小麦双引物RAPD分析方法的研究

ＲＡＰＤ标记是近几年迅速发展的一种新型分子标记,标准的ＲＡＰＤ的反应是以１０个寡聚核苷酸作引物,通过ＰＣＲ反应扩增出基因组的部分片段,我们在研究外源ＤＮＡ导入小麦后外源遗传物质的追踪时,对这个方法进行了改进,采取了双引物进行扩增,结果双引物反应能够比单引物反应扩增出更多的多态性片段。分子杂交结果表明,双引物扩增出的新片段与单引物扩增片段无同源性,并对双引物扩增出的多态性片段产生的可能原因进行讨论。     

RAPD和SSR两种标记构建的中国对虾遗传连锁图谱

利用RAPD和SSR分子标记结合拟测交策略,对中国对虾(Fenneropenaeuschinensis)“黄海1号”雌虾与野生雄虾作为亲本进行单对杂交产生的F1代,采用RAPD和SSR两种分子标记技术初步构建了中国对虾雌、雄遗传连锁图谱。对460个RAPD引物和44对SSR引物进行筛选,共选出61个RAPD引物和20对SSR引物,用于对父母本和82个F1个体进行遗传分析。共得到母本分离标记146个(RAPD标记128个,微卫星标记18个)和父本分离标记127个(RAPD标记109个,微卫星标记18个)。雌性图谱包括8个连锁群、9个三联体和14个连锁对,标记间平均间隔为11·28cM,图谱共覆盖1173cM,覆盖率为59·36%;雄性图谱包括10个连锁群、12个三联体和7个连锁对,标记间平均间隔为12·05cM,图谱共覆盖1144·6cM,覆盖率为62·01%。中国对虾遗传图谱的构建为其分子标记辅助育种、比较基因组作图及数量性状位点的定位与克隆奠定了基础。     

烟草RAPD双引物与单引物PCR扩增多态性效率的比较分析

以两个亲缘关系较远的烟草（Nicotiana tabacum）品种台烟7号和白肋21为材料（GS=0.58）,研究了单引物扩增和双引物聚合扩增对RAPD-PCR分子标记多态性的影响。结果显示：双引物反应能够比单引物反应扩增出更多的多态性片段,双引物在白肋21和台烟7号中扩增出的多态性片段总数是单引物反应扩增出的多态性片段总数的6.6倍。因此,双引物RAPD的运用有助于提高烟草多态性分子标记的有效性。     

中国植物遗传连锁图谱构建研究进展

遗传连锁图谱构建是基因组研究中的重要环节,是基因定位与克隆乃至基因组结构与功能研究的基础上。近十几年来,分子生物学特别是分子标记技术的飞速发展,为构建高饱和的植物遗传连锁图谱和利用分子标记进行辅助育种奠定了基础。综述了我国在植物遗传连锁图谱构建研究方面的进展及发展动态,列举了我国利用DNA分子标记构建的34张植物遗传连锁图谱实例,且讨论了当前我国在该领域研究中存在的问题并提出了解决途径。     

利用RAPD标记构建美洲黑杨×欧美杨分子标记连锁图谱

本文利用RAPD标记和美洲黑杨(Populusdeltoides)×欧美杨(P.euramericana)的F1 群体 ,构建了美洲黑杨×欧美杨的分子标记连锁图谱。实验过程中对1040个寡核苷酸随机引物进行了重复筛选 ,共选出127个引物用于作图群体(包括双亲共92个无性系)的随机扩增 ,这127个引物产生229个多态基因座 ,其中符合“拟测交”1∶1分离的有214个。利用多点连锁分析 ,形成19个连锁群及6个三连体和14个连锁对。由19个连锁群构成的图谱含标记129个 ,总图距为1914 2cM ,覆盖杨树基因组约73 62 %。标记间的平均间距为14 84cM。本研究获得了中等密度的美洲黑杨×欧美杨的一个连锁框架。     

利用RAPD标记构建美洲黑杨×欧美分子标记连锁图谱

本文利用RAPD标记和美洲黑杨(Populus deltoides)×欧美杨(P.euramericana)的F1群体,构建了美洲黑杨×欧美杨的分子标记连锁图谱。实验过程中对1040个寡核苷酸随机引物进行了重复筛选,共选出127个引物用于作图群体(包括双亲共92个无性系)的随机扩增,这127个引物产生229个多态基因座,其中符合“拟测交”1∶1分离的有214个。利用多点连锁分析,形成19个连锁群及6个三连体和14个连锁对。由19个连锁群构成的图谱含标记129个,总图距为1914.2cM,覆盖杨树基因组约73.62％。标记间的平均间距为14.84cM。本研究获得了中等密度的美洲黑杨×欧美杨的一个连锁框架。 Abstract:A molecular linkage map was constructed for the parents of a P.deltoides × P.euramericana F1 family based on random amplified polymorphic DNA(RAPD)markers.A set of 1040 random oligonucleotide primers were screened and 127 primers were selected to generate RAPD markers within a sample of 90 F1 progenies.A total of 229 segregating loci were identified.Among the 229 loci,15 loci were found distorted from the normal 1∶1 ratio.Using multiple analysis,the 214 markers formed 19 main Linkage groups (including 129 markers)and b triples and 14 pairs.The resulting Linkage map of Populus deltoides × P.euramericana (including 129 markers)spanned 1914.2cM(73.62％ coverage of genome length)with an average distance of 14.84cM between markers.     

利用RAPD标记构建美洲黑杨×欧美分子标记连锁图谱

本文利用RAPD标记和美洲黑杨(Populus deltoides)×欧美杨(P.euramericana)的F1群体,构建了美洲黑杨×欧美杨的分子标记连锁图谱。实验过程中对1040个寡核苷酸随机引物进行了重复筛选,共选出127个引物用于作图群体(包括双亲共92个无性系)的随机扩增,这127个引物产生229个多态基因座,其中符合“拟测交”1∶1分离的有214个。利用多点连锁分析,形成19个连锁群及6个三连体和14个连锁对。由19个连锁群构成的图谱含标记129个,总图距为1914.2cM,覆盖杨树基因组约73.62％。标记间的平均间距为14.84cM。本研究获得了中等密度的美洲黑杨×欧美杨的一个连锁框架。 Abstract:A molecular linkage map was constructed for the parents of a P.deltoides × P.euramericana F1 family based on random amplified polymorphic DNA(RAPD)markers.A set of 1040 random oligonucleotide primers were screened and 127 primers were selected to generate RAPD markers within a sample of 90 F1 progenies.A total of 229 segregating loci were identified.Among the 229 loci,15 loci were found distorted from the normal 1∶1 ratio.Using multiple analysis,the 214 markers formed 19 main Linkage groups (including 129 markers)and b triples and 14 pairs.The resulting Linkage map of Populus deltoides × P.euramericana (including 129 markers)spanned 1914.2cM(73.62％ coverage of genome length)with an average distance of 14.84cM between markers.     

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。     

RAPD技术及其在动物遗传育种中的应用

ＲＡＰＤ技术是在ＰＣＲ基础上发展起来的一种ＤＮＡ多态性检测技术,已广泛应用于基因组研究的种个领域。本文概述了ＲＡＰＤ反应的原理、特点,总结了其在遗传多样性检测、亲缘关系鉴定、遗传连锁分析和数量性状的辅助标记选择等方面的应用,并肯定了ＲＡＰＤ在动物遗传户种领域的应用前景。     

食用菌遗传连锁图谱研究进展

食用菌遗传连锁图谱的构建工作虽然较晚于动植物研究,但随着基础研究不断完善,连锁图谱构建工作陆续开展。文中综述了双孢蘑菇(Agaricus bisporus)、香菇(Lentinula edodes)、双色蜡蘑(Laccaria bicolor)、草菇(Volvariella volvacea)、糙皮侧耳(Pleurotus ostreatus)、杏鲍菇(Pleurotus eryngii)、金针菇(Flammulina velutipes)、斑玉蕈(Hypsizigus marmoreus)等食用菌的遗传连锁图谱研究进展,展望了其应用现状及前景,并对笔者所在的国家食用菌产业技术体系黑木耳育种与菌种繁育团队的黑木耳遗传连锁图谱研究进展进行了简介。     

中国白菜RAPD分子遗传图谱的构建

以芜菁（Ｂｒａｓｓｉｃａ ｃａｍｐｅｓｔｒｉｓ Ｌ．ｓｓｐ．ｒａｐｉｆｅｒａ Ｍｅｔｚｇ）和结球白菜（Ｂ．ｃａｒｎｐｅｓｔｒｉｓ Ｌ．ｓｓｐ ．Ｐｌｋｉｎｅｎｓｉｓ（Ｌｏｕｒ．）Ｏｉｓｓｏｎ）杂交的Ｆ２群体为试材,采用ＲＡＰＤ标记,用８４个１０核苷酸随机引物构建了白菜的ＲＡＰＤ遗传图谱。该图谱覆盖基因组的１６３２．４ｃＭ（ｃｅｒｔｉ Ｍｏｒｇａｎ）标记间的平均间隔为１６．５ｃＭ。其中最长的连锁群为     

中国白菜RAPD分子遗传图谱的构建

A molecular genetic map of Brassica campestris L. (syn. B. rapa) was constructed based on the segregation of 99 RAPDs (random amplified polymorphic DNAs) markers from eighty-four 10-base random primers using DNA samples extracted from F2 population of turnip (B． campestris L. ssp. rapifera Metzg) × Chinese cabbage (B． campestris L. ssp. pekinensis Lour. Olsson). This genetic map covered 1 632.4 cM (centiMorgan) genome (Kosambi Function) with 16.5 cM mean intervals between flanking markers and defined thirteen linkage groups, in which the longest linkage group is 267.5 cM with 20.6 cM mean interval and the shortest linkage group is 62.2 cM with 15.6 cM mean interval. The size and distribution of linkage groups in this map is similar to other RFLP maps and karyotype data in B. campestris.     

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.     

RAPD标记构建家蚕分子连锁图

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。     

梨遗传连锁图谱的构建及其与苹果图谱的比较

以‘丰水’为母本、‘砀山酥梨’为父本杂交所得的F1代104株单体为作图群体,利用SSR分子标记进行遗传连锁分析,应用Jionmap 3.0作图软件,构建了一张包含104个SSR分子标记,分属于18个连锁群的梨遗传连锁图谱,覆盖梨基因组总长831.8cM,平均图距为8.0cM。根据定位到该图谱上的SSR标记与苹果‘Fiesta’图谱进行比较,25个共有的SSR标记将该图谱和苹果图谱各连锁群连接起来,这些标记不仅呈现良好的共线性而且它们之间的相对遗传距离也很相近。研究认为,SSR标记作为锚定引物,可以与不同物种的遗传图谱相比较整合,为不同物种之间遗传信息的转移提供参考依据;同时该研究为梨树相关性状的基因定位、分离以及克隆奠定了基础。     

Comparison of RAPD and RFLP markers for mapping F2 generations in maize (Zea mays L.)

The F₂ generations from two maize crosses were used to compare the ability of RAPD and RFLP marker systems to create a genetic linkage map. Both RFLPs and RAPDs were shown to provide Mendelian-type markers. Most of the RFLPs (80%) could be placed with a good level of certainty (LOD>4) on the genetic linkage map. However, because of their dominant nature, only between 37% and 59% of the RAPDs could be placed with such a LOD score. The use of combined data from RFLPs and RAPDs increases the level of information provided by RAPDs and allows the creation of a combined RFLP/RAPD genetic linkage map. Thus, the RAPD technique was found to be a powerful method to provide improved probes coverage on a previously created RFLP map and to locate markers linked to chromosomal regions of interest.     

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.     

利用RAPD标记分析大麦种质资源的遗传多样性

利用RAPD标记对19份西藏近缘野生大麦材料、33份我国不同省市的地方品种以及8份国外引进大麦品种共60份大麦种质资源的遗传多样性进行检测.结果表明材料间遗传差异明显.32个RAPD引物中,有25个引物(占78.13%)可扩增出清晰且具多态性的条带,另外7个引物能扩增出1～3条清晰但无多态性的条带.每个引物可扩增出1～8条多态性带,平均为3.72条.32个引物共产生119条DNA片段,其中87条具有多态性,多态性比率(PPB)为73.11%,平均多态信息量(PIC)为0.434;每个位点平均有效等位基因数(Ne)为2.304;材料间遗传相似系数GS变化范围为0.757～0.981,平均值为0.871.19份来源于西藏的近缘野生大麦材料间GS值变幅为0.818～0.969,平均为0.892;33份我国栽培大麦地方品种间的GS值变化范围为0.783～0.981,平均为0.879;8份分别来自8个国家的栽培大麦品种间的GS值变幅为0.820～0.956,平均为0.882.根据RAPD标记分析的结果,对60份大麦种质资源进行聚类分析,在平均GS值0.871水平上60份大麦材料可聚为5类,聚类结果能在一定程度上反应材料的地理分布关系,但某些相同地理来源的材料也较分散地分布在整个聚类树中.本研究从分子水平上进一步证明了我国栽培大麦丰富的遗传多样性,是世界栽培大麦的遗传多样性中心之一.     

The linkage maps of Dendrobium species based on RAPD and SRAP markers

Dendrobium plants are used commonly as tonic herbs and health food in many Asian countries,especially in China.Here we report the genetic map construction of two Dendrobium species with a double pseudo-testcross strategy using random amplified polymorphic DNA (RAPD) and sequence-related amplified polymorphism (SRAP) markers.A F1 mapping population of 90 individuals was developed from a cross between D.officinale and D.hercoglossum.A total of 307 markers,including 209 RAPD and 98 SRAP,were identified and used for genetic linkage group (LG) analysis.The D.officinale linkage map consisted of 11 major linkage groups and 3 doublets,which covered 629.4 cM by a total of 62 markers with an average locus distance of 11.2 cM between two adjacent markers.The D.hercoglossum linkage map contained 112 markers mapped on 15 major and 4 minor linkage groups,spanning a total length of 1,304.6 cM with an average distance of 11.6 cM between two adjacent markers.The maps constructed in this study covered 92.7% and 82.7% of the D.hercoglossum and D.officinale genomes respectively,providing an important basis for the mapping of horticultural and medicinal traits and for the application of marker-assisted selection in Dendrobium breeding program.