三角鲂×翘嘴鲌、团头鲂×翘嘴鲌两种杂交后代微卫星遗传结构分析

为了指导三角鲂(Megalobrama terminalis)、团头鲂(Megalobrama amblycephala) 与翘嘴鲌(Erythroculter ilishaeformis)的杂交育种工作, 利用筛选出的16对微卫星引物, 比较分析了团头鲂、三角鲂、翘嘴鲌、团头鲂♀×翘嘴鲌♂、三角鲂♀×翘嘴鲌♂后代群体的遗传结构; 结果显示, 平均等位基因数(N_a)分别为3.56、3.63、3.44、4.00和4.31, 平均观测杂合度(H_o)分别为0.3510、0.3757、0.3175、0.3818和0.4079, 平均期望杂合度(H_e)分别为0.6182、0.6290、0.5921、0.6490和0.6825, 平均多态信息含量(PIC)分别为0.5354、0.5367、0.5258、0.5785和0.6067。杂交群体的平均多态信息含量均大于他们的亲本团头鲂、三角鲂和翘嘴鲌, 表明杂交亲群体的遗传多样性较高。聚类分析显示团头鲂与三角鲂首先聚类, 团头鲂♀×翘嘴鲌♂与三角鲂♀×翘嘴鲌♂首先聚类, 然后这2大类聚为一支, 最后与翘嘴鲌聚类。其中团头鲂与翘嘴鲌遗传距离最远, 为0.5204, 团头鲂和三角鲂遗传距离最近, 为0.0853, 结合遗传相似度分析表明2种杂交子代均具有母本效应。基因型分析表明, 2种杂交后代的等位基因均来自于父母本。引物TTF3、TTF4、TTF10以及Mam25在5个群体中均可产生特异性条带, 可区分5个群体。研究结果对三角鲂×翘嘴鲌和团头鲂×翘嘴鲌的良种选育、种质资源保存以及种群鉴定具有重要意义。     

鲂属鱼类形态度量学研究

鲂属（Ｍｅｇａｌｏｂｒａｍａ）鱼类是鲤科（Ｃｙｐｒｉｎｉｄａｅ）■亚科（Ｃｕｌｔｒｉｎａｅ）中一群分布广的较大型经济鱼类[１］,有的种已作为淡水养殖对象,如著名的武昌鱼。多年以来,由于过度捕捞,广泛的移殖和人工繁殖等诸多因素,鲂属鱼类的种质资源正受到衰退和混杂的威胁[２］。以中国科学院水生生物研究所淡水鱼类标本馆收藏的标本为基础,对鲂属鱼类的广东妨、厚颌鲂、鲂、团头鲂和鳊的标本进行了测量和分析,希望对鲂属鱼类的种间差异、物种鉴定特征、物种分化过程有较全面的了解,并为鲂属鱼类资源的利用和保护提供参考…     

厚颌鲂种群遗传结构及哑基因问题

厚颌鲂Megalobramapellegrini（Tchang）属鲤形目、鲤科、鳇亚科、鲂属,仅分布于长江上游特别是四川境内[1],体高,肉肥,为分布地重要的经济鱼类之一,因其体色偏黑而被称为“乌鳊”。随着团头鲂养殖业的发展,团头鲂的天然种质资源遭到严重破坏[2],因此对其它鳊鲂鱼类的开     

利用RAPD分析披碱草属、鹅观草属和猬草属模式种的亲缘关系

利用随机扩增多态性DNA（RAPD）技术对小麦族披碱草属、鹅观草属和猬草属3个属的模式种进行了基因组DNA多态性分析。42个引物产物的290条谱带中,257条（88.62%)表现出多态性,说明披碱草属、鹅观草属和猬草属3个属的模式种间具有丰富的遗传多样性。利用290个RAPD标记,计算材料间Nei氏遗传相似性系和遗传距离,在NTSYS程序中利用UPGMA进行聚类。结果表明,Elymus sibiricus种不同居群间的遗传差异较小,遗传距离在0.097-0.180之间。E.sibiricus,Roegneria caucasica和Hystrix patula的种间遗传差异明显,遗传距离在0.458-0.605之间。H.patula与E.sibiricus的亲缘关系较近。R.caucasica与E.sibiricus的亲缘关系较远。     

鲂属鱼类杂交子代肌间骨的形态学比较

本研究对团头鲂(Megalobrama amblycephala)与鲂属其他3种鱼类三角鲂(M.skolkovii)、广东鲂(M.terminalis)和厚颌鲂(M.pellegrini)正反交及其亲本自交共10种组合子代的肌间骨数目、形态、分布和长度进行了比较分析。10种不同繁育组合子代个体的肌间骨总数范围为96~134,平均值为119,各组合肌间骨的数目无显著性差异(P0.05),其中,厚颌鲂自交组合子代个体的肌间骨数目最少(平均值为114)。不同杂交组合子代体重和体长与肌间骨的长度均呈线性相关。不同部位肌间骨长度存在差异,其中躯干轴上肌的肌间骨长度显著大于躯干轴下肌、尾部轴上肌和尾部轴下肌三个部位的肌间骨长度(P0.05)。躯干轴下肌的肌间骨数目显著少于其他部位(P0.05),尾部轴上肌和尾部轴下肌的肌间骨数目无显著性差异(P0.05)。杂交鲂肌间骨形态复杂多样,分别为"1"形、"卜"形、"y"形、一端多叉形、两端多叉形和"("形6种类型,肌间骨越靠近鱼体前端形态越复杂,且与存在部位有一定的关系,躯干轴上肌的肌间骨形态最复杂。研究结果表明,鲂属鱼类近缘杂交子一代的肌间骨相关性状特征无显著改变。     

鲌鲂F1、F2及其亲本肌间骨的比较分析

为研究新型鲌鲂杂交鱼的肌间骨,采用常规测量和解剖法对鲌鲂F1 (female Culter alburnus Basilewskymale Megalobrama amblycephala Yih)、鲌鲂F2 (self-crossing of F1 hybrid of female C. alburnusmale M. amblycephala)及其母本翘嘴鲌(C. alburnus)、父本团头鲂(M. amblycephala)肌间骨的数目、形态和分布进行统计分析,结果表明,翘嘴鲌肌间骨数目为134-139,平均为136.8根; 团头鲂的肌间骨数目为121-129,平均为124.2根; 鲌鲂F1肌间骨数目为129-134,平均为131.6根; 鲌鲂F2肌间骨数目为127-134,平均为130.1根; 鲌鲂F1、F2与翘嘴鲌、团头鲂之间肌间骨数目差异显著(P0.05); 鲌鲂F1每一肌节所含肌间骨数目最多,为0.8024; 鲌鲂F2最少,为0.7744; 翘嘴鲌和团头鲂介于鲌鲂F1和F2之间,分别为0.7953和0.7763。4种鱼均含有I形、卜形、Y形、一端多叉形、两端两分叉形、两端多叉形和树枝形7种类型肌间骨,髓弓小骨比脉弓小骨多且复杂; 鱼体左右两侧肌间骨的数目不完全相等,形态也不完全对称,但较为接近,且肌间骨越靠前端,形态越复杂; 研究获得的鲌鲂F1和F2在肌间骨总数、复杂型肌间骨数目和躯体轴下肌肌间骨数目均较母本有所减少,鲌鲂F2较F1还呈下降的趋势,且每一肌节所含肌间骨最少,表现出一种有利于提高食用品质和精深加工的优势。研究结果为鲌鲂属间远缘杂交培育少肌间骨新品种提供了基础资料。     

用RAPD分子标记探讨沙拐枣属的种间关系

利用随机扩增多态性DNA（RAPD）技术分析了14种沙拐枣属（Calligonum L.)植物,通过对16个Sangon公司十聚体随机引物进行PCR扩增,3个引物能产生多态性带。对3个引物扩增产生的45条扩增产物,计算单匹配系数,应用UPGMA方法构建亲缘关系树状图。分析结果表明：（1）物种间遗传差异明显,具有丰富的遗传多样性;（2）14种沙拐枣属植物明显聚为4类,与传统的形态学分类结果基本一致。     

“太湖鲂鲌”及其亲本肌肉营养成分的分析与评价

为评价新型杂交鱼“太湖鲂鲌”(翘嘴鲌Culter alburnus♀×三角鲂Megalobrama terminalis♂)的肌肉营养价值, 采用生化测定法比较分析了“太湖鲂鲌”、翘嘴鲌和三角鲂的肌肉营养成分, 结果表明: (1)“太湖鲂鲌”肌肉水分含量显著低于翘嘴鲌和三角鲂(P<0.05), 而肌肉粗蛋白质含量较高(P<0.05)。(2)“太湖鲂鲌”的必需氨基酸(EAA)含量显著高于三角鲂(P<0.05); 3种鱼的第一限制性氨基酸均为含硫氨基酸(蛋氨酸+胱氨酸)。(3)“太湖鲂鲌”肌肉脂肪酸中的不饱和脂肪酸(UFA)含量显著高于翘嘴鲌和三角鲂(P<0.05)。(4)3种鱼肌肉矿物质元素铁、铜、锰、锌含量均无显著差异(P>0.05)。综上所述, “太湖鲂鲌”肌肉营养继承并综合了双亲的优良性状, 是一种富含蛋白质、EAA和UFA的优良养殖品种, 具有推广价值。     

26种冬青属植物遗传多样性分析

以26种冬青属植物种质资源为研究材料,利用RAPD和AFLP技术对基因组DNA进行扩增,以研究其物种间遗传多样性以及亲缘关系.结果表明:在RAPD分析中,从400条10个碱基的寡核苷酸引物中筛选出反应稳定、扩增性强、重复性好的引物20个,共扩增出312条多态性条带,多态率为95.41%;聚类分析显示26种冬青属植物间,布利奥特夫人枸骨叶冬青和黄果在AFLP分析中,10对选择性引物组合均扩增出了丰富的多态性片段,共扩增出350条谱带,其中336条具有多态性,占95.96%.综合RAPD和AFLP聚类结果,枸骨、无刺枸骨和日拉斯纳尔逊枸骨的亲缘关系较近,钝齿冬青、金宝石钝齿冬青和龟甲冬青三者的亲缘关系较近,可为冬青属植物的杂交育种与种质创新提供理论依据.     

大豆不同亲本类型F2主要性状遗传参数分析

对大豆3个不同亲本类型的杂交组合F2群体主要性状遗传变异系数、遗传力和遗传进度进行了估算分析,结果表明：这些遗传参数因性状或亲本类型的不同而有明显差异。含有野生亲缘关系的组合,类型间、性状间F2遗传力有较大差异,性状变异幅度增大,相对遗传进度也较大,从而有较广泛的选择基础。     

团头鲂卵壳蛋白的分离与纯化

报道了一种从团头鲂卵壳中提取卵壳蛋白的有效方法。该提取淮经SDS－PAGE电泳鉴定后,证明有三条主要的蛋白带,其分子量分别为64KDa,56KDa和52KDa,借助于制备型SDS－PAGE电泳和电洗脱分离纯化技术,获得了三种高纯度的印壳蛋白。     

Evolutionary distances and identification of Candida species in clinical isolates by Randomly Amplified Polymorphic DNA (RAPD)

Fast and reliable identification of different species of the genus Candida is important to define adequate therapeutic decisions, because the different species have highly variable susceptibilities to antifungal drugs; azoles and amphothericin B. Accurate statistical records on case history and epidemiological studies also depend on effective identification. To address this problem we established a RAPD method that enabled direct identification of five very common species of Candida. Initially, reference band patterns were established for C. albicans, C. tropicalis, C. parapsilosis, C. glabrata and C. krusei. One of the primers, M2, showed remarkably conserved intra-specific patterns of approximately 10 bands each, ranging in size from 2.0 to 0.1 kb. These patterns were significantly different and species-specific. Few bands were conserved between different species of Candida, which was assumed to be consistent with their phylogenetic relatedness. In addition, band patterns were constant and reproducible and DNA isolated from single colonies yielded sufficient DNA for identification. The reference band patterns were then used, in blind experiments, to identify species of Candida in 50 randomly chosen samples, including clinical isolates and ATCC strains. RAPD results were 100% consistent with results obtained by conventional diagnostic methods and were achieved in one day instead of several days taken by conventional methods. Because ideal identification methods should be consistent with phylogeny and taxonomy we tested whether RAPD could be used to calculate genetic distances. Comparison of RAPD phylogenetic trees with 18S rRNA trees showed significant differences in tree topologies which indicated that RAPD data could not accurately measure the relative distances between different species. Also, computer simulations of RAPD random patterns were used to test whether the observed degree of RAPD band pattern similarities could occur at random. These simulations suggested that the level of inter-specific band pattern similarities observed in our data could be obtained at random, while intra-specific pattern similarities could not. RAPD would be helpful to discriminate between isolates but not to quantitate the differences. We suggest that the inaccurate estimate of genetic distances from RAPD is a general limitation of the technique and not a specific problem of our identification method. Because of the repetitive character of the target sequences, genetic distances calculated from RAPD could be affected by paralogy, namely, recombination and duplication events not parallel with speciation events. This revised version was published online in June 2006 with corrections to the Cover Date.     

Random amplified polymorphic DNA (RAPD) markers reveal genetic homogeneity in the endangered Himalayan species Meconopsis paniculata and M. simplicifolia

Random amplified polymorphic DNA (RAPD) marker-based analysis was carried out to study the extent of genetic polymorphism between populations of the two endangered Himalayan poppy species, Meconopsis paniculata and M. Simplicifolia. Of the 90 primers tested, 38 revealed marked inter-species genetic polymorphism between individuals of the two species from geographically isolated populations. However, intra-species genetic homogeneity was also evident with respect to a number of primers both within and between populations. A comprehensive analysis incorporating data from RAPDs, DNA fingerprinting and isozyme pattern was carried out and, based on the presence or absence of bands, three matrices of similarity indices were estimated. These matrices were subsequently utilized in cluster analysis. In order to compare the three clusters generated using these three different marker systems, a Mantel matrix-correspondence test was carried out on the basis of comparisons of co-phenetic values. The overall representation of relationships by cluster analysis was similar for all three marker systems and this was substantiated by high correlations among the three analyses revealed by the Mantel matrix-correspondence test. Our results point to very low or absence of, genetic polymorphism in M. paniculata and M. simplicifolia, and are in broad agreement with our previous observations on genetic diversity of Meconopsis species which point to a genetic basis for the possible extinction of this economically important genus.     

Genetic heterogeneity in natural populations of Microphallus piriformes and M. pygmaeus parthenites (Trematoda: Microphallidae)

The random amplified polymorphic DNA (RAPD) technique was applied for the studies of genetic heterogeny between several natural populations of trematodes belonging to the Microphallida family. Initially, the metacercariae from the daughter sporocysts infestating Littorina saxatilis and Littorina littorea periwinkles were used. Comparison of the banding patterns obtained for the different metacercariae within one sporocyst gave an unpredicted results. For two of three studied species (M. pygmaeus and M. pseudopygmaeus), the considerable differences in RAPD patterning was detected. According to the classical point of view, the process of cercariae (metacercariae in case of the "pygmaeus" group of microphallides) formation does not include DNA recombination. Because of that, all metacercariae within one single sporocyst should be genetically identical. However, data obtained clearly shows that at least in some cases it is not so. We can hardly believe that such result could be a methodological artifact, for not single difference in a RAPD patterns was recorded between the metacercariae within sporocysts of M. piriformes. Moreover, even the 100 fold dilution of the total DNA used for PCR amplification does not change the banding patterns. Hence, our results can not be explained by slight fluctuations in the DNA concentrations between the samples. The most evident conclusion is that we came across yet strange, but real phenomenon--some degree of genetic difference within the progeny of each of the sporocysts--metacercariae. However, the detailed study is needed to understand and interpret these observations correctly. Amplification of the total DNA extracted from the whole sporocysts (containing metacercariae) never showed any differences in RAPD patterns between the parasites been derived from one infestated snail (local parasite hemipopulation). That allowed us to compare different parasite populations referring the RAPD pattern of one sporocyst from a snail to as a representative of one local hemipopulation. Analysis of the RAPD-loci frequencies showed a considerable genetic differences between the subpopulations of M. piriformes, infestating different paraxenic intermediate hosts--L. saxatilis and L. obtusata. This phenomenon was statistically significant for 2 localities of 3 studied. No heterogeny within populations was recorded for M. pygmaeus. Both M. piriformes and M. pygmaeus are characterized by the genetic differentiation in the microgeographic scale (within the Chupa bay of the White sea, the longest distance between the analyzed localities is 20 km). According to the frequencies of the RAPD-loci, parasites from the sheltered locality differ significantly from the parasites of other two localities exposed to the open sea. For both species the degree of genetic similarity between the populations correlates positively with the distance between the localities. We can suppose that the population structure of microphallids depended mainly upon the population structures of their intermediate hosts, definitive hosts and geographical structure of the areal. However, taking into consideration the low motility of snails, we believe that the distribution, migration and species composition of the definitive hosts play the key role in the genetic structuring of M. pygmaeus and M. piriformes hemipopulations. As an addition, the RAPD analysis of the parasite populations from the Barents Sea (East cost of Murman peninsula) and North Sea (Western cost of Sweden) revealed no significant genetic differences between the worms from those places and from the White Sea. However in case of this macrogeographic comparison, insufficient number of samples does not allow us to draw any final conclusions.     

Comparison of RFLP and RAPD markers to estimating genetic relationships within and among cruciferous species

Restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) markers are being used widely for evaluating genetic relationships of crop germplasm. Differences in the properties of these two markers could result in different estimates of genetic relationships among some accessions. Nuclear RFLP markers detected by genomic DNA and cDNA clones and RAPD markers were compared for evaluating genetic relationships among 18 accessions from six cultivated Brassica species and one accession from Raphanus sativus. Based on comparisons of genetic-similarity matrices and cophenetic values, RAPD markers were very similar to RFLP markers for estimating intraspecific genetic relationships; however, the two marker types gave different results for interspecific genetic relationships. The presence of amplified mitochondrial and chloroplast DNA fragments in the RAPD data set did not appear to account for differences in RAPD- and RFLP-based dendrograms. However, hybridization tests of RAPD fragments with similar molecular weights demonstrated that some fragments, scored as identical, were not homologous. In all these cases, the differences occurred at the interspecific level. Our results suggest that RAPD data may be less reliable than RFLP data when estimating genetic relationships of accessions from more than one species.     

利用RAPD对稻蝗属昆虫亲缘关系的研究

通过20个随机引物的PCR扩增,得到了日本主要稻蝗的随机扩增多态性DNA（RAPD）图谱,根据扩增结果,计算了种间相似系数和遗传距离,建立了UPGMA系统树。结果表明,分布没有重叠、种间容易交配、能产生杂种的中华稻蝗台湾亚种与小翅稻蝗的亲缘关系最近;分布重叠的日本稻蝗与中华稻蝗台湾亚种、日本稻蝗和小翅稻蝗的亲缘关系较近。小稻蝗与其它3种稻蝗的亲缘关系较远。     

RAPD Analysis Reveals Geographic Differentiations within Allium schoenoprasum L. (Alliaceae)

Abstract: Random amplified polymorphic DNA (RAPD) analysis was used to study the phylogenetic relationships between species in Allium section Schoenoprasum and for the investigation of the intraspecific differentiation of A. schoenoprasum. RAPD analysis of 39 samples representing eight species of sect. Schoenoprasum and one sample of A. atrosanguineum (sect. Annuloprasum ) resulted in 233 interpretable RAPD bands. The analysis clearly distinguishes the species of section Schoenoprasum. The arrangement of the accessions of A. schoenoprasum in all dendrograms mirrors the geographical distribution, with a clear differentiation between an Asian and European subgroup. Within the European group, Scandinavian material is clearly distinct from S and E European material. Informally described morphological types of A. schoenoprasum could not be confirmed by RAPD analysis but represent recurrent ecological adaptations. A combination of phenetic (UPGMA, neighbour-joining analysis), cladistic (parsimony analysis), and statistical (PCA) methods of data analysis resulted in clearer phylogenetic interpretations than each of the methods facilitates when used separately.     

Genetic differentiation of Helicoverpa armigera (Hübner) and H. assulta (Guenée) (Lepidoptera: Noctuidae) based on AFLP markers

Abstract Here we use amplified fragment length polymorphism (AFLP) to assess genetic differentiation of Helicoverpa armigera and H. assulta . The results indicated that both species-specific fingerprints and cluster analysis showed the ability of AFLP technique to discriminate the two sibling species; among a total 1963 AFLP markers amplified from nine primer combinations: 777 (39.6%) were H. armigera -specific, 602 (30.7%) were H. assulta -specific, and 584 (29.7%) were common bands. The mean number of H. armigera -specific bands was significantly more than that of H. assulta -specific bands for nine primer combinations ( P < 0.05); the intraspecific distance of H. armigera and H. assulta was 0.123 0 and 0.110 7 respectively, and the interspecific distance was 0.178 3. In addition, the percentage of polymorphic loci and estimated average heterozygosity were used to estimate genetic diversity of the two species. This study therefore demonstrates that AFLP analysis is a sensitive and reliable technique to study genetic differentiation and genetic relationships between species and provides sufficient molecular markers for future linkage map construction, location and eventual cloning of genes involved in traits differentiation.     

Qualitative and quantitative characterization of RAPD variation among snap bean (Phaseolus vulgaris) genotypes

Ten snap bean (Phaseolus vulgaris) genotypes were screened for polymorphism with 400 RAPD (random amplified polymorphic DNA) primers. Polymorphic RAPDs were scored and classified into three categories based on ethidium bromide staining intensity. An average of 5.19 RAPD bands were scored per primer for the 364 primers that gave scorable amplification products. An average of 2.15 polymorphic RAPDs were detected per primer. The results show that primer screening may reduce the number of RAPD reactions required for the analysis of genetic relationships among snap-bean genotypes by over 60%. Based on the analysis of the distribution of RAPD amplification, the same number of polymorphic RAPDs were amplified from different genotypes for all RAPD band intensity levels. A comparison of RAPD band amplification frequency among genotypes for the three categories of bands classified by amplification strength revealed a measurable difference in the frequencies of RAPDs classified as faint (weakly amplifying) compared to RAPD bands classified as bold (strongly amplifying) indicating a possible scoring error due to the underscoring of faint bands. Correlation analysis showed that RAPD bands amplified by the same primer are not more closely correlated then RAPD bands amplified by different primers but are more highly correlated then expected by chance. Pairwise comparisons of RAPD bands indicate that the distribution of RAPD amplification among genotypes will be a useful criterion for establishing RAPD band identity. For the average pairwise comparison of genotypes, 50% of primers tested and 15.8% of all scored RAPDs detected polymorphism. Based on RAPD data Nei's average gene diversity at a locus was 0.158 based on all scorable RAPD bands and 0.388 if only polymorphic RAPD loci were considered. RAPD-derived 1 relationships among genotypes are reported for the ten genotypes included in this study. The data presented here demonstrate that many informative, polymorphic RAPDs can be found among snap bean cultivars. These RAPDs may be useful for the unique identification of bean varieties, the organization of bean germplasm, and applications of molecular markers to bean breeding.     

矮牡丹与紫斑牡丹RAPD分析初报

用10个任意序列的寡核苷酸片段作为引物,将采自陕西、山西、甘肃等地矮牡丹与紫斑牡丹基因组DNA,在毛细管气浴式PCR热循环仪上随机扩增。在对所有引物扩增条件严格标准化的条件下,这些引物可产生清楚的、可重复的与扩增产物相应的琼脂糖凝胶电泳区带。这10个有效扩增的引物在矮牡丹平均每一个体中扩增出71条带,其中多态的带16条(22．5％),在紫斑牡丹平均每一个体中扩增出76条带,其中多态的带2l条(27．6％)。对每两个个体,每条多态带进行成对比较,累加后求分子标记差异的平均值。在矮牡丹3个居群间的平均差异是7．9,在紫斑牡丹4个居群间的平均差异是8．7,在矮牡丹与紫斑牡丹2个种间的平均差异是10.3。显然,若用来扩增的植物个体数目和引物数目增加,将会得到更满意的结果。初步结果表明濒危植物矮牡丹与紫斑牡丹种内低水平的DNA多态性。RAPD技术用于检测野生牡丹居群内与居群间的遗传变异是有用的与可行的;用于研究种间的进化和亲缘关系也有潜力。