AFLP和RFLP标记检测水稻亲本遗传多样性比较研究

 利用AFLP和RFLP技术对 2 0个水稻品种 (系 )进行了遗传多样性分析。在AFLP分析中 ,用 6 4对引物组合进行了初筛 ,然后用其中适合水稻的 15对引物进行了详细研究 ,每对AFLP引物组合扩增出了 4 7～ 118条带 ,其中多态性带为 5～ 32条 ,15个引物组合共得到多态性条带 2 99条 ;在RFLP分析中 ,4 9个RFLP探针共检测出 10 7条多态性带。聚类分析表明 ,两种标记均能将 2 0个材料分成籼粳两大类群 ,但AFLP标记比RFLP标记更能反映品种间系谱亲缘和地理生态上的差异。两种标记检测出的遗传距离的趋势是一致的 ,但AFLP标记相对于RFLP而言提高了品种间的遗传距离的估计 ,降低了亚种间遗传距离的估计 ,因此 ,AFLP更适应于多样性的研究 ,RFLP更适应于籼粳分类及度量籼粳分化程度     

部分菊花品种遗传多样性的AFLP分析

利用AFLP分子标记技术研究了26个菊花品种的遗传多样性,7个引物组合共扩增出385条带,其中多态性条带285条,多态性占74.03%,平均每个引物组合扩出多态性条带40.7条,每一个引物组合都可将26个品种完全分开。利用AFLP数据,进行聚类分析,将26个菊花品种聚为5类。结果表明:AFLP技术在菊花品种分类中的应用是可行的。     

文心兰种质遗传多样性的SRAP分析

利用SRAP标记分析文心兰部分种质资源的遗传多样性,采用30对引物组合对33份文心兰种质进行扩增,从中筛选出19个多态性较高的引物组合,共产生277条多态性条带,平均每个引物组合产生14．6个多态性条带,相似系数在0．280-0．927。通过聚类分析发现,在值为0．37时,可将33份文心兰种质划分为4大类群。第1类厚叶大花品种,第1I类为厚叶小花品种,第III、第Ⅳ类均为薄叶品种。结果表明,文心兰种质的聚类与其遗传背景有很大的关系。     

地方鸡种微卫星DNA指纹图谱建立与遗传多样性研究*

 利用20个微卫星标记对我国19个地方鸡种保种群进行了遗传检测，构建了各个品种的微卫星DNA指纹图谱，通过计算各群体的等位基因频率、平均基因杂合度、平均多态信息含量及各群体间的遗传距离，并用类平均法进行聚类分析，分析了所研究鸡种的遗传关系。研究结果表明：20个微卫星标记在19个地方鸡种保种群共检测到184个等位基因，平均为9.2个,基因频率分布在0.013～0.838之间。19个地方鸡种平均杂合度在0.5824～0.7432之间。其中藏鸡最高，白耳鸡最低。20个微卫星座位的平均多态信息含量在0.5238～0.7023之间，均大于0.5，表现为高度多态性；19个鸡种聚为6类。各鸡种的遗传距离及聚类结果与所保存的地方鸡种的地理分布、现实状况是相吻合的，从而表明用该方法分析品种间的亲缘关系是可行的。     

7种木兰科常绿植物亲缘关系的ISSR和AFLP分析

利用ISSR和AFLP 2种分子标记方法对7种木兰科常绿植物遗传多态性和亲缘关系进行了分析,建立了木兰科常绿植物ISSR和AFLP的优化反应体系,构建了UPGMA系统树.利用筛选的11个引物对7份样本进行ISSR分析,共获得131条带,其中多态性带86条,平均每个引物产生11.10条带,平均多态性检出率为70.37%,说明供试的7种材料种质间有较高等位多态性.用22对EcoRⅠ/MseⅠ引物组合对7份样本进行了AFLP分析,共扩增出1 310条带,其中多态性带1 204条,平均每对引物扩增带数59条,多态性率92.5%.通过2种标记发现,木兰属与含笑属聚类在一起,他们之间存在的遗传差异通过分子标记可以鉴定出来,而木莲属的川滇木莲在聚类分析中差异性最明显,尤其与含笑属,虽然他们的外部形态相似,但遗传距离较远.研究还发现,在2组试验的遗传相似系数结果中,均有超出0～1范围的数值出现,根据分析为偏分离现象,初步推断出现此种结果应与材料来源有关.     

利用SRAP和RGA标记对新疆海岛棉品种的聚类分析

[目的]对新疆自育的新海系列品种进行遗传多样性研究.[方法]利用SRAP和RGA标记,检测出多态性位点;利用NTSYSpc2.1软件,分别计算两种分子标记数据的系数矩阵,采用UPGMA法对所选材料进行聚类分析.[结果]SRAP共检测出了134个多态性位点,每组合的多态性条带数从2～8不等,平均每个引物组合产生2.91个多态性位点,表明SRAP能检测出较多的遗传位点,能够较好地反映海岛棉的遗传多样性.RGA标记共扩增出了46个多态性位点,每组合的多态性条带数从2～5不等,平均每个引物组合产生2.08个多态性位点,表明RGA作为一种分子标记也能检测出较多的遗传位点,能够较好地反映海岛棉在抗病方面的遗传多样性.[结论]SRAP聚类结果基本与品种系谱来源一致,RGA聚类基本与材料的抗病水平一致.分子标记在鉴别品种和品种遗传多样性研究方面具有重要作用.     

半番鸭羽色性状AFLP和RAPD标记分析

利用RAPD和AFLP技术对半番鸭2个羽色池DNA进行检测,探讨两种技术对半番鸭羽色性状标记的效果。结果显示：(1)RAPD方法扩增带和特异性条带都较少,37个引物共扩增到297个清晰条带(平均每个引物4条)；在37个引物中,7个引物含有羽色特异性条带,共15条,其中自羽半番鸭特有的条带为10条。(2)AFLP方法荧光图谱清晰度高,扩增带丰富,16对引物组合共检测1000多条扩增带(平均每个引物组合30多)；在16引物中,4个引物组合检测到羽色特异性条带,共90条,仅白羽半番鸭特有的条带可达37条。可见,采用AFLP技术对半番鸭羽色标记效果更佳。     

羽扇豆种间遗传差异AFLP分析

 利用AFLP对羽扇豆 (LupinusL .) 3个种 (L .angustifoliusL .,L .albusL .,L .pilosusMurr.)的 18个品种指纹图谱进行了分析 ,4个引物组合共产生 46 2条带、多态性带为 44 2 ,平均每个引物组合产生 110 .5多态性带 ,多态性百分率为 95 .7%。总体平均遗传多样性指数为 0 .781。种间遗传差异大于品种间的遗传差异 ,4个引物组合种间和品种间的平均鉴定效率分别为 10 0 %和 84.7%。按UPGMA方法进行聚类分析 ,所有的供试材料明显地分为 3个组 ,每一个组由同一个种的品种构成。     

马铃薯品种遗传多样性的AFLP分析

利用AFLP分子标记技术对19份克新系列马铃薯品种进行了遗传多样性分析,用30对引物组合进行了初筛,选出7对有多态性的引物组合进行了详细研究。每对AFLP引物组合扩增出54￣90条带,共获得495条带,其中多态性条带为302条。AFLP分析表明19个材料的遗传距离介于0.2091￣0.7679之间,平均值为0.4811。聚类分析将19份材料划分为4类,其中第2类包括14个品种,占总数73.86%,表明多数品种亲缘关系较近,但有少数品种亲缘关系较远,说明克新系列马铃薯的遗传基础有所拓宽。研究表明:AFLP指纹分析技术具有很高的分辨率,适于进行马铃薯遗传多样性研究。     

3种分子标记分析罗非鱼选育群体 的多态性效率比较

应用RAPD、AFLP、SSR等3种分子标记,对吉富品系尼罗罗非鱼选育群体的多态性进行了分析.在40条RAPD引物、19对SSR引物、64对AFLP引物中,具有多态性、重复性好的引物分别为17条、19对和8对,扩增出多态性条带数分别为35、92、181条.结果表明,RAPD、AFLP、SSR多态性条带比例分别为20.35％、79.64％和58.77％,平均每个(对)多态性引物可以扩增出多态性片段数目分别为2.06、4.84和22.63.说明SSR和AFLP是研究吉富罗非鱼选育群体更为有效的分子标记.     

利用RAMP标记研究24份优质抗稻瘟病水稻种质资源的遗传多样性

对24份优质抗稻瘟病水稻种质资源进行了RAMP(random amplified microsatellite polymorphism)标记分析。结果表明,80个RAMP引物组合中,有22个引物组合可扩增出清晰且具多态性的条带。这22个引物组合共扩增出108条带,其中80条具有多态性,占74.1%,每个引物组合可扩增出1~9条多态性带,平均3.6条。RAMP标记遗传相似性系数(GS)平均值为0.462(0.138~0.900)以平均GS值0.462为阈值,可将所有供试材料划分为6类。表明这24份优质抗稻瘟病种质资源具有较丰富的遗传多样性。这些优质抗源材料对选育高产、优质、抗稻瘟病新品种(组合)具有重要利用价值。     

The power of microsatellite markers and AFLPs in revealing the genetic diversity of Hashemi aromatic rice from Iran

Hashemi, a popular aromatic rice among Iranians, is famous for its fragrance and taste. Such features are major reasons for its higher price compared to non-aromatic varieties available in Iran. Therefore, the knowledge of genetic diversity of this profitable crop is a fundamental ineterst for plant breeders in future breeding programs. In the present research, genetic diversity among 35 genotypes of Hashemi aromatic rice(Oryza sativa L.) from Guilan and Mazandaran provinces of Iran was estimated using simple sequence repeat(SSR) and amplified fragment length polymorphism(AFLP) markers. Out of 21 SSR and 12 Eco RI-Mse I AFLP marker combinations, only 16 SSRs and 10 AFLPs exhibited polymorphic patterns while others were monomorphic. The 10 AFLP primer combinations produced a total of 142 of bands and 20 were polymorphic(14.08%). Moreover, 40 out of 47 bands amplified with 16 SSR markers showed polymorphism(85.1%). The average number of alleles identified by SSR primers was 2.56 alleles per locus with a range of 2 to 4. The average value of polymorphic information content(PIC) was 0.393 and 0.468 for AFLP and SSR markers, respectively. However, the genetic similarity values ranged from 0.26 to 1 for SSRs and 0.21 to 1 for AFLPs. Later, a unweighted pair group method with arithmetic mean(UPGMA) dendrogram was generated and genotypes were clustered into four groups with SSRs at similarity coefficient of 0.55 while AFLPs clustered them into six groups at similarity coefficient of 0.41. Cluster analysis revealed a narrow genetic diversity and low correlation between geographical differentiation and genetic distance within cultivars.     

Assessment of genetic diversity among Chinese upland cottons with Fusarium and/or Verticillium wilts resistance by AFLP and SSR markers

Genetic diversity among 95 Chinese upland cottons with Fusarium and/or Verticillium wilts resistance was estimated using Amplified Fragment Length Polymorphism (AFLP) and Simple Sequence Repeat (SSR) markers. Twenty EcoRI-MseI AFLP and 19 SSR primers with polymorphism were selected to perform the fingerprinting. The results showed that 20 AFLP primer pairs produced a total of 1 480 major bands among 95 genotypes, and 214 were polymorphic bands. The number of total bands per primer pair ranged from 47 to 109, with an average of 74.0. The polymorphism information content (PIC) values for the 20 primer pairs varied from 0.01 (E-ACT/M-CAT) to 0.24 (E-ACA/M-CTA), and the average value was 0.09. Nineteen SSR primers generated 89 DNA bands, of which 61 were polymorphic. The total number of alleles per locus varied from 3 to 8, with an average of 4.7. The average PIC value for the SSR amplification products was 0.69. Genetic similarity estimates for the entire data set ranged from 0.978 to 0.998 based on AFLP and SSR bands. It was proved that the close genetic relationship and narrow genetic diversity existed in the tested cultivars. The clustering patterns could not be correlated to the geographic origin, the pedigree and common parentage of the cultivars.     

Comparison Between AFLP and RFLP Markers in Detecting the Diversity of Rice (Oryza sativa L.)

AFLP and RFLP were used to study the diversity of 20 rice cultivars. 15 primer combinations were used in the AFLP analysis and 47 - 118 bands were amplified in each lane. A total of 107 polymorphic bands were detected in the RFLP analysis using 49 RFLP probes. The cluster analysis based on RFLP data showed that 20 rice cultivars could be divided into an indica group and a japonica group, as did the AFLP data; however AFLP is more suitable in detecting the difference of pedigree and ecotype among the 20 cultivars.The genetic distance based on AFLP and RFLP data showed the same tendency, but AFLP markers increased the measure of genetic distance among intra-subspecific cultivars and decreased the measure of genetic distance among inter-subspecific cultivars relative to RFLP markers. That indicated that AFLP is more suitable than RFLP in the diversity study and RFLP is more suitable to study the indica-japonica differentiation of cultivars.     

利用ISSR和AFLP标记分析石斛种质资源的遗传多样性

本研究利用ISSR和AFLP两种分子标记对51个药用石斛材料（包括35个铁皮石斛种质资源）进行遗传多样性研究。利用ISSR标记筛选出6条引物,共扩增出79条带,平均每条引物扩增条带数约为13.17条,多态条带比率都达到100%。利用AFLP标记筛选出5对引物,共扩增207条带,其中206条具多态性,多态性比率高达99.52%。两标记分析获得的群体种系间关系与变异基本一致,且群体内的分化较小,群体间的分化较大,群体间的基因流动较小。所得到的聚类图表明：ISSR和AFLP标记技术能有效地将研究材料分开,且基本反映了它们之间的亲缘关系,说明两种标记均适合用于石斛植物的遗传多样性研究。     

江苏3个秀丽白虾野生群体遗传多样性的AFLP分析

采用AFLP分子标记技术对江苏省内3个秀丽白虾野生群体的遗传多样性水平及遗传结构进行了分析.采用3个引物组合(E-AAC/M-CAA、E-AAC/M-CAG、E-AAC/M-CAT),共扩增出102个清晰位点,其中93个为多态性位点.结果显示,3个秀丽白虾野生群体在物种水平和群体水平上均蕴含着较为丰富的遗传多样性.在物种水平上,秀丽白虾多态性位点百分率(PPL)为91.18%,Nei's基因多样性指数(h)为0.391,Shannon信息指数(I)为0.562.群体水平上的PPL为58.82%～70.59%,平均为65.4%,其中骆马湖群体最高,为70.6%;长江群体最低,为58.8%.群体间Nei's多样性指数为0.268～0.320,平均为0.292;Shannon信息指数为0.381～0.454,平均为0.417.长江南京段群体与太湖群体间的分化系数较低(0.141),长江南京段群体与骆马湖群体间的遗传分化系数较高(0.234).建议加强秀丽白虾引种、育种工作中的管理力度,建立详实的引育种档案,推动秀丽白虾养殖工作的发展.     

Assessment of genetic diversity in glandless cotton germplasm resources by using agronomic traits and molecular markers

Seventy-one glandless cotton germplasm resources were firstly evaluated genetically by using nine agronomic traits, 33 simple sequence repeat (SSR) primers and ten amplified fragment length polymorphism (AFLP) primer combinations. Principal component analysis (PCA) of the agronomic traits showed that the first six principal components (PCs) explained a total of 86.352% of the phenotypic variation. A total of 329 alleles were amplified for 33 SSR primers, and 232 polymorphic bands in a total of 389 bands were obtained by using ten AFLP primer combinations. The average polymorphic information content (PIC) value was 0.80 and 0.18 for SSR primers and AFLP primer combinations, respectively. The DIST (average taxonomic distance) and DICE (Nei and Li’s pairwise distance) coefficients ranged from 0.373 to 3.164 and 0.786 to 0.948, respectively, for agronomic traits and SSR&AFLP data based on UPGMA analysis. This suggested that there was a higher diversity in the evaluated population for both agronomic traits and molecular markers. The Mantel’s test showed that the correlation between the dendrograms based on agronomic traits and SSR&AFLP data was non-significant.     

部分牡丹品种遗传多样性的AFLP分析

【目的】研究牡丹遗传多样性为合理利用牡丹种质资源、培育观赏价值高的牡丹新品种提供依据。【方法】利用荧光标记AFLP技术，采用8对M+3和P+3引物组合对30份牡丹栽培品种进行了总基因组DNA水平上的多态性检测。【结果】检测结果共获得1 123条可统计的条带，其中965条呈多态性，多态性带百分率达86%。揭示了牡丹栽培种质丰富的遗传多样性。8组引物在30个品种中检测到数目不等的品种特异带型，这些品种的特异带型对供试牡丹品种具有一定的鉴别价值。【结论】8对引物能将30个牡丹品种完全区分开。聚类分析结果表明多数来源地相同的牡丹种质表现出较为密切的亲缘关系。