华山新麦草自然居群沿海拔梯度的的遗传分化

华山新麦草为我国特有种,仅分布于陕西华山,是国家一级珍稀濒危植物和急面保护有农作物野生亲缘种。应用RAPD技术,选取8条引物对华山新麦草自然分布分布区的3个山峪（种群）11个样方（亚居群）79个华山新麦草个体的总DNA进行随机扩增,共得到65个RAPD位点。统计分析表明,在华山的3个山峪（居群）中,黄埔峪（居群）与其它2个山峪（居群）发生较显著的遗传分化,华山峪6个亚居群的个体平均RAPD位点数有随海拔的升高而下降的趋势,6个亚居群间的相似性系数也有随海拔的升高而下降的趋势,华山峪的高海拔亚居群和低海拔亚居群间表现出明显差异。主成分分析结果进一步证明了黄埔峪居群与华山峪居群和仙峪居群间、以及华山峪的的高海拔亚居群与低海拔亚居群间已发生一定程度的分化。研究结果暗示海拔差异是导致华山新麦草自然居群遗传分化的主要因素,海拔差异造成的有限的基因流可能才是影响因居群和亚居群遗传分化的主要因子,而亚居群内遗传变异程度则与该亚居群的所处的特定生境有关。     

用RAPD检测华山新麦草自然居群的遗传结构和居群分化

华山新麦草特产于我国陕西华山,为国家一级重点保护植物。应用RAPD技术,用2 0条随机引物对黄埔峪、仙峪、华山峪3个居群不同海拔的13个亚居群共2 6 6个华山新麦草取样个体进行扩增,共得12 2个扩增片段。平均每个引物得到的扩增片段数为6 .1(2～10 )。实验数据的统计分析表明:黄埔峪居群的多态性位点比率为6 0 .6 6 % ,仙峪居群的多态性位点比率为90 .98% ,华山峪居群的多态性位点比率为95 .0 8% ,总的多态性位点比率为95 .0 8% ,说明华山新麦草具有较高的遗传变异性。Shannon多样性指数(0 .330 6 )和基因分化系数(GST=0 .32 6 3)揭示了华山新麦草居群遗传变异多存在于亚居群内。华山新麦草亚居群间的基因流N m=1.0 32 2 ,低于一般风媒传粉植物(N m=5 .2 4 )的基因流水平。亚居群间平均的遗传距离为0 .15 71(变化范围:0 .0 0 2 2～0 .2 90 1) ,最大的遗传距离值出现在黄埔峪亚居群(hp1)和华山峪高海拔亚居群(h8)之间,仙峪亚居群与华山峪高海拔亚居群之间以及华山峪高海拔亚居群与低海拔亚居群之间的遗传距离值也较大。遗传距离与海拔垂直距离之间的相关性分析表明二者有显著的相关性(p<0 .0 1)。聚类分析和主成分分析也显示华山新麦草自然居群已发生明显分化,主要表现为黄埔峪居群、仙峪居群与华山峪居群     

华山新麦草自然居群的遗传结构和种内遗传多态性研究

华山新麦草为我国特有种,公分布在陕西华山,应用5种酶系统共13个基因位点对采自华山13个亚居群的华山新麦草进行等位酶分析,研究结果发现,多态性位点的比例（P）为69％,个位点等位基因的平均数（A）为1．8,平均每个位点的预期杂合性（He)为0．344,Simpson指数为0．998,证明华山新麦草居群内有较高的遗传多态性和克隆多样性,固定指数（F）显示有意义的负值（－0．252),预期的随机交配相比较,居群内有过多的杂合体,这可能与华山新麦草的交酸系统和繁育方式有关,平均的遗传距离为0．046（变化范围：0－0．139）,大约有91％的遗传变异存在于亚居群内,应用间接法测得华山新麦草自然居群间的基因流（Nm)为2．77,明显你于一般风媒传粉植物（5．24）,暗示华山新麦草自然居群的基因流水平似乎正处于一种临界状太,有进一步分化的潜能,但受多种因素的影响。     

应用RAPD技术研究阿尔卑斯山黄花茅居群内的遗传分化

应用RAPD技术,沿一个海拔梯度研究了阿尔卑斯山黄花茅自然居群的遗传变异和分化。实验 表明,虽然在亚居群间有很少的亚居群独有遗传标记的存在,但通过RAPD资料的聚类分析、主因子分 析以及相关分析观察到遗传分化沿海拔梯度发生,而且亚居群间的遗传分化和它们的海拔高度(地理距 离)呈有意义的正相关。研究结果暗示,在阿尔卑斯山的高山-亚高山过渡区,亚居群间的海拔高度差别 可能导致黄花茅开花和生长物候期的变化,而后者限制了亚居群间的基因流,从而引起居群内的遗传分化。     

华山新麦草种质资源研究进展

华山新麦草属禾本科小麦族新麦草属,是一个多年生二倍体物种,因只分布于陕西华山而得名,属于普通小麦的三级基因源;因具有抗逆境、抗病虫、早熟、优质等特点,成为小麦遗传改良的重要外源基因资源。近年来,国内外对华山新麦草的研究已经取得了较大的进展。本综述从遗传多样性、形态学、生物化学与分子生物学、遗传学及其开发与利用等方面,首次对该种质资源的研究进展进行了总结,旨在提高对华山新麦草的整体认识,为对其进一步深入研究和高效利用提供参考。     

华山新麦草胚和胚乳的发育研究

采用常规石蜡切片法,观察了华山新麦草胚和胚乳的发育过程,结果表明,华山新麦草胚和胚乳的发育过程与一般禾本科植物基本相同,胚胎发生属紫宛型,顶细胞和基都参与胚体的形成,胚胎发育经过二细胞原胚,多细胞原胚,球形原胚,梨形原胚,分化胚和成熟胚阶段,成熟胚具有胚根,胚芽,盾片,胚牙鞘,胚根鞘,外胚叶等典型禾本科植物成熟胚的结构,胚乳发育类型为核型,包括游离核阶段,细胞化阶段和生长成熟阶段,待大量游离核形成之后才形成细胞壁,紧贴胚囊的一层胚乳细胞最后形成种子的糊粉层,其余的胚乳细胞最后充满淀粉粒,其特点为：（1）有双球形原胚的现象;（2）反足细胞解体较早;（3）胚乳游离核时期和细胞时期胚乳细胞核的核仁多样。     

华山新麦草大孢子发生和雌配子体的形成

华山新麦草大孢子发生始于孢原细胞。大孢子母细胞减数分裂形成直线型四分体。功能大孢子位于合点端,大孢子发育为蓼型胚囊。成熟胚囊为八核七细胞,卵细胞极性明显,助细胞在极核融合前已退化一个,另一个宿存,宿存助细胞丝状器明显,并保留退化助细胞的痕迹。极核排列方向不定,反足细胞在胚囊发育早期增殖,并液泡化。此外,在华山新麦草中发现有双孢原细胞及双胚囊的现象。     

华山新麦草开花物候期观测和自然种群基因流的间接估测

华山新麦草（Psathyrostachys huashanica keng)为我国特有种,仅分布在陕西华山。通过对自然种群传粉物候期观测发现,华山新麦草的传粉高峰期与海拔有一定关系。海拔每升高200m,传粉物候期就推迟2－3d;海拔差异超过500m的亚种群,传粉物候期就不会出现重叠。同时应用等位酶分析技术和E－统计分析间接估测华山新麦草自然种群的基因流（Nm）,其值较一般风媒传粉植物低。通过华山新麦草资源状况、生长环境的调查以及移栽试验的观察,认为该物种稀的主要原因之一 可能是其生存竞争能力弱,最终被迫生长在其它物种所不能生存的环境。     

华山新麦草濒危原因及种群繁殖对策

华山新麦草特产于我国华山,为国家一级保护植物。由于华山新麦草和小麦属于亲缘种,并且具有很强的抗旱性和耐盐性,所以对其的研究就 很有实践意义。从生境特征和繁殖对策的角度对其濒危原因进行了探讨。华山新麦草生长在恶劣和不连续的生境中,每个居群个体数量都很少。极低的繁殖分配造成其有性繁殖水平低下。脆弱的竞争力和沉重的环境压力共同作用使华山新麦草侧重选择了营养繁殖方式,在生活史上策略上选择上偏重于K－策略。     

华山新麦草叶片的光合生理特性

对华山新麦草(Psathyrostachys huashanica Keng ex P.C.Kuo)营养叶的净光合速率(Pn)和蒸腾速率(Tr)的日变化曲线进行了分析,并在对叶温(Tl)、气孔阻力(Rs)、光合有效辐射强度(PAR)和气温(Ta)的日变化曲线进行测定的基础上分析了它们对华山新麦草Pn的影响规律。结果表明:华山新麦草Pn的日变化曲线呈"三峰"型,峰值分别为6.5、6.2和9.0μmol.m-2.s-1,依次出现在9:30、11:30和16:30,而且具有明显的"午降"现象;Tr的日变化曲线呈"单峰"型,最大值为1.7 mmol.m-2.s-1,出现在13:30;Tl、Rs、PAR和Ta的日变化曲线均呈"单峰"型,峰值分别出现在12:30、11:30、12:30和13:30。华山新麦草的Pn对Tl、PAR和Ta的响应曲线均呈"抛物线"型,Pn在一定范围内与Tl、PAR和Ta呈正相关,随着Tl、PAR和Ta的升高逐渐增加至最大值后逐渐降低;而Pn与Rs则呈负相关,Pn在一定范围内随Rs的增大逐渐降低。根据拟合方程,华山新麦草营养叶的光补偿点和光饱和点分别为1.1和531.5μmol.m-2.s-1,说明该种类具有很强的喜光性,且对光照强度的适应范围较广。研究结果表明:较大的气孔阻力是造成华山新麦草叶片净光合速率偏低的主要原因。     

Subpopulation differentiation of Anthoxanthum alpinum (Poaceae) along an altitudinal gradient detected by random amplified polymorphic DNA

Random amplified polymorphic DNA (RAPD) phenotypes generated by 15 primers were scored in 5 subpopulatious of Anthoxanthum alpinum along an altitudinal gradient. Although few subpopulation-specific markers were found, a significant population differentiation was revealed by the use of principal factor analysis and Ward' s cluster analysis. The result was also confirmed by the significant positive correlation when comparing beth the matrix of similarity between the individuals within subpopulations and the matrix among subpopulations, as well as the matrix of distances and the differences of altitude between the subpopulations (Mantel test). This study suggests that the differences of altitude between subpopulation sites may result in the differences of phenologicalperiod for flowering and growth which restrict gene flow. Decontaminated thianthrene disproportion. Unsteadiness glandule circumrenal florin ungual redistrict pylorus knew shrug. 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Genetic structure and differentiation of Psathyrostachys huashanica populations detected with RAPD markers

Psathyrostachys huashanica Keng is a perennial grass and belongs to genus Psathyrostachys under Triticeae.sathyrostachys is found in the center of Middle Asia and the Caucasus Mountain,while P.huashanica,a species endemic to China,is only located in Mt.Hua in the Shaanxi province,China.At present,the population of this species is decreasing,and reaching the edge of extinction.Due to the limitation in distribution and the importance as breeding material for germplasm storage,it has been considered as first class among the national protected rare plants.For this reason,the present study is significant in probing plant flora,origin and evolution of Triticeae,and crop breeding.Randomly amplified polymorphic DNA (RAPD)markers were used to analyze the genetic structure and differentiation of P.huashanica populations sampled in three valleys (Huangpu,Xian,and Huashan Valleys)in Mr.Hua.One hundred and twenty-two RAPDfragments were obtained in all 266 individuals with 20 primers with a mean of 6.1 (2-10)fragments per primer.The percentage of polymorphic loci (PPB)was 60.66%in Huangpu Valley,90.98%in Xian Valley,95.08%in Huashan Valley,and the total PPB was 95.08%,which indicated a highly genetic variability of P.huashanica.The Shannon's Information index and GST were 0.3306 and 0.3263,respectively,indicating that there were more genetic variations within the subpopulations than those among the subpopulations.The gene flow among the subpopulations of P.huashanica (Nm=1.0322)was much less than that of the common anemophytes (Nm=5.24).Mean genetic distance is 0.1571(range:0.0022-0.2901).The highest value of genetic distance was found between the subpopulation (hp1)of Huangpu Valley and the highest altitude subpopulation (h8)of Huashan Valley.Correlation analysis detected significant correlation between genetic distance and vertical distance of altitude.Clustering analysis and principal coordinate analysis revealed the genetic differentiation among the populations of P.huashanica.Differentiation mainly occurred between the higher altitude subpopulations and the lower altitude subpopulations,suggesting that altitude might be the major factor that restricted the gene flow between different altitude subpopulations and resulted in differentiation of subpopulations.     

Identification and genetic homogeneity of Trichophyton tonsurans isolated from several regions by random amplified polymorphic DNA

Trichophyton tonsurans is an anthropophilic dermatophyte mostly causing tinea capitis and tinea corporis. This study was carried out to identify T. tonsurans and to clarify whether it has any genetic differences depending on the phenotype or region of isolation by random amplified polymorphic DNA (RAPD) analysis with three random primers. The assay was performed in 11 Korean, 2 Japanese, 2 Taiwanese, 5 Brazilian and 1 American isolates of T. tonsurans together with the other 10 anamorphic species of dermatophytes and 3 Arthroderma spp. All tested species of dermatophytes showed distinct bands and T. tonsurans was differentiated from other dermatophytes. It was most clearly ifferentiated from T. mentagrophytes by using primer 5-GAAGGCTCCC-3 (OPAO-15). No difference was found in RAPD band patterns in all strains of T. tonsurans with these random primers. It was considered that T. tonsurans is a genetically homogeneous species regardless of its isolation regions, morphologic or physiologic characteristics.This revised version was published online in October 2005 with corrections to the Cover Date.     

Characterization of Zebu cattle breeds in Tanzania using random amplified polymorphic DNA markers

A total of 141 short primers, of arbitrary nucleotide sequence, were used singly in poly-merase chain reactions to amplify DNA fingerprints in pools of DNA representing three Zebu cattle breeds. Two primers, which discriminated between the breed-specific DNA pools were used further to amplify individual pool components in order to establish band frequencies of the amplified fingerprints. One of the primers (ILO 1127) amplified a RAPD fingerprint in 61%of TSZ animals but less than 6% in the other breeds, while another primer (ILO 1065) revealed a DNA sequence common to 89% of the Boran animals and less than 30% in the other two breeds. Bandsharing and mean average percentage difference calculated within and between the three breeds using RAPD fingerprint data showed a higher degree of homogeneity within than across the breeds and indicated measurable divergence between the three breeds. It is concluded that RAPD polymorphisms are useful as genetic markers for cattle breed differentiation.     

Population differentiation in randomly amplified polymorphic DNA of red-cockaded woodpeckers Picoides borealis

Random amplified polymorphic DNA (RAPD) phenotypes generated by 13 primers were scored for 101 individuals in 14 populations of the endangered red-cockaded woodpecker Picoides borealis. Although no population-specific markers were found, the frequencies of several markers differed significantly among populations. Application of the recently developedamova method (analysis of molecular variance; Excoffier, Smouse & Quattro 1992) showed that more than 90% of phenotypic variance occurred among individuals within populations; of the remaining variance, half was attributed among groups of geographically adjacent populations and half among populations within those groups. The statistical significance of these patterns was supported by Monte Carlo sampling simulations and permutation tests. Estimation of allele frequencies from phenotypes provided somewhat weaker evidence for population structure, although among-population variance in allele frequencies was detectable (F_st= 0.19; x²₁₆₉= 509.3, P < 0.0001). Upgma cluster analyses based on Rogers' (1972) genetic distance revealed grouping of some geographically proximate populations. A Mantel test indicated a positive (r = 0.16), although not significant, correlation between geographic and genetic distances. We compared a subset of our RAPD data with data from a previous study that used allozymes (Stangel, Lennartz & Smith 1992). RAPD (n= 75) and allozyme (n= 245) results based on samples from the same ten populations showed similar patterns. Our study indicates that RAPDs can be helpful in differentiating populations at the phenotypic level even when small sample sizes, estimation bias, and inability to test for Hardy-Weinberg equilibrium complicate the genotypic interpretation. Lack of large differences among populations of red-cockaded woodpeckers may allow flexibility in overpopulation translocations, provided factors such as habitat preference, latitudinal direction of translocation, and status of donor populations are considered.     

应用RAPD技术对我国不同地域红色毛癣菌分离株的遗传多样性研究

目的探讨我国不同地域红色毛癣菌分离株的遗传多样性。方法采用随机扩增DNA多态性(RAPD)方法对来源于我国不同地域(江苏南京,山东济南,广东广州)的32株红色毛癣菌临床分离株进行DNA多态性分析。结果红色毛癣菌种内差异明显,根据遗传相似性分成三大聚类群,与地域差异及取材部位无明显相关性,而与表型具有一定相关性。结论随机扩增DNA多态性方法可用于红色毛癣菌的DNA分型,其DNA带型具有一定的遗传变异性,与菌株表型有一定关系,与地域差异、侵犯部位无明显相关性。     

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。     

三种摇蚊幼虫RAPD扩增条件的优化及在昆虫系统发育分析中的应用

为了研究摇蚊科昆虫种群遗传的多样性,以促进对其资源的合理保护,以萨摩亚摇蚊Chironomus samoensisEdwards基因组DNA为模板,对摇蚊幼虫的RAPD扩增条件进行优化,建立了摇蚊幼虫RAPD扩增反应的最佳体系:按照利用优化的RAPD扩增条件进行研究,实验有着良好的重现性。用16个随机引物对3种摇蚊幼虫类群各10个个体进行RAPD扩增,其中萨摩亚摇蚊共扩增出78个条带,多态座位率为41.03%,Shannon遗传多样性指数为0.2570,群体内相似度为0.8730;红裸须摇蚊Propsilocerus akamusi(Tokunaga)共75个条带,多态座位率为44.0%,Shannon遗传多样性指数为0.2472,群体内相似度为0.8731;刺铗长足摇蚊Tanypus punctipennis(Fabricius)共67个条带,多态座位率为41.79%,Shannon遗传多样性指数为0.1943,群体内相似度为0.9066。聚类分析结果表明,刺铗长足摇蚊与红裸须摇蚊的亲缘关系较近。