中华鳖黄河群体选育世代F1、F2及F3遗传变异微卫星分析

以中华鳖黄河群体的选育基础群体（F₀）和太湖群体为对照群体,采用微卫星标记技术对黄河群体的选育世代F₁、F₂及F₃的遗传变异进行了测定和分析。结果表明：（1）从所筛选的10个微卫星位点中,5个测试群体共检测出240个等位基因;平均等位基因数(N_a) 为4.60～5.30,平均有效等位基因数(N_e)为3.24～3.59,平均观察杂合度(H_o)为0.483 3～0.530 0,平均期望杂合度(H_e)为0.594 9～0.645 9,平均多态信息含量(C_PI)为0.534 0～0.568 4,这些遗传参数表明中华鳖存在丰富的可供选育用的遗传多样性基础。（2）F₁、F₂及F₃选育世代的遗传多样性参数H_o、H_e及C_PI一致地显示出随着选育代数的增加而降低的明显趋势,证明以生长、体色、体型等表型指标为直观选育指标的群体选育,对遗传型指标产生了可检测到的影响。（3）黄河群体的选育基础群体和3个选育世代在聚类分析中聚为一支,而太湖群体单独为一支,显示中华鳖的黄河、太湖两大地理群体间存在较大遗传分化,3代选育并未改变这一历史性格局。     

日本沼虾太湖和鄱阳湖群体及其F1的遗传结构分析

采用12对微卫星标记分析了日本沼虾太湖、鄱阳湖两个野生群体及其双列杂交的F₁的遗传结构和遗传多样性。结果显示,所用引物的平均等位基因数(N_a)为17.67个,有效等位基因数(N_e)为12.48个,大小在112～427 bp之间。平均观测杂合度(H_o)为0.693 8,平均期望杂合度(H_e)为0.913 4。2个亲本及4个子代组合的两两遗传分化指数F_ST值在0.020 5~0.042 7之间,基因流(N_m)数值在5.607 5~11.957 0之间,表明日本沼虾组合间分化不明显。聚类分析结果表明,TP（太湖♀?鄱阳湖♂）组合与PT（鄱阳湖♀?太湖♂）组合,TT（太湖♀?太湖♂）组合与TH（太湖亲本）组合,PP（鄱阳湖亲本）组合与PY（鄱阳湖♀?鄱阳湖♂）组合分别先聚在一起,之后是TP,PT,TT,TH组合聚在一起,最后再与PP,PY组合聚合在一起。试验结果显示,杂交组合的观测杂合度有明显提高;杂交组合与其它组合分化不明显;杂交子代与母本的遗传结构更相似。本试验通过杂交来分析亲本及子代各个组合之间遗传多样性及遗传结构的变化情况,为日本沼虾资源保护和开发利用提供理论依据。     

舟山群岛四个养殖獐种群遗传多样性和遗传结构

基于ISSR分子标记技术,对来自舟山群岛4个獐(Hydropotes inermis)养殖种群的遗传多样性和遗传结构进行了分析。26条ISSR引物共扩增出286个可分析位点,多态位点百分比(PPL)为64.34%。獐物种水平上的Nei''s遗传多样性指数(H)为0.210,Shannon''s多态信息指数(I)为0.318,各种群的H介于0.157-0.190之间,I在0.228-0.278之间,与已报道的ISSR标记在其它动物物种中的应用结果相比,其遗传多样性较为丰富。Structure软件分析结果显示,所有个体根据遗传信息的不同可以被分为4个组群,每个养殖场内的个体基本上都在各自独立的组群中,这与地理区域的划分相似。另外发现各养殖场獐种群间已表现出了较大的遗传分化(Gst=0.163)。同时,为了解各种群间的遗传关系,计算了各种群间的Nei''s遗传距离,结果显示舟山种群(ZS)和秀山种群(XS)之间的遗传距离最近,为0.045,而岱山种群(DS)和朱家尖种群(ZJJ)间的遗传距离最大,为0.066。基于以上结果,建议加强不同岛屿种群间的个体交流,特别是朱家尖种群(ZJJ)与其它种群间的交流。     

土壤氧气可获得性对双季稻田温室气体排放通量的影响

为探讨土壤氧气可获得性(SOA)对双季稻田温室气体排放的影响,利用静态箱气相色谱法对多种管理措施影响下稻田温室气体排放通量和土壤氧化还原电位(Eh)、pH值及田间淹水深度(H)等3种SOA因子进行了观测。结果表明,甲烷(CH₄)排放最集中的Eh值、pH值和H范围分别为-100-0mV、5 < pH < 6和1-5cm,3个范围内分别观测到48.8%、61.1%和77.0%的CH₄排放,其中H对CH₄排放影响最明显,单独由其就可解释37.8%的CH₄排放通量(P < 0.0001)。对于氧化亚氮(N₂O),观测到较多的负通量,其纯排放最密集的3种SOA因子的范围分别是:0-100mV、5 < pH < 6和1-5cm,而200-300mV是其排放的临界Eh范围,高于此范围N₂O排放极少。厌氧的反硝化过程是双季稻田N₂O产生的主导过程。可为水稻田温室气体排放机理研究提供基础数据。     

西太公鱼微卫星的筛选与分析

利用磁珠富集法从西太公鱼（Hypomesus nipponensis）基因组中筛选到14对多态性微卫星引物,并对来自黑龙江镜泊湖的西太公鱼24个样本进行检测：扩增的等位基因数在3～25个之间,平均为8.4个;观察杂合度（H_O）和期望杂合度（H_E）分别为0.266 7～1.000 0和0.282 5～0.932 2;多态信息含量（PIC）为0.239 2～0.910 6;位点HN-075和位点HN-250显著偏离哈迪-温伯格平衡（P<0.05）,这些多态性微卫星位点可用于公鱼属鱼类种群遗传结构及其相关研究。     

中国部分乌鸡品种血清酶蛋白多态性研究

采用聚丙烯酰胺凝胶电泳法测定鸡血清脂酶(E_s-1),碱性磷酸酶(Akp)、碱性磷酸酶-2(Akp-2)的多态性结果表明,E_s-1由常染色体上等显性等位基因E_s-1^A,E_s-1^B所支配,其中E_s-1^B基因在我国药用和肉用乌鸡中出现频率较高(0.7768～0.8700),而E_s-1^A基因在蛋用型莱航和罗斯鸡中占优势(0.6169～0.6818).Akp、Akp-2均由显隐性基因所控制,其出现频率亦因鸡种不同而表现一定的差异.试验中发现有些个体E_s-1^A基因可进一步区分为E_s-1^A1和E_s-1^A2,从而使所研究鸡种中E_s-1的等位基因数目扩展至3个.     

基于EST-SSR标记的陕西茶树种质资源遗传多样性研究

【目的】明确陕西省茶树种质资源的遗传多样性,为茶树育种提供依据。【方法】以陕西地方和野生种质资源、陕西临近省份以及西南茶区共8个省份的118份茶树资源为材料,利用43对高质量EST-SSR引物,用PROSize 2.0软件统计扩增条带,用Powermarker V3.25分析软件计算主等位基因频率（MAF）、基因型、等位基因数（N_a）、观测杂合度（Ho）、Nei基因多样性指数（H）和多态信息含量（PIC）,用MEGA 6软件进行UPGMA聚类分析,对陕西茶树资源的遗传多样性进行分析。【结果】从118份茶树材料中共检测到310个等位基因和832个基因型。主等位基因频率（MAF）、每个位点等位基因数（N_a）、扩增基因型和多态信息含量（PIC）的平均值分别为0.336 8,7.21个,19.35和0.727 2。Nei基因多样性指数（H）在0.232 3～0.911 0,平均值为0.760 0;观察到的杂合度（Ho）在0.105 7～0.991 8,平均值为0.811 2。聚类分析结果将陕西省茶树种质资源分为3大类群,同一地区的陕西茶树资源多聚在一起;对不同地区茶树资源的聚类结果表明,陕西省茶树资源聚为一支,且与湖北的茶树资源关系较近。【结论】陕西省茶树资源具有丰富的遗传多样性和相对独立的遗传背景,且与湖北省茶树资源遗传关系较近。     

乐清湾和三沙湾缢蛏群体遗传多样性的微卫星分析

利用微卫星标记分析了我国浙江乐清湾翁垟(ZWY)、乐成(ZYC)、南岳(ZNY)、南塘(ZNT)、清江(ZQJ)、湖雾(ZHW)、坞根(ZWG)、海山(ZHS)和芦浦(ZLP) 9个群体和福建三沙湾漳湾(FZW)、梅田(FMT)、三都(FSD)、渔江(FYJ)、白招(FBZ)、霞塘(FXT)、长春(FCC)、沙江(FSJ)和溪尾(FXW)9个缢蛏群体的遗传多样性和遗传分化,以期评价乐清湾和三沙湾内缢蛏群体的遗传结构差异,为我国缢蛏主要原产地种质资源保护和遗传育种提供基础资料。遗传多样性结果表明,浙江乐清湾和福建三沙湾内的缢蛏群体中各位点平均有效等位基因数N_e变化范围分别为8.146~10.457和7.457~9.947,平均期望杂合度H_e分别为0.872~0.909和0.846~0.894,Nei’s基因多样性指数分别为0.863~0.899和0.836~0.886,显示乐清湾和三沙湾缢蛏群体的遗传多样性仍处于较高水平。群体间遗传分化指数（F_ST）结果表明,缢蛏各群体间F_ST为0.000 1~0.052 3。基于Nei’s遗传距离（D_A）构建非加权组平均法（UPGMA）系统树结果显示,18个群体聚类为2支,除ZWY和ZWG群体外,ZYC、ZNY、ZNT、ZQJ、ZHW、ZHS 和ZLP浙江群体单独聚为一支;而ZWY、ZWG群体与FZW、FMT、FSD、FSJ、FBZ、FXT、FCC、FSJ 和FXW 9个福建群体聚类在一起,因此推测浙江的ZWY、ZWG群体可能来自福建,揭示在缢蛏的养殖过程中可能存在引苗现象。     

我国特有植物青檀遗传结构的ISSR分析

青檀是我国特有的第三纪残遗植物,是宣纸纤维来源的重要原材料,现已被列入国家Ⅲ级保护植物。采用简单序列重复区间扩增多态性(ISSR)标记技术,对采自27个地理种群的628个青檀个体的遗传多样性和遗传结构进行了检测。8对引物共得到66个清晰的扩增位点,其中多态性位点63个,多态性位点百分率为95.45%。分析结果表明,青檀在物种水平上具有很高的遗传多样性(PPB=95.45%、Ao=1.9545、Ae=1.5729、He=0.3335、I=0.4980、Hb=0.3437)。在种群水平上的遗传多样性(PPB=69.98%、Ao=1.6998、Ae=1.4449、He=0.2561、I=0.3793、Hb=0.2656)和种群间遗传分化水平(Gst=0.23,Ф_ST=0.25)均处于中等水平。华北地区(30°-42°N)和华南地区(22°-30° N)青檀的遗传多样性和遗传结构的差异并不显著。古老的起源历史、较广的分布范围、异交的繁育系统、风媒种子、长命的生活史以及华南和华北地形和植被的差异可能是导致青檀目前遗传格局的主要原因。结合叶绿体序列(cpDNA)序列的遗传结构特征对青檀的保护策略进行了探讨。     

晴空条件下光合有效辐射中散射辐射的测量模型

为了解决散射辐射地面测量中难以摆脱遮光装置的问题,本研究通过调整传统的直接测量方法,拟建立晴空条件下的散射辐射估算模型。具体方法是,选择晴朗无云的天气条件,在美国华盛顿州普罗瑟市郊无绿色植物覆盖的空旷场地上测试07:30—19:30(间隔15 min)的北向局部散射辐射数据(PAR_f-N)和总体散射辐射数据(PAR_f)。测试同时拍摄天空图像,并计算其灰度特征值(H)和颜色特征值(B)。结果分析表明:1)PAR_f-N与PAR_f之间呈指数函数关系,决定系数r²为0.768;2)天空图像的H值与PAR_f之间呈三次函数关系,r²为0.688;3)结合PAR_f-N、H和B值,及常见气象因素,构建多元线性回归模型,最优模型引入了7个变量,其校正和预测相关系数及校正和预测均方根误差分别为0.951、0.958和27.897、25.526。研究表明该估算模型可在较大程度上提高预测性能及其稳定性。     

显微介导远缘基因鲤家系的建立及遗传多样性分析

运用显微介导远缘杂交技术,将未经遗传修饰的中国对虾(Fenneropenaeus chinensis)总DNA直接导入受体鲤(Cyprinus carpio L·)内,以期通过这种基因组水平的远缘杂交技术改善受体鲤的品质。创建的远缘基因鲤提高了鲤的品质,丰富了鲤的遗传基础。本研究对48尾显微介导远缘基因鲤亲鱼进行标记取样,通过20个具有多态的鲤微卫星引物分析,选出在中度遗传距离(0.5～0.6)的亲本进行繁殖,从子代群体中选出品质性状优良的3个家系为研究对象。利用微卫星标记,分别对3个家系进行了遗传背景分析。结果表明,3个家系的平均有效等位基因数(Ne)分别为2.153 5、2.384 0和2.411 0;平均观测杂合度(Ho)分别为0.482 5、0.430 0和0.445 0;平均期望杂合度(He)分别为0.519 5、0.572 0和0.570 0;平均多态信息含量(PIC)分别为0.430 0、0.471 5和0.494 5。其遗传多样性处于中度多态(0.25PIC0.5)偏高水平。3个家系的基因分化系数(GST)为0.074,家系之间属于中度分化水平。     

Microsatellites reveal strong genetic structure in the common cutworm,Spodoptera litura

The common cutworm, Spodoptera litura(Lepidoptera: Noctuidae), is a voracious agricultural pest. To increase understanding of the migration patterns and genetic diversity of different geographic populations of this species, we analyzed genetic variation in nine microsatellite loci among 576 individuals collected from 17 locations in China and one in Myanmar during 2011–2012. We successfully identified 162 alleles, with an average of 18 alleles per locus and a range of 5 to 34. The mean observed heterozygosity of the 18 populations ranged from 0.18 to 0.98, and the expected heterozygosity ranged from 0.19 to 0.89. For the nine microsatellite markers studied, polymorphism information content ranged from 0.18 to 0.88(mean=0.64). We found low genetic differentiation among the 18 populations(mean F-statistics(F_(ST))=0.05) and high genetic diversity among individuals. Principle coordinates analysis indicated no significant correlation between geographic and genetic distance(r=0.04). The value of N_m(N_m4) estimated using coalescent-based methods suggests strong gene flow with migration. The nine microsatellite markers identified in this study will be beneficial for further investigation of migration patterns and genetic diversity in S. litura.     

Genetic potential analysis of German mirror carp (Cyprinus carpio L.) using microsatellite markers

Using 30 microsatellite markers and combining quantifiable characteristics such as body weight, body length and body width, we evaluated the genetic potential of 3 German mirror carp (Cyprinus carpio L.) populations. Number of effective alleles (Ae), observed (Ho) and expected (He) heterozygosity values and polymorphic information contents (PIC) were all calculated. Two hundred and eighty-seven alleles and 559 genotypes were detected. The DNA fragment length was 109–400 bp. The Hardy-Weinberg Equilibrium was checked and the phenomenon of some disequilibrium was studied according to the χ² test. The results showed that the level of genetic variability was moderate, but genetic potential of Shuanglai population was much lower than that of Huanxin and Songpu breeding populations. PIC of the three populations of German mirror carp were between 0.08787 and 0.5377, both highly and moderately polymorphic markers were 13. The number of the Ae was between 1.1014 and 6.4665. The Ho and He heterozygosity values were 0.0968–0.9892 and 0.0926–0.8554, respectively. The linkage correlation was analyzed using the data of body weight, body length and body width, and 30 loci. The result showed that there existed 2 loci, HLJ319 and HLJ693, associated with body length. The HLJ693 locus was significantly correlated with body weight trait. The HLJ677 locus was linked with body width. And then the result was verified in Recombinant Inbred Lines (RIL) of common carp. It showed that the HLJ319 locus was significantly linked with body length, the same as the result of quantitative trait loci (QTL) location for common carp. __________ Translated from Hereditas (Beijing), 2007, 29(12): 1509–1518 [译自: 遗传 (北京)]     

Genetic diversity analysis of black locust (Robinia pseudoacacia L.) distributed in China based on allozyme markers approach

Using starch gel and polyacrylamide gel electrophoresis, the genetic diversity of 19 populations of black locust (Robinia pseudoacacia L.) distributed in China has been evaluated based on seven allozyme systems. Among the 14 enzyme loci studied, 12 of them showed polymorphic, giving a total of 40 alleles. The average allelic number (A) and the average effective allelic number (Ae) were 3.004 and 2.178, whereas the average of expected heterozygosity (He) and the observed heterozygosity (Ho) were 0.525 and 0.440, respectively. The total genetic differentiation (H _T ) in the 19 populations tested was 0.536, including 0.515 within population (H _S ) and 0.021 among populations (D _ST ). The mean coefficient of genetic differentiation (G _ST ) was 0.038, indicating that 3.8% of genetic differentiation was resulted from the populations, and the rest from those within the population. The genetic distance among the 19 populations was small, ranging from 0.015 to 0.065. Cluster analysis based on AVERAGE showed that populations with closer geographical distribution had higher genetic similarity than that in populations with far away distribution. There was no evident geographic differentiation pattern in Chinese black locust populations at a biochemical level.     

长江下游放流鲢群体遗传多样性的微卫星标记分析

利用12个微卫星分子标记对长江下游4个放流鲢群体进行了遗传多样性分析。在12个基因座位中,共检测到56个等位基因,每个座位检测到的等位基因数为1～7个,其中有10个基因座位具有多态性,多态位点百分率为83.33%,4个群体的平均等位基因数A为3.98,平均有效等位基因数Ne为2.384 0,观测杂合度Ho平均值为0.467 6,期望杂合度He平均为0.490 6,多态信息含量平均值为0.381 2。4个鲢群体的遗传多样性较丰富,与文献报道的下游野生群体大致相当,但明显低于上游野生群体;放流鲢群体的平均观测杂合度均低于各自相对应的平均期望杂合度,表现出一定程度的近交现象。4个放流鲢群体间遗传相似系数为0.937 1～0.971 8,遗传距离为0.028 2～0.062 8,基因分化系数Fst为0.027 5～0.050 6,表明群体间的遗传分化程度较弱。     

菊黄东方鲀野生和人工选育群体遗传变异的SSR和ISSR分析

利用SSR和ISSR两种分子标记技术,对菊黄东方鲀的人工选育群体(SH)和野生群体(YS)的遗传变异和遗传结构进行了联合分析。SSR分析的结果显示,14个微卫星位点总共获得了146个等位基因,选育和野生群体的平均期望杂合度(He)分别为0.730 0和0.832 3,平均多态信息含量(PIC)分别为0.705 1和0.813 6;ISSR结果显示,选育和野生群体的平均Nei’s基因多样性指数(H)分别为0.267 2和0.282 3,平均Shannon信息指数(I)为0.407 2和0.425 6,表明菊黄东方鲀选育群体的遗传变异程度低于野生群体。此外,两种分子标记遗传分化的结果显示,两个群体的遗传分化指数分别为0.061 9(SSR)和0.061 5(ISSR),表明两个群体已经产生了中等程度的遗传分化。综上,本单位菊黄东方鲀的人工选育群体仍然具有相对较丰富的遗传变异以供进一步的遗传改良。     

Analysis of the Genetic Diversity and Origin of Some Chinese Domestic Duck Breeds

Twelve fluorescence-labeled microsatellite markers were used to analyze the genetic diversity of 12 domestic duck breeds and 2 wild duck breeds to determine the relationship and origin of Chinese domestic duck breeds. Gene frequency, effective number of alleles （Ne）, expected heterozygosity （He）, polymorphism information contents （PIC）, inbreeding coefficient in population （Fis）, standard genetic distance （Ds）, and genetic distance （DA） were calculated by FSTAT and distance and phylogenetic analysis after the dates which were output from the Microsatellite-Toolkit software. Genetic distances between 12 domestic duck breeds and 2 wild duck breeds were analyzed by variance analysis. Unweighted pair group method with arithmetic mean （UPGMA） and phylogenetic trees used for cluster analysis were structured. The results indicated that 11 loci had medium- or high-level genetic diversity among the 12 loci, which could be efficiently used in the detection of the genetic parameters of each population. The values of He were 0.5414 to 0.7343, those of PIC proved similar, and those of Fis were 0.1101 to 0.3381 among all populations. All breeds were clustered into three groups by UPGMA phylogenetic trees. Banzui duck was clustered into a separate group. Differences of the DA were analysed by t-test. The results showed that difference in DA between the 12 domestic duck breeds and Lvtou duck and the Banzui duck were very significant （P〈0.01）, indicating that these 12 domestic duck breeds originated from Lvtou wild duck, but not Banzui duck.     

Development of SSR Markers for a Phytopathogenic Fungus,Blumeria graminis f.sp. tritici,Using a FIASCO Protocol

Simple sequence repeats (SSR) have been widely used as molecular markers due to their abundance and high polymorphism. However, up to now, the SSR markers had not been developed in the obligate biotrophic phytopathogenic fungus, Blumeria graminis f.sp. tritici. From (AC)₁₀ and (AG)₁₀ enriched genomic libraries for Bgt, 25 primer pairs were designed using the FIASCO (fast isolation by AFLP of sequences containing repeats) protocol. Five primer pairs exhibited polymorphism with allelic diversity from two to seven alleles and produced 29 alleles in a survey of 90 isolates collected from six provinces (cities) in China, while the others displayed monomorphic. Levels of observed heterozygosity ranged from 0.000-0.044 (mean 0.025) and expected heterozygosity ranged from 0.297-0.816 (mean 0.538). These molecular markers provide a novel source to genetic diversity assays and to genetic and physical mapping of Bgt. SSR markers of Bgt need to be further explored.     

利用微卫星标记分析长鳍鲤、锦鲤和龙凤鲤的遗传多样性

利用20对微卫星标记埘长鳍鲤、锦鲤和龙凤鲤3个鲤鱼群体的遗传多样性和遗传结构进行分析.结果表明:3个群体在20个位点上分别检测到108、164、154个等位基因,各群体等位基因数2～16个不等,平均有效等位基因数分别为4.496、5.695和5.606.3个群体平均期望杂合度分别为0.769、0.806、0.801,多态信息含量分别为0.703、0.761和0.757,遗传多样性指数分别为0.764、0.803和0.799,群体间遗传分化系数为0.071.结果说明3个群体遗传多样性较为丰富.     

长江口江豚的遗传多样性现状及种群动态

长江口地处长江与东海、黄海的交汇处,是窄脊江豚东亚亚种和长江亚种的重叠分布区和死亡率高发区。筛选出16个多态性微卫星位点,对采自长江口的36头窄脊江豚东亚亚种样本作了遗传多样性和种群动态分析。结果显示,除了2个微卫星位点显著偏离了Hardy-Weinberg平衡外,其他14个位点均未偏离Hardy-Weinberg平衡。16个微卫星位点共获得了129个等位基因,平均等位基因数达8. 1个。计算获得的平均观察杂合度(Ho)为0. 733,平均期望杂合度(He)为0. 758,多态性信息含量(PIC)均在0. 5以上。其遗传多样性水平与印度洋江豚和海豹等海洋哺乳动物相近,但明显高于大熊猫、东北虎等陆生珍稀哺乳动物。杂合度检验和Mode-shift模型分析显示,该种群未经历过近期的遗传瓶颈效应。Msvar软件的4次模拟均未发现该种群在历史上发生过明显的有效数量波动,估算的当前有效种群大小约为5 623头。研究显示,在长江口发现的窄脊江豚东亚亚种个体,属于一个大而稳定的种群。