黑龙江省南部小飞鼠种群mtDNA控制区的遗传结构

小飞鼠（Pteromys volans）为树栖夜行滑行类啮齿动物,受森林采伐与栖息地丧失等因素影响,其种群数量与分布在不断减少。为增强保护与管理的针对性,亟需对其种群的遗传变化,特别是遗传多样性与基因流等种群遗传信息开展进一步评价。本研究在黑龙江省南部的方正、宾县、苇河和穆棱4个重点研究区域,分别采集到20、26、15和27只小飞鼠个体的毛发样本,采用mt DNA控制区全序列（1 070 bp）评估其种群遗传多样性与遗传结构。研究发现,在88个小飞鼠控制区全序列中,共检测到155个变异位点和67个单倍型,单倍型多样性（H_d）为0.992（0.945～0.986）,核苷酸多样性（P_i）为1.772%（1.221%～1.974%）,仅单倍型H＿20为宾县与苇河种群共享,只有1只个体的单倍型有52个,其稀有单倍型高达77.16%（52/67）;种群之间遗传分化系数(F_ST)为0.051 3～0.135 7（P<0.05）,基因流（N_m）为3.19～9.24;种群之间遗传分化程度与地理距离的Mantel检验呈现不显著的正相关（r=0.413,P>0.05）。结果表明,小飞鼠种群遗传多样性丰富,其中,苇河和穆棱种群最高、宾县种群次之、方正种群最低,高比例稀有单倍型提示,未来种群存在遗传多样性下降的风险;4个地理种群之间均存在显著的中等水平遗传分化,距离隔离模型不能有效地解释小飞鼠种群目前遗传格局的形成;系统进化树与单倍型网络图没有呈现出明显的系统地理格局,其种群间基因流的限制可能是近期出现的。研究结果提示,需开展栖息地修复和生态廊道构建,从而实现该物种的种群增长与扩散。     

An AFLP analysis of genetic diversity and structure of Caragana microphylla populations in Inner Mongolia steppe, China

Genetic diversity and structure of five natural populations of Caragana microphylla from the Inner Mongolia steppe were estimated using AFLP markers. Five pairs of primers generated a total of 312 bands among 90 individuals, with percentage of polymorphic bands (PPB) being 63% at the population level and 76% at the species level, respectively. The genetic diversity within populations was correlated significantly with the soil N:P ratio. AMOVA analysis revealed high genetic variations within populations (95.5%). The estimated number of migrants per generation (Nm) was 10.72, indicating a high level of gene flow among populations. There was no significant correlation (r = 0.36) between genetic distance and geographical distance. These results were discussed in terms of eco-geographical variations among populations, together with the life history traits and breeding system of the species. The knowledge obtained may have important implications for better conservation and wise use of the vegetation dominate by C. microphylla.     

Population genetic structure of striped mullet, Mugil cephalus, along the coast of China, inferred by AFLP fingerprinting

The striped mullet, Mugil cephalus, is an economically important species for both aquaculture and commercial fisheries in China. In this study, the amplified fragment length polymorphism (AFLP) technique was employed to analyze population genetic diversity and genetic distance between different populations with the aim of elucidating the population structure and gene flow of M. cephalus along the coast of China. A total of 230 fragments with 150–500 bp were identified from 118 individuals by five AFLP primer combinations. The polymorphic loci within populations varied from 46.52% to 64.78%, with an average of 53.91%, and the average heterozygosity from 0.1829 to 0.2282, with an average of 0.2074. The UPGMA phenograms of 118 individuals were constructed based on Dice similarity coefficients and four clusters were recognized. AMOVA analysis revealed that 60.7% of genetic variations were within populations and 39.3% between populations. The estimated genetic distance (φ_ST) value over all polymorphic loci across the six populations was 0.393 (p < 0.0010), indicating a strong population structuring. The pairwise φ_ST value ranged from 0.1112 to 0.5358, with an average of 0.3693. The population pairwise gene flows (average N_m = 0.73) are low. In addition, the result of the Mantel test showed that there was a significant correlation between geographic and genetic distances (r = 0.5434, p = 0.0050). It was speculated that there exist at least four distinct geographic populational subdivisions of M. cephalus along the Chinese coast. This research has provided new molecular data which will aid our understanding of the genetic structure of this species.     

Genetic variation amongst viviparid snails in the genus Mekongia (Prosobranchia: Viviparidae) in Thailand

Allozyme variation in viviparid snails from the genus Mekongia in Thailand were examined across the different species, subspecies and geographical locations (river drainage systems). Using horizontal starch gel electrophoresis, 11 presumed allozyme loci (eight polymorphic) from eight enzyme systems were screened. Heterozygosity was moderately low (Hexp = 0.000–0.109, mean = 0.037). One population of Mekongia pongensis (Nong Khai) was monomorphic at all 11 examined loci in contrast to the other two populations, suggesting bottleneck within this population. Populations were more differentiated in the Mekongia sphaericula complex (F_ST = 0.587) than in either the Mekongia swainsoni complex (F_ST = 0.161) or M. pongensis species (F_ST = 0.073). Mekongia sphaericula sphaericula and Mekongia sphaericula extensa exhibited fixed allele differences at two loci, a high genetic distance (D = 0.265–0.300) and a potential polyphyletic relationship, suggesting two distinct lineages (species).     

Comparisons of microsatellite variability and population genetic structure of two endangered wild rice species, <Emphasis Type="Italic">Oryza rufipogon</Emphasis> and <Emphasis Type="Italic">O. officinalis</Emphasis>, and their conservation implications

Conserving endangered wild rice species requires a thorough understanding of their population genetic structure and appropriate approaches. We applied six and seven microsatellite loci to study the genetic structure of six populations throughout the range of Chinese Oryza rufipogon and Oryza officinalis, respectively. The results showed that O. rufipogon possesses higher levels of genetic diversity but lower differentiation (R_S = 3.2713, P = 100.0%, H_O = 0.1401, H_S = 0.5800, F_ST = 0.271) than O. officinalis (R_S = 2.0545, P = 57.14%, H_O = 0.0470, H_S = 0.2830, F_ST = 0.554). Mean population F_IS was slightly larger for O. officinalis (F_IS = 0.844) than that for O. rufipogon (F_IS = 0.755), indicating that O. officinalis has slightly higher departures from Hardy–Weinberg expectations and heterozygosity deficits than O. rufipogon. In addition to different origins and evolutionary histories, O. officinalis has restricted gene flow, high inbreeding, isolated small populations and fewer opportunities of hybridization with other taxa, which may determine major differences in population genetic structure from O. rufipogon. Our results suggest the adoption of a plan of involving fewer populations but more individuals within populations for O. rufipogon, while both the number of populations and the individuals for a sampled population should be almost equally considered for O. officinalis. The known high degree of inbreeding in the populations of both species implies that conservation and restoration genetics should particularly focus on the maintenance of historically significant processes such as high levels of outbreeding, gene flow and large effective population sizes. We finally proposed to further estimate the role of rice gene flow in the conservation of O. rufipogon, and to perform detailed analysis of mating systems in both species for better conservation perspectives of their ecological and evolutionary processes.     

Utilisation of a tropical bay as a nursery area by sharks of the families Carcharhinidae and Sphyrnidae

Synopsis At least eight species of sharks of the families Carcharhinidae and Sphyrnidae use Cleveland Bay in northern Australia as a communal nursery area.Carcharhinus dussumieri, C. fitzroyensis, C. limbatus andC. tilstoni use the bay as a seasonal primary nursery, with juveniles occurring in it for only a few months each year immediately after birth. Alternatively,Carcharhinus sorrah, Rhizoprionodon acutus andR. taylori use the bay as a year-round primary and secondary nursery, with juveniles remaining in it up to the size at maturity. AdultR. taylori also persist in the bay, a behavioural pattern possibly explained by their small maximum size. While present immediately after birth the type of utilisation pattern displayed bySphyrna lewini could not be clarified in this study. Although diets of these species in the bay are similar, there is probably little direct competition for food due to the highly productive habitats in the bay supporting an abundance of food resources. The highest numbers of juveniles occur when prey species are the most abundant, and when temporal separation of some seasonally-occurring species of sharks in effect.     

Automated multiple development method for determination of glycerol produced by wine yeasts

A rapid and efficient analytical method for the determination of glycerol in wines is described. This method utilizes high-performance thin layer chromatography (HPTLC) plates coupled with an automated multiple development system with an elution gradient based on acetonitrile–acetone–hexane on silica gel layers. The absence of clean-up procedures, sometimes only centrifugation, makes this method suitable also for the large-scale control of alcoholic beverages. In particular the capacity of different wine yeast species (Saccharomyces cerevisiae, Zygosaccharomyces bailii, Kloeckera apiculata and Saccharomycodes ludwigii) to produce glycerol was determined. Generally, the strains of S. cerevisiae produced elevated amounts of glycerol together with Z. bailii, whereas K. apiculata strains formed the lowest amounts of glycerol, exhibiting also a great strain variability.     

基于线粒体控制区的嘉陵裸裂尻鱼种群遗传结构分析

嘉陵裸裂尻鱼为青藏高原特有鱼类,近年来随自然地理气候的变迁和人类活动的影响,种群数量急剧减少。为了解嘉陵裸裂尻鱼的遗传背景以便更好的保护其遗传资源,本研究采用线粒体控制区部分序列变异,分析了嘉陵裸裂尻鱼6个地理种群的遗传结构和分布动态。在147尾个体中共发现17个变异位点,定义了14种单倍型,群体总的单倍型多样性较高为0.810,核苷酸多样性低为0.00698。AMOVA分析显示,44.29%的分子差异源于群体间,55.71%的分子差异源于群体内,群体间遗传分化极显著。Fst值统计检验表明,除宕昌群体和舟曲群体之间差异不显著外,其余两两群体之间Fst值统计检验均显著。基因流估计显示各群体间的基因流水平较高,遗传交流较频繁。Mantel test检验表明,嘉陵裸裂尻鱼种群之间遗传分化程度与地理距离存在显著相关。系统树和单倍型网络进化图显示,6个地理群体的单倍型按照嘉陵江水系和渭河水系形成两个大的类群。错配分布和中性检验表明嘉陵裸裂尻鱼群体在近期历史上群体大小保持稳定,未出现显著的种群扩张。根据本文所揭示的嘉陵裸裂尻鱼种群遗传结构特征,建议将分布在嘉陵江水系的嘉陵裸裂尻鱼作为一个整体进行保护,嘉陵裸裂尻鱼渭河种群属高度分化的单倍型类群,且遗传多样性极低,需对该种群进行优先保护。     

Assessment of genetic diversity and population structure of Chinese wild almond, Amygdalus nana, using EST- and genomic SSRs

Amygdalus nana L., commonly known as wild almond, is an endangered wild relative of cultivated almond, which has great potential in almond crop breeding. In this study, we used microsatellite (SSR) loci derived from both expressed sequence tag (EST) and anonymous genomic sequence to explore the genetic diversity and population structure of A. nana in Xinjiang of China. Seven natural populations were collected across the whole distribution of A. nana in China, including populations from both inside (four populations) and outside (three populations) the established protected areas. A total of 22 and 19 alleles were detected from the seven pairs of EST and genomic SSR loci, respectively. Generally, the genomic SSRs showed lower levels of variation than EST-SSRs, which may partially due to the higher cross-species transferability in EST-SSRs than in genomic SSRs. The population-level genetic diversity (A = 1.84, P = 50.00%, H_o = 0.3491, H_E = 0.2271) was lower than cultivated almond and several wild fruit species with similar breeding system. Most of the genetic variation (82.16%) was partitioned within populations. In particular, the population collected from Tacheng County (outside the protected areas) had the highest levels of genetic diversity and had significantly different genetic constitution from other populations.     

Genetic diversity and population differentiation of chestnut blight fungus, Cryphonectria parasitica, in China as revealed by RAPD

Seventeen Cryphonectria parasitica populations sampled from six regions in China were investigated using RAPD. Across all 169 isolates from the 17 populations evaluated, 52 of the 71 markers (73%) were polymorphic, total genetic diversity (h) was 0.1463, and Shannon’s index was 0.2312. Diversity within populations accounted for 74% of total genetic diversity, and genetic differentiation among populations was 0.26 (G _ST = 0.26). Gene flow was 1.4 among the populations; higher gene flow was found among populations within regions and among regions [N _m (G _SR) = 2.8 and N _m (G _RT) = 3.5]. The unweighted pair group mean analysis (UPGMA) dendrogram revealed two distinct clusters: the northern China group and the southern China group. The spatial autocorrelation analysis revealed that the variation at most loci was randomly distributed and lacked spatial structure, but several loci and closer distances were spatially structured. Human activity and habitat could also be important factors affecting genetic structure among C. parasitica populations in China. Genetic diversity was highest in Southwest China, descending in an orderly fashion to Northeast China. This pattern indicated that Southwest China might be the center of origin of C. parasitica in China. The present study provides useful information for understanding the origin and spread of chestnut blight fungus in China and valuable data for formulating relevant strategies for controlling the disease in China.     

Genetic polymorphisms of Echinococcus tapeworms in China as determined by mitochondrial and nuclear DNA sequences

The genetic polymorphisms of Echinococcus spp. in the eastern Tibetan Plateau and the Xinjiang Uyghur Autonomous Region were evaluated by DNA sequencing analyses of genes for mitochondrial cytochrome c oxidase subunit 1 (cox1) and nuclear elongation factor-1 alpha (ef1a). We collected 68 isolates of Echinococcus granulosus sensu stricto (s.s.) from Xinjiang and 113 isolates of E. granulosus s. s., 49 isolates of Echinococcus multilocularis and 34 isolates of Echinococcus shiquicus from the Tibetan Plateau. The results of molecular identification by mitochondrial and nuclear markers were identical, suggesting the infrequency of introgressive hybridization. A considerable intraspecific variation was detected in mitochondrial cox1 sequences. The parsimonious network of cox1 haplotypes showed star-like features in E. granulosus s. s. and E. multilocularis, but a divergent feature in E. shiquicus. The cox1 neutrality indexes computed by Tajima’s D and Fu’s Fs tests showed high negative values in E. granulosus s. s. and E. multilocularis, indicating significant deviations from neutrality. In contrast, the low positive values of both tests were obtained in E. shiquicus. These results suggest the following hypotheses: (i) recent founder effects arose in E. granulosus and E. multilocularis after introducing particular individuals into the endemic areas by anthropogenic movement or natural migration of host mammals, and (ii) the ancestor of E. shiquicus was segregated into the Tibetan Plateau by colonising alpine mammals and its mitochondrial locus has evolved without bottleneck effects.     

Population structure and genetic diversity in two species of Hawaiian picture-winged Drosophila

Over the last several decades many picture-winged Drosophila have become less common in both geographical distribution and local population size (pers. obs., Foote pers. comm., Montgomerey pers. comm.). Here we report on a study of two Hawaiian Drosophila species, D. engyochracea, and D. hawaiiensis, to determine the impact that changes in population sizes over the past thirty years have had on the genetic diversity of these species. D. engyochracea is known from only two locations on the Island of Hawai'i (Kipuka Ki and Kipuka Pua'ulu), while D. hawaiiensis is currently more wide spread across Hawai'i Island. We collected 65 D. hawaiiensis and 66 D. engyochracea from two forest patches (kipuka) isolated by a 400 year old volcanic ash deposit. DNA sequence data for 515 bases of the mitochondrial gene COII was analyzed for both species to estimate relative total genetic diversity as well as inter-kipuka gene flow. The more wide spread species, D. hawaiiensis, has more genetic diversity (23 vs. 11 unique haplotypes) than the rarer species, D. engyochracea. The distribution of haplotypes in the kipuka is consistent with more gene flow in D. engyochracea than in D. hawaiiensis. Phylogenetic analysis indicates a small number of individuals morphologically identified as one species but have DNA sequence diagnostic for the other species. These results are consistent with these individuals being descendant from hybrids between species.     

Genetic variation and population structure in Korean populations of sand dune speciesSalsola komarovi (Chenopodiaceae)

Salsola komarovi lljin is a herbaceous annual native to the sand dunes and beaches of Japan, northern China, Sakhalln and Korea. Starch-gel electrophoresis was conducted on leaves and stems collected from 300 plants in eight Korean populations. The mean number of alleles per locus (A _p=1.51), mean expected heterozygosity (He _p=0.116), and total genetic diversity (H _T=0.279) were comparable with those for species with similar life history and ecological traits. A general conformance of genotype frequencies to Hardy-Weinberg expectations (meanF _IS=−0.030) indicates thatS. komarovi is an outcrossing species. Slightly more than 20% of the genetic variation was found among populations (F _ST=0.204). In addition, significant differences in allele frequency were detected between populations at all 11 polymorphic loci (P<0.001). Nei's genetic identities range from 0.885 to 0.985 with a mean of 0.942. However, indirect estimates of the number of migrant per generation (0.97, calculated fromF _ST and 0.31, calculated from seven private alleles) indicate that the levels of gene flow is low among Korean populations. Although the species maintains a moderate level of genetic variation within populations, the small, isolated natural populations of the species have been severely destructed by human activities, particularly in summer season. If this is true, conservation efforts should be focused on those populations that currently maintain the most genetic diversity (e.g., populations of Cheju Island and coast of the southwestern Korean Peninsula).