茄子耐盐种质资源遗传多样性的SRAP和ISSR分析

利用SRAP和ISSR分子标记,研究了14份耐盐茄子种质资源的遗传多样性,结果表明,2种标记均能揭示材料间较高的遗传多样性,其中ISSR标记多态性略高于SRAP标记。在SRAP分析中,每对引物组合可扩增出8-15条DNA片段,平均为12．12条：26对SRAP引物组合共扩增出315条DNA片段,其中263条具有多态性,多态性比率为83．49％;材料间遗传相似系数变化范围为0．212～0．923,平均值为0．755。在ISSR分析中,每个引物可获得5～16条DNA片段,平均为10．87条;15个ISSR引物共扩增出163条DNA片段,其中141条具有多态性,多态性比率为86．50％;材料间遗传相似系数变幅为0．333-0．957,平均值为0．736。聚类分析表明,2种标记都能将供试材料完全区分开来,聚类结果具有一定的相似性,但也存在明显差异。Mantel相关分析表明,SRAP分析与ISSR分析的相关性达到极显著性水平（r=0．904,P〈0．01）。     

西南地区乡土杨树遗传变异的SRAP分析

采用SRAP分子标记技术对西南地区11种乡土杨树共333份样本的遗传变异进行分析,7对引物组合共扩增出215条带,其中多态性条带158条,多态性条带百分率为73.49%,表明11种乡土杨树间存在广泛变异。AMOVA分析结果显示,种间遗传变异分量为10.84,占总变异的48.70%,遗传差异达极显著水平(P0.001)。种间的遗传相似系数变幅在0.8199~0.9607之间,平均遗传相似系数为0.8983。聚类结果表明,昌都杨和藏川杨之间的遗传差异最小,大叶杨和三脉青杨之间的遗传差异最大。本研究结果为西南地区乡土杨树基因资源的保护、开发和利用提供了一定的科学理论依据。     

SSR与SRAP标记在玉米品种鉴定中的比较研究

利用SSR标记和SRAP标记对19个玉米品种及8份莱农15样品进行了分析,比较了2种标记的分辨能力及在亲缘关系和杂交种纯度鉴定中的表现.与SSR标记相比,SRAP标记用于玉米品种鉴定扩增住点数量更多,PIC值更高,具有更高的分辨率;在亲缘关系分析方面,SSR检测的遗传距离变幅更大,2种标记计算的遗传距离呈极显著正相关,分类结果基本一致;在杂交种纯度检测中,SRAP标记的期望位点在杂交种群体中检测率高于SSR标记相应位点,检测杂交种纯度结果更接近田问种植鉴定,因而准确度更高.SRAP在玉米品种鉴定中具有一定的优势,可作为SSR标记技术的有益补充,特别是在杂交种纯度鉴定中应用.     

棉花遗传多样性SCoT和SRAP标记的研究及比较分析

利用SCoT和SRAP两种分子标记技术对30份彩色棉与白色棉种质资源,进行遗传多样性研究。用29对SRAP引物组合和26个SCoT引物分别对供试棉花的基因组DNA进行扩增。SCoT引物共扩增出163条带,多态性比率为61.96%,遗传相似系数GS值变化范围为0.5405～0.9972。SRAP引物组合共扩增条带1067条,多态性比率仅为14.1%,遗传相似系数GS值变化范围为0.5415～0.9109。两种标记系统得到了相似但并不完全相同的聚类图,2种标记方法间存在显著相关性(r=0.5518,P<0.05)。结果表明,SRAP与SCoT标记均适用于棉花种质的遗传多样性分析,且SCoT的标记指数MI高于SRAP标记,为SCoT这种新兴的标记技术在棉花育种中的应用提供了重要的依据。     

浙南柚类地方资源遗传多样性分析和鉴定

利用SRAP标记对13份浙南柚类地方资源和琯溪蜜柚及芽变进行遗传多样性分析和鉴定。结果表明:平均每个引物组合可扩增出15.7条谱带,14对SRAP引物共产生220条谱带,其中多态性谱带为122条,多态率为55.4%,表明15份材料间检测到的SRAP位点多态性不高,不同引物组合可将11个基因型完全分开。聚类分析结果显示,15份柚类种质在遗传相似系数0.97处可以分为8大类,第1类群为四季柚7个优异株系,第2类群包括琯溪蜜柚及其芽变,而平阳文旦、早香柚、处红柚、红心1号土柚、红心2号土柚、酸柚分别单独为第3、4、5、6、7、8类群。四季柚选系中务城1号、务城3号、马站红心四季柚的指纹图谱有可区分的差异,说明DNA水平发生轻微变异。     

应用SRAP标记分析黄瓜的遗传差异

利用49对SRAP引物对4种不同类型28份黄瓜种质资源进行了遗传差异分析.结果表明,有35对引物扩增出多态性,在28份资源间共产生724条扩增带,平均每对引物组合产生20.69条;共检测出337个多态性位点,多态性比率为46.5%,每对引物平均为96.3个.利用NTSYS软件分析遗传相似系数,UPGMA方法聚类分析表明,28份资源可聚为两大类.试验结果表明SRAP标记位点多,重复性好,可以揭示不同类型黄瓜种质之间的遗传基础.     

基于SRAP和TRAP标记的河北野生狗牙根种质遗传多样性分析

以SRAP和TRAP 2种标记技术对36份狗牙根材料的遗传多样性及亲缘关系进行了分析,其中包含34份河北省野生狗牙根种质资源。分别由238对SRAP和85对TRAP引物组合中筛选获得具有多态性的SRAP和TRAP引物组合各10对,PCR扩增总条带分别为186和161条,多态性条带156和132条,平均每对引物扩增出多态性条带各15.6和13.2条,多态性位点比率分别为83.4%和81.0%。2种标记合并进行聚类分析,所有供试的36份狗牙根材料遗传相似系数GS=0.519~0.983,平均为0.7。当GS=0.68时,可将36份供试材料分为4个类群。本研究结果表明河北野生狗牙根种质资源存在较丰富的遗传多样性,可为种质资源保护和选育优良狗牙根新品种提供科学依据。     

Genetic diversity of endangered Manglietia patungensis assessed by inter simple sequence repeat and sequence-related amplified polymorphism markers

Manglietia patungensis Hu is an endangered plant native to China. Knowledge of its genetic diversity and structure would aid its conservation. This study assessed nine natural populations of M. patungensis using two methods: inter simple sequence repeat (ISSR) and sequence-related amplified polymorphism (SRAP) markers. Using 10 ISSR primer pairs, 334 bands were generated, and 10 SRAP primer pairs generated 276 bands. The percent of polymorphic bands (91.32% and 93.48%), Nei's genetic diversity (0.3448 and 0.3323), and Shannon's information index (0.5075 and 0.4935) revealed a high level of genetic diversity at the species level. Total heterozygosity was 0.3439 by ISSR and 0.3281 by SRAP. The mean heterozygosity was 0.2323 by ISSR and 0.2521 by SRAP. The coefficient of genetic differentiation among natural populations was 0.3245 by ISSR and 0.2316 by SRAP. These data indicated higher levels of genetic diversity of M. patungensis within, rather than among, populations. Estimates of gene flow among natural populations were 1.0411 and 1.0589, which implied a certain amount of gene exchange among populations. A Mantel test revealed no significant correlation between genetic and geographic distance. ISSR and SRAP markers are both effective for genetic diversity research in M. Patungensis. Based on these results, conservation of M. patungensis should be performed both in situ and ex situ.     

Genetic diversity in Vicia amoena (Fabaceae) germplasm resource in China using SRAP and ISSR markers

The genetic diversity in 17 germplasms of Vicia amoena L. from north China was analyzed using SRAP and ISSR markers. Three hundred and sixty-eight (94.11%) polymorphic bands (211 and 157 obtained from 20 pairs of SRAP and ISSR primers, respectively) were scored. Although SRAP was more effective than ISSR markers with higher PIC, RP and larger variation range of genetic distance, both the markers were useful for assessing V. amoena genetic diversity. Cluster analysis showed that the 17 germplasms were clustered into 5 groups. The results of principal coordinate analysis supported UPGMA clustering. The germplasms from source areas where the annual average temperature ranged from −1.0 to 5 °C exhibited the highest level of genetic diversity with the highest PPI, I and H. These results have important implications in genome mapping, breeding purposes, and germplasm conservation.     

基于SRAP分子标记的冰草遗传连锁图谱构建

构建高密度遗传连锁图谱是冰草抗性、品质、产量等重要性状QTL精细定位及标记辅助育种研究的基础。该试验以四倍体杂交冰草F2群体的202个分离单株及其亲本为材料,利用SRAP分子标记技术和Join Map 4.0作图软件对冰草的遗传连锁图谱进行了构建。结果表明:(1)共筛选出22对多态性好、标记位点清晰稳定的SRAP适宜引物,对冰草杂种F2分离单株的基因组DNA进行PCR扩增,共获得510个SRAP多态性标记位点,其比率占88.2%。(2)偏分离分析表明,偏分离标记比率仅为14.12%,符合遗传作图的要求。(3)成功构建了冰草的SRAP分子标记遗传连锁图谱,该图谱有14个连锁群、510个标记,连锁群间长度范围86.4～179.0cM,覆盖基因组总长度1 912.9cM,标记间平均间距3.75cM,为高密度遗传图谱。     

辣椒31个优良自交系的亲本类群分析

以包含我国重要尖椒品种的亲本材料在内的31份优良自交系为材料, 利用SRAP标记和基因型值分析技术开展了辣椒自交系间遗传差异的分析与类群划分研究。结果表明: 在30个引物组合中, 27个引物组合可以 在自交系间扩增出多态性条带, 共扩增出310个多态性条带, 平均每个引物组合产生11.5个多态性条带, 显示出SRAP技术具有较强的分析效率; 基于SRAP标记和Yule相似系数对这些自交系进行的聚类分析中, 可以基本区分辣椒的2个变种(C. annuum var. grossum和C. annuum var. longum), 而且可以反映出自交系间的亲缘及系谱关系; 在相似系数为0.67处, 可将这31个自交系分为4个类群; 基于基因型值和标准Euclidean距离对这些自交系进行的聚类分析可成功地将辣椒的两个变种完全区分; 在遗传距离约4.5处, 可将这31个自交系分为4个类群; 自交系间基于SRAP标记与基因型值的遗传距离存在一定的相关性。     

Genetic diversity analysis among and within populations of Pogostemon cablin from China with ISSR and SRAP markers

The diversity and genetic relationship among and within 16 populations of Pogostemon cablin from China were analyzed using ISSR and SRAP markers. High percentage of polymorphism observed by these two markers indicated a high level of genetic diversity existed among P. cablin populations. The percentage of polymorphic bands revealed by these two markers showed genetic variations were much higher between populations of P. cablin than within. The UPGMA clustering results using ISSR, SRAP or ISSR + SRAP combination showed most accessions from the same or adjacent regions were classified together. However, cluster results based on the absolute contents of patchouli alcohol and pogostone did not keep in good accordance with their geographic distributions, which revealed that the forming of patchouli alcohol and pogostone were relevant to the cultivation conditions except for genetic factors and environmental conditions. Finally, an appropriate strategy for conserving the patchouli germplasm was proposed.     

29个香蕉基因型遗传多样性的SRAP分析

采用SRAP分子标记技术对29个香蕉品种(系)的多样性进行研究,结果显示,64对SRAP引物中筛选出25个多态性较高的引物组合,共扩增出324条条带;UPGAM聚类图显示所有供试的29个香蕉品种(系)可分为2个类群且与基因型相一致;实验结果与形态、农艺性状标记分类基本一致。研究表明,SRAP技术可有效运用于香蕉基因型的遗传和育种研究。     

Sequence-related amplified polymorphism (SRAP) marker as a new method for identification of endophytic fungi from Taxus

A total of 20 endophytic fungi stains were classified into four groups using traditional morphological identification method, and were studied for genetic diversity by sequence-related amplified polymorphism (SRAP) technique. Genomic DNA (deoxyribonucleic acid) of these strains was extracted with CTAB method. SRAP analysis was done with 24 pairs of primers. All strains could be uniquely distinguished with 584 bands and 446 polymorphism bands which generated 76.4% of polymorphic ratio. Unweighted pair-group method with arithmetical averages cluster analysis enabled construction of a dendrogram for estimating genetic distances between different strains. All strains, which were just divided into four groups by traditional morphology identification, were clustered into four major groups at GS = 0.603 and further separated into eight sub-groups at GS = 0.921. Dendrogram also revealed a large genetic variation in 20 strains; different primer combinations allowed them distinctly distinguished one from others with relatively low genetic similarity. The results show that the SRAP technology is more efficient than traditional morphology identification. It is found that SRAP markers could more really reflect the genetic diversity of endophytic fungi strains from Taxus, and also could be used as a method for identification of endophytic fungi from Taxus. It also suggests that SRAP can be used to establish foundation for further screening of taxol-producing endophytic fungi strains which can produce high levels of paclitaxel.     

珍稀濒危植物金毛狗的SRAP分析

利用SRAP分子标记技术,对7个居群79株金毛狗进行遗传多样性分析。结果表明:10对SRAP引物组合共得到107条扩增条带,多态性条带比率为85.98%,Nei’s基因多样性指数为0.229 6,Shannon’s多样性指数为0.358 6,表明金毛狗居群水平具有较高的遗传多样性;金毛狗7个居群的总基因多样度为0.229 6,居群内遗传多样度为0.135 4,居群间的遗传分化指数为0.410 6,表明有41.06%的变异存在于居群间,有58.94%的变异存在于居群内;居群间基因流为0.717 8,表明居群间基因交流频率较低;遗传一致度和UPGMA聚类分析结果显示,生境条件相似的居群优先聚集,说明金毛狗种质亲缘关系与地理分布相关性不显著。     

Genetic molecular analysis of Coffea arabica (Rubiaceae) hybrids using SRAP markers

In Coffea arabica (arabica coffee), the phenotypic as well as genetic variability has been found low because of the narrow genetic basis and self fertile nature of the species. Because of high similarity in phenotypic appearance among the majority of arabica collections, selection of parental lines for inter-varietals hybridization and identification of resultant hybrids at an early stage of plant growth is difficult. DNA markers are known to be reliable in identifying closely related cultivars and hybrids. Sequence Related Amplified Polymorphism (SRAP) is a new molecular marker technology developed based on PCR. In this paper, sixty arabica-hybrid progenies belonging to six crosses were analyzed using 31 highly polymorphic SRAP markers. The analysis revealed seven types of SRAP marker profiles which are useful in discriminating the parents and hybrids. The number of bands amplified per primer pair ranges from 6.13 to 8.58 with average number of seven bands. Among six hybrid combinations, percentage of bands shared between hybrids and their parents ranged from 66.29% to 85.71% with polymorphic bands varied from 27.64% to 60.0%. Percentage of hybrid specific fragments obtained in various hybrid combinations ranged from 0.71% to 10.86% and ascribed to the consequence of meiotic recombination. Based on the similarity index calculation, it was observed that F1 hybrids share maximum number of bands with the female parent compared to male parent. The results obtained in the present study revealed the effectiveness of SRAP technique in cultivar identification and hybrid analysis in this coffee species.     

SRAP标记技术在甜瓜品种亲缘关系和种子纯度鉴定上的应用

利用SRAP分子标记对12份甜瓜材料进行亲缘关系鉴定和F1代杂交种进行纯度检测。结果表明:从40对引物中筛选出13对多态性好的引物,共扩增出78条带,多态性带的比例为50%;聚类分析结果显示,12份甜瓜材料的相似系数在0.263～0.921之间,可将该12份甜瓜材料分为2大类,进一步细分为5类。利用引物Me15-Em7和Me3-Em16对‘红绿早脆’100S杂交种进行纯度检测,杂交种纯度为77%。研究为甜瓜品种间亲缘关系及种子纯度检测提供了可靠的理论依据。     

Molecular diversity and population structure of the forage grass Hemarthria compressa (Poaceae) in south China based on SRAP markers

Hemarthria compressa is one of the most important and widely utilized forage crops in south China, owing to its high forage yield and capability of adaptation to hot and humid conditions. We examined the population structure and genetic variation within and among 12 populations of H. compressa in south China using sequence-related amplified polymorphism (SRAP) markers. High genetic diversity was found in these samples [percentage polymorphic bands (PPB) = 82.21%, Shannon's diversity index (I) = 0.352]. However, there was relatively low level of genetic diversity at the population level (PPB = 29.17%, I = 0.155). A high degree of genetic differentiation among populations was detected based on other measures and molecular markers (Nei's genetic diversity analysis: G(ST) = 54.19%; AMOVA analysis: F(ST) = 53.35%). The SRAP markers were found to be more efficient than ISSR markers for evaluating population diversity. Based on these findings, we propose changes in sampling strategies for appraising and utilizing the genetic resources of this species.     

Genetic diversity and population structure of Butea monosperma (Lam.) Taub.- a potential medicinal legume tree

Three molecular marker systems, Random Amplified Polymorphic DNA (RAPD), Inter-Simple Sequence Repeats (ISSR) and Sequence-Related Amplified Polymorphism (SRAP) were employed to investigate the genetic structure and diversity among the 14 natural populations of Butea monosperma collected from different geographical regions of India. Detected by 17 RAPD, 15 ISSR and 11 SRAP primer combinations, the proportions of polymorphic bands were 84.2 %, 77.2 % and 91.9 %, respectively, and the mean Nei’s genetic distances among the populations were 0.13, 0.10 and 0.13, respectively. Partitioning of genetic variability by Analysis of molecular variance (AMOVA) revealed that the high genetic diversity was distributed within the populations. AMOVA also revealed that the coefficient of gene differentiation among populations based on F_ST was very high irrespective of markers used. The overall gene flow among populations (Nm) was very low. Cophenetic correlation coefficients of Nei’s distance values and clustering pattern by Mental test were statistically significant for all three marker systems used but poor fit for ISSR data than for RAPD, SRAP and combined data set of all three markers. For all markers, a high similarity in dendrogram topologies was obtained, although some differences were observed with ISSR. The dendrogram obtained by RAPD, SRAP and combined data set of all three markers reflect relationship of most of the populations according to their geographic distribution.     

Genetic diversity assessment of a germplasm collection of Salvia miltiorrhiza Bunge. based on morphology,ISSR and SRAP markers

Salvia miltiorrhiza is one of the most important traditional Chinese medicinal plants for its therapeutic effects. In the present study, morphological traits, ISSR (inter-simple sequence related) and SRAP (sequence-related amplified polymorphism) markers were used to analyze the genetic diversity of 59 S. miltiorrhiza phenotypes. Out of the 100 ISSR primers and 100 SRAP primer combinations screened, 13 ISSRs and 7 SRAPs were exploited to evaluate the level of polymorphism and discriminating capacity. The results showed that the 13 ISSRs generated 190 repeatable amplified bands, of which 177 (93.2%) were polymorphic, with an average of 13.6 polymorphic fragments per primer. The 7 SRAPs produced 286 repeatable amplified bands, of which 266 (93.4%) were polymorphic, with an average of 38.1 polymorphic fragments per primer. Cluster analysis readily separated different morphological accessions, wild and cultivated controls based on morphological traits, ISSR and SRAP markers. The study indicated that morphological traits, ISSR and SRAP markers were reliable and effective for assessing the genetic diversity of phenotypic S. miltiorrhiza accessions. The overall results suggested that the introduction of genetic variation from morphology-based germplasms enlarged the genetic base for the collection, conservation and further breeding program of S. miltiorrhiza germplasm.