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菜薹雄性不育相关基因的ISSR分子标记筛选 总被引：5，自引：3，他引：2
菜薹(Brassica campestris L.ssp.Chinensis var.utilis Tsenet Lee)属于十字花科芸薹属,原产中国,是华南地区重要的特产蔬菜之一.利用分子标记技术进行菜薹雄性不育相关基因的定位对菜蕞品质创新和选育新品种具有重要意义.目前尚未见有利用ISSR分子标记技术进行菜薹雄性不育分析的研究报道.利用ISSR技术对菜薹雄性不育系及其保持系的基因组DNA进行比较分析:结果显示100个ISSR引物中,只有引物UBC843的扩增结果在两系之间表现出了差异性,找到了不育系和保持系间的特异扩增片段.回收该特异扩增片段并进行克隆测序,测序结果表明该片段全长为462 bp.与白菜其它育性相关序列比较,同源性均低于50%.根据测序结果设计了一对特异引物,将其转化为更稳定的SCAR标记.经F2群体验证,ISSR引物扩增得到的特异片段很有可能来自核DNA. 相似文献
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来源于长穗偃麦草的基因Lr24对小麦叶锈病具有很高的抗性,本研究旨在开发用于Lr24基因分子标记辅助育种的新的分子标记。从定位于小麦3D染色体的22对SSR、EST-SSR引物中筛选出4对揭示TcLr24多态性的引物,用468株F2抗感群体对这4对引物进一步检测,得到1个与Lr24共分离的EST-SSR标记Xcwem17。对该标记进行测序,并设计了STS引物。用该STS引物及已知的Lr24SCAR引物对试验群体进行验证,两对引物在该F2群体中均表现共分离,且Xcwem17可在TcLr24单基因系和已知含Lr24的农家品种泰山1号中可扩增出180bp单一条带,感病对照及其余7个近等基因系无扩增。该EST-SSR标记可直接用于分子标记辅助选择。 相似文献
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不同类型抗虫棉品种基于RAPD的遗传多样性分析 总被引：1，自引：0，他引：1
以97014、 BR-S-10和sGK321等14份不同类型的抗虫棉花品种(系)为基础,利用RAPD分子标记技术,对这些棉花品种(系)的遗传多样性进行了研究.结果表明,从210个随机引物中筛选出21个引物能在14份抗虫棉品种(系)间扩增出稳定性较好的多态性片段,筛选的21个引物在该品种(系)群体中共扩增了137个RAPD标记,其中多态性标记57个,占总标记数的42.2%,平均每条引物扩增6.52条带.利用Jaccard遗传相似系数计算了14份抗虫棉品种(系)的RAPD数据的距离矩阵,按UPGMA方法进行聚类,发现14个品种(系)间的遗传距离在0.0351～1.056之间.聚类分析表明,14个棉花品种可分为2大类4亚类,揭示了抗虫棉花品种种质资源的多样性,大多数品种的遗传基础比较狭窄. 相似文献
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RAPDs和RFLPs分析甘蓝型杂交油菜亲本的遗传多样性 总被引：12，自引：0，他引：12
利用RAPD和RFLP分子标记技术对甘蓝型杂交油菜亲本遗传多样性的分析结果表明:(1)20个亲本间具有丰富的遗传多样性,40个引物扩增出277条多态性带,其中21条带为10个亲本所特有,12个探针得到了117条多态性杂交带,其中7条带为5个亲本所特有;(2)不育系与恢复系之间的遗传差异大于不育系内和恢复系内的遗传差异;(3)依RAPDs与RFLPs估 相似文献
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首批海岛棉基因组来源的微卫星标记的分离、评价和定位 总被引：1，自引：1，他引：0
海岛棉(Gossypium barbadense)是世界上最重要的栽培棉种之一。海岛棉纤维品质优良,是优质棉的重要产源。为了研究海岛棉的遗传多样性,为海岛棉育种提供参考依据,从海岛棉遗传标准系中分离基因组来源的微卫星标记用于海岛棉遗传评价。采用两种方法分离微卫星标记,一是用ISSR (inter simple sequence repeat) 引物扩增Pima3-79,克隆测序后从中开发微卫星标记;二是利用简并引物扩增Pima3-79,克隆测序后从中开发微卫星标记。共挑选1 447个克隆,筛选出239个独立克隆。测序后得到214个单一序列,其中包含微卫星并可用于引物设计的序列70个,获得86对引物。86对引物用于扩增56个海岛棉材料和4个陆地棉材料,16对引物没有扩增,43对引物在所有材料中没有多态性;27对引物在海岛棉和陆地棉之间有多态性,19对引物在海岛棉中表现多态性。利用Jaccard相似系数和UPGMA方法进行聚类分析可以明显区分陆地棉和海岛棉,并且将海岛棉分为4类。14对引物在BC1群体中表现多态性,产生14个位点。9个位点整合到BC1连锁图的7个染色体上,4个位于A亚基因组,5个位于D亚基因组。海岛棉微卫星标记扩展了棉花微卫星标记,有助于海岛棉遗传多样性的研究,有利于棉花遗传图谱的进一步丰富。 相似文献
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小麦雌蕊和雄蕊突变体与川麦28之间特异性ISSR标记筛选 总被引：1，自引：0，他引：1
利用42条ISSR标记对小麦三雌蕊突变体(TP)和雄蕊同源转化为雌蕊突变体(HTS-1)与川麦28之间特异性的ISSR分子标记进行筛选,为利用ISSR标记定位Pis1基因和控制雄蕊同源转化成雌蕊的hts1和hts2基因奠定基础.结果表明:42条引物共扩增403个条带,有9条引物能在TP和CM 28之间扩增出差异条带,占所用引物的21.4％.有20条引物能在HTS-1和CM 28之间扩增出差异条带,占所用引物的47.6％.有21条引物能在TP与HTS-1之间扩增出差异条带,占所用引物的50％.每条引物最多能扩增出4条差异条带,大部分产生1～2条差异性条带.差异条带大小主要集中在250～750 bp之间.这也证明了ISSR标记是一种简单有效的分子标记,可作为SSR的一种重要补充标记用于遗传图谱的构建. 相似文献
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Random amplified polymorphic DNA (RAPD) markers were used to distinguish between several Cichorium intybus genotypes, comprising four white witloof inbred lines, three red witloof experimental inbred lines and a number of F1 hybrids derived from two white parents. Amplification conditions and reproducibility of RAPD patterns were examined. Comparison of polymerase chain reaction (PCR) products obtained by using 100 10-mer arbitrary primers allowed identification of all the lines analysed. With several primers, we defined line-specific RAPD markers, while with others polymorphism was more extensive, revealing several RAPD markers for several lines. All the differences were confirmed both on individual heads and young seedlings for each genotype. Because of the Mendelian segregation of these molecular markers, this method was applied to evaluate the genetic purity of F1 hybrid seed samples. 相似文献
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利用RAPD标记鉴定大白菜杂交种纯度的研究 总被引：13，自引：0，他引：13
应用随机扩增多态性ＤＮＡ（ＲＡＰＤ）标记鉴定大白菜杂种商品种子的纯度。用５０个随机引物检测了２个杂交种北京５７号和北京１０６号及其亲本,引物ＯＰＥ－２０在北京５７号杂交种中产生了有别于双亲的特殊标记,引物ＯＰＨ－０６及ＯＰＨ－０７在北京１０６杂交种中产生了有别于双亲的特殊标记,能清楚地区分杂交种及其双亲,并将这些引物应用在北京５７号和北京１０６号杂交种样品的纯度检测中。这个结果显示了ＲＡＰＤ标记在大白菜杂交种商品种子纯度检测上的实际用途。 相似文献
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利用RAPD标记鉴定大豆种质 总被引：49，自引：4，他引：49
本研究以５７个中国大豆祖先吕系及育成品种和１８个美国大豆祖先品系为ＤＮＡ样品来源，通过随机引物ＰＣＲ扩增基因组ＤＮＡ的多态性，探索利用ＲＡＰＤ标记鉴定和相关种质的可能性。研究结果表明，５０个１０摩尔随机引物共扩增可分辩产物２４６个，其中８２．４％的随机引物可产生多态性产物，所扩增产物的５４．４％至少在两个基因毒草境存在差异。上ＰＣＲ扩增产物分别以１和０记录存在与否。扩增产物间的成对比较可产生非相似 相似文献
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黑麦重复序列在检测小麦品种中外源染色体的应用 总被引：2，自引：0，他引：2
本研究根据RAPD引物OPH20在黑麦中扩增出的特异序列pSc20H.2设计一对PCR引物pSc20ht-23/24,以来源于黑麦的小麦抗叶锈近等基因系材料TcLr45及感病对照Thatcher为亲本进行PCR扩增。并对42个小麦抗叶锈近等基因系及103个小麦品种材料进行检测。引物pSc20ht23/24在TcLr45中扩增出一条约750bp的条带,而在Thatcher中无扩增条带。对该特异片段回收、克隆测序为734bp。42个小麦抗叶锈近等基因系检测在TcLr26中扩增出与TcLr45相同的条带,而在同样来源于黑麦的小麦抗叶锈近等基因系TcLr25中未扩增出该条带;中国春-Imperial黑麦附加系1R-7R中除5R外均扩增出该条带;13个1B/1R易位系小麦品种也扩增出该条带;90个地方小麦品种中有16个扩增出该条带,6个品种经系谱分析具有黑麦遗传背景,表明该标记可用于检测小麦中含有的除黑麦5R染色体之外的外源染色质。 相似文献
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Genetic diversity of hexaploid wheat cultivars estimated by RAPD markers, morphological traits and coefficients of parentage 总被引：3，自引：0，他引：3
S. Mari&#; S. Bolari&#; J. Martin&#;i&#; I. Peji&#; V. Kozumplik 《Plant Breeding》2004,123(4):366-369
Estimates of genetic diversity can be based on different types of data. The aim of this research were to study genetic diversity among Croatian wheat cultivars by random amplified polymorphic DNA (RAPD) markers, morphological traits and pedigree records; to analyse differences between wheat cultivars from two breeding centres; and to evaluate usability of RAPD markers for estimation of genetic diversity among wheat cultivars in comparison with morphological traits and pedigree record data. Studies were conducted on 14 wheat cultivars and breeding lines from two breeding centres in Croatia. For the RAPD analysis 36 primers were screened and the 14 most polymorphic ones yielded 341 polymorphic bands. Twelve morphological traits were used for morphological analysis. Pedigrees were composed of seven generations of ancestors. RAPD markers showed a high level of polymorphism among the cultivars examined and the breeding lines. No significant correlations were observed among the methods tested. 相似文献
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Randomly amplified polymorphic DNA (RAPD) markers were applied in purity control of hybrid seed production of tomato (Lycopersicon esculentum Mill.). DNA from three commercial F1-hybrid cultivars and their parental lines was subjected to RAPD screening with 50 primers. Two of four primers which detected polymorphism between the parents tested, generated paternal-specific RAPDs, enabling a clear distinction to be made between hybrids and their maternal parents. In addition, combination of the polymorphic DNA products generated by these primers exhibited hybrid-specific patterns, enabling each cultivar to be identified. This result indicates the practical usefulness of RAPD markers in hybrid-tomato-seed purity-control tests and cultivar identification. The approach is advantageous in its rapidity and simplicity, particularly as an alternative for those cultivars for which lengthy and costly phenotypic tests are currently used. 相似文献
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RAPD and SCAR markers linked to the sex expression locus M in asparagus 总被引：13，自引：0，他引：13
Bulk segregant analysis (BSA), random amplified polymorphic DNA (RAPD) and sequence characterized amplified region (SCAR)
methods were used to map molecular markers to the sex locus M of asparagus. Two parents, A19 (male, Mm) and MW25 (female,
mm), and 63 progeny were used for the study. Two DNA bulks, one male and one female, were made by pooling equal amounts of
DNA from 10 randomly selected progeny of each sex type. A total of 760 arbitrary decamer oligonucleotide primers were used
for RAPD analysis. Primer OPC15 produced two RAPD markers, OPC15-98 and OPC15-30, both of which were linked to the M locus
at a distance of 1.6 cM. Subsequently, amplified RAPD fragment OPC15-98 was cloned and sequenced. The sequence was then used
to design flanking 24-mer oligonucleotide SCAR primers SCC15-1 and SCC15-2. Both of these SCAR primers amplified a single
980 bp fragment; the same size as the cloned RAPD fragment. However, the SCAR marker was dominant as was the original OPC15-98
band from which it was derived. These RAPD and SCAR markers could be used for scoring male and female progeny in the mapping
population, but were not found to be applicable to other asparagus germplasm studied.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
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Genetic diversity of Tainan-white maize inbred lines and prediction of single cross hybrid performance using RAPD markers 总被引：8，自引：0，他引：8
To evaluate the genetic diversity of 13 maize inbred lines, and to determine the correlation between genetic distance and
single cross hybrid performance, we employed the randomly amplified polymorphic DNA(RAPD)- a PCR-based technique. Six of these
lines came from the Taichung population, and others derived from seven different sites. Forty different primers were used
to give a total of 646 reproducible amplification products, 547 (84.7%) of them being polymorphic. Genetic divergence was
determined using Jaccard's similarity coefficient, and a final dendrogram was constructed by UPGMA (unweighted pair-group
method with arithmetical averages) cluster analysis in the CLUSTER procedure of the SAS system. The RAPD analysis was a useful
tool in determining the extent of genetic diversity among Tainan-white maize inbred lines in the present case. Cluster analysis
showed that the 13 inbred lines could be classified into distinct heterotic groups. There was no significant linear regression
of grain dry weight heterosis value and mean performance of hybrids on genetic distance. And their coefficients of determination(R2) are small, so that predictive value is limited. The present results showed that the Jaccard's similarity coefficients based
on RAPD data cannot be used to precisely predict the F1 hybrids yield performance and heterosis value.
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
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Twenty two RAPD and 22 ISSR markers were evaluated for their potential use in determination of genetic relationships in chickpea
(Cicer arietinum L.) cultivars and breeding lines. We were able to identify six chickpea cultivars/breeding lines by cultivar-specific markers.
All of the cultivars tested displayed a different phenotype generated either by the RAPD or ISSR primers. Though ISSR primers
generated less markers than RAPD primers, the ISSR primers produced higher levels of polymorphism (% of polymorphic markers
per primer) than RAPD primers. A high level of within cultivar homogeneity was observed in chickpea. Cultivars/breeding lines
originating from a common genetic background showed closer genetic relationship. Chickpea lines with similar seed type(kabuli
or desi) had a tendency to cluster together.
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献