer sample size was also much larger than that of the wild boars. We therefore repeated the analysis of genetic structure in the red deer with randomly sub-sampled data sets of decreasing size. The power to detect the genetic structure using clustering methods decreased with decreasing sample size.     

山西省及邻近地区中华稻蝗5个种群RAPD分析及其亲缘关系

采用10条10bp随机引物对来自山西省临猗县(山西南部)、屯留县、太原市近郊(山西中部)、代县(山西北部)、内蒙呼和浩特市郊5个不同种群的中华稻蝗(Oxya chinensis)和广西南宁日本稻蝗(Oxya japonica)共64个个体进行了RAPD分析,探讨了中华稻蝗5个不同种群的亲缘关系。结果表明：10条随机引物对供试的64个个体共产生115条谱带。所获片段的分子量大小为200～2500bp,单个引物扩增的谱带数是5～15条,平均为12条。根据扩增出的RAPD图谱,运用SPSS软件包中的Between-groups linkage法进行聚类分析,聚类结果显示,中华稻蝗不同种群的个体优先聚为1类;供试中华稻蝗5个种群的系统树分为两大分支,山西临猗种群、屯留种群亲缘关系最近,首先相聚,然后与太原种群聚为一支,山西代县和内蒙呼和浩特2种群聚为另一支,两支相聚后与日本稻蝗相聚。表明供试中华稻蝗5个种群存在一定程度的差异,其亲缘关系的远近与地理距离之间存在一定的正相关关系。     

Wildlife genetics and disease: allozyme evolution in the wild boar (Sus scrofa) caused by a swine fever epidemy

Enzyme polymorphism at 42 loci was compared before and after a major epidemy of swine fever in wild boars from northern Vosges (France). No change was observed in the 38 monomorphic loci, but allele frequencies at the phosphoglucomutase locus PGM-2* changed significantly. Possible causes for this observation are discussed, and it appears that PGM-2 locus could be a genetic marker of resistance to this viral disease.     

Limitations of the RAPD technique in phylogeny reconstruction in Drosophila

In this study the limitations of the RAPD technique for phylogenetic analysis of very closely related and less related species of Drosophila are examined. In addition, assumptions of positional homology of amplified fragments in different species are examined by cross-hybridization of RAPD fragments. It is demonstrated that in Drosophila the use of RAPD markers is very efficient in identification of species. For assessment of phylogenetic relationships, however, the method is limited to sibling species, and reliable measures for genetic distances cannot be obtained. Hybridization experiments demonstrate that fragments of similar length amplified from different species are not always derived from corresponding loci, and that not all RAPD fragments within the same amplification pattern are independent.     

Use of RAPD analysis in devising conservation strategies for the rare and endangered Grevillea scapigera (Proteaceae)

Extensive human impact in south western Australia has resulted in a high incidence of rarity throughout the highly endemic flora of the region. Grevillea scapigera (Proteaceae) is a typical example, with 27 plants (represented by four extant populations) remaining in the wild. In order to devise an appropriate strategy for the conservation of this species, its population genetics were studied using RAPD analysis, which enabled the discrimination of individual plants and the detection of a relatively high amount of variability (V = 0.32) within G. scapigera. This variability was found to be evenly distributed within the plants analysed despite the clear distinction between most populations (87% of the variability being attributable to single plant difference and 13% to population difference). Finally, RAPD analysis was used to select a small group of plants that captured maximum genetic variability to be used in the recovery program of the species. Because of the low genetic difference between populations, the mixing of these selected plants during the recovery process should not create genetic imbalances. The methods used in this study provide a useful model for future projects involving the recovery of rare flora.     

Analysis of genetic variability of South American wild rice populations (Oryza glumaepatula) with isozymes and RAPD markers

The knowledge of population structure and genetic diversity of wild relatives of rice is needed to investigate their evolutionary history and potential use in breeding programs. Very little is known about the wild rice species ( Oryza spp.), particularly those that are native to South America. A study using isozyme and RAPD markers was conducted to estimate the level of genetic diversity of four South American wild rice populations ( Oryza glumaepatula ) recently collected in the Amazon forest and western Brazil rivers. F -statistics and genetic diversity parameters calculated from isozyme and RAPD markers indicated high values for inbreeding coefficients and differentiation among the four populations. In agreement with this, a pattern of greater variation between than within populations was observed with both types of markers. These findings were corroborated by an AMOVA analysis, which indicated that a large portion of the total genetic variation was attributed to regional divergence. The partition of the AMOVA analysis among populations showed that most of the genetic diversity was due to differences among populations. This distribution pattern of genetic variation of O. glumaepatula populations is in agreement with the expectation for an autogamous species and provides important baseline data for conservation and collection strategies for this species.     

水稻RAPD反应体系的正交优化

以焦旱1号总DNA为材料,首先对影响RAPD-PCR反应的模板DNA、Mg~(2+)、dNTP、引物和Taq DNA聚合酶浓度等因素进行了初步优化,分析了各因素对RAPD-PCR扩增结果的影响.在此基础上对影响RAPD-PCR反应的Mg~(2+)、dNTP、引物和Taq DNA聚合酶浓度等4个主要因素进行正交优化,研究结果表明:在25 μl RAPD-PCR反应体系中,模板DNA 20 ng;Mg~(2+)浓度1.5mmol/L;dNTP的浓度0.2mmol/L;引物量15 pmol;Taq DNA聚合酶1.0 U.在此最佳条件下,利用引物B8对18个北方粳稻品种进行了成功的扩增.     

荔枝DNA提取及RAPD扩增条件优化

为应用RAPD技术开展对荔枝种质资源的分析,以S43(GTCGCCGTCA)为引物,通过试验设计,分别研究了退火温度、模板浓度、引物浓度、dNTP浓度、Taq DNA聚合酶用量对荔枝RAPD-PCR反应的影响。建立并优化了适宜荔枝RAPD分析的扩增体系:20μL的反应体系,30ng的模板DNA度,0.25μmol/LRAPD引物、1.0UTaqDNA聚合酶,0.2μmol/LdNTP为荔枝适宜的RAPD-PCR扩增条件。     

Relationship between population genetic structure and riparian habitat as revealed by RAPD analysis of the rheophyte Acorus gramineus Soland. (Araceae) in Taiwan

Acorus gramineus Soland. (Araceae) is a rheophyte and is distributed in southeastern Asia. Its populations are restricted to riparian habitats. The discontinuous distribution might result in high genetic diversification among plants of different river systems. In the present study, leaf samples were collected from populations along six river systems in western Taiwan and the genetic variation was investigated by employing RAPD markers. A total of 255 samples from 17 sampling sites was studied. Eighty random 10-mer primers were screened and six of them, which showed better amplification results, were selected to analyse all of the samples. Data of 34 high-intensity and highly reproducible polymorphic fragments were used in statistical analyses. The results of AMOVA analyses indicated that, of the total variation, 46.84% was attributable to differences among river systems, 16.88% to differences among sampling sites within river systems, and 36.28% to differences among individuals within sampling sites. The results of cluster analysis and principal coordinate analysis revealed that sampling sites of each river system formed distinct clusters and the sampling sites of six river systems were clustered into three main groups according to latitudinal relationships. The results of the present study indicated that the population genetic structure of the plants of different river systems is highly diversified, which seems to imply that the gene flow among them is very limited.     

Impact of scoring error and reproducibility RAPD data on RAPD based estimates of genetic distance

RAPD band reproducibility and scoring error were evaluated for RAPDs generated by 50 RAPD primers among ten snap bean (Phaseolus vulgaris L.) genotypes. Genetic distances based on different sets of RAPD bands were compared to evaluate the impact of scoring error, reproducibility, and differences in relative amplification strength on the reproducibility of RAPD based genetic distance estimates. The measured RAPD data scoring error was 2%. Reproducibility, expressed as the percentage of RAPD bands scored that are also scored in replicate data, was 76%. The results indicate that the probability of a scored RAPD band being scored in replicate data is strongly dependent on the uniformity of amplification conditions between experiments, as well as the relative amplification strength of the RAPD band. Significant improvement in the reproducibility of scored bands and some reduction in scoring error was achieved by reducing differences in reaction conditions between replicates. Observed primer variability for the reproducibility of scored RAPDs may also facilitate the selection of primers, resulting in dramatic improvements in the reproducibility of RAPD data used in germplasm studies. Variance of genetic distances across replicates due to sampling error was found to be more than six times greater than that due to scoring error for a set of 192 RAPD bands. Genetic distance matrices computed from the RAPD bands scored in replicated data and RAPD bands that failed to be scored in replicated data were not significantly different. Differences in the ethidium bromide staining intensity of RAPD bands were not associated with significant differences in resulting genetic distance matrices. The assumption of sampling error as the only source of error was sufficient to account for the observed variation in genetic distance estimates across independent sets of RAPD bands.     

Primer Design and Optimization for RAPD Analysis of Nepenthes

Primers with higher G+C content produced better random amplified polymorphic DNA (RAPD) profiles in Nepenthes. The occurrence of clustered G's and C's in the center of the primer seemed also to influence the banding patterns. It was also observed that for certain polymerases, the use of different buffers other than that recommended by the manufacturer provided a better amplification profile for Nepenthes.     

Comparison of Thoroughbred and Arabian horses using RAPD markers

We compared pools of DNA from 10 Thoroughbred horses and 10 Arabian horses for the presence of randomly amplified polymorphic DNA (RAPD) markers which might be useful in distinguishing between the breeds. Using 212 decamer oligonucleotides and our polymerase chain reaction (PCR) conditions, 173 of the primers produced scoreable bands. The number of bands ranged from 0 to 9 with an average of 3·6. In family studies using 11 arbitrarily selected primers, five of the 11 primers produced polymorphic bands which exhibited Mendelian inheritance as dominant markers. When comparing the pooled DNA from Thoroughbred and Arabian horses we found 10 primers which identified markers present in the pooled DNA from one breed but absent in the pool from the other breed. Testing individual horses revealed that only two markers were wholly absent for one group while being present among members of the other. Primer UBC-85 (5′-GTGCTCGTGC-3′) detected a pair of markers absent in Thoroughbred horses but present among 11 of 31 Arabian horses. These markers were 1500 and 1700 base pairs (bp) long and designated UBC-85^C and UBC-85^D, respectively. Primer UBC-126 (5′-CTTTCGTGCT-3′) detected a 1000 bp marker (designated UBC-126^C) absent in 20 of 20 Thoroughbred horses but present in 31 of 31 Arabian horses. UBC-126^C would be particularly effective for breed comparisons, especially if the DNA band were cloned, sequenced and an allelic marker present in Thoroughbred horses but rare or absent among Arabian horses was identified. The distribution of such markers among other horse breeds might be useful to infer relationships among breeds. These kinds of markers may also be useful in detecting unwanted crossbreeding between two horse breeds.     

The distribution of random amplified polymorphic DNA (RAPD) diversity amongst populations of Isotoma petraea (Lobeliaceae)

RAPDs were generated from plants of six populations of Isotoma petraea F. Muell. The species occurs on rock outcrops in southern and western Australia, with populations exhibiting different breeding systems, including complete autogamy, varying levels of outbreeding and complex hybridity. Non-metric multidimensional scaling (nMDS) analysis of the random amplified polymorphic DNA (RAPD) data set clearly resolved all populations. The Pigeon Rock population, which is home to both complex hybrid and structural homozygote plants, was divided into those two groups by the nMDS analysis. There was little diversity in highly autogamous populations, but levels were higher in the outbred Yackeyackine population. All complex hybrid populations and plants possessed numerous genetic system-specific RAPDs, some of which were shown to be held in fixed heterozygosity. Estimating G _ST using RAPDs has been problematical due to their dominance, and analytical methods usually rely on knowledge of the selfing rate or assume Hardy–Weinberg equilibrium. This assumption does not hold when populations exhibit fixed heterozygosity, and an alternative method, Shannon's Index, was used to partition genetic diversity. The distribution of genetic diversity fit expectations for an inbreeding species, with most of the variation (87.5%) occurring between populations. This compares to an average RAPD-based G _ST of 59.6% for inbreeding species generally and 15.5% for outbreeding species.     

Rapid evolution and gene-specific patterns of selection for three genes of spermatogenesis in Drosophila

Hybrid males resulting from crosses between closely related species of Drosophila are sterile. The F1 hybrid sterility phenotype is mainly due to defects occurring during late stages of development that relate to sperm individualization, and so genes controlling sperm development may have been subjected to selective diversification between species. It is also possible that genes of spermatogenesis experience selective constraints given their role in a developmental pathway. We analyzed the molecular evolution of three genes playing a role during the sperm developmental pathway in Drosophila at an early (bam), a mid (aly), and a late (dj) stage. The complete coding region of these genes was sequenced in different strains of Drosophila melanogaster and Drosophila simulans. All three genes showed rapid divergence between species, with larger numbers of nonsynonymous to synonymous differences between species than polymorphisms. Although this could be interpreted as evidence for positive selection at all three genes, formal tests of selection do not support such a conclusion. Departures from neutrality were detected only for dj and bam but not aly. The role played by selection is unique and determined by gene-specific characteristics rather than site of expression. In dj, the departure was due to a high proportion of neutral synonymous polymorphisms in D. simulans, and there was evidence of purifying selection maintaining a high lysine amino acid protein content that is characteristic of other DNA-binding proteins. The earliest spermatogenesis gene surveyed, which plays a role in both male and female gametogenesis, was bam, and its significant departure from neutrality was due to an excess of nonsynonymous substitutions between species. Bam is degraded at the end of mitosis, and rapid evolutionary changes among species might be a characteristic shared with other degradable transient proteins. However, the large number of nonsynonymous changes between D. melanogaster and D. simulans and a phylogenetic comparative analysis among species confirms evidence of positive selection driving the evolution of Bam and suggests an yet unknown germ cell line developmental adaptive change between these two species.     

吉林省长白山区野猪种群资源现状调查

于2007年至2009年的两个冬季,采用样带法对长白山区的野猪资源现状进行了调查.根据野猪的生物学特性以及长白山地区的自然环境特征,随机布设样带189条.对样带内野猪活动新鲜足迹链进行记录,并对其它痕迹进行综合分析,转化成实体数,计算样带密度,运用DPS7.05软件进行统计分析,求出吉林省东部长白山区野猪的种群数量,结...