THE ORIGIN OF WEST EUROPEAN SUBSPECIES OF HONEYBEES (APIS MELLIFERA): NEW INSIGHTS FROM MICROSATELLITE AND MITOCHONDRIAL DATA

Apis mellifera is composed of three evolutionary branches including mainly African (branch A), western and northern European (branch M), and southeastern European (branch C) populations. The existence of morphological clines extending from the equator to the Polar Circle through Morocco and Spain raised the hypothesis that the branch M originated in Africa. Mitochondrial DNA analysis revealed that branches A and M were characterized by highly diverged lineages implying very remote links between both branches. It also revealed that mtDNA haplotypes from lineages A coexisted with haplotypes M in the Iberian Peninsula and formed a south-north frequency cline, suggesting that this area could be a secondary contact zone between the two branches. By analyzing 11 populations sampled along a France-Spain/Portugal-Morocco-Guinea transect at 8 microsatellite loci and the DraI RFLP of the COI-COII mtDNA marker, we show that Iberian populations do not present any trace of “africanization” and are very similar to French populations when considering microsatellite markers. Therefore, the Iberian Peninsula is not a transition area. The higher haplotype A variability observed in Spanish and Portuguese samples compared to that found in Africa is explained by a higher mutation rate and multiple and recent introductions. Selection appears to be the best explanation to the morphological and allozymic clines and to the diffusion and maintenance of African haplotypes in Spain and Portugal.     

Simple sequence repeats for the genetic analysis of apple

 The development of highly informative markers, such as simple sequence repeats, for tagging genes controlling agronomic characters is essential for apple breeding. Furthermore the use of these markers is fundamental both for variety identification and for the characterisation and management of genetic resources. We have developed 16 reliable simple sequence repeat (SSR) markers that amplify all alleles from a panel of 19 Malus x domestica (Borkh.) cultivars or breeding selections and from Malus floribunda 821. Those markers show a high level of genetic polymorphism, with on average 8.2 alleles per locus and an average heterozygosity of 0.78. Due to this high level of polymorphism, it was possible using two selected SSRs to distinguish all cultivars except Starking and Red Delicious. Ten of the markers we developed have been mapped on a RAPD linkage map, proving their Mendelian segregation as well as their random distribution in the apple genome. Finally, we discuss the importance of using co-dominant markers in outbreeding species. Received: 8 October 1997 / Accepted: 9 December 1997     

基于SSR标记的越橘亲缘关系分析及品种鉴定

利用SSR标记对22个越橘栽培品种进行遗传差异及亲缘关系分析,并建立一套稳定的越橘品种鉴定体系,以期为越橘种质资源评价、鉴定、管理及越橘新品种培育奠定基础。本试验优化了一套越橘SSR-PCR反应体系,筛选出15个条带清晰、重复性好、多态性高的SSR标记,对22个越橘栽培品种进行亲缘关系分析,聚类结果与各品种的遗传背景以及表型呈现高度的一致性。从以上15个SSR标记中筛选出可用于越橘品种鉴定的SSR核心引物NA961和NA1040,核心引物组合能够完全区分22个越橘品种。用核心引物制作了参照分子量标记,构建了22个越橘品种的指纹图谱,建立了一套完整的越橘品种鉴定体系。实践验证SSR标记用于越橘品种鉴别的方法可行、结果可靠。     

金线兰及近缘种植物遗传多样性ISSR分子标记分析

利用ISSR分子标记技术对我国不同地区收集的金线兰及其近缘种种质资源进行亲缘关系及遗传多样性分析。结果显示:9条ISSR引物在50份材料中共扩增条带156条,其中多态性条带155条,多态性比率达99.36%;UPGMA树状图分析显示,样本总体相似系数介于0.56~0.90之间,在相似系数0.65处可分4大类群,其中金线兰主要集中于A大类群。A大类群又可分为3个亚类5个小亚类,亚类中不同地区来源的金线兰种质没有明显的界限。去除部分地区来源不明材料,46份材料依不同地理来源分成8个种群,其中福建野生金线兰4个种群,二者Nei's基因多样性指数(H)分别为0.322和0.295,处兰科植物中等水平;种群遗传分化系数(Gst)分别为0.328和0.240,处兰科植物较低水平。AMOVA分析显示样本种群内变异指数87%,种群间变异指数13%,表明种群间可能存在基因交流。     

应用SSR标记分析鹰嘴豆种质资源遗传多样性

种质资源的遗传多样性是育种工作的基础,本研究利用SSR标记对鹰嘴豆资源进行遗传多样性分析,旨在发掘鹰嘴豆资源中丰富的遗传变异。从48对鹰嘴豆SSR引物中筛选出18对核心多态性引物,对96份不同来源的鹰嘴豆种质资源进行SSR标记的遗传多样性分析。结果表明,18对SSR引物共检测到115个等位变异,占总检测位点的52.99%,每对SSR引物可检测出3~10个等位变异,平均6.39个;平均每个位点多态信息量(PIC)为0.8107,变化范围为0.6661~0.8984。Shannon's信息指数平均为1.4769,变化范围为0.0607~1.9584。PGMA聚类结果表明,在遗传相似系数0.59处,可将96份鹰嘴豆资源分为6个类群,类群Ⅰ包含9份资源,类群Ⅱ包含2份资源,类群Ⅲ包含28份资源,类群Ⅳ包含4份资源,类群Ⅴ包含40份资源,类群Ⅵ包含13份资源。本研究对我国鹰嘴豆种质资源的评价与利用、优异基因的挖掘、育种亲本材料的选择等提供科学依据。     

基于SSR和ISSR的鄱阳湖流域野生菰资源的遗传多样性分析

用SSR和ISSR标记对鄱阳湖流域30个野生菰居群的遗传多样性与遗传结构进行了分析。筛选出的19对SSR引物共扩增出多态性条带253条,平均多态性条带比率(PPB)为91.67%,Nei's基因多样性指数(He)和平均Shannon信息指数(I)分别为0.2712和0.4144,遗传相似性系数(GS)为0.5590~0.8368,遗传距离(GD)的变化范围为0.1632~0.4410;筛选出的14个ISSR标记引物共扩增出83条条带,平均多态性条带比率(PPB)为78.29%,Nei's基因多样性指数(He)和平均Shannon信息指数(I)分别为0.2386和0.4174,遗传相似性系数(GS)为0.5132~0.9342,遗传距离(GD)变化范围为0.0658~0.4868。根据SSR和ISSR基因型数据,采用UPGMA法分别在阈值为0.698和0.728时可将30个野生菰居群聚为3类。可能受人为、水流、动物活动、风等多种因素的影响,居群间的亲缘关系与地理分布无明显相关性。本研究表明,鄱阳湖流域野生菰居群间SSR和ISSR基因型的多样性丰富,居群间的这种遗传差异或变异,对该地区乃至更大范围内野生菰的遗传进化、基因资源的开发利用和种质资源的保护有着重要意义。     

新疆野生油菜胞质雄性不育系1193A对应恢复基因的SSR标记分析

以油菜细胞质雄性不育系1193A和恢复系1193R2为亲本构建F2分离群体,并运用BSA法构建了可育和不育基因池。利用1521对SSR引物进行了多态性分析,结果表明有36对引物在亲本和基因池间都表现多态性,用F2单株验证表明有11对引物与恢复基因连锁,离恢复基因较近的2个标记CB10316和Bn GMS171分布在恢复基因Rf的两侧,遗传距离分别为3.9 c M和5.7 c M,可作为恢复系标记辅助育种的候选标记。     

Low genetic variation detected within the widespread mangrove species Nypa fruticans (Palmae) from Southeast Asia

Genetic diversity within and among six natural populations of Nypa fruticans from China, Vietnam, and Thailand was assessed using SSR and ISSR analysis. Our results showed an extremely low level of genetic diversity of N. fruticans (at the species level, P = 11.76% and 2.88%, He = 0.0279 and 0.0113, I = 0.0470 and 0.0167 by SSRs and ISSRs, respectively) across a total of 183 individuals. No genetic variation was detected within any population except for the Thailand population by SSRs (P = 11.76%, He = 0.0417; I = 0.0622). The bottlenecks during glacial epochs, founder effects, and propagation pattern may be responsible for the extremely low level of genetic diversity of N. fruticans.     

Assessment of the genetic diversity of wild hops (Humulus lupulus L.) in Europe using chemical and molecular analyses

The goal of this study was to characterize European wild hops (Humulus lupulus L.) by chemical and molecular genetic analyses in comparison to cultivated hops and North American wild hops. The contents of alpha and beta and acids varied from 0.45% to 5.55% and from 1.22% to 5.73% in European wild hops, respectively. Low bitter acid contents, alpha/beta acid ratios of lower than 1.0 and cohumulone content not exceeding 30% were typical as well as for traditional European cultivars. The lower myrcene content, the presence of farnesene and high selinene content were typical for European wild hops. We evaluated molecular genetic diversity in European wild hops by microsatellite and gene-specific markers and found that this variability did not correlate with the chemical characteristics. Our phylogenetic analysis confirmed overlapping variability and close genetic relationships in Europe, the separation of wild hops from the Caucasus region and the high diversity of North American wild hops.     

利用SSR标记与毛细管电泳对甘蔗属进行的遗传分析

为探讨甘蔗属内不同种之间的遗传多样性,利用SSR标记与毛细管电泳技术,对来自甘蔗属3个不同种的12个材料19对引物进行检测,共检测到229个DNA多态性条带,19对引物扩增的DNA条带范围集中在100～260bp之间。12个甘蔗材料的Jaccard遗传相似度,最小0.09,最大0.65,平均为0.26。通过遗传相似性系数分析,UPGMA聚类图内12个甘蔗材料可分为两个群,三个割手密种材料分为一个亚群,甘蔗栽培品种与甘蔗热带种合为一个亚群。结果表明:热带种比割手密种具有和甘蔗栽培品种更亲近的遗传关系;SSR分子标记与毛细管技术结合,相比别的分子标记技术或电泳技术,具有更准确、简便、自动化等优点。