限制性位点扩增多态性（restriction site amplification polymorphism，RSAP）是根据基因组内普遍存在的酶切位点来设计特殊的引物，通过简单的梯度PCR反应来产生多态性。本研究首次将RSAP标记技术成功应用于紫菜遗传多样性检测和种质...限制性位点扩增多态性（restriction site amplification polymorphism，RSAP）是根据基因组内普遍存在的酶切位点来设计特殊的引物，通过简单的梯度PCR反应来产生多态性。本研究首次将RSAP标记技术成功应用于紫菜遗传多样性检测和种质鉴定。利用所建立的适合于紫菜RSAP分析的PCR反应体系和反应条件，使用30对引物对15个紫菜系DNA进行了PCR扩增，经过3次重复验证，有12对引物能够扩增出清晰且稳定的条带。这12对引物共扩增出413条带，其中408条为多态性条带，多态性比例96％，平均每对引物产生34条多态性条带，片段大小在50-500bp之间。利用这413条带进行聚类分析，产生了这15个紫菜系的进化树。15个紫菜系在0.69相似系数水平上分为两大类：一类包括坛紫菜；另一类包括条斑紫菜和少精紫菜。选2对引物R1／R6和R3／R4所扩增的10条稳定且重复性好的条带，构建了这15个紫菜系的RSAP-DNA指纹图谱。在该指纹图谱中，每个紫菜系都有其独一无二的指纹模式，能够很容易地与其它紫菜系相区分。展开更多
初步探讨限制性位点扩增多态性（Restriction site amplified polymorphism，RSAP）分子标记技术在龙须菜遗传多样性检测和种质鉴定上的应用。利用所优化的适合于龙须菜RSAP分析的PCR反应体系，对青岛湛山湾野生群体和栽培品系981、07-...初步探讨限制性位点扩增多态性（Restriction site amplified polymorphism，RSAP）分子标记技术在龙须菜遗传多样性检测和种质鉴定上的应用。利用所优化的适合于龙须菜RSAP分析的PCR反应体系，对青岛湛山湾野生群体和栽培品系981、07-2进行遗传多样性分析和比较。试验采用15对引物组合在3个群体14个龙须菜个体中共扩增出669个位点，其中多态位点数为146个，多态性比例22％，平均每对引物产生10条多态性条带，片段大小在100～1000bp之间。湛山湾野生群体的Na（1．0075）、Ne（1．0071）、H（O．0036）、I（0．0051）与栽培品系981的Na（1．003O）、NP（1．0021）、H（0．0012）、I（0．0018），以及栽培品系07-2的Na（1．009O）、Ne（1．0063）、H（0．0037）、I（0．0054）相比较可知3个群体的遗传多样性是有差异的。种群间的遗传多样性分析表明，在所有检测的样品中，遗传多样性多来自不同群体间的多样性。群体遗传结构分析表明，2个栽培品系981、07—2间遗传距离不大，但栽培品系与湛山湾野生群体间的遗传距离相对较大，而UPGMA聚类分析也明显的将湛山湾野生群体与栽培品系981、07—2区分开来，表明野生群体与栽培品系间已产生一定的遗传隔离。通过分析湛山湾野生群体及栽培品系981、07—2的RSAP指纹图谱，从中筛选栽培品系981的特异条带，并将其转化为稳定性好、特异性高的序列特征化扩增区（Sequence characterized amplified regions，SCAR）分子标记。展开更多
[Objective] This study aimed to analyse the genetic diversity of and genet- ic relationship between 33 Dendrobium species by using RSAP （Restriction Site Amplification Polymorphism） marker technique. [Method] A PCR-...[Objective] This study aimed to analyse the genetic diversity of and genet- ic relationship between 33 Dendrobium species by using RSAP （Restriction Site Amplification Polymorphism） marker technique. [Method] A PCR-reaction system was established, by which 33 pairs of primers screened from 45 pairs were used to con- duct the PCR amplification. [Results] The 33 pairs of primers produced 2 047 ampli- fied fragments, of which 2 044 were polymorphic, with a polymorphism rate of 99.8%, and each pair of primers produced 61.9 polymorphic fragments on average. The genetic similarity coefficients between species ranged from 0.081 to 0.442, sug- gesting that the experimental materials used in this study possess a wide genetic background. According to the result of RSAP, the 33 Dendrobium species were di- vided into 6 groups by the cluster analysis （UPGMA）. The result showed that the genetic relationship between D. acinaciforme and D. guangxiense was the closest, and that between D. sinense and either D. lituiflorum or D. hancockii was the fur- thest. [Conclusion] The result suggested that Dendrobium germplasms have a great genetic diversity. This study provides a molecular basis for the classification and breeding of Dendrobium plants, as well as the selection of hybridization parents in the future.展开更多
限制性位点扩增多态性(restriction site amplified polymorphism,RSAP)是一种新型的分子标记技术,为了明确该标记技术在苎麻上利用的可行性。对限制性位点扩增多态性(RSAP)-PCR反应体系利用正交试验进行优化。结果表明:25μL体系中,其...限制性位点扩增多态性(restriction site amplified polymorphism,RSAP)是一种新型的分子标记技术,为了明确该标记技术在苎麻上利用的可行性。对限制性位点扩增多态性(RSAP)-PCR反应体系利用正交试验进行优化。结果表明:25μL体系中,其最优条件为TaqDNA聚合酶1U,dNTPs 0.3mmol/L,引物0.25μmol/L,DNA 10ng。展开更多
目的:研究重楼属植物DNA指纹图谱,初步探讨RSAP分子标记技术在重楼属植物遗传多样性研究上的应用。方法:采用限制性位点扩增多态性(restriction site amplification polymorphism,RSAP)技术对重楼属8个种进行基因组DNA多态性分析。结果:...目的:研究重楼属植物DNA指纹图谱,初步探讨RSAP分子标记技术在重楼属植物遗传多样性研究上的应用。方法:采用限制性位点扩增多态性(restriction site amplification polymorphism,RSAP)技术对重楼属8个种进行基因组DNA多态性分析。结果:用10条RSAP引物即选出18对引物组合构建了8个种的DNA指纹图谱。通过遗传距离构建系统进化树,重楼属8个种的进化关系得到了很好的分辨。结论:RSAP标记能够对重楼属植物进行准确的分子鉴定,为重楼属中药材的种类鉴定和种间的分类地位提供分子生物学依据。展开更多
文摘限制性位点扩增多态性（restriction site amplification polymorphism，RSAP）是根据基因组内普遍存在的酶切位点来设计特殊的引物，通过简单的梯度PCR反应来产生多态性。本研究首次将RSAP标记技术成功应用于紫菜遗传多样性检测和种质鉴定。利用所建立的适合于紫菜RSAP分析的PCR反应体系和反应条件，使用30对引物对15个紫菜系DNA进行了PCR扩增，经过3次重复验证，有12对引物能够扩增出清晰且稳定的条带。这12对引物共扩增出413条带，其中408条为多态性条带，多态性比例96％，平均每对引物产生34条多态性条带，片段大小在50-500bp之间。利用这413条带进行聚类分析，产生了这15个紫菜系的进化树。15个紫菜系在0.69相似系数水平上分为两大类：一类包括坛紫菜；另一类包括条斑紫菜和少精紫菜。选2对引物R1／R6和R3／R4所扩增的10条稳定且重复性好的条带，构建了这15个紫菜系的RSAP-DNA指纹图谱。在该指纹图谱中，每个紫菜系都有其独一无二的指纹模式，能够很容易地与其它紫菜系相区分。
基金Supported by the Hainan Natural Science Foundation(312024)Special Fund for Scientific Research in the Public Interest,China(201203071)~~
文摘[Objective] This study aimed to analyse the genetic diversity of and genet- ic relationship between 33 Dendrobium species by using RSAP （Restriction Site Amplification Polymorphism） marker technique. [Method] A PCR-reaction system was established, by which 33 pairs of primers screened from 45 pairs were used to con- duct the PCR amplification. [Results] The 33 pairs of primers produced 2 047 ampli- fied fragments, of which 2 044 were polymorphic, with a polymorphism rate of 99.8%, and each pair of primers produced 61.9 polymorphic fragments on average. The genetic similarity coefficients between species ranged from 0.081 to 0.442, sug- gesting that the experimental materials used in this study possess a wide genetic background. According to the result of RSAP, the 33 Dendrobium species were di- vided into 6 groups by the cluster analysis （UPGMA）. The result showed that the genetic relationship between D. acinaciforme and D. guangxiense was the closest, and that between D. sinense and either D. lituiflorum or D. hancockii was the fur- thest. [Conclusion] The result suggested that Dendrobium germplasms have a great genetic diversity. This study provides a molecular basis for the classification and breeding of Dendrobium plants, as well as the selection of hybridization parents in the future.
文摘目的:研究重楼属植物DNA指纹图谱,初步探讨RSAP分子标记技术在重楼属植物遗传多样性研究上的应用。方法:采用限制性位点扩增多态性(restriction site amplification polymorphism,RSAP)技术对重楼属8个种进行基因组DNA多态性分析。结果:用10条RSAP引物即选出18对引物组合构建了8个种的DNA指纹图谱。通过遗传距离构建系统进化树,重楼属8个种的进化关系得到了很好的分辨。结论:RSAP标记能够对重楼属植物进行准确的分子鉴定,为重楼属中药材的种类鉴定和种间的分类地位提供分子生物学依据。
