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。     

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.     

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     

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。     

三系杂交稻亲本随机扩增多态性DNA(RAPD)分析

选用9个随机引物对31份杂交水稻亲本材料进行了RAPD分析, 共检测到60条多态性带。聚类分析结果表明,所有供试材料可以被明确地区分。在9个随机引物中,有8个具有较高的多态性检测能力。以这8个引物为基础,选用任两个引物即可在任一对材料中检测出多态性的频率在96.13%以上,而选用任3个引物则该频率在99.21%以上。这显示了运用RAPD鉴定稻种具有简便、灵敏、高效的优点,在鉴定杂交稻种的实践中有着良好的应用前景。 Abstract:Seven rice sterile lines,12 maintainer lines and 12 restorer lines were analyzed by RAPD with 9 primers.Altogether,118 fragments were generated,of which 60 detected polymorphisms among rice marker.Eight of nine primers can detect high polymorphism.The frequencies of polymorphism in any primers were used,the frequencies would be higher than 99.21%.The eight primers were therefore recommended as candidates for the identification of hybrid rice seeds.     

用双单倍体群体构建水稻的分子连锁图

本研究以窄叶青８号（籼稻）×京系１７（粳稻）的Ｆ１花培株系──ＤＨ群体为基础建立了１个水稻的ＲＦＬＰ连锁框架图,该图含ＲＦＬＰ标记、同功酶标记等共１０８个位点,标记间的平均间距为８．６ｃＭ。该图谱与已发表的用其他群体构建的图谱有很高的可比性。利用该框架图定位了２个未知位点的同功酶标记基因和１个籼稻亲本的抗稻瘟病基因。研究表明,目前的群体可进一步扩大成为一个永久性的作图群体,并应用于水稻基因定位和基因组研究     

Genetic Molecular Linkage Map Construction and Genome Analysis of Rice Doubled Haploid Population

A high density genetic linkage map comprised of aA. 4 loci was constructed from a doubled haploid population derived from a inter-subspecific cross between an Oryza satire L. ssp. Indica vari.t.v ("Zhaiyeqing 8") and a japonica variety ("Jingxi 17"). The genetic map consisted of 276 RFLP markers, 34 RAPD markers, 89 microsatellite markers, 10 AFLP markers, 26 markers based on telomeric repetitive associated sequence (TAS) and 9 isozyme markers. This genetic map was highly comparable with other high density rice genetic maps and had its unique feature which meritted it suitable for sustained genetic analysis.     

甘蓝分子连锁图的构建与品质性状的QTL定位

以两个不同生态型甘蓝(Brassica oleracea var．capitata)品种杂交得到的F2代为作图群体,用RAPD标记构建甘蓝分子连锁图。通过对520个随机引物进行筛选,236个引物在两亲本间表现多态性,多态性比例为47．7％。选取111个引物对群体进行分析,构建了一张含有135个标记位点,9个连锁群,覆盖长度为1023．7cM的分子连锁图。利用该图谱对甘蓝叶球紧实度和中心柱长两性状进行了QTL定位分析。检测到3个与叶球紧实度相关的QTL,总贡献率为62．5％;检测到4个与中心柱长相关的QTL,总贡献率为59．1％。     

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。     

水稻双单倍体群体的分子标记图示基因型分析

利用窄叶青８号（籼稻）／京系１７（粳稻）花培产生的双单倍体群体建立了一个包含１６０个分子标记的遗传连锁图,在此基础上利用ＨＹＰＥＲＧＥＮＥ软件建立了５２个ＤＨ系的图示基因型,并对ＤＨ系的亲本基因组比率和染色体的交换重组频率进行了比较分析。结果表明本实验所用的ＤＨ群体没有显著偏离正态分布,籼粳稻杂交后代中植株的籼粳表现与同工酶、形态指数和基因组比率的分析结果一致,此外还发现ＤＨ群体中出现了大量的交换罕见染色体。利用图示基因型分析发现株高和分子标记ＲＺ９７８和ＲＧ４Ａ相关,生育期和ＲＲＫ０８－１、ＲＧ４７７和ＲＧ５１１相关。本文还就图示基因型分析技术在ＤＨ群体的遗传分析和选择育种中的应用进行了讨论．     

利用两个测序水稻品种构建微卫星连锁图谱

利用已完成基因组测序的两个水稻品种日本晴和931l的数据库成功开发出水稻微卫星新标记,并利用由90个单株组成的日本晴×9311 F2作图群体,构建了一张包含152个SSR标记位点、覆盖基因组总长度2 455.7 cM的连锁图谱,有46个SSR新标记为自主开发,该图谱标记间的平均遗传距离为16.16 cM;并将未能在Temnykh等人(2001)构建的图谱上定位的微卫星标记RM345和RM494定位在第6染色体上.通过与Temnykh等人(2001)和兰涛等人(2003)所构建的图谱从作图群体的类型和大小、标记的类型和数量、标记在染色体上的线性排列顺序等几个方面进行比较,所绘制的图谱其标记在染色体线性排列上与Temnykh等人绘制的图谱具有很高的一致性,达93.81%.     

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。     

水稻叶绿体基因文库的构建和精细限制图谱的制作

水稻幼叶在加有高浓度抗坏血酸的缓冲液中匀浆,以获得完整的叶绿体,从中分离到ctDNA得率高达100μg/100g叶,纯度足以用于限制性核酸内切酶分析。ctDNA经Mbo I部分酶解得到的片段克隆到载体pcos 2 EMBL的Bam HI位点,重组DNA经体外包装后感染宿主菌,筛选表型Tc~5Km~R的重组子,通过计数克隆有效率达5×10~4重组菌落/1微克插入DNA。用λ-末端酶对重组环状双链DNA在cos位点切成线性分子,产生两个(ON-L及ON-R)可供标记和杂交的末端,线性Cosmid DNA经限制酶部分消化,凝胶电泳分离,干燥凝胶放射自显影,得到了6种限制性核酸内切酶的限制图谱。水稻ctDNA全长为129.5kb,在ctDNA上Pvu Ⅱ、Sal Ⅰ、Pst Ⅰ、Hind Ⅲ、Eco RI及Bam HI的切点分别为11、12、17、37、67和44个,1R A和B为21.7kb,LSC为73.7kb,SSC为12.4kb。     

用微卫星标记构建两系稻培矮64s/E32的分子遗传连锁图 Construction of a Microsatellite Linkage Map in a DH Population

用两系杂交稻强优组合培矮64s/E32的一个加倍单倍体（DH）群体,共86个株系,构建了水稻的SSR标记遗传连锁图。选用美国康耐尔大学公布的302对SSR引物,共有127对在两个亲本间检测到多态性,比率为4205%。建成的水稻染色体的图谱（记为PEMAP）共包含122个SSLP标记座位,总长度为1213.4 cM。PEMAP与Temnykh等发表的图谱（记为CUMAP）具有很高的可比性,绝大多数标记都被定位于相同的染色体上,且排列顺序一致。该DH群体的偏分离情况较严重,122个标记座位中有34个发生显著偏分离,比例达27.8%。值得注意的是,在第1、3、10、11染色体上的标记全部偏向培矮64s,第4、6、7、8、9染色体上的标记则全部偏向E32。 Abstract:A doubled haploid population (DH) consisting of 86 lines derived by anther culture of Peiai64s/E32,a two-line hybrid rice variety with high heterosis,was used to construct a microsatellite or SSLP linkage map of rice chromosomes.A total of 302 PCR primers for SSLP analysis on these chromosomes were chosen from a map published by Cornell University (designated CUMAP) and 127 (42.05%) of them were found polymorphic between the two parents.Those polymorphic PCR primers were used for population genotyping.The map (designated PEMAP) comprises 122 microsatellite maker loci,covering a total length of 1213.4 cM.The PEMAP is highly comparable with the CUMAP.Most of the markers were mapped onto the same chromosomes and aligned in the same order.Serious segregation distortion was observed in this DH population,with 34 (27.8%) markers showing significant deviation.It is noted that all markers on chromosomes 1,3,10 and 11 were biased to Peiai64s,while those on chromosomes 4,6,7,8 and 9 were opposite.     

Construction of a Microsatellite Linkage Map with Two Sequenced Rice Varieties

Based on the successful development of new microsatellite markers from the data of two whole-sequenced rice varieties, japonica variety Nipponbare and indica variety 9311, an F₂ population of 90 lines, which was derived from a single cross between Nipponbare and 9311, was applied to construct a genetic linkage framework map. The map covered 2 455.7 cM of total genomic length, and consisted of 152 simple sequence repeats (SSRs) loci including 46 pairs of new SSR primers developed by our research institute. The average genetic distance between two markers was 16.16 cM. In addition, markers RM345 and RM494, which have not been mapped on the Temnykh's map et al. (2001) were anchored on the sixth chromosome of this map. We compared this research with maps of Temnykh et al.(2001) and LAN et al. (2003) regarding the aspects of type and size of population, type and quantity of markers, and the marker arrangement order on chromosome, etc. Results indicated that the similarity of marker linear alignment was 93.81% between this map and T-map, Finally, the important significance of using sequenced rice varieties to construct linkage map was also discussed.     

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。     

SNP Discovery and Linkage Map Construction in Cultivated Tomato

Few intraspecific genetic linkage maps have been reported for cultivated tomato, mainly because genetic diversity within Solanum lycopersicum is much less than that between tomato species. Single nucleotide polymorphisms (SNPs), the most abundant source of genomic variation, are the most promising source of polymorphisms for the construction of linkage maps for closely related intraspecific lines. In this study, we developed SNP markers based on expressed sequence tags for the construction of intraspecific linkage maps in tomato. Out of the 5607 SNP positions detected through in silico analysis, 1536 were selected for high-throughput genotyping of two mapping populations derived from crosses between ‘Micro-Tom’ and either ‘Ailsa Craig’ or ‘M82’. A total of 1137 markers, including 793 out of the 1338 successfully genotyped SNPs, along with 344 simple sequence repeat and intronic polymorphism markers, were mapped onto two linkage maps, which covered 1467.8 and 1422.7 cM, respectively. The SNP markers developed were then screened against cultivated tomato lines in order to estimate the transferability of these SNPs to other breeding materials. The molecular markers and linkage maps represent a milestone in the genomics and genetics, and are the first step toward molecular breeding of cultivated tomato. Information on the DNA markers, linkage maps, and SNP genotypes for these tomato lines is available at http://www.kazusa.or.jp/tomato/.     

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     

玉米RFLP连锁图谱构建及大斑病QTL定位

玉米大斑病菌存在有生理小种分化的现象,目前5个已定名的生理小种在我国均已发现,还有一些尚未定位名的新类群也出出现,提高玉米对大斑病的抗性,只有提高数量抗性才能达到目的,为了弄清楚玉米对大斑病数量抗性的基因数目及效效应,利用抗病自交5系P138和感病自交系缩3为亲本构建了F2：3家系群体,采用RFLP标记构建了包含了124个标记的玉米RFLP连锁图,覆盖玉米基因组1999．8cM,标记间平均距离为16．5cM,定位了玉米大斑病的病斑长,病斑宽和病斑面积的QTL分别为3、3、2个,其联合贡献率分别为58．1％、71．5％和27．5％,没有检测到病斑数／叶的QTL,其表现为单基因或者寡基因控制的性状,研究结果增加了对玉米大斑病的认识,对玉米抗大斑病育种具有重要的指导意义。     

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