激光诱导沙田柚变异的RAPD研究

选用经初步筛选的２０个短序列引物（１０ｎｔ）和随机抽取的２个随机长序列引物,对激光诱导产生的无核沙田柚（处理组）及未经激光处理的有核沙田柚（对照组）进行了ＲＡＰＤ（随机扩增多态性ＤＮＡ）分析。结果有６个引物扩增出了共２０条多态性片段,从而表明在沙田柚的处理组与对照组之间,存在ＤＮＡ分子水平上差异,也就是说在激光诱导沙田柚无核化的过程中,已经引起了沙田柚后代基因组的显著变异,且这种诱导所引起的性状改     

皖西白鹅遗传多样性的RAPD分析

用随机扩增多态性ＤＮＡ（ＲＡＰＤ）技术对皖西白鹅产区内１１个不同地点的２４只个体一进行遗传多样性分析。从３４个随机引物中筛选出的１２个引物对所有个体的总基因组ＤＮＡ进行了ＰＣＲ扩增。共扩增出８７个ＤＮＡ片段,其中的２２个多态性片段,计算了各多态性片段在群体中分布的频率及各个体间的遗传距离指数。同时根据遗传距离指数对它们进行了聚类分析。作为家养动物的个品种,皖西白鹅的遗传多样性程度较高。     

ISSR 和 RAPD PCR技术对东北地区主推玉米品种的鉴别

运用现代分子生物学简单重复区间序列扩增多态性（ISSR）和随机扩增片段多态性DNA（RAPD）两种分子标记技术， 对吉林省主推6个玉米杂交种及其父本、 母本共计18份基因图谱进行鉴别. 以ISSR为主要检测方法、 RAPD为验证方法，分别从30条ISSR引物中选出4条、 从30条RAPD引物中筛选出2条扩增效果明显、 特异性高的引物. 结果表明， 运用ISSR和RAPD-PCR技术提高了玉米品种鉴定和纯度鉴定的准确性.     

草鱼基因组随机扩增多态性引物及多态性位点的筛选

为了建立草鱼基因组DNA多态性分析的指标体系,应用RAPD技术,从180条10碱基随机引物中筛选出了31条能扩增出多态性DNA片段的引物。用这31条多态性引物共扩增出了327条重复性好、带型清晰、分辨率高的谱带。扩增产物的片段大小范围在400—2000bp之间。单一引物扩增条带为5—14条。用这些多态性引物在草鱼基因组DNA中检测到了93个多态性位点,并对这些多态性位点上的等位基因频率进行了统计分析。这31条多态性引物和所检测到的93个多态性位点初步为草鱼基因组的多态性分析提供了可靠的分析指标体系。     

浙江有尾两栖动物分类与演化的RAPD研究

选用２０个随机短引物和６个随机长引物对来自３个科５个属７个种的有尾两栖动物进行ＲＡＰＤ分析，１２个引物得到清晰的ＤＮＡ扩增图谱，共计２０７条多态性片段。     

草鱼RAPD指纹的初步研究

运用２０个随机引物对草鱼进行了随机扩增多态ＤＮＡ分析。结果表明,所用引物都能对草鱼扩增出稳定的ＲＡＰＤ谱带,扩频的谱带数在２￣１１之间,扩增的片段大小在０．１ｋｂ￣３．０ｋｂ之间．其中,ＯＰＭ－９等６个引物能在草鱼个体间扩增出多态性片段,占引物总数的３３．３％,ＯＰＭ－１等１４个引物对草鱼不同个体扩增的谱带是一致的,占引物总数的６６．７％,为草鱼种质资源及遗传多样性的研究奠定了基础。     

中国鹅5个品种随机扩增多态DNA(RAPD)分析

目的 用随机扩增多态ＤＮＡ（ＲＡＰＤ）技术分析中国鹅的５个品种一皖西白鹅、太湖鹅、浙江白鹅、四川白鹅和狮头鹅的随机扩增多态ＤＮＡ（ＲＡＰＤ）。方法 从３４个１０－寡核苷酸随机引物中筛选出的１４个引物对每一品种的总基因组ＤＮＡ进行了单引物ＰＣＲ扩增,结果 ５个品种共扩增出１７４个ＤＮＡ片段,其中４８个（占２７．６％）在５个品种间表现多态性,根据扩增ＤＮＡ片段的异同计算了各品种间的遗传距离指皖西白鹅和     

花椰菜自交不亲和性的分子标记研究

运用随机扩增多态性DNA(RAPD)和inter-简单重复序列(ISSR)等分子标记技术,分别对5组花椰菜自交不亲和系和对应的自交亲和系的基因组进行指纹差异分析.采用10个10 mer随机引物和5个ISSR引物进行PCR扩增,结果表明,扩增片段分子量在0.3～5 kb之间,指纹图谱的稳定性和重复性较好.经过3次以上重复发现,ISSR4引物扩增图谱中,自交亲和系比不亲和系多扩增出3 800 bp片段;ISSR6引物扩增图谱中,除第3组外,不亲和系比亲和系多扩增出1 100 bp片段.结果表明,花椰菜自交不亲和系与自交系基因组DNA之间存在差异,扩增出的差异片段与花椰菜自交不亲和性相关,并可作为自交不亲和系和自交亲和系育种筛选的分子标记.初步探讨了花椰菜自交不亲和性的遗传机制及RAPD和ISSR技术在自交不亲和系选育过程中的应用前景.     

龙柚芽变系的RAPD分析鉴定

利用100个10bp的随机引物对龙柚的4个芽变系的基因组DNA进行RAPD扩增分析,探讨芽变系遗传物质的变化．在这100个随机引物中,筛选出能在4个芽变系中扩增出稳定多态性片段的引物11个,一共扩增出了63条片段,其中多态性片段有19个,占30．16％．聚类分析的结果,芽变系2和4的遗传距离最小（0．1001）,相似系数最大（0．9048）．表明了芽变引起的遗传物质的改变能够通过无性繁殖的方式存在,用RAPD的方法来分析不同芽变系的遗传关系是可行的．     

果梅RAPD标记不同电泳指纹比较及

利用11个碱基引物, 在对退火温度等反应条件进行严格优化的基础上,以不同品种果梅为材料,同时利用琼脂糖凝胶和聚丙烯酰胺凝胶(PAGE)电泳检测了RAPD扩增产物。结果表明：PAGE电泳检测出的谱带总数量以及多态性谱带的数量分别为204和145, 以琼脂糖凝胶检测的为91和58。根据PAGE电泳系统获得的标记信息对16个果梅品种进行聚类分析,表明所有品种都能被很好地区分,并在一定程度上反映了果梅品种的亲缘关系。通过对20个随机挑选的扩增片段进行克隆与测序发现,20个片段都是对应引物的RAPD扩增产物,在不同的16个序列中有4条是编码蛋白的基因片段,说明了RAPD不仅扩增基因组上的非编码蛋白序列,同时可以扩增编码蛋白的基因片段,是对基因组不同区域的随时机扩增,所以能够客观地反映不同果梅品种间的遗传差异。     

Phenotypic and genetic variation between two populations of the Chinese yellow pond turtle, Mauremys mutica （Cantor, 1842）

Mauremys mutica (Cantor, 1842) is an endangered species in China. Main phenotypic variations in body color, body weight, body shape, clutch size, egg size, and hatchling size were revealed between the southern and northern populations. Both populations have the phenomenon of "larger male" sexual size dimorphism (SSD), especially in the southern population. Furthermore, genetic variations between the two populations were analyzed by RAPD band patterns of 30 random individuals in each population. The average genetic distance was 0.299±0.108 among the samples of two populations. The average genetic distance between southern and northern populations was 0.305±0.046. Cluster analysis indicated that all the individuals from the southern and northern populations were clustered among themselves to form two distinct clades. A total of 20 population-specific RAPD fragments were scored from 16 primers, and could be used as RAPD markers for distinguishing the southern and the northern population. Based on the nucleotide sequences of two RAPD markers, two pairs of SCAR primers (SC1-S and SC2-S) were designed, which could be used as SCAR markers for the southern population. According to the significant phenotypic and genetic variations, we suggested that the northern population and southern population might be considered as two separate taxa, the "northern taxon" and the "southern taxon", and the conservation should be respectively conducted on the two taxa.     

金华地区野生大豆小种群的分子生态学研究

为了阐明小地区范围内野生大豆种群的遗传分化情况,应用随机扩增多态性ＤＮＡ（ＲＡＰＤ）方法,对浙江金华地区５个一年生野生大豆种群,进行了分子生态学研究。用根据ＲＡＰＤ数据计算得 Ｓｈａｎｎｏｎ指数估计了５个野生大豆种群的遗传多样性。发现大部分的遗传变异存在于野生大豆种群间,只有少部分的遗传变异存在于种群内。     

Genetic relationships among Chinese and American isolates of Phytophthora sojae assessed by RAPD markers

The genetic diversity of three geographic populations of Phytophthora sojae from China and the United States was determined using random amplified polymorphic DNA （RAPD）. The purpose was to explore genetic relationships among Chinese and American isolates of the organism. 21 random primers were selected among 200 random primers screened. A total of 223 reproducible RAPD fragments were scored among 111 individuals, of which 199 （89.23%） were polymorphic. Analysis of genetic variation showed that there existed higher genetic variation in the United States population in comparison to the Chinese populations. Nei＇s genetic identity and principal component analysis indicated that the populations of Fujian and United States are closer to each other than to Heilongjiang populations. Shannon-Wiener diversity index revealed that the United States populations have a higher genetic di- versity than that of Chinese populations. These data are in support of the hypothesis that P. sojae in the United States might not have been introduced from China.     

裙带菜种质类群的RAPD遗传分析

利用随机扩增多态生ＤＮＡ（ＲＡＰＤ）技术对裙带菜９个品系的种质类进行分析,共筛选出１８个引物扩增出２２１条清晰可重复的带。进一步分析表明,９个裙带菜品都保持着一定的遗传离距,且有６个品系具有可作为遗传标记的特征带。上述结果在ＮＤＡ水平上提供了裙带菜品进行杂交组配时远缘亲本选择的理论依据,并初步证明了利用ＲＡＰＤ技术对裙带菜进行种质来源分析和种质差异鉴定的有效性。     

油蒿种群遗传分化的RAPD分析

采用随机扩增多态性DNA(RAPD)方法对油蒿(Artemlsia ordosica Krasch．)的5个种群进行了研究．用15个随机引物扩增出297条清晰谱带，其中285条为多态性谱带．利用POPGENE 32软件对数据进行处理，结果如下：(1)油蒿有着丰富的遗传多态性，多态位点百分率达96％，各种群多态位点百分比在84．6％～90．9％之间．(2)油蒿的种群间分化较小Gat=0．1364，86．36％的遗传变异存在于种群内，各种群的遗传一致度都在96％以上(3)聚类分析显示，地理分布近的种群被聚到了一起，反映了油蒿种群的遗传分化和地理距离有着一定的相关性．     

Population Genetic Structure and Genetic Differentiation of the Pufferfish Takifugu Rubripes and Takifugu Pseudommus Revealed by RAPD Analysis

RAPD analysis is used to assay the population genetic structure and genetic differentiation of pufferfish T.rubripes and T.pseudommus.One hundred and twenty fragments are aplified with 21 random sequence 10-mer primers.The proportion of polymorphic fragments of T.rubripes populations from the coast of China(TRC),the coast of Japan(TRJ),general population of T.rubripes both from China and Japan(TRCJ),and the population of T.pseudommus are 31.7%,33.3%,35.0%and 39.2% respectively.The mean expected heterozygosities of the four populations are 0.116,0.125,0.126,and 0.144,respectively.Low genetic distances(ranging from 0.0118 to 0.0309)and Fst estimates(0.020 to 0.024) among the populations indicated that there is no significant differentiation between T.rubripes and T.pseudommus and suggested that only one species is involved.     

Influence of long-period pesticide force on genetic polymorphism of wolf spider Pardosa pseudoannulata (Lycosidae: Araneae)

Wolf spiders are predators in large quantities in the fields. How wolf spiders keep their dominant species status under long-period pesticide force? To answer this question, eight geographical populations of Pardosa pseudoannulata were used as materials to test the influence of geographical habitats on their genomic DNA polymorphism. The RAPD pattern showed polymorphic variations among and within different populations. Total 84 bands amplified by 10 random primers, of which 62 (73.81%) are polymorphic, were generated from 55 individuals of eight geographical populations. Meanwhile, Shannon’s index (Ho= 0.5177) showed a rich genetic diversity of P. pseudoannulata, and most of the genetic variation (64.24%) was found within populations. Multiple regression analysis suggested that it is the climatic variation (such as annual average temperature etc.) that results in adaptive eco-geographic differentiation, and it is the long-period pesticide force that speeds up the genetic differentiation of P. pseudoannulata which changed the genetic diversity of the population.     

Clones identification of Sequoia sempervirens (D. Don) Endl. in Chile by using PCR-RAPDs technique

A protocol of polymerase chain reaction-random amplified polymorphic DNAs (PCR-RAPDs) was established to analyse the gene diversity and genotype identification for clones of Sequoia sempervirens (D. Don) Endl. in Chile. Ten (out of 34) clones from introduction trial located in Voipir-Villarrica, Chile, were studied. The PCR-RAPDs technique and a modified hexadecyltrimethylammonium bromide (CTAB) protocol were used for genomic DNA extraction. The PCR tests were carried out employing 10-mer random primers. The amplification products were detected by electrophoresis in agarose gels. Forty nine polymorphic bands were obtained with the selected primers (BG04, BF07, BF12, BF13, and BF14) and were ordered according to their molecular size. The genetic similarity between samples was calculated by the Jaccard index and a dendrogram was constructed using a cluster analysis of unweighted pair group method using arithmetic averages (UPGMA). Of the primers tested, 5 (out of 60) RAPD primers were selected for their reproducibility and high polymorphism. A total of 49 polymorphic RAPD bands were detected out of 252 bands. The genetic similarity analysis demonstrates an extensive genetic variability between the tested clones and the dendrogram depicts the genetic relationships among the clones, suggesting a geographic relationship. The results indicate that the RAPD markers permitted the identification of the assayed clones, although they are derived from the same geographic origin.     

浙江省及其临近地区益母草的RAPD分析

运用8个有效的10mer-寡核苷酸引物对浙江省及其邻近地区益母草(Leonurus japonicus)7个居群的基因组DNA进行了RAPD(random amplified polymorphic DNA)扩增，共检测到65个位点，其中多态位点45个，占69．23％。应用Jaccard公式计算了7个居群在65个位点上的相似性，并以此为基础，应用主坐标排序法作出了7个居群遗传分化的三维排序图，结果表明，浙江省及其邻近地区益母草的遗传性状总体较为接近，7个居群的遗传分化与其地理分布有一定的联系。