RAPD标记构建水稻分子连锁图

利用随机扩增多态性ＤＮＡ（ＲＡＰＤ）,在一个水稻（Ｏｒｙｚａ ｓａｔｉｖａ Ｌ．）的双单倍体（ＤＨ）群体中发展分子标记,仅用５２ 个ＲＡＰＤ标记建成了一个水稻ＲＡＰＤ分子连锁图。该图覆盖基因组的总长度为８９８．４ ｃＭ （ｃｅｎｔｉｍ ｏｒｇａｎ）,标记间的平均间距为１７．３ ｃＭ,它能与用同一群体构成的ＲＦＬＰ图谱互相补充     

RAPD标记构建家蚕分子链锁图

以大造、Ｃ１０８及其Ｆ２群体构建了１个家蚕的ＲＡＰＤ连锁框架图,该图含ＲＡＰＤ标记位点１８２个,来自大造的１０３个标记分属前２３个连锁群,来自Ｃ１０８的７９个标记分属后１６个连锁群,覆盖基因组的总长度为１１４８．３ｃＭ（ｃｅｎｔｉｍｏｒｇａｎ）,它能与本室用同一群体构建的ＳＡＤＦ图谱相整合,亦可与相应的ＲＦＬＰ图谱相互补充。     

结合SADF与RAPD标记构建家蚕连锁图

用40个选择性扩增多态性引物和137个随机性扩增多态性引物对家蚕Bombyx mori F₂群体进行分子标记的筛选。获得544个符合遗传分离比的多态性位点,并用Mapmaker软件进行连锁分析。在最小LOD值为4,最大重组值为0.2条件下拆分连锁群,并对处于同一连锁群的位点进行合理排序,计算出位点间的重组值后转换为图谱距离,构建了一个家蚕的分子连锁图。     

利用RAPD标记和单株树大配子体构建马尾松的分子标记连锁图谱

利用300个随机的寡核苷酸引物和马尾松(Pinus massoniana Lamb.)种子的胚乳(大配子体)产生的随机扩增多态性DNA(RAPD)分子标记,进行马尾松标记连锁图谱的构建。经筛选共获得64个稳定的RAPD标记,利用多点连锁分析,其中的48个标记分属于13个不同的连锁群(连锁对),该图谱覆盖的基因组总长度约为692.5cM(centimorgan),标记间平均距离约为14.7cM,它为马尾松连锁图谱的构建提供了一个连锁框架。     

利用RAPD标记构建响叶杨和银白杨分子标记连锁图谱

利用ＲＡＰＤ标记响叶杨（ Ｐｏｐｕｌｕｓ ａｄｅｎｏｐｏｄａ Ｍａｘｉｍ ．） ×银白杨（ Ｐ． ａｌｂａ Ｌ．） 的Ｆ１ 群体,按照拟测交的作图策略,分别构建了响叶杨和银白杨的分子标记连锁图谱。实验过程中对６００ 个随机的寡核苷酸引物进行了重复筛选,共选出１２８ 个引物用于作图群体的随机扩增,选择符合１∶１ 分离的拟测交位点。作图群体大小为８２ 个单株（ 包括双亲） 。结果获得了３２６ 个拟测交分离位点和７ 个３∶１ 分离位点。拟测交位点中有２３８ 个位点来源于银白杨,有８８个位点来源于响叶杨。经偏分离检测,用于银白杨作图的位点共计２１２ 个（２６ 个位点偏分离） ,用于响叶杨作图的位点共计７８ 个（１０ 个位点偏分离） 。利用多点连锁分析,银白杨中有１８９ 个连锁标记构成了２０ 个不同的连锁群（４个以上标记）,６ 个三连体和１６ 个连锁对,连锁标记覆盖的总图距为２４０２ ．４ ｃＭ（ｃｅｎｔｉｍｏｒｇａｎ） 。而响叶杨有４１ 个连锁标记分属于１２ 个不同的连锁群（２ 个以上标记） ,标记覆盖的总图距为４７９ ．４ ｃＭ。获得了中等密度的银白杨连锁图谱和响叶杨图谱的一个连锁框架     

白桦RAPD遗传连锁图谱的构建

以80个来自欧洲白桦(Betula pendula Roth)×中国白桦(Betula platyphylla Suk)的F1个体为作图群体。利用2个亲本和10个F1个体对1,200个10 bp的随机寡核苷酸引物进行筛选, 确定了208个多态性引物。利用RAPD标记, 按照拟测交的作图策略, 分别构建了欧洲白桦和中国白桦的分子标记连锁图谱。对2个亲本和80个F1代作图群体进行随机扩增, 共获得了364个多态性位点。χ2检验结果表明有307个位点符合1∶1分离的拟测交分离, 26个位点符合3∶1分离, 31个位点属偏分离位点。拟测交位点中有145个位点来自欧洲白桦, 有162个位点来自中国白桦。利用2点连锁分析, 欧洲白桦中的145个连锁标记构成了14个不同的连锁群(4个以上标记), 6个三连体和6个连锁对, 37个为非连锁位点, 连锁标记覆盖的总图距为955.6 cM (centimorgan), 平均图距14.9 cM。而来自中国白桦的162个标记构成了15个连锁群(4个以上标记), 4个三连体和6个连锁对, 21个为非连锁位点, 连锁标记覆盖的总图距为1,545.8 cM (centimorgan), 平均图距15.2 cM。该图谱的建立为进一步将两个图谱整合为一个高密度图谱及重要基因的定位奠定了基础。     

香菇品种遗传多样性RAPD分子标记的研究

对32个中国栽培香菇品种和2个野生子实体的遗传多样性进行了RAPD标记研究,9个引物共扩增出113条DNA带,83.19%具有多态性。结果显示,34个香菇菌株间均有遗传上的差异,但遗传相关性极高,表明中国栽培菌株间的遗传背景单一。     

大鼠RAPD标记的观察

采用随机扩增多态ＤＮＡ（ＲＡＰＤ）技术,分析ＳＤ和Ｗｉｓｔａｒ二种大鼠的基因多态性,探讨用ＲＡＰＤ标记鉴别二种大鼠及其血标本实验中的认证,结果表明,二种大鼠表现出了各自不同的多态性ＲＡＰＤ标记,作为大鼠的分子标记,可在基因水平区别二种大鼠,故认为是一种大鼠研究的分子依据。     

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C100和大造的回交一代BC,群体进行连锁图谱的构建。经28对引物组合的选择性扩增,共获得了3956条带,平均每对引物产生141．3条带,获得多态性带只有1018条,多态性带的比率为25．7％。其中693(68．1％)个多态性位点符合1：1孟德尔分离比例。利用Mapmaker／Exp3．0软件进行连锁分析,构建了一张含有408个标记位点、33个连锁群、总图距为3676．7cM的连锁图谱,并将绿茧基因定位在该图谱的22连锁群上,表明该连锁群与家蚕经典遗传学的第15染色体相对应。     

Construction of Rice RAPD Molecular Linkage Map

A rice (Oryza sativa L. ) molecular linkage map has been constructed form over 52 random amplified polymorphic DNA (RAPD) markers with the double haploid (DH) population. It covered a total genetic distance of over 898.4 cM (centimorgan) with 17.3 cM between markers and was complemental with RFLP linkage map.     

Construction of Molecular Linkage Map in Masson Pine Using RAPD Markers and Megaga metophytes from a Single Tree

A set of 300 random ohgonucleotide primers was screened for random amplified polymorphic DNA (RAPD) fragments within a sample of 79 megagametophyte DNAs of a single masson pine ( Pinus massoniana Lamb. ) to generate RAPD markers and construct the molecular linkage map for masson pine. 64 repeatable RAPD markers segregated in a 1 present to 1 absent ratio. Through multipoint linkage analysis, 47 markers were assigned to 13 different linkage groups(pairs). It covered a tdtal genetic distance of over 692.5 cM (centimorgan). The average distance between markers was 14.7 cM. It supplied a linkage framework for a more saturated linkage map of masson pine.     

家蚕分子生物学研究进展

近年来,应用RAPD技术,通过构建近等基因系的方法,已筛选出家蚕性别、抗性等连锁的分子标记。日本已成功地构建了家蚕28个连锁群的RAPD分子连锁图,并有7个连锁群和经典的遗传图相对应．我国也公布了RAPD和AFLP分子连锁图,由于密度不够出现28个以上的连锁群;家蚕生物反应器研究和开发已近20年历史,表达了数百种外源基因,由于表达量不高及产物分离纯化难度和成本问题,至今未能进入产业化;家蚕转基因有过比较好的尝试,改进转基因技术提高外源基因的整合率是今后主攻方向。     

Molecular Identification of Tropical Tasar Silkworm (Antheraea mylitta) Ecoraces with RAPD and SCAR Markers

The tropical tasar silkworm, Antheraea mylitta, has several ecoraces, 10 of which are commercially exploited for the production of tasar silk. These ecoraces are identified by morphological markers that are greatly influenced by photoperiod, humidity, altitude, and host plants. The DNA markers, random amplification of polymorphic DNA (RAPD), and sequence-characterized amplified region (SCAR) are identified to complement the existing morphological markers. Seven RAPD bands are selected that identify 8 of the 10 ecoraces. These identified RAPD fragments are sequenced and primers are designed for SCAR markers. Of the seven sets of primers, a single primer pair produced polymorphic SCAR bands that diagnose 5 of the 10 ecoraces. All 10 ecoraces are identified by the use of RAPD and SCAR markers together.     

Molecular Variability of Mycosphaerella graminicola as Detected by RAPD Markers

A total of 90 isolates of Mycosphaerella graminicola, the cause of septoria tritici leaf blotch of wheat, were tested for DNA polymorphism using 15 decamer random primers. There was a high level of genetic variability among isolates. In 131 random amplified polymorphic DNA (RAPD) fragments, which were produced, 96% were polymorphic. Based on multilocus analysis, 40 different molecular phenotypes were detected. These molecular phenotypes were randomly distributed among sampling sites, suggesting that no clonal structure existed in the population. Cluster analysis showed that the maximum similarity value among isolates was approximately 81% and no identical isolates were detected, indicating that every isolate was a unique genotype. The high degree of DNA polymorphism, the large number of different molecular phenotypes, their random distribution and the results of the cluster analysis all suggested that sexual reproduction has a major role in the genetic structure of M. graminicola in western Canada. The presence of sexual reproduction provides the opportunity for development of new virulent genotypes in the population and suggests that the pathogen may adapt rapidly to any race‐specific sources of resistance. Therefore, when breeding for resistance to M. graminicola, emphasis should be placed on use of non‐race‐specific resistance.     

RAPD analysis of natural populations of Acanthopanax brachypus

Random amplified Polymorphic DNA(RAPD) analysis is a new technology of molecular marking which has proved very powerful in detecting genetic diversity at the level of population.The genomic DNAs used in our experiment were extracted from fresh leaves taken from 59 individuals sampled from three natural populations in Yan an,Shanxi Province.Through more than 2,000 PCRs,deep-going RAPD analysis was carried out on DNA samples from 49 inviduals.The percentage of polymorphic RAPD loci found in these three populations were respectively 27.2%,18.6% and 5.4%;the average genetic distances within population,0.055,0.026 and 0.008;the average genetic distances between populations (I-II),(I-III) and (II-III),0.105,0.096 and 0.060.The genetic diversity of A.brachypus within and between populations was found,for the first time,to be rather poor,thus revealing innate factors as the cause contributing to its endangered status.In addition,our work also provides basic materials for elucidating the underlying cause of its endangerment and for its protection biology.     

分子标记技术在蛛形学研究中的应用

DNA分子标记作为新发展起来的一种遗传标记形式,凭借其可靠有效等优点,在动植物研究中的应用已越来越广泛。简述了DNA分子标记技术的种类、原理和特点,并综述了分子标记技术在蜘蛛亲缘关系界定和系统进化等方面的研究进展,对有关问题进行了初步讨论和展望。     

RAPD技术的特点及其在昆虫分类中的应用

随机扩增的多态性DNA(RAPD)技术,是近年来发展起来的一项DNA分子水平上 的大分子多态检测手段。由于它具有简捷、灵敏、对材料要求不高,取材少、成本低等优点, 备受人们青睐,在遗传学、分子进化、生物分类等领域被广泛地运用。在昆虫分类的过程中, 由于昆虫的种类繁多,形态、生态差异很大,许多在其它领域被广泛运用的分子生物学技术不能在昆虫分类中充分发挥作用。同时在昆虫分类中出现的一些问题却又需要用涉及遗传本质的分子生物学技术进行探讨和研究。本文就RAPD技术的特点及其在解决昆虫分类中的问题时的优势作一简介。