Molecular genetic basis for the rhino mouse from Chinese KM subcolony

Both the rhino mouse and hairless mouse resulted from hairless gene mutation, but they show different phenotypes of skin physiology. The rhino mouse has more similar histological characters to human papular alopecia. Therefore rhino mouse is a good experimental animal model for human papular alopecia. This study reports a hairless mouse named rhino KIZ, arose from KM colony in Kunming Institue of Zoology, by systematic studies on morphology, skin histopathology, gene sequence, pedigree and protein domain analysis. The results demonstrate that a C-to-T transition in exon 11 of hr gene (The mutant gene has been applied for a Chinese patent (patent No. 03135280)) results in the rhino KIZ. The rhino KIZ with clear genetic mechanism will be a useful animal model.

Molecular phylogenetics and biogeography of Lepus in Eastern Asia based on mitochondrial DNA sequences

In spite of several classification attempts among taxa of the genus Lepus, phylogenetic relationships still remain poorly understood. Here, we present molecular genetic evidence that may resolve some of the current incongruities in the phylogeny of the leporids. The complete mitochondrial cytb, 12S genes, and parts of ND4 and control region fragments were sequenced to examine phylogenetic relationships among Chinese hare taxa and other leporids throughout the World using maximum parsimony, maximum likelihood, and Bayesian phylogenetic reconstruction approaches. Using reconstructed phylogenies, we observed that the Chinese hare is not a single monophyletic group as originally thought. Instead, the data infers that the genus Lepus is monophyletic with three unique species groups: North American, Eurasian, and African. Ancestral area analysis indicated that ancestral Lepus arose in North America and then dispersed into Eurasia via the Bering Land Bridge eventually extending to Africa. Brooks Parsimony analysis showed that dispersal events followed by subsequent speciation have occurred in other geographic areas as well and resulted in the rapid radiation and speciation of Lepus. A Bayesian relaxed molecular clock approach based on the continuous autocorrelation of evolutionary rates along branches estimated the divergence time between the three major groups within Lepus. The genus appears to have arisen approximately 10.76 MYA (+/- 0.86 MYA), with most speciation events occurring during the Pliocene epoch (5.65 +/- 1.15 MYA similar to 1.12 +/- 10.47 MYA). (c) 2005 Elsevier Inc. All rights reserved.

PCR-RFLP analysis of mitochondrial DNA ND5/6 region among 3 subspecies of common carp (Cyprinus carpio L.) and its application to genetic discrimination of subspecies

Mitochondrial DNA ND5/6 region was studied by PCR-RFLP analysis among ten representative strains belonging to three subspecies (Cyprinus carpio carpio, Cyprinus carpio haematopterus and Cyprinus carpio rubrofuscus) of common carp (Cyprinus carpio L.). A total of 2.4 kb fragment was amplified and subjected to restriction endonuclease analysis with nine restriction endonucleases subsequently. The results indicated that each subspecies owned one hyplotype and four restriction enzymes (Dde I, HaeIII, Taq I and Mbo I) produced diagnostic restriction sites which could be used for discriminating the three subspecies and as molecular genetic markers for assistant selective breeding of common carp.

Molecular biotechnology of marine algae in China

Molecular biotechnology of marine algae is referred to as the biotechnology on the identification, modification, production and utilization of marine algal molecules. It involves not only the manipulation of macromolecules such as DNA, RNA and proteins, but also deals with low molecular weight compounds such as secondary metabolites. In the last decade, molecular systematic researches to investigate the relationship and to examine the evolutionary divergence among Chinese marine algae have been carried out by Chinese scientists. For example, RAPD has been widely used in several laboratories to elucidate genetic variations of the reds, such as Porphyra, Gracilaria, Grateloupia and the greens such as Ulva and Enteromorpha. Some important data have been obtained. The study on molecular genetic markers for strain improvement is now in progress. In 1990s, genetic engineering of economic seaweeds such as Laminaria, Undaria, Porphyra, Gracilaria and Grateloupia has been studied in China. For Laminaria japonica, the successfully cultivated kelp in China, a model transformation system has been set up based on the application of plant genetic techniques and knowledge of the algal life history. Progress has been made recently in incorporating a vaccine gene into kelp genome. Evidence has been provided showing the expression of gene products as detectable vaccines. In the present paper, the progress of molecular biotechnological studies of marine algae in China, especially researches on elucidating and manipulating nucleic acids of marine algae, are reviewed.

Silver carp, Hypophthalmichthys molitrix, in the Poyang Lake belong to the Ganjiang River population rather than the Changjiang River population

The Poyang Lake is the largest lake and the main nursery area in the middle basin of the Changjiang (Yangtze) River. We compared molecular genetic markers of silver carp among populations of the Changjiang River, the Ganjiang River and the Poyang Lake using the ND5/6 region of mtDNA. Analysis of restriction fragment length polymorphisms (RFLPs) of this region revealed distinct variation between the Ganjiang River and the Changjiang River populations. The Poyang Lake is linked with the Ganjiang River and the Changjiang River. Shared RFLP fragments between the Ganjiang River population and the Poyang Lake population are as high as 61.4%. The value is 47.74% between the populations of the Changjiang River and that of the Poyang Lake. Frequencies of bands peculiar to the Ganjiang River population are the same as in the Poyang Lake population. We conclude that the Poyang Lake silver carp population consists mainly of the Ganjiang River population. The water level of the Poyang Lake outlet, which is higher than that of the Changjiang River in the silver carp spawning season, supports this conclusion.

中国海域斑海豹种群资源和分子遗传特征研究

斑海豹，西北太平洋广泛分布的冷水性海洋哺乳动物，为我国的二类保护野生动物，属于濒危物种，也是唯一能在我国海域自然繁殖的鳍脚类动物。辽东湾结冰区是斑海豹在世界上8个繁殖区中最南端的一个。为了能够更好地保护斑海豹资源，对渤海海域斑海豹的栖息地、种群动态及分布时间、重金属体内积累、以及斑海豹的分子遗传特性进行了研究。结果如下： 斑海豹在渤海海域的栖息地有辽宁双台子河口水域、大连虎平岛和山东庙岛群岛海域三处，其中的上岸点分别为河口泥沙滩、海里的浅石滩和海岛周围的小岛礁三种类型。斑海豹出现在三个栖息地的时间为每年的3~5月，2002~2008年间各栖息地的斑海豹的数量变化不明显。 对斑海豹肌肉、肝和肾脏组织重金属元素含量的分析结果显示，汞（Hg）、镉（Cd）、铅（Pb）和砷（As）等有毒元素在斑海豹体内的积累远未达到致死浓度。 对斑海豹的部分mtDNA 序列分析发现，辽东湾斑海豹群体的遗传距离、控制区DNA的单元型多样度和核苷酸多样度均远小于日本群体，辽东湾群体的遗传多样性水平较低。 微卫星引物标记对辽东湾斑海豹群体的遗传多样性研究显示，平均有效等位基因数（2~4个）和期望杂合度（0.24~0.72）等指标均较低，表明辽东湾斑海豹群体遗传多样性水平下降，可能曾出现过一定程度的瓶颈效应。 对斑海豹的41个MHC-I基因的序列分析，得到40个等位基因。斑海豹MHC-I基因多态性水平高，说明辽东湾斑海豹种群MHC-I基因的丰富性。 以上研究结果对于我国辽东湾斑海豹种群的保护和管理具有一定意义。

牙鲆Paralichthys olivaceus（♀）×夏鲆Paralichthys dentaus（♂）分子遗传学及数量遗传学研究

本论文采用扩增片段长度多态性（AFLP）技术对牙鲆Paralichthys olivaceus群体和夏鲆Paralichthys dentaus 群体进行了种群鉴定和遗传结构分析。共用8 对引物对两个群体（每个群体30 个个体）进行了分析。总共产生了379 个条带，条带大小在60－1000bp。牙鲆与夏鲆的遗传多态性分别为53.83%和22.22%。牙鲆群体的多态位点数显著低于夏鲆群体。牙鲆群体和夏鲆群体种的特异条带分别为27.3% 和 29.61%。两群体平均杂和度分别为0.0701 和 0.1556，香农氏多态指数分别为0.1044 和0.2387。群体内平均遗传距离分别为0.0705 （0.0214 到0.1377）和0.1656 （0.0629 到 0.2338），两群体间的平均遗传距离为0.6328。 AFLP 技术是进行种间群体结构分析的一个很好的分子标记方法，对牙鲆和夏鲆亲本群体进行遗传背景析为今后的杂交育种阐明种间杂交的遗传机理奠定基础。 在本论文中我们采用5×8 因子交配设计建立了牙鲆P. olivaceus（♀）× 夏鲆P. dentaus（♂）杂交家系，对生长相关性状的遗传力进行分析。所有家系都混合养殖在同一养殖池中，40 日龄，在养殖池中随机采集600 个个体，测量体长，体宽，体重等数量性状；然后提取杂交子代DNA 进行家系的鉴定。首先从牙鲆的微卫星中筛选了在牙鲆、夏鲆以及杂交子代扩增多态性较好的10 对微卫星引物（Po1, Po13, Po20, Po35, Po42, Po48, Po56, Po58, Po91, Poli 23TUF）。我们采用了三对引物对600 个个体进行了家系鉴定，共有400 多个体鉴定出自己的亲本，成功率达到80％以上。基于以上结果我们认为微卫星标记可以做为一个有效的标记来代替现实的物理标记，并且可以在子代的早期进行亲本的鉴定。 在养殖过程中，我们对自交和杂交家系生长状况进行了跟踪分析，在181刘清华 牙鲆（♀）×夏鲆（♂）分子遗传学及数量遗传学研究 博士学位论文II日龄之前杂交鲆体长和体重均为未表现出杂种优势，杂种优势率值始终为负值，但是绝对值在逐渐减小。从196 日龄之后杂交鲆杂种优势开始表现出来，并且在196 日龄之后杂种优势率显著的增加。256 日龄体长杂种优势率为14.29%，体重杂种优势率为59.78%, 271 日龄体长杂种优势率达到27.36%，体重杂种优势率为 102.32%。 本研究对杂交鲆在40 日龄的体长，体高、尾柄长、尾柄高，以及全长和体重的遗传力进行了估计，5 个性状的体长半同胞遗传力h2 S 为0.00146－0.719，5个性状的全同胞遗传力h2D 为0.00121－0.632，5 个性状的半同胞和全同胞的平均遗传力h2SD 为0.001335－0.6755。其中全长的遗传力最大。实验结果说明对早期幼鱼进行体长、体重等性状实施选育策略可能会显著影响后期杂交鲆的生长。

Permanent Genetic Resources added to Molecular Ecology Resources Database 1 May 2009-31 July 2009

This article documents the addition of 512 microsatellite marker loci and nine pairs of Single Nucleotide Polymorphism (SNP) sequencing primers to the Molecular Ecology Resources Database. Loci were developed for the following species: Alcippe morrisonia morrisonia, Bashania fangiana, Bashania fargesii, Chaetodon vagabundus, Colletes floralis, Coluber constrictor flaviventris, Coptotermes gestroi, Crotophaga major, Cyprinella lutrensis, Danaus plexippus, Fagus grandifolia, Falco tinnunculus, Fletcherimyia fletcheri, Hydrilla verticillata, Laterallus jamaicensis coturniculus, Leavenworthia alabamica, Marmosops incanus, Miichthys miiuy, Nasua nasua, Noturus exilis, Odontesthes bonariensis, Quadrula fragosa, Pinctada maxima, Pseudaletia separata, Pseudoperonospora cubensis, Podocarpus elatus, Portunus trituberculatus, Rhagoletis cerasi, Rhinella schneideri, Sarracenia alata, Skeletonema marinoi, Sminthurus viridis, Syngnathus abaster, Uroteuthis (Photololigo) chinensis, Verticillium dahliae, Wasmannia auropunctata, and Zygochlamys patagonica. These loci were cross-tested on the following species: Chaetodon baronessa, Falco columbarius, Falco eleonorae, Falco naumanni, Falco peregrinus, Falco subbuteo, Didelphis aurita, Gracilinanus microtarsus, Marmosops paulensis, Monodelphis Americana, Odontesthes hatcheri, Podocarpus grayi, Podocarpus lawrencei, Podocarpus smithii, Portunus pelagicus, Syngnathus acus, Syngnathus typhle,Uroteuthis (Photololigo) edulis, Uroteuthis (Photololigo) duvauceli and Verticillium albo-atrum. This article also documents the addition of nine sequencing primer pairs and sixteen allele specific primers or probes for Oncorhynchus mykiss and Oncorhynchus tshawytscha; these primers and assays were cross-tested in both species.

Optimal feedback control of two-photon fluorescence based on genetic algorithm

An optimal feedback control of two-photon fluorescence in the ethanol solution of 4-dicyanomethylene-2-methyl-6-p-dimethyl-amiiiostryryl-4H-pyran (DCM) using pulse-shaping technique based on genetic algorithm is demonstrated experimentally. The two-photon fluorescence of the DCM ethanol solution is enhanced in intensity of about 23%. The second harmonic generation frequency-resolved optical gating (SHG-FROG) trace indicates that the effective population transfer arises from the positively chirped pulse. The experimental results appear the potential applications of coherent control to the complicated molecular system.

Optimal feedback control of two-photon fluorescence in Coumarin 515 based on genetic algorithm

An optimal feedback control of two-photon fluorescence in the Coumarin 515 ethanol solution excited by shaping femtosecond laser pulses based on genetic algorithm is demonstrated experimentally. The two-photon fluorescence intensity can be enhanced by similar to 20%. Second harmonic generation frequency-resolved optical gating traces indicate that the optimal laser pulses are positive chirp, which are in favor of the effective population transfer of two-photon transitions. The dependence of the two-photon fluorescence signal on the laser pulse chirp is investigated to validate the theoretical model for the effective population transfer of two-photon transitions. The experimental results appear the potential applications in nonlinear spectroscopy and molecular physics. (c) 2005 Elsevier B.V. All rights reserved.

GENETIC DIVERSITY IN THE CHINESE PANGOLIN (MANIS-PENTADACTYLA) INFERRED FROM PROTEIN ELECTROPHORESIS

We examined protein polymorphism of Chinese pangolins (Manis pentadactyla) from Yunnan Province of China, including two forms of three brown and nine dusky Chinese pangolins. Sixty-two genetic loci were screened; 12 loci were found to be polymorphic. The percentage of polymorphic loci (P) is 0.194, the mean individual heterozygosity (H) is 0.078, and the mean number of alleles (A) is 1.258. Furthermore, we calculated the genetic distance (D) between the two forms and found a low level of genetic divergence (D = 0.0206) between them, which indicates an almost-indistinguishable divergence at the level of proteins.

Genetic diversity and population history of the red panda (Ailurus fulgens) as inferred from mitochondrial DNA sequence variations

The red panda (Ailurus fulgens) is one of the flagship species in worldwide conservation and is of special interest in evolutionary studies due to its taxonomic uniqueness. We sequenced a 236-bp fragment of the mitochondrial D-loop region in a sample of 5