基于mtDNA-COII基因部分序列探讨蟋蟀科五属的进化关系(直翅目:蟋蟀科)

采用DNA测序技术并比较了蟋蟀科5个属:斗蟋属Velarifictorus、棺头蟋属Loxoblemmus、油葫芦属Teleogryllus、灶蟋属Gryllodes和蟋蟀属Gryllus以及蛉蟋科针蟋亚科Allonemobiussocius的mtDNA-COII基因部分序列,依据分子数据建立了5个属的系统发育关系。结果显示A、T、C和G碱基含量分别为34.8%、35.3%、19.3%和10.6%,AT含量(70.1%)仅略高于半翅目锥猎蝽和缨尾目。分子系统树显示,油葫芦属黄脸油葫芦与蟋蟀属家蟋亲缘关系较近。显示油葫芦属与斗蟋属的亲缘关系较近。并且,棺头蟋属的多伊棺头蟋和窃棺头蟋先聚合,这与形态学特征的结果相符。     

基于mtDNA-COⅡ基因部分序列探讨蟋蟀科五属的进化关系（直翅目：蟋蟀科）

采用DNA测序技术并比较了蟋蟀科5个属：斗蟋属Velarifictorus、棺头蟋属Loxoblemmus、油葫芦属Teleogryllus、灶蟋属Gryllodes和蟋蟀属Gryllus以及蛉蟋科针蟋亚科Allonemobiussocius的mtDNA-COⅡ基因部分序列,依据分子数据建立了5个属的系统发育关系。结果显示A、T、C和G碱基含量分别为34．8％、35．3％、19．3％和10．6％,AT含量（70．1％）仅略高于半翅目锥猎蝽和缨尾目。分子系统树显示,油葫芦属黄脸油葫芦与蟋蟀属家蟋亲缘关系较近。显示油葫芦属与斗蟋属的亲缘关系较近。并且,棺头蟋属的多伊棺头蟋和窃棺头蟋先聚合,这与形态学特征的结果相符。     

九种蟋蟀mtDNA-16S rRNA基因序列及其系统进化(直翅目:蟋蟀科)

蟋蟀科5属9个种的线粒体16S rRNA基因部分序列被测定或从Gen Bank获得,比较其同源性,计算核苷酸使用频率,并构建NJ和MP分子系统树。在获得的449bp的序列中A、T、C和G碱基含量分别为31．8％、36．9％、9．9％和21．4％,A T平均含量为68．7％。研究结果表明：所研究的5属9种蟋蟀聚成3个聚类簇,斗蟋属先与灶蟋属汇合,再与棺头蟋属构成聚类簇Ⅰ;油葫芦属黑脸油葫芦和北京油葫芦与蟋蟀属的家蟋相聚构成聚类簇Ⅱ;蟋蟀属的田蟋单独构成聚类簇Ⅲ。     

中国油葫芦属昆虫的RAPD分析（直翅目：蟋蟀总科：蟋蟀科）

用RAPD技术研究了我国油葫芦属Teleogryllus 5种蟋蟀的系统发生关系。研究中使用了15个标本,在所试用的54种随机引物中,有9种引物能扩增出清晰而稳定的多态性片断,多态性片断共计167条。应用UPGMA法对多态性片断进行聚类分析,构建树状图,推测其系统发生关系。每一种蟋蟀先各自聚为一类,聚类结果所呈现的属内种间关系与传统分类研究结果基本一致,且支持黑脸油葫芦与黄脸油葫芦为独立种的观点。     

运用RAPD技术对棺头蟋属昆虫亲缘关系的研究

用RAPD技术研究了棺头蟋属Loxoblemmus 9个种的亲缘关系。研究中每种使用了3个标本,试验所用的54种随机引物中,有9种引物能扩增出清晰而稳定的多态性片断,多态性片断共计193条。根据扩增结果,计算了个体间及种间扩增片断共享度和遗传距离,用UPGMA法进行聚类分析,构建系统树。每个种均各自聚为一类,聚类结果所呈现的属内种间关系与传统分类研究基本一致。     

一种蟋蟀寄生螨的生物学观察

Leplothrombiumsp．是属绒螨科的一种寄生螨,可寄生于北京油葫芦、银川油葫芦、大扁头蟋、斗蟋、长颚蟋等多种农田蟋蟀。其寄生率一般在10％～30％,个别达80％以上。单头寄生螨数通常1～5头,多者达20头以上。该寄生螨1年1代,7至9月幼螨寄生于蟋蟀虫体,8月下旬离开蟋绊进入土壤中,变为若螨。若螨在20cm以下的土壤中越冬,第2年5月变为成螨。6～7月成螨出土活动,卵产于蟋蟀体上。7月中旬,田间开始出现被寄生蟋蟀,8月为奇生盛期,主要寄生姐蟀的中、老龄着虫和成虫,以螫胶吸附在蟋蟀虫体背面、后胸及腹部第亚、2腹节的节间、足基部…     

中国蔗蝗属三种之间亲缘关系的RAPD分析（直翅目：蝗总科：斑腿蝗科）

用RAPD技术对中国蔗蝗属3种蝗虫基因组DNA的多态性进行研究。在事先优化的反应条件下用12个随机引物扩增,共得到179条清晰稳定的多态性片段,片段长度为200—2000bp。统计这些片段,根据扩增片段的共享度计算出相对遗传距离指数,然后用NJ和UPGMA聚类方法对其进行分析,构建系统树,以确定这3种间的亲缘关系。结果表明：等歧蔗蝗与异歧蔗蝗二者的亲缘关系较近,而与斑角蔗蝗的关系较远。     

利用RAPD分析杜氏藻属（Dunaliella）嗜盐种间遗传多样性

本研究首次运用随机引物对杜氏藻属（Dunaliella）中6个嗜盐种的基因组DNA进行RAPD分析。筛选获得的6个有效引物共扩增出98个可重复的DNA片段,其中95条带具有多态性,多态性条带的频率为96．9％。根据系统进化树图将杜氏藻属6个种分别归于亲缘关系相对较远的2个类群中,且6个种与它们各自的生态分布联系不紧密。     

稻蝗属部分种类RAPD及其分子系统学关系

用经筛选的8个随机引物对稻蝗属(Oxya Audient-Serville)中的中华稻蝗(Oxya chinensis)、日本稻蝗(Oxyajaponica)、山稻蝗(0xya agaxisa)、短翅稻蝗(Oxya brachyptera)和小稻蝗(Oxya intricata)等5个种41个个体进行扩增,均扩增出特异性产物,在96条清晰稳定的扩增片段中,多态性片段为95条,种间共有片段1条(引物S362扩增的约850 bp的片段),所获片段的长度大小为200—2500bP,单个引物扩增的条带数在8-16之间,平均为12。根据扩增结果,应用SPSS软件包中的Between-groups linkage法进行聚类分析,得到5种稻蝗的分子系统树。结果显示：中华稻蝗与短翅稻蝗关系密切,日本稻蝗与山稻蝗关系较近,而小稻蝗与它们的亲缘关系较远。聚类图所呈现的种间关系与传统的形态及细胞分类学结果基本一致,5种稻蝗亲缘关系的远近程度得以清楚显示,说明RAPD技术在稻蝗属种间分子系统学研究中具有很好的适用性。     

棺头蟋属六种常见蟋蟀鸣声特征分析与种类鉴定(直翅目:蟋蟀总科)

本文对蟋蟀科Ｇｒｙｌｉｄａｅ棺头蟋属Ｌｏｘｏｂｌｅｍｍｕｓ６种常见种类的鸣声特征进行了较为系统的分析研究。从其频域特征和时域特征上明显地显示了种间差异,并将其鸣声特征用于分类。     

陕西省常见四种实蝇的RAPD研究初报(双翅目:实蝇科)

应用31种随机引物对陕西省常见4种实蝇的基因组DNA进行扩增,筛选出5种引物S61、S107、S126、S275、S1142,可以对4个种扩增出稳定清晰的多态性片段,其中引物S126可以把南瓜实蝇和具条实蝇,具条实蝇和三点棍腹实蝇分开;利用UPGMA法聚类构建的系统树与传统分类完全一致。     

二十八份玉米自交系的RAPD亲缘关系分析

采用RAPD技术,对28份玉米自交系的亲缘关系进行分析。旨在DNA水平上揭示玉米自交系之间的亲缘关系,为进一步提高玉米杂种优势利用水平提供有益的信息从100个10bp随机引物中筛选出24个多态性较好的引物,对28份玉米自交系DNA进行扩增,扩增出24张DNA指纹图谱,其中多态性DNA谱带106条,占总扩增带数的64％。利用DNA扩增结果进行聚类分析,建立了28个玉米自交系的亲缘天系树状图,将供试材料划分为五个类群,RAPD分析结果与已知系谱的亲缘关系基本一致。     

白杨派树种亲缘关系的RAPD分析

运用RAPD技术对白杨派中7个树种和3个人工杂交种共计50个无性系的亲缘关系进行分析.结果表明:筛选出的26条随机引物共扩增产生222条谱带,其中200条为多态性谱带,多态性带达90.09%,且种间遗传变异大于种内无性系间的遗传变异.聚类分析结果显示,参试的50个材料可分为2大类:第一类由山杨、响叶杨、河北杨组成;第二类由毛白杨、银毛杨、84K杨、银白杨、银灰杨、新疆杨和银新杨组成;3个人工杂交种与白杨亚派树种的亲缘关系较近;人工杂交种银毛杨与84K杨间亲缘关系最近.     

五种索科线虫RAPD亲缘关系分析

采用RAPD技术构建了索科线虫4属5种的指纹图谱。从47个引物中筛选出12个稳定性好、多态性高的引物,共扩增出161条谱带,其中150条谱带具遗传多态性,占93·17%。所获片段长度大小为200~3200bp,单个引物扩增的条带数在11~16之间,平均为13·42条。采用RAPDistance软件及MEGA程序,计算Nei氏相似系数和遗传距离,建立UPGMA和NJ聚类图,两个聚类图拓扑结构相同,将5种索科线虫分为两大分支:同属于蚊幼寄生罗索属线虫的食蚊罗索线虫(Romanomermisculicivorax)与武昌罗索线虫(R.wuchangensis)亲缘关系最近,先聚在一起,再与同翅目(Homoptera)寄生长沙多索线虫(Agamermischang-shaensis)聚为一支;鳞翅目(Lepidoptera)寄生中华卵索线虫(Ovomermissinensis)和同翅目寄生两索属线虫(Amphimermissp·)亲缘关系较近,两者聚为一支。5种索线虫属内种间的遗传距离较小,食蚊罗索线虫与武昌罗索线虫之间遗传距离仅为0·1789;而属间遗传距离较大,在0·4471~0·5488之间。上述结果表明:RAPD技术可以应用于索科线虫亲缘关系的分析,能够反映出不同线虫间的遗传差距,从而成功地进行属、种的分类及进化问题研究。     

Evolutionary distances and identification of Candida species in clinical isolates by Randomly Amplified Polymorphic DNA (RAPD)

Fast and reliable identification of different species of the genus Candida is important to define adequate therapeutic decisions, because the different species have highly variable susceptibilities to antifungal drugs; azoles and amphothericin B. Accurate statistical records on case history and epidemiological studies also depend on effective identification. To address this problem we established a RAPD method that enabled direct identification of five very common species of Candida. Initially, reference band patterns were established for C. albicans, C. tropicalis, C. parapsilosis, C. glabrata and C. krusei. One of the primers, M2, showed remarkably conserved intra-specific patterns of approximately 10 bands each, ranging in size from 2.0 to 0.1 kb. These patterns were significantly different and species-specific. Few bands were conserved between different species of Candida, which was assumed to be consistent with their phylogenetic relatedness. In addition, band patterns were constant and reproducible and DNA isolated from single colonies yielded sufficient DNA for identification. The reference band patterns were then used, in blind experiments, to identify species of Candida in 50 randomly chosen samples, including clinical isolates and ATCC strains. RAPD results were 100% consistent with results obtained by conventional diagnostic methods and were achieved in one day instead of several days taken by conventional methods. Because ideal identification methods should be consistent with phylogeny and taxonomy we tested whether RAPD could be used to calculate genetic distances. Comparison of RAPD phylogenetic trees with 18S rRNA trees showed significant differences in tree topologies which indicated that RAPD data could not accurately measure the relative distances between different species. Also, computer simulations of RAPD random patterns were used to test whether the observed degree of RAPD band pattern similarities could occur at random. These simulations suggested that the level of inter-specific band pattern similarities observed in our data could be obtained at random, while intra-specific pattern similarities could not. RAPD would be helpful to discriminate between isolates but not to quantitate the differences. We suggest that the inaccurate estimate of genetic distances from RAPD is a general limitation of the technique and not a specific problem of our identification method. Because of the repetitive character of the target sequences, genetic distances calculated from RAPD could be affected by paralogy, namely, recombination and duplication events not parallel with speciation events. This revised version was published online in June 2006 with corrections to the Cover Date.     

RAPD and internal transcribed spacer sequence analyses reveal Zea nicaraguensis as a section Luxuriantes species close to Zea luxurians

Genetic relationship of a newly discovered teosinte from Nicaragua, Zea nicaraguensis with waterlogging tolerance, was determined based on randomly amplified polymorphic DNA (RAPD) markers and the internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA using 14 accessions from Zea species. RAPD analysis showed that a total of 5,303 fragments were produced by 136 random decamer primers, of which 84.86% bands were polymorphic. RAPD-based UPGMA analysis demonstrated that the genus Zea can be divided into section Luxuriantes including Zea diploperennis, Zea luxurians, Zea perennis and Zea nicaraguensis, and section Zea including Zea mays ssp. mexicana, Zea mays ssp. parviglumis, Zea mays ssp. huehuetenangensis and Zea mays ssp. mays. ITS sequence analysis showed the lengths of the entire ITS region of the 14 taxa in Zea varied from 597 to 605 bp. The average GC content was 67.8%. In addition to the insertion/deletions, 78 variable sites were recorded in the total ITS region with 47 in ITS1, 5 in 5.8S, and 26 in ITS2. Sequences of these taxa were analyzed with neighbor-joining (NJ) and maximum parsimony (MP) methods to construct the phylogenetic trees, selecting Tripsacum dactyloides L. as the outgroup. The phylogenetic relationships of Zea species inferred from the ITS sequences are highly concordant with the RAPD evidence that resolved two major subgenus clades. Both RAPD and ITS sequence analyses indicate that Zea nicaraguensis is more closely related to Zea luxurians than the other teosintes and cultivated maize, which should be regarded as a section Luxuriantes species.     

Use of RAPD technique in evolution studies of four species in the family Canidae

The RAPD-PCR technique was applied to identify genetic markers able to distinguish between four canid species: the arctic fox (Alopex lagopus), red fox (Vulpes vulpes), Chinese raccoon dog (Nyctereutes procyonoides procyonoides) and six breeds of the domestic dog (Canis familiaris). A total of 29 ten-nucleotide arbitrary primers were screened for their potential use in the differentiation of these species. Ten primers amplified RAPD profiles that made it possible to distinguish between the investigated taxa. A number of species-specific bands was scored within RAPD profiles produced by these primers: 35.6% of all the polymorphic bands were unique to the Chinese raccoon dog, 29.6% were unique to the domestic dog, 21.2% were diagnostic for the red fox and 13.6% for the arctic fox. No breed-specific fragments were amplified from canine DNA; however, three primers produced bands characteristic for the dog, but not present in all of the investigated breeds. A Neighbor-Joining tree constructed on the basis of the analysis of RAPD profiles amplified by six primers revealed that the phylogenetic distance between the dog and the arctic fox is larger than the distance between the dog and the red fox. The phylogenetic branch of the Chinese raccoon dog was the most distinct on the dendrogram, suggesting that this species belongs to a different phylogenetic lineage. Obtained results make it possible to conclude that RAPD analysis can be a powerful tool for developing molecular markers useful in distinguishing between species of the family Canidae and for studying their phylogenetic relations.     

鹅膏菌属真菌RAPD分析及亲缘关系的研究

本文对采自湖南莽山的２６种鹅膏菌属（Ａｍａｎｉｔａ）真菌进行了随机扩增多态性ＤＮＡ（ＲＡＰＤ）分析,４０个随机引物中筛选出扩增效果较好的６个引物,每个引物能产生１～１０条ＤＮＡ条带,获得的ＲＡＰＤ谱带清晰并呈现多态性ＯＰＧ１５、ＯＰＨ０４两个引物扩增的ＲＡＰＤ谱带能将２６种鹅膏菌完全区分开来,通过６个引物的ＲＡＰＤ分析获得的平均相似性系数表明种与种之间的相关系数在２０－６０％之间,平均链锁聚类分析可将２６种鹅膏菌分为二大类,且一些具菌环和苞状菌托的种类聚在一起,与形态分类基本相吻合。     

用RAPD技术探讨抗病草鱼F_7(83-2系鱼F_2)与原代双亲的亲缘关系

用RAPD技术对抗病草鱼F7(83-2系鱼F2)与原代亲本草鱼、兴国红鲤进行亲缘关系的研究.在事先优化的反应条件下,在使用的60个随机引物中,有10个引物扩增出清晰稳定的片段,共计161条,大小在200 bp～2 500 bp之间.3种鱼均有其特异性扩增片段,可作为种类鉴别的依据.根据BAPD data analyzer 1.3v软件所得数据,用UPGMA和NJ程序构建的分子系统树显示:抗病草鱼F2与草鱼的亲缘关系较近、与兴国红鲤的亲缘关系较远.两种聚类的结果相一致.研究结果与借助形态学和生化特征进行的传统系统分析具有一致性.在3个群体之间遗传距离矩阵中,抗病草鱼F,与草鱼的平均遗传相似度为0.6801,与兴国红鲤的平均遗传相似度为0.5961.通过分析,认为RAPD技术对分析杂交选育鱼的亲缘关系具有一定的适用性.