Characterization of 10 new tetranucleotide microsatellite markers for the European eagle owl,Bubo bubo: Useful tools for conservation strategies

Bubo bubo is the largest owl in the world, showing a wide geographical distribution throughout the Palaearctic region. It underwent a demographic decline in many European countries during the last century and was considered “vulnerable” (Annex II of the CITES). Nowadays, it is classified as “Least Concern” according to IUCN.Despite its ecological importance and conservation status, few polymorphic molecular markers are available to study its diversity and population genetics. We report on the isolation and development of 10 new microsatellites for the Eagle owl, B. bubo. All loci (10 tetra-nucleotide) are characterized by high polymorphism levels. Number of alleles ranged from 5 to 13 and expected heterozygosity varied from 0.733 to 0.840. These microsatellites would be very useful to assess the genetic diversity, connectivity patterns and parentage of B. bubo. This information will allow to establish new conservation strategies and improve the management of the species.     

南天竹微卫星标记开发及特性分析（英文）

南天竹(Nandina domestica)是一种具有较高观赏价值的常绿灌木。本研究从南天竹基因组中开发和筛选出10个微卫星位点,能在该物种中进行稳定PCR扩增且具一定的多态性。采用4个南天竹群体24个个体进行检测发现,每个位点的等位基因数为2~6,期望杂合度和表观杂合度分别为0.153~0.778和0~1。本研究结果为后续对南天竹遗传多样性的研究打下了基础。另外,这10个位点在小檗科4个其他物种中也有一定的通用性。     

Microsatellite markers for the common vole (<Emphasis Type="Italic">Microtus arvalis</Emphasis>) and their cross-species utility

Fragmentation of natural habitats of the common vole (Microtus arvalis) provides an excellent model system to study the consequences of restricted gene flow and small population sizes for isolated populations. Here we describe the isolation and characteristics of 10 autosomal and one X-linked microsatellite marker. These new markers were tested in 24 voles from a natural population in eastern Germany. Loci were highly polymorphic with numbers of alleles per locus ranging from three to 26 and expected heterozygosities from 0.51 to 0.97. All loci except for the X-linked locus Mar105 followed Hardy–Weinberg expectations. Cross-species amplifications revealed that most loci were polymorphic as well in M. agrestis, M. thomasi, and M. pennsylvanicus.     

Development of twenty-one polymorphic tetranucleotide microsatellite loci for Schizothorax o&#x27;connori and their conservation application

Schizothorax o'connori, of the subfamily Schizothoracinae, is an endemic fish in the Yarlung Tsangpo River in Tibet, China. To evaluate the population genetic diversity as a means of devising conservation strategies, a total of 21 polymorphic tetranucleotide microsatellite loci were isolated from S. o'connori using the fast isolation by AFLP of sequences containing repeats (FIASCO) protocol. The characterization of these loci was assessed against a panel of 42 individuals. The number of alleles per locus ranged from 4 to 29 with an average of 17.76. Expected locus-specific heterozygosity and the multilocus Shannon–Wiener diversity index ranged from 0.114 to 0.951 and from 0.280 to 3.131, with an average of 0.851 and 2.338, respectively. The cross-species amplification and applicability of these loci were tested against the two other Schizothorax species from the same river. These microsatellite loci will be valuable for further studies of population genetic diversity and genetic structure, and assessments of the artificial propagation release effect of S. o'connori and other related species.     

Predation by Moses perch, Lutjanus russelli, and blue-spotted trevally, Caranx bucculentus, on juvenile brown tiger prawn, Penaeus esculentus: effects of habitat structure and time of day

The effects of habitat structure and time of day on predation success of Moses perch, Lutjanus russelli , and blue-spotted trevally, Caranx bucculentus , feeding on juvenile brown tiger prawn, Penaeus esculentus , were tested in the laboratory. Our experiments demonstrated that both softsubstratum, in which prawns can burrow, and seagrass habitats reduce predation rates by fish during daytime when compared to hard-substratum habitats. However, the protection offered by burrowing in a soft substratum during daytime disappears at night when prawns become active and are more available to fish. Seagrass structure offers the same protection during both night and day. This is attributed to the structural complexity of the seagrass habitat, as well as the behaviour and disruptive coloration of the prawns. The results suggest that fish predation on juvenile P. esculentus may not be very important in seagrass nurseries but could be very high in other habitats.     

Isolation and characterization of tri‐ and tetranucleotide microsatellite loci in the Asian elephant,Elephas maximus

Asian elephants (Elephas maximus) are an endangered species. Their future survival depends on intensive conservation and management, based on in‐depth knowledge of particular populations. Molecular genetic methods, especially microsatellite analysis through noninvasive sampling, provides an effective means of obtaining such information. The use of tri‐ and tetranucleotide microsatellite markers is advantageous in noninvasive sampling through dung analysis. Here we describe the isolation and characterization of five tri‐ and tetranucleotide markers in the Asian elephant. All five loci were found to be polymorphic in a sample of 20 Asian elephants from Sri Lanka.     

Characterization of 21 microsatellite marker loci in the ring-tailed lemur (<Emphasis Type="Italic">Lemur catta</Emphasis>)

Ring-tailed lemur (Lemur catta) is the only species in the Genus Lemur, distributed in the deciduous and spiny forests of southwestern Madagascar. This species is listed as endangered due to the loss and fragmentation of its natural habitat, a consequence of deforestation. Twenty-one nuclear microsatellite loci were isolated from a genomic DNA derived from a free-ranging ring-tailed lemur population from the Tsimanampetsotsa National Park, Madagascar. We report various parameter estimates and measures to establish the utility of this marker suite as screened among individuals this single forest fragment.     

中国特有单种属血水草的微卫星分子标记开发与评价

血水草(Eomecon chionantha Hance)是中国亚热带地区特有的单种属植物。本研究采用(AG)15、(AC)15两种生物素探针及磁珠富集法从血水草基因组中开发并筛选出13个微卫星位点,且在该物种中能进行稳定的PCR扩增并表现出多态性。利用筛选出的13个SSR多态性标记对血水草4个野生居群24个个体进行PCR扩增,结果显示每个微卫星位点的等位基因数为2~7个,观测杂合度(HO)和期望杂合度(HE)分别为0.125~0.792和0.299~0.691。说明这些SSR标记可用于血水草遗传多样性的后续研究,并揭示该物种地理分布格局形成的原因,有助于保护和利用中国亚热带地区的血水草野生资源。     

Early Developmental Stages of the Green Tiger Prawn, Penaeus semisulcatus de Haan (Crustacea, Decapoda, Penaeidae)

Gravid females of Penaeus semisulcatus were spawned in the laboratory by natural means. The embryos were documented and the larvae were reared from hatching to postlarval stage at 28.2–30.0 °C and 33.5–34.5 g kg⁻¹ salinity for about 10 days (223 h 55 min). Six naupliar stages, three protozoea stages, three mysis stages and the first postlarval stage were described and illustrated. The larvae were fed only with microalgae Tetraselmis tetrathele and Chaetoceros gracilis from first protozoea until the second mysis, with about 90% survival rate; from the third mysis until the first postlarva they were fed with similar microalgae coupled with rotifer Brachionus plicatilis and Artemia nauplii. The embryonic and larval stages of P. semisulcatus are generally similar to those of other closely related species in the family Penaeidae, such as Melicertus canaliculatus, Fenneropenaeus merguiensis, and Marsupenaeus japonicus, except for the size and structure of diagnostic characters, setation of appendages and duration of metamorphoses. The change in the feeding habit during ontogeny was related to morphological transformation of the feeding apparatus of larvae and postlarvae. This paper is the first comprehensive and complete account of the early developmental stages of P. semisulcatus.     

湖栖鳍虾虎鱼微卫星DNA标记的开发与群体遗传多样性分析

为研究雷州半岛湖栖鳍虾虎鱼(Gobiopterus lacustris)遗传多样性,对湖栖鳍虾虎鱼雌雄性腺进行转录组测序,采用MISA软件进行微卫星分析,共获得25 452个微卫星标记,其中包含14 708个单碱基重复类型,6 175个2碱基重复类型,3、4、5和6碱基重复类型数目分别为4 223、327、15和4。随机选取50个微卫星位点设计引物,能够扩增出清晰稳定条带的有39对,其中,11个位点表现出不同程度的多态,28个位点呈现单态。利用11个具有多态性的微卫星位点分析湖栖鳍虾虎鱼在廉江市高桥镇红树林保护区、湛江东海岛、雷州市附城镇和雷州市九龙山红树林国家湿地公园4个野生群体中的遗传多样性及遗传结构,11个微卫星位点呈现出不同程度的多态性,等位基因数(Na)2～15,平均等位基因为(6?3.9)个,有效等位基因数(Ne)、平均观测杂合度(Ho)和期望杂合度(He)分别在1.919～2.485、0.343～0.465和0.381～0.483之间,平均多态信息含量(PIC)为0.348～0.465。Hardy-Weinber平衡分析显示,4个群体的大部分位点未偏离平衡。在每个群体中进...     

Microsatellite Polymorphism and the Population Structure of the Black Tiger Shrimp (Penaeus monodon) in Thailand

: Genetic variation and differentiation of Thai Penaeus monodon from five geographic locations (Chumphon, Trad, Phangnga, Satun, and Trang) were investigated using five microsatellite loci (CUPmo18, Di25, Di27, CSCUPmo1, and CSCUPmo2). The number of alleles across the five loci ranged from 19 to 30, and heterozygosities ranged from 0.49 to 0.95. The mean number of alleles and effective number of alleles per locus were 21.0 to 26.6 and 13.1 to 20.4, respectively. The average heterozygosity across all investigated samples was 0.78, indicating high genetic diversity in this species. Geographic heterogeneity analysis of the results from two of the loci, CUPmo18 and Di25, showed significant differences among the Gulf of Thailand (Trad and Chumphon) but not the Andaman samples. Comparison between regions revealed significant heterogeneity of the Andaman and Trad P. monodon (P < .001), whereas those from Chumphon and the Andaman were genetically similar (P > .05). Significant genetic differentiation was consistently observed between the Andaman-Trad samples (FST = 0.0101, P < .0001) and the Chumphon-Trad samples (FST = 0.0101, P < .0001). On the basis of our analyses, the investigated samples from five geographic locations were allocated to three distinct populations composed of the Andaman Sea (A), Chumphon (B), and Trad (C).     

Variation of Microsatellite Size Homoplasy Across Electromorphs, Loci, and Populations in Three Invertebrate Species

Size homoplasy was analyzed at microsatellite loci by sequencing electromorphs, that is, variants of the same size (base pairs). This study was conducted using five interrupted and/or compound loci in three invertebrate species, the honey bee Apis mellifera, the bumble bee Bombus terrestris, and the freshwater snail Bulinus truncatus. The 15 electromorphs sequenced turned out to hide 31 alleles (i.e., variants identical in sequence). Variation in the amount of size homoplasy was detected among electromorphs and loci. From one to seven alleles were detected per electromorph, and one locus did not show any size homoplasy in both bee species. The amount of size homoplasy was related to the sequencing effort, since the number of alleles was correlated with the number of copies of electromorphs sequenced, but also with the molecular structure of the core sequence at each locus. Size homoplasy within populations was detected only three times, meaning that size homoplasy was detected mostly among populations. We analyzed population structure, estimating F _st and a genetic distance, based on either electromorphs or alleles. Whereas little difference was found in A. mellifera, uncovering size homoplasy led to a more marked population structure in B. terrestris and B. truncatus. We also showed in A. mellifera that the detection of size homoplasy may alter phylogenetic reconstructions. Received: 21 July 1997 / Accepted: 29 January 1998     

Characterization of 20 microsatellites marker loci in the golden-crowned sifaka (<Emphasis Type="Italic">Propithecus tattersalli)</Emphasis>

The golden-crowned sifaka (Propithecus tattersalli) is one of the most critically endangered species of Propithecus endemic to a limited area in the Daraina region of north-eastern Madagascar. This species is endangered due to habitat loss and fragmentation, a consequence of deforestation. Twenty nuclear microsatellite loci were isolated from genomic DNA derived from a Propithecus tattersalli from the Daraina area in north-eastern Madagascar. Population genetic parameters were estimated on 20 individuals from two Daraina forest fragments to determine the potential utility of this marker suite for future studies on the golden-crowned sifaka.     

Characterization of 22 microsatellite marker loci in the Madagascar rousette (Rousettus madagascariensis)

Twenty-two nuclear microsatellite loci were isolated from a genomic DNA library derived from Madagascar’s Rousettus madagascariensis. Marker characteristics were determined from a single population (37 individuals) from Fort Dauphin (southeastern Madagascar). Sixteen of the 22 loci were within Hardy–Weinberg expectations. These loci are highly informative with polymorphic information content values ranging between 0.757 and 0.916. These loci will provide valuable information for the study of population genetics and gene flow within this species of bats. Due to the dramatic reduction and alteration of their habitat, data generated utilizing this marker suite will potentially provide additional information for the effective long-term management of this near-threatened bat species.     

Microsatellite DNA in Actinidia chinensis: isolation, characterisation, and homology in related species

 We have isolated and sequenced 263 microsatellite-containing clones from two small insert libraries of Actinidia chinensis enriched for (AC/GT) and (AG/CT) repeats, respectively. Primer pairs were designed for 203 microsatellite loci and successfully amplified from both plasmid and A. chinensis genomic DNA. In this paper we report the sequences of 40 primer pairs for which we have demonstrated Mendelian segregation in the progeny from controlled crosses. The polymorphism of ten microsatellites of each type was evaluated in four diploid and six tetraploid genotypes of A. chinensis. All microsatellites proved to be polymorphic, the number of alleles per locus detected in polyacrylamide sequencing gels ranging from 9 to 17. The high degree of polymorphism in Actinidia renders these markers useful either for mapping in A. chinensis or for fingerprinting cultivars of both domesticated kiwifruit species (A. chinensis and A. deliciosa). While most primer pairs produced single amplification products, about 20% generated banding patterns consistent with the amplification of two different loci. This supports the hypothesis that diploid species of Actinidia (2n=2x=58) are polyploid in origin with a basic chromosome number x=14/15 and that chromosome duplication may have occurred during the evolution of the genus. Finally, we have assayed the cross-species transportability of primer pairs designed from A. chinensis sequences and have found extensive cross-species amplification within the genus Actinidia; 75% of primer pairs gave successful amplification in the eight species assayed (A. arguta, A. rufa, A. polygama, A. chrysantha, A. callosa, A. hemsleyana, A. eriantha, and A. deliciosa), which are representative of the four sections into which the genus is currently split. Received: 14 February 1998 / Accepted: 26 May 1998     

Characterization of 21 microsatellite marker loci in the silky sifaka (Propithecus candidus)

The silky sifaka, Propithecus candidus, considered one of the rarest and most endangered primates in the world, exists in only a few fragmented forests in northeastern Madagascar. This species faces increasing pressures as a direct result from loss of habitat in the form of tavy (slash and burn agriculture), illegal logging and mining along with hunting for subsistence, even within protected areas. We report a marker suite of 21 loci developed from genomic DNA from a silky sifaka collected in 2003 from Anjanaharibe-Sud Special Reserve. Polymorphism of each locus evaluated in 18 individuals pooled from Marojejy National Park and Anjanaharibe-Sud Special Reserve. The number of observed alleles per locus ranges from 2 to 7. The observed and expected heterozygosity were 0.389–0.889 and 0.322–0.789, respectively. The information revealed in this study will provide useful tools for further study of the social structure and population dynamics of the silky sifaka to facilitate conservation management in the imminent future.     

Microsatellite variability in grapevine cultivars from different European regions and evaluation of assignment testing to assess the geographic origin of cultivars

Nine microsatellite markers (VVMD5, VVMD7, VVS2, ssrVrZAG21, ssrVrZAG47, ssrVrZAG62, ssrVrZAG64, ssrVrZAG79 and ssrVrZAG83) were chosen for the analysis of marker information content, the genetic structure of grapevine cultivar gene pools, and differentiation among grapevines sampled from seven European vine-growing regions (Greece, Croatia, North Italy, Austria and Germany, France, Spain and Portugal). The markers were found to be highly informative in all cultivar groups and therefore constitute a useful set for the genetic characterization of European grapevines. Similar and high levels of genetic variability were detected in all investigated grapevine gene pools. Genetic differentiation among cultivars from different regions was significant, even in the case of adjacent groups such as the Spanish and Portuguese cultivars. No genetic differentiation could be detected between vines with blue and white grapes, indicating that they have undergone the processes of cultivar development jointly. The observed genetic differentiation among vine-growing regions suggested that cultivars could possibly be assigned to their regions of origin according to their genotypes. This might allow one to determine the geographical origin of cultivars with an unknown background. The assignment procedure proved to work for cultivars from the higher differentiated regions, as for example from Austria and Portugal. Received: 10 April 1999 / Accepted: 25 May 1999     

林麝微卫星座位的分离和鉴定

利用改进的磁珠富集法从林麝(Moschus berezovskii)的基因组中分离到10个新的(AC)n重复微卫星座位,并分析其在24个林麝个体(取样于中国四川金凤山群体)中的多态性。其中,5个位点在24个林麝个体中具有4—13个等位基因,观察杂合度和期望杂合度分别是0.429—0.957和0.587—0.902,平均多态信息含量是0.730。表明这5个微卫星位点具有高度的多态性,可以用于林麝遗传多样性研究,对林麝的保护具有很重要的意义。     

克毒丹防治斑节对虾Penaeus monodon Fabricius苗种病毒的试验

全球气候异常,“埃厄尔诺”现象的频频发生,在一定程度上造成生态不平衡,至使１９９３年世界范围虾病的大流行,而我国尤为严重,产量大幅度下降。在斑节对虾ＰｅｎａｅｕｓｍｏｎｏｄｏｎＦａｂｒｉｃｉｕｓ育苗期间,发生虾病毒而引起苗种大量死亡的现象,使用抗生素仍无法控制其死亡。使用克毒丹０２×１０－６～０６×１０－６浸洒苗池,以便改变其微生态环境,又相间使用虾丹特号药物。结果苗种成活率比对照组平均提高１１％以上,从而达到控制病毒的试验效果。     

21个野生,培育及地方猪种免疫及生化遗传标记基因位点的遗传分化研究

分析了亚,欧,美,非２１个野生,培育及地方猪种４个免疫遗传标记基因位点（Ｅ,Ｆ,Ｇ,Ｌ系统）和３人生化遗传标记基因位点（Ｃｐ,Ａｍ和Ｔｆ）的遗传多态性,认为Ｅ＾ａｅｇ,Ｆ＾ａ,Ｔｆ＾ｃ基因在亚洲猪种与欧美猪种间存在显著差异,是亚洲猪种３个重要的遗传标记基因,根据Ｆ＾ａ基因频率在亚,欧,美,非猪种的变化规律分析了其基因流向特点,证明欧美猪的育种过程中利用了中国地方猪种的血缘,计算了所测猪种间的Ｎｅｉ