Genetic Variation in <i>Salamandra</i><i>infraimmaculata</i>from Different Habitats Using Amplified Fragment Length Polymorphism

The purpose of the present study was to examine the genetic variation in Salamandra infraimmaculata from different breeding site habitats using the Amplified Fragment Length Polymorphism (AFLP) method. The results of the dendogram from a hierarchical cluster analysis show that the grouping of S. infraimmaculata as cluster 5 differs from all the other clusters, including the St1 (Tel-Dan stream) population, which was the most predictable. Five Haplogroups (Hg) were characterized. The mean number of alleles per locus in each population (Ne) ranged from 10.566 (Sp1) to 2.720 (Po6). An average estimated heterozygosity (He) by population ranged from 0.100 (Po6) to 0.186 (St1). Population St1, a permanent breeding site where water was available all year round, exhibited the highest level of polymorphism, while population Po6, from the ephemeral breeding site, exhibited the lowest level of polymorphism. Gene flow between clusters showed that clusters 3 and 4 are sources of migrants and also receive gene flow, while clusters 1 and 2 may be a source of migrants but may not receive much gene flow. A phylogenetic analysis, based on clustering using Nei’s genetic distance, demonstrated that the Tel-Dan population is located on a separate branch within its sub-population. The conclusion of the present study shows that the genetic divergence among isolated populations is not correlated to distance but is affected by the variation of habitats.

A Polymerase Chain Reaction Based DNA Fingerprinting Technique: Amplified Fragment Length Polymorphism for Molecular Typing of Perchlorate Reducing Bacteria

Genetic profiling of environmentally important organisms is very essential for easy identification of biodegrading bacteria. In the previous study, we have reported the perchlorate biodegrading bacteria and characterized them by biochemical analysis and 16 S sequencing. We have observed a very similar isolates of Arthrobacter (Actinobacteria) degrading 4.1 mM and 4.7 mM of ammonium perchlorate [1] 08D0C9EA79F9BACE118C8200AA004BA90B02000000080000000E0000005F005200650066003300390038003100390037003300340037000000 . In this study, we report PCR based DNA fingerprinting technique to generate the genomic signature of these closely related group of Arthrobacter species. This study also effectively generates unique genomic signature for each of these isolates that has potential for use in molecular monitoring as well as for tracking genomic variation and rearrangements.

Genetic Mapping of Laminaria japonica and L. Iongissima Using Amplified Fragment Length Polymorphism Markers in a ＂Two-Way Pseudo- Testcross＂ Strategy

With a ＂two-way pseudo-testcross＂ mapping strategy, we applied the amplified fragment length polymorphism （AFLP） markers to construct two moderate density genetic linkage maps for Laminaria. The linkage maps were generated from the 60 progenies of the F1 cross family （Laminaria iongissima Aresch. × L. Japonica Miyabe） with twenty pairs of primer combinations. Of the 333 polymorphic loci scored in 60 progenies, 173 segregated in a 1：1 ratio, corresponding to DNA polymorphisms heterozygous in a single parent, and the other 58 loci existing in both parents followed a 3：1 Mendelian segregation ratio. Among the loci with 1：1 segregating ratios, 79 loci were ordered in 14 linkage groups （648.6 cM） of the paternal map, and 72 loci were ordered in 14 linkage groups （601.9 cM） of the maternal map. The average density of loci was approximately 1 per 8 cM. To Investigate the homologies between two parental maps, we used 58 loci segregated 3：1 for further analysis, and deduced one homologous linkage group. The linkage data developed in these maps will be useful for detecting loci-controlling commercially important traits for Laminaria.

基于AFLP分子标记的鸭绿江茴鱼遗传多样性分析

为探讨鸭绿江茴鱼群体遗传结构及分化水平,为其种质资源保护提供遗传学依据,利用扩增片段长度多态性(AFLP)方法,对分别采自吉林省内鸭绿江上游、松花江上游和临江金鲨养殖场3个群体的总计90尾鸭绿江茴鱼的遗传多样性进行了比较研究。结果显示,鸭绿江上游、松花江上游和临江金鲨养殖场3个群体的观测等位基因数(Na)分别为1.340 9、1.251 7和1.269 1,有效等位基因数(Ne)分别为1.153 8、1.130 0和1.144 1,Nei’s遗传多样性指数分别为0.093 2、0.078 2和0.086 3,Shannon’s信息指数分别为0.144 7、0.119 8和0.131 9,遗传分化系数(G_(st))为0.176 7~0.254 7,平均值为0.219 4,遗传相似性为0.820 6~0.936 3。UPGMA聚类分析结果显示,松花江上游群体单独聚为一支,鸭绿江上游群体和临江金鲨养殖场群体出现了聚群情况。结果表明,采样的3个鸭绿江茴鱼群体均具有较高的遗传多样性水平,3个群体发生了显著的遗传分化。

Inheritance of Powdery Mildew Resistance in Cucumber (Cucumis sativus L.) and Development of an AFLP Marker for Resistance Detection

Cucumber powdery mildew is one of the most destructive diseases of cucumber throughout the world. In the present study, inheritance of powdery mildew resistance in three crosses, and linkage of resistance with amplified fragment length polymorphism （AFLP） markers are studied to formulate efficient strategies for breeding cultivars resistant to powdery mildew. The joint analysis of multiple generations and AFLP technique has been applied in this study. The best model is the one with two major genes, additive, dominant, and epistatic effects, plus polygenes with additive, dominant, and epistatic effects （E-l-0 model）. The heritabilities of the major genes varied from 64.26% to 97.82%, and susceptibility was incompletely dominant for the two major genes in the three crosses studied. The additive effects of the two major genes and the dominant effect of the second major gene were high, and the epistatic effect of the additive-dominant between the two major genes was the highest in cross I . In cross II, the absolute value of the additive effect, dominant effect, and potential ratio of the first major gene were far higher than those of the second major gene, and the epistatic effect of the additive-additive was the highest. The genetic parameters of the two major genes in cross III were similar to those in cross II. Correlation and regression analyses showed that marker E25/M63-103 was linked to a susceptible gene controlling powdery mildew resistance. The marker could account for 19.98% of the phenotypic variation. When the marker was tested on a diverse set of 29 cucumber lines, the correlation between phenotype and genotype was not significant, which suggested cultivar specialty of gene expression or different methods of resistance to powdery mildew. The target DNA fragment was 103 bp in length, and only a small part was found to be homologous to DNA in the other species evaluated, which indicated that it was unique to the cucumber genome.

Development of Sequence Characterized Amplified Region (SCAR) Primers for the Detection of Resistance to Sporisorium reiliana in Maize

Head smut of maize （Zea mays L.）, which was caused by Sporisorium reiliana, occurred in most of the maize growing areas of the world. The purpose of this study was to develop SCAR markers for map-based cloning of resistance genes and MAS. Two sets of BC3 progenies, one （BC3Q） derived from the cross Qi319 （resistance）×Huangzao 4 （susceptible）, the other （BC3M） from Mol7 （resistance）× Huangzao 4 （susceptible）, were generated. Huangzao 4 was the recurrent parent in both progenies. A combination of BSA （bulked segregant analysis） with AFLP （amplified fragment length polymorphism） method was applied to map the genes involving the resistance to S. reiliana, and corresponding resistant and susceptible bulks and their parental lines were used for screening polymorphic AFLP primer pairs. One fragment of PI3M61-152 was converted into SCAR （sequence charactered amplified fragment） marker S130. The marker was mapped at chromosome bin 2.09, the interval of a major QTL region previously reported to contribute to S. reiliana resistance. Furthermore, S130 was highly and facilitate map-based cloni associated with resistance to S. reiliana, and could be useful for marker-assisted selection ng of resistance genes.

Genetic Relationships among Prunus mume var. pendula Using AFLP Markers

Genetic relationships among Prunus mume var. pendula were studied by using AFLP markers. 18 accessions representing 14 cultivars of Prunus mume var. pendula were selected from the germplasm collection at the Research Center of China Mei Flower. Seven Mse I-EcoR I AFLP primer combinations revealed 450 legible bands, and 269 of which were polymorphic markers. A similarity matrix was prepared using the simple matching coefficient of similarity and Neis (72) distance coefficient. A UPGMA dendrogram demonstrated the genetic relationships of the cultivars. The information given by AFLP markers was basically consistent with the morphological classification and the evolutionary history of the morphotypes, and roughly supported the new revised classification system for Chinese Mei Cultivars. But there were still several exceptions: 1) the Guhong Chuizhi inserted between the Tiaoxue Chuizhi and the Danfen Chuizhi; 2) the Wufu Chuizhi kept off the Pink Pendant Form, and the Moshan Chuizhi was removed from Viridiflora Pendant Form; 3) the Danbi Chuizhi and the Shuangbi Chuizhi of Viridiflora Pendant Form got together well but fell within the Pink Pendant Form.

An AFLP marker linked to the leaf rust resistanc gene LrBi16 and test of allelism with Lr14a on chromosome arm 7BL

Leaf rust(LR), caused by Puccinia triticina, is one of the most widespread diseases of common wheat(Triticum aestivum L.) worldwide. The LR resistance gene Lr Bi16 has been mapped on chromosome arm 7BL in Chinese wheat cultivar Bimai 16 and was closely linked to SSR loci Xcfa2257 and Xgwm344 with genetic distances of 2.8 c M and 2.9 c M, respectively. In the present study, a total of 304 AFLP primer pairs were used to screen Bimai 16 and Thatcher and resistant and susceptible DNA bulks. The polymorphic AFLP marker P-ATT/M-CGC173 bp was used to genotype F2 and F3populations to identify markers more closely linked to Lr Bi16. Marker P-ATT/M-CGC173 bp was tightly linked to Lr Bi16 with a genetic distance of0.5 c M. As Lr Bi16 was mapped near the Lr14 a locus, 809 F2 plants from the Bimai 16/RL6013(Lr14a) cross were inoculated with the Pt pathotype FHNQ to test the allelism of Lr14 a and Lr Bi16. All of the F2 plants were resistant to FHNQ(IT between; and 2), suggesting that Lr14 a and Lr Bi16 are allelic.

Development of a SCAR marker for male gametophyte of Gracilariopsis lemaneiformis based on AFLP technique

The red alga Gracilariopsis lemaneiformis(Bory) is an economically valuable macroalgae. As a means to identify the sex of immature Gracilariopsis lemaneiformis, the amplifi ed fragment length polymorphism(AFLP) technique was used to search for possible sex- or phase-related markers in male gametophytes, female gametophytes, and tetrasporophytes, respectively. Seven AFLP selective amplifi cation primers were used in this study. The primer combination E-TG/M-CCA detected a specifi c band linked to male gametophytes. The DNA fragment was recovered and a 402-bp fragment was sequenced. However, no DNA sequence match was found in public databases. Sequence characterized amplifi ed region(SCAR) primers were designed from the sequence to test the repeatability of the relationship to the sex, using 69 male gametophytes, 139 female gametophytes, and 47 tetrasporophytes. The test results demonstrate a good linkage and repeatability of the SCAR marker to sex. The SCAR primers developed in this study could reduce the time required for sex identifi cation of Gracilariopsis lemaneiformis by four to six months. This can reduce both the time investment and number of specimens required in breeding experiments.

利用AFLP指纹技术研究中国沿海真鲷群体的遗传变异和趋异

用AFLP指纹技术对中国沿海真鲷群体进行遗传变异和趋异分析 ,使用了 6对引物 :E -ACC/M -CAT、E -AGG/M -CTG、E -AGG/M -CTT、E -AAC/M -CTA、E -ACA/M -CTG、E -AAC/M -CAA ,在威海、厦门和北部湾海区三个野生群体共 42个体中检出了 6 46个扩增位点 (片段大小 5 0～ 40 0bp) ,其中 70 %位点在群体内或群体间呈现多态。三个群体的多态位点比例变化在 5 8.4%～ 6 4.0 % ,遗传相似系数变化在 0 .842 5～0 .82 0 0 ,以威海群体的遗传变异量最大 ,北部湾群体的最小。三群体间遗传距离和UPGMA谱系图显示 ,厦门海区与北部湾的真鲷差异较小 ,可以认为是同一个亚种群的不同群体 ,黄海真鲷与前二个群体差异比较大 ,属于另一个不同的亚种群。

AFLP法构建人参、西洋参基因组DNA指纹图谱

目的 采用扩增片段长度多态性DNA(AFLP)分子遗传标志技术 ,分析人参、西洋参基因组DNA多态性。方法 人参、西洋参干燥根基因组DNA ,经EcoRI/MseI酶切并与其相应的人工接头连接后 ,使用选择性引物进行PCR扩增。结果 经变性聚丙烯酰胺凝胶电泳检测 ,成功构建出多态性丰富和重复性好的人参、西洋参DNA指纹图谱。结论 AFLP法有望成为一种独立的切实可行的手段 ,将在人参、西洋参等药用植物的鉴定、生物进化、系统发育研究及指导道地性药材的科学栽培等方面发挥重要作用。

AFLP分析中多态性扩增产物的回收、克隆及鉴定

本研究在摸索和优化了水稻ＡＦＬＰ分析体系的基础上，发展了多态性ＡＦＬＰ产物的高效克隆方法。特异ＡＦＬＰ扩增产物直接从变性聚丙烯酰胺凝胶上分离纯化，再经过一至二轮ＰＣＲ扩增，即可高效地克隆于ｐＧＥＭ－Ｔｅａｓｙｖｅｃｔｏｒ系统中。本实验利用该方法成功地克隆了水稻温敏核不育等位突变系５４６０Ｓ和５４６０Ｆ间的４个多态性ＡＦＬＰ产物，Ｓｏｕｔｈｅｒｎｂｌｏｔｉｎｇ分析证明其中３个产物在水稻基因组中为单拷贝序列，另一个为低拷贝序列。ＡＦＬＰ技术强有力的多态性检出能力再结合多态性扩增产物的高效克隆方法，为寻找与目标基因紧密连锁的分子标记提供了有力工具。

黄姑鱼群体遗传多样性的AFLP分析

对青岛和厦门黄姑鱼群体的遗传多样性进行了AFLP分析,5对选择性引物在两个群体47个个体中,共扩增出461个位点,多态位点265个。青岛和厦门群体的多态位点比例、Nei遗传多样性指数和Shannon遗传多样性指数分别为51.70%、51.99%,0.1022、0.0996,0.1643、0.1622;两个群体遗传多样性在同一水平上。基因分化系数Gst、Shannon遗传多样性指数和AMOVA分析均显示黄姑鱼的遗传变异主要来源于群体内个体间,而群体间无明显的遗传分化。群体的显性基因型频率分布和位点差异数分布显示两个群体有基本相同的群体遗传结构。结果表明,黄姑鱼青岛和厦门群体间无明显的遗传差异,群体间有明显的基因交流。

基于AFLP分子标记的核桃核心种质的构建

【目的】构建核桃核心种质以便更好地保存、评价和利用丰富的核桃种质资源。【方法】利用AFLP分子标记,对131份核桃原始种质采用逐步聚类法建立候选核心种质,比较不同候选核心种质的多态性位点数、多态性位点百分率,结合形态学指标及地理来源,最终确定核桃核心种质,并进行评价。【结果】建立的核桃核心种质保留了原始种质10%的样品,分别为河北的天桥1号、陕西的西洛2号和西林1号、山西的晋龙1号、山东的丰辉、辽宁的辽宁8号和辽73013、新疆的温185、河南的绿波、北京的北京746、美国引进品种维纳、日本引进品种清香和朝鲜的品种安边1号。根据遗传多样性评价结果,与原始种质相比,核心种质的多态性位点保留率为75.4%。【结论】构建的核桃核心种质能较大程度地代表原始种质的遗传信息,符合核心种质要求。

家蚕AFLP分子连锁图谱的构建及绿茧基因定位

利用改进的AFLP技术 ,对家蚕品系C10 0 和大造的回交一代BC1群体进行连锁图谱的构建。经 2 8对引物组合的选择性扩增 ,共获得了 395 6条带 ,平均每对引物产生 14 1 3条带 ,获得多态性带只有 10 18条 ,多态性带的比率为 2 5 7%。其中 6 93(6 8 1% )个多态性位点符合 1∶1孟德尔分离比例。利用Mapmaker/Exp 3 0软件进行连锁分析 ,构建了一张含有 4 0 8个标记位点、33个连锁群、总图距为 36 76 7cM的连锁图谱 ,并将绿茧基因定位在该图谱的 2 2连锁群上 ,表明该连锁群与家蚕经典遗传学的第

荔枝AFLP分析体系的建立

以39份荔枝种质资源为试材,建立起荔枝AFLP分析体系。对AFLP分析过程中的影响因子(酶切与连接、预扩增、标记)进行了研究。双酶切必须完全彻底。选出适于荔枝AFLP分析的引物组合3对,分别为EcoRIAAC+MseICTG,EcoRIAGC+MseICAT,EcoRIACC+MseICAT。应用其中2对引物(EcoRIAAC+MseICTG,EcoRIACC+MseICAT),共在106个位点扩增出条带,多态性带92个(86.7%)。对荔枝AFLP分析的影响因子进行了分析。

丹参不同栽培农家类型的AFLP鉴定

鉴定丹参种内不同类型间的遗传差异,为丹参的进一步系统选育研究提供一定的理论依据。采用扩增片段长度多态性DNA(AFLP)分子遗传标记技术,对不同类型丹参的基因组DNA进行酶切、连接、预扩增后采用选择性引物进行PCR扩增。7对引物扩增后得到899条清晰的带,61条为特异条带,其中丹参种内四种不同类型的特异位点分别占总的特异位点的22.95%,29.51%,26.23%,21.31%,说明丹参种内不同类型间的遗传差异较大,其遗传多样性是丰富的。聚类分析结果显示,四个类型中小叶型丹参(XY)与其它3个类型间遗传距离较远,皱叶型丹参(ZY)与单叶型丹参(DY)遗传距离最近,由此可初步将小叶型丹参确定为一个变种,丹参原型(ZC)与皱叶型丹参也可初步定为丹参的栽培变种。结论:AFLP技术可以作为鉴别丹参种内不同类型间遗传差异的手段,丹参种内存在较为丰富的遗传变异。

福建近海蓝圆鲹种群遗传多样性的AFLP分析

以往研究表明,福建近海的蓝圆鲹分属2个地理种群,即东海西部种群和闽南—粤东近海地方种群。为研究这两个群系的遗传结构,对蓝圆鲹闽东(30尾)和闽南(32尾)种群进行了AFLP分析,8对选择性引物在2个种群62个个体中,共扩增出563个位点,其中多态位点364个。闽东和闽南种群的多态位点比例、Nei遗传多样性指数和Shannon遗传多样性指数分别为62.70%、58.97%,0.1875、0.1809和0.2878、0.2763。与其他鱼类对比显示,福建近海蓝圆鲹种群的遗传多样性水平高,说明种群遗传结构尚未遭到明显破坏;基因分化系数GST、Shannon遗传多样性指数和AMOVA分析均显示蓝圆鲹的遗传变异主要来源于种群内,而种群间无明显的遗传分化。Nm显示2个种群间基因交流频繁。种群的显性基因型频率分布和位点差异数分布显示2个种群有基本相同的群体遗传结构。结果表明,蓝圆鲹闽东和闽南种群间无明显的遗传差异,因此可将福建海域的蓝圆鲹划归同一个管理保护单元。较强的扩散能力及海洋环流可能是造成福建近海蓝圆鲹种群间遗传同质性较高的原因。

鳡鱼群体遗传多样性的AFLP分析

利用AFLP分子标记技术对长江下游苏州段野生和野生F1代人工养殖的2个鳡鱼(Elopichthys bambusa)群体遗传多样性进行分析研究.6对选择性引物在2个群体47个个体中,共扩增出313个位点,多态位点47个.鳡鱼野生和野生F1代人工养殖2个群体的多态位点比率和Shannon’s指数分别为13.42%、8.31%,0.0544、0.0320;前者遗传多样性较后者略高.基因分化系数Gst和Shannon’s指数分析均显示2个鳡鱼群体之间出现一定遗传分化.鳡鱼UPGMA系统树有分支出现且依据群体分别聚类,表现出一定的遗传趋异.结果分析表明,鳡鱼群体的遗传多样性相对贫乏;野生F1代人工养殖群体尚未形成自己独立的遗传结构,但2个群体间已经产生了一定的遗传分化,经过较多世代的人工繁育有可能形成自己独立而稳定的遗传结构.

石斛属4种植物的AFLP分析

目的 研究石斛属植物 DNA指纹图谱 ,初步探讨 AFL P分子标记技术在石斛地道性鉴别上的应用。方法采用扩增片段长度多态性 (AFL P)技术对石斛属内石斛组 4个种和一个外类群种进行基因组 DNA多态性分析。结果 从 6 4对引物组合中选出 5对引物组合构建了 5个种的 DNA指纹图谱。通过聚类分析 ,石斛属 4种植物聚成一个大类 ,彼此关系得到了很好的分辨 ,并获得 bootstrap校验的有力支持。结论 AFL