野生斑鳠西江地理群的遗传多样性分析

为了解斑鳠(Mystus guttatus)不同地理种群之间的遗传分化程度,基于RAPD技术,分析了珠江水系西江段野生斑鳠的遗传多样性。利用20个随机引物对30个斑鳠个体的基因组DNA进行了PCR扩增,共扩增出3210条DNA片段,平均每个个体扩增出107条带。在检测到的107个位点中,多态位点数为48个,占44.9%,仅有一个引物S30没有扩增出多态带。个体间最大的遗传距离0.2804,最小的遗传距离0.0467,平均遗传距离0.1526±0.037,种群内个体间平均的相似率为84.7±3.7%。     

窄冠型杨树杂交子代SRAP鉴定及其遗传变异研究

为鉴定杨树杂交子代真实性及阐明子代群体的遗传关系,本研究以窄冠型杨树的7个杂交子代群体和亲本为试验材料,并采用SRAP分子标记技术对其进行子代真实性鉴定及遗传变异研究。从110对引物中筛选出了8对多态性高,主条带清晰且稳定的引物,对7个杂交子代群体扩增,共产生126条谱带,其中多态性带为118条,多态带比例(PPB)达93.65%;基于Pop Gen 32软件对7个杂交组合的遗传多样性分析表明,派内杂交时,黑杨派内杂交子代的遗传多样性水平低于白杨派内的遗传多样性水平;派间杂交时,白杨做母本的子代遗传多样性比黑杨做母本的遗传多样性丰富。对其中4个杂交组合的当年杂种苗的生长量、形质指标与其亲本间遗传距离进行相关性分析,结果表明,亲本间遗传距离与苗木的地径、苗高成正相关,与其分枝角度成负相关。70个杂交子代均具有父本特征带,且经鉴定全部为真杂种,本试验结果为杨树杂交亲本的筛选及杂交子代早期鉴定提供了一定的理论依据和技术参考。     

雁鹅遗传多样性的ISSR研究*

本试验以来自安徽省郎溪县雁鹅原种鹅场的雁鹅为研究材料,应用ISSR技术对雁鹅种群的遗传多样性进行研究。筛选出43个ISSR引物,对94个供试样本进行了扩增,共扩增出216个位点,每条引物扩增出的谱带数在2~7之间,平均为5.0条;多态条带比率在16.67~100%之间,平均为65.60%,材料间遗传距离为0.2～0.45。用不加权算术平均组对法（UPGMA）对94个个体间的关系进行聚类分析,构建了雁鹅种群的遗传系谱图。     

小麦杂种优势群研究III普通小麦和斯卑尔脱小麦微卫星分子标记遗传差异的研究

选用１５份我国不同生态区的普通小麦品种（系）及９份不同国家的斯尔脱小麦品种（系）,利用微卫星分子标记对小麦种间,品种（系）间遗传差异进行了研究,探讨扩大杂交小麦育种亲本遗传基础的途径。所利用的１８对微卫星引物的２４份材料中均有搁增产物,共扩增出分子量小于５００ｂｐ的条带４９５条,其中４６８条带（占９６．７８％）具有多态性,平均每个引物可拉增出２６．６条多态性带。研究发现,９份斯卑尔脱小麦微卫星多态     

月鳢、斑鳢Cyt b基因序列分析与比较

Cyt b基因在进化过程中受到的选择压力较小，具有较快的进化速率，加上容易使用一些通用引物进行扩增和测序，非常适合进行种间系统进化的分析，近年来被广泛应用于鱼类的系统分化和分子分类的研究。月鳢和斑鳢均属鲈形目，鳢科鱼类，是我国南方名贵的水产养殖品种，有着类似的形态特征和相似的生活习性，在生产上人们常将它们统称为乌鱼。在分类上曾将月鳢和斑鳢分属在不同的属，月鳢为月鳢属(Channa)，斑鳢为鳢属(Ophiocephalus)。为了从分子水平研究这两种鱼的亲缘关系，本研究测定了月鳢和斑鳢线粒体Cyt b全基因测序，并进行了比较和分析。     

两个罗非鱼杂交组合亲本群体与子一代的AFLP分析

运用AFLP指纹技术分别对尼罗罗非鱼和奥利亚罗非鱼及它们的杂交子一代、橙色莫桑比克罗非鱼和荷那龙罗非鱼及它们的杂交子一代的遗传结构进行分析。从90对选扩引物组合中筛选出8对进行PCR扩增，8对引物组合共检出250个扩增位点（片段大小100～500bp）。每一个体的每对引物扩增条带数在20～47之间。群体内遗传相似度从高到低依次为：奥利亚罗非鱼（0.913），尼罗罗非鱼（0.871），荷那龙罗非鱼（0.829）和橙色莫桑比克罗非鱼（0.784）。奥利亚罗非鱼和尼罗罗非鱼间的遗传相似度是0.811，遗传距离为0.189；奥利亚罗非鱼和尼罗罗非鱼与杂交子一代的遗传距离分别为0.165、0.142。橙色莫桑比克罗非鱼和荷那龙罗非鱼间的遗传相似度0.756，遗传距离为0.244；橙色莫桑比克罗非鱼和荷那龙罗非鱼与杂交子一代的遗传距离分别为 0.148、0.162。引物组合E-AGG/M-CTT在尼罗罗非鱼、E-ACA/M-CAC在尼奥鱼中扩增出差异的DNA片段，可作为区分奥利亚罗非鱼和尼罗罗非鱼与杂交子一代的分子标记。     

用ISSR分析四川苎麻品种(系)间的遗传关系及雄性不育分子标记的建立

本文从100条ISSR引物中筛选出21条引物,对8个四川苎麻品种(系)间的遗传关系进行了分析。PCR扩增结果表明,21条引物在8份材料中共扩增出86条带,平均每条引物扩增出4.1条,其中多态性位点71个,各引物扩增出的位点数3~8个不等,平均每条引物可以检测到3.4个多态性位点。聚类分析和遗传距离分析结果表明,供试品种川7、川8与其亲本遗传距离较远。3个杂交品种(川7、川8、川9)均表现特有的偏父本遗传现象。此外,本研究用ISSR引物U835筛选到了1个雄性不育分子标记,并将其扩增产物克隆测序,结果表明该序列大小为658bp。根据该序列,此标记被转化成稳定的SCAR标记,可用于苎麻雄性不育分子标记辅助育种。     

RAPD分析野生和养殖太湖秀丽白虾的遗传多样性

摘要：用60个随机引物对野生和养殖的太湖秀丽白虾进行随机扩增多态DNA(RAPD)分析,筛选的23个随机引物共检测到105个位点（200～2000bp）。用PopGen1.32分析结果表明：野生种群多态位点比例为50.47%,养殖种群多态位点比例为43.33%;野生种群和养殖种群的种群内遗传相似系数分别为0.8635和0.8795;野生种群和养殖种群之间的遗传距离为0.0361;野生种群平均Shannon多样性指数h0为0.2830,略高于养殖种群的0.272。     

泰国产方斑东风螺养殖群体遗传多样性的RAPD分析

用随机扩增多态性DNA(RAPD)技术对泰国产方斑东风螺养殖群体的遗传多样性进行检测,从100个随机引物中筛选出21个引物对方斑东风螺的DNA进行扩增,结果表明:21个引物共检测到222条清晰且重复性好的条带,每个引物可扩增出4~16条带,分子量在200~2200bp之间,其中多态位点为156个,占70.27%;群体的Shannon多样性指数为0.2818,Nei基因多样性指数为0.2491;个体间最大遗传距离为0.291,最小遗传距离为0.066。通过与其他贝类遗传多样性的研究结果比较,可初步判断泰国产方斑东风螺养殖群体的遗传多样性比较丰富。     

硬叶兜兰SRAP-PCR体系建立及亲缘关系研究

采用CTAB法对硬叶兜兰叶片总DNA进行提取,通过正交实验设计,用SRAP正向引物me1（5＇-TGAGTCCAAACCGGATA-3＇）和反向引物em2（5＇-ACACACACACACACACT-3＇）对反应体系进行优化;并应用其进行种群遗传差异分析。结果表明：优化的20μL的PCR反应体系中DNA模板为90 ng,Taq DNA聚合酶1.6 U,dNTPs 0.30 mmol/L,MgCl22.0 mmol/L和引物各0.50μmol/L,经重复电泳验证适合进行硬叶兜兰的SRAP分析;利用该体系对7个不同种源的167份硬叶兜兰进行SRAP-PCR扩增,对筛选的10个引物扩增,共得到288条条带,其中多样性条带234条,多态位点百分率（PPB）为81.25%;种群的遗传距离值在0.065 9～0.191 6,UPGMA聚类结果显示供试种群在遗传相似度为0.863时,可以分为2支,即第一分支由马固、斗咀和杨柳井居群组成,第二支由古林箐、夹寒箐、小坝子和田坝居群组成。SRAP标记可较好地适用于硬叶兜兰种群遗传差异的鉴定。     

鰤鱼诺卡氏菌对乌鳢血液指标的影响

应用浓度为106 cfu/mL的鰤鱼诺卡氏菌（Nocardia seriolea）菌悬液腹腔注射乌鳢,人工感染诺卡氏菌病.在感染后3 d、6 d、9 d、12 d、18 d和24 d时抽取乌鳢血液,检测乌鳢的血细胞数、血细胞脆性、溶菌酶、血清总蛋白、碱性磷酸酶、乳酸脱氢酶和尿素氮等指标.结果表明,与对照组相比,感染鰤鱼诺卡氏菌后乌鳢血细胞数呈先升高后降低的趋势,而血细胞脆性在各实验时间点均高于对照组;血清碱性磷酸酶呈先降低后升高趋势,血清溶菌酶、血清总蛋白和血清尿素氮等均呈先降低后增高再降低的趋势,血清乳酸脱氢酶活力显著增加.说明鰤鱼诺卡氏菌感染会引起乌鳢血液指标明显变化.     

四个鲤鱼种群遗传多样性的AFLP分析

本文采用AFLP技术对黑龙江野鲤、黄河鲤、建鲤和荷包红鲤4个鲤鱼种群共96个个体进行了遗传多样性分析。结果显示,8对选择性扩增引物共扩增得到502个位点,其中多态性位点273个,多态性比率为54．38％。同时对4个种群的Shannon多样性指数,Nei’s基因多样性等参数进行了分析,结果表明,4个种群的Shannon多样性指数分别为0．2114±0．2705,0．1825±0．2694,0．1888±0．2587和0．1600±0．2426,Nei’s基因多样性指数分别为0．1398±0．1872,0．1225±0．1863,0．1235±0．1774和0．1036±0．1636;总基因多样性（H4）平均值为0．1721±0．0350;种群内基因多样性（Hs）平均值为0．1224±0．0190;基因分化系数（Gst）为0．2892,种群内的基因多样性占总群体的71．08％,种群间为28．92％,而基因流系数（Nm）为1．2291。另一方面,分子方差分析（AMOVA）结果表明,种群平均近交系数（Fst）为0-31191,变异31．19％来自种群间,68．81％来自种群内。4个种群中黑龙江野鲤的种内多态性比例最高,而荷包红鲤种群最低,并且4个鲤鱼种群当前的种质资源良好,具有一定的种群稳定性;建鲤已经开始分化,与亲本荷包红鲤亲缘关系逐渐分化,逐步形成自己稳定的遗传结构。本研究为探讨鲤鱼种群的遗传特性和遗传分化提供参考,也为其种质资源的保护及合理利用提供科学依据。     

应用RAPD分析川西北高原老芒麦自然居群的遗传多样性

利用RAPD标记对来自青藏高原东南部川西北高原的8个老芒麦自然居群的遗传多样性和群体遗传结构进行了分析和评价。从150个RAPD引物中筛选出25个能扩增出高度重复性条带的引物。这25个引物共扩增出370条可分辨的条带,其中291条(占78.65%) 具有多态性,表明供试居群在物种水平上存在较高水平的变异。同时各居群的多态性位点比率(PP)在46.49%到53.78%之间变化,表明群体水平的变异较低。居群的平均基因多样性(HE)为0.176(变幅为0.159~0.190),而物种水平的平均基因多样性达0.264。基于Nei’s基因多样性、Shannon指数和贝叶斯方法的群体分化系数分别为32.0%、33.7%和33.5%。AMOVA 分析表明居群内遗传达到总变异的59.9%,而居群间变异仅有40.1%,但二者均达到极显著水平(P < 0.001)。居群间每世代迁入个体数(Nm)达到0.503个。各居群间存在较高的Nei’s遗传一致度。本研究获得的老芒麦的遗传结构不同于已报导的大多数披碱草属物种。另外,基于聚类分析及AMOVA的结果均表明各居群间存在较为明显的地理分化,8个居群分化为采集地的南部和北部2个分支。总之,研究结果表明来自青藏高原东南部的老芒麦居群具有较高水平的遗传变异。在该地区应尽量选择遗传多样性高的老芒麦居群实施就地保护。     

Assessment of genetic diversity in Amomum tsao-ko Crevost & Lemarié, an important medicine food homologous crop from Southwest China using SRAP and ISSR markers

Amomum tsao-ko Crevost & Lemarié is an important crop that has been widely used in traditional Chinese medicine and daily diets for a long time. In this study, the genetic diversity and relationships of eight cultivated populations of A. tsao-ko grown in Southwest China were examined using sequence-related amplified polymorphism (SRAP) and inter-simple sequence repeat (ISSR) markers. The results showed that 139 (99.29%) of 140 and 185 (99.46%) of 186 bands were polymorphic by SRAP and ISSR primers amplification, respectively. The polymorphic information content of detected bands were 0.270 (SRAP) and 0.232 (ISSR), respectively. The average Nei’s gene diversity (H?=?0.217) and Shannon’s information index (I?=?0.348) at the species level generated by SRAP primer were higher than those by ISSR analysis (H?=?0.158, I?=?0.272). Genetic differentiation coefficients and molecular variance analysis (AMOVA) indicated that the genetic variance of A. tsao-ko mainly occurred within populations rather than among populations. The high genetic identity among populations was revealed by SRAP (0.937) and ISSR (0.963). Using UPGMA cluster analysis, principal coordinate analysis, and population structure analysis, the accessions were categorized into two major groups. Overall, results obtained here will be useful for A. tsao-ko germplasm characterization, conservation, and utilization.

     

Genetic diversity of <Emphasis Type="Italic">Guizotia abyssinica</Emphasis> (L. f.) Cass. (Asteraceae) from Ethiopia as revealed by random amplified polymorphic DNA (RAPD)

Genetic diversity of 70 populations of niger (Guizotia abyssinica) representing all its growing regions in Ethiopia was investigated using random amplified polymorphic DNA (RAPD) to reveal the extent of its populations genetic diversity. Ninety-seven percent of the loci studied was revealed to be polymorphic for the whole data set. The within population diversity estimated by Shannon diversity index and Nei gene diversity estimates was revealed to be 0.395 and 0.158, respectively. The extent of genetic variation of populations from major niger producing regions was significantly lower than that of populations from other regions; however, it is distributed regardless of altitude of growth. Genetic differentiation between populations was estimated with Shannon index as G^′ _ST (0.432), Nei’s G _ST (0.242) and AMOVA based F _ST (0.350) and appears to be equivalent to the average values calculated from various RAPD based studies on outcrossing species. Higher proportion of the variation detected by AMOVA resided within populations (64.58%) relative to the amount of variation among populations (35.42%). UPGMA cluster analysis showed that most of the populations were clustered according to their region of origin. However, some populations were genetically distant from the majority and seem to have unique genetic properties. It is concluded that the crop has a wide genetic basis that may be used for the improvement of the species through conventional breeding and/or marker assisted selection. Collection of germplasm from areas not yet covered and/or underrepresented is the opportunity to broaden the genetic basis of genebank collection.     

Genetic diversity of anchote (Coccinia abyssinica (Lam.) Cogn.) from Ethiopia as revealed by ISSR markers

Anchote (Coccinia abyssinica) is plant endemic in Ethiopia with a high calcium content grown for its edible tuberous roots. In spite of its importance as food security crop, there is no information available on molecular genetic diversity of this crop. Therefore, the aim of this study was to assess genetic diversity within and among 12 populations of anchote using ISSR markers. Using nine ISSR primers, a total of 87 scorable bands was generated of which 74 were polymorphic. Within population diversity based on polymorphic bands ranged from 13.8 to 43.53 % with a mean of 33.05 %, Nei’s genetic diversity of 0.04–0.156 with a mean of 0.12, Shannon information index of 0.07–0.23 with a mean of 0.175 and analysis of molecular variation (AMOVA) of 51.4 % were detected. With all diversity parameters, the highest diversity was obtained from Gimbi, Bedele and Ale populations, whilst the lowest was from Manna. AMOVA showed a 48.56 % between populations variability and significantly lower than that of within population variation. Population differentiation with F_ST was 48.56 %. From Jaccard’s pairwise similarity coefficient, Decha and Nedjo were most related populations exhibiting 0.76 similarity and Manna and Nedjo were the most distantly related populations with similarity of 0.52. The only pentanucliotide primer used in the study, Primer 880 (GGAGA)3, showed a unique band in some individuals that appeared to be associated with morphological quantitative traits (lowest seed number, high protein content, largest fruit size and smallest vine length). Illubabor and Gimbi populations exhibited highest genetic diversity so that the populations should be considered as the primary sites in designing conservation areas for this crop.     

Analysis of genetic diversity in Turkish sesame (Sesamum indicum L.) populations using RAPD markers⋆

The genetic diversity of 38 cultivated populations of Sesamum indicum L. from four different regions of Turkey was estimated at the DNA level with the random amplified polymorphic DNA (RAPD) technique. Sixty-one bands were obtained for all populations 78% of which were polymorphic. Analysis of molecular variance (AMOVA) was used to investigate the genetic diversity of the populations which yielded highly significant differences among populations within regions (91.9% of the total genetic diversity). According to AMOVA and Shannon's index that were performed separately for each region, the highest value of genetic variation was observed among Northwest region populations (CV = 7.7; H₀ = 0.304) and lowest in the Southeast regions' populations (CV = 2.6; H₀ = 0.068). Nei and Li's similarity index was calculated and phylogenetic tree was established using the neighbor-joining algorithm. This phenetic analysis grouped 35 of 38 accessions in six groups leaving three highly diverse accessions outside. Wagner phylogenetic method was used to assess the phylogenetic relationships among the populations. In the majority-rule consensus tree, only 7 of the 32 forks showed above 60% occurrence. Using Principal Coordinate Analysis (PCO) of the RAPD data set, the groups were clearly separated along the first three axis. These results indicate that RAPD technique is useful for sesame systematics, and should be valuable for the maintenance of germplasm banks and the efficient choice of parents in breeding programs.     

华东竹黄菌不同居群遗传分化的RAPD分析

为了解竹黄地理居群间的遗传分化,本研究采用随机引物扩增多态性DNA分子标记技术对江苏、安徽和浙江3省的8个居群共32个竹黄样本进行了遗传多样性分析。从50个RAPD引物中筛选得到了5个随机引物,对供试材料的DNA进行扩增,共检测出77个位点,其中多态性位点52个,多态性位点比率为67.53%。UPGMA聚类分析结果表明,这8个居群分为三类：安吉居群、临安居群、宜兴居群、广德居群和泾县居群聚为一类;宁国居群和休宁居群聚为一类;淳安居群单独为一类。遗传多样性分析表明8个竹黄居群中,淳安居群的遗传多样性水平最高,安吉居群的遗传多样性水平最低。Nei＇s基因多样性指数和Shannon信息指数均表明竹黄物种水平的遗传多样性高于居群水平。     

细基江蓠繁枝变种（Gracilaria tenuistipitata var.Liui）群体遗传多样性的ISSR分析

运用ISSR标记对来自深圳、北海和防城港的三个细基江蓠繁枝变种群体进行遗传多样性检测,筛选出的16条ISSR引物共扩增出111条带,引物平均多态位点比例为54.95%。三个群体的多态位点比例深圳群体最高为23.42%,北海群体最低为4.50%;群体平均多态位点比例为11.12%,平均基因多样性（Hs）为0.051 3,表明细基江蓠繁枝变种群体遗传多样性较低。群体遗传分化指数为0.756 5,说明遗传变异主要存在于群体间且群体间的遗传分化水平较高。Mantel检验发现遗传距离与地理距离无关。适应生态环境选择无性繁殖模式可能是细基江蓠繁枝变种群体遗传多样性较低、群体间遗传分化大的重要因素。     

青藏高原野生老芒麦种质醇溶蛋白遗传多样性分析

采用酸性聚丙烯酰胺凝胶电泳(A-PAGE) 对采集自青藏高原的54份野生老芒麦（Elymus sibiricus L.）种质进行遗传多样性分析,获得下述结果:（1）供试材料共分离出42条带纹,多态率达92.86%。4个电泳分区（α、β、γ、ω）的平均Shannon指数为0.4627,Nei-Li遗传相似系数（GS）变异范围为0.2424~0.9767,平均值为0.5822。说明供试野生老芒麦材料具有较为丰富的醇溶蛋白遗传多样性。（2）对所有材料的聚类分析和主成分分析发现,在GS值为0.562的水平上供试材料可聚成4个大类,绝大部分来自于相同或相似生态地理环境的材料聚成一类,主成分分析显示了相似的结果,即醇溶蛋白图谱类型与材料的生态地理环境具有一定的相关性。（3）基于Shannon 多样性指数估算了5个老芒麦地理类群内和类群间的遗传分化,发现类群内遗传变异占总变异的68.17%,而类群间的遗传变异占总变异的31.83 %。（4）对各地理类群基于Nei氏无偏估计的遗传一致度的聚类分析表明,各地理类群间的遗传分化与其所处的地理生态环境具有较高的相关性。