濒危植物夏蜡梅花的形态变异

研究了不同地理位置的3个夏蜡梅自然种群间以及种群内花部形态变异,并对3个种群进行聚类分析。结果表明：夏蜡梅花部表型性状在种群间和种群内都存在一定程度的变异;各性状总的平均变异系数在6.293（内瓣长）~46.90（花柄长）之间,种群间总的平均变异系数在13.430（DMS）~15.720 8（DLS）,因此认为夏蜡梅花的表型性状无论是在物种水平还是在种群水平都具有较丰富的表型多样性,其中又以大雷山种群最高;单因素方差分析显示：部分指标分化显著或极显著,13个指标在种群内的变异占了总变异的82.9%,这说明夏蜡梅种群内变异是表型变异的主要来源,由此可以推测遗传上的多样性是形成夏蜡梅花部形态变异的主导因素;种群间平均变异组分占了17.1%,表明花部形态在种群间存在一定程度的表型分化;聚类分析显示距离较近的大明山种群和龙须山种群先聚在一起,表型差异与地理距离之间存在显著正相关,经分析认为微生境的异质性和种群隔离以及遗传漂变均可能导致夏蜡梅种群间的表型分化;相关性分析显示部分性状之间存在显著的相关性。     

濒危植物金花茶果实、种子形态分化

对濒危植物金花茶5个天然居群和1个人工居群的果实、种子形态进行了研究.结果表明:6个金花茶居群按果实、种子大小进行排序为:广西植物所(GXIB)＞那子山中海拔2(NZSH2)＞大王江(DWJ)＞庚毛山(GMSH)＞那子山高海拔3(NZSH3)＞那子山低海拔1(NZSHI);表型性状在居群内和居群间均存在一定程度的变异,不同形态性状在居群间变异系数的平均值从0.429(种子质量/果,SWPF)到0.069(果实整体形态,FT/FD),果实整体形态最为稳定,各居群形态性状变异系数的平均值从0.287(NZSH3)到0.155(GXIB)不等;在调查的15个性状中,果实直径、果实质量等3个性状在居群间的差异达到显著水平(P＜0.05),种长、种宽等7个性状在居群间的差异达极显著水平(P＜0.01),多数居群已产生较明显的形态分化;聚类分析结果表明,在居群水平上,形态分化并不与居群间的地理距离远近相关联.影响果实、种子表型性状变异的主导因子为海拔高度、土壤有机质、土壤含水量和乔木层透光率.     

缙云山草珊瑚(Sarcandra glabra)天然居群结实特征的变异性研究

对缙云山草珊瑚8个天然居群结实特征的20项表型性状的变异性进行了研究,结果表明:各表型性状在居群内与居群间都存在很大的变异性,其中种实产量、果序数和果序分枝的变异程度较高,而果实和种子的特征指数变异程度较低,同类性状果实的变异程度要大于种子;平均变异系数最大的性状是四分枝果序百分比(1.477),最小的是种子长宽比(0.041);变异程度最大的居群是居群6(0.564),最小的是居群4(0.292);各居群间除果序分枝总数差异不显著、果序数和二分枝果序百分比差异显著外,其它17项表型指标均达到极显著水平,表明缙云山草珊瑚各居群间已经发生了明显的形态分化;居群内和居群间变异分别占总变异的60.7%和39.3%,说明居群内变异是表型变异主要来源,但居群间变异也不容忽视;欧氏距离聚类将所有居群划归为两大类群,主要与居群的群落类型与干扰程度有关;表型性状的Pearson相关性聚类显示草珊瑚同类性状在表型分化过程中具有很强的相关性。     

珍稀濒危植物夏蜡梅遗传多样性的ISSR分析

利用ISSR分子标记技术,对濒危植物夏蜡梅10个居群的遗传多样性进行了分析.结果表明:夏蜡梅种的遗传多样性较高,多态位点百分率为73.08%,Shannon指数为0.3097,Nei指数为0.1987;而居群水平的遗传多样性较低,多态位点百分率平均为23.65%,Shannon指数平均为0.1251,Nei指数平均为0.0839.AMOVA分子差异分析显示:居群间遗传分化程度高,57.11%的变异存在于居群间,42.89%存在于居群内,基因分化系数(G_st)为0.5779;居群间的基因流为0.3651.生境的片断化使居群间的基因流受阻,可能是导致居群间高遗传分化和居群水平低遗传多样性的主要原因.10个居群间的平均遗传距离为0.1471.利用UPG-MA法对10个居群进行聚类,结果是天台县内的2个居群、临安市内的8个居群各组成一大类群.     

濒危植物夏蜡梅黄酮化合物薄层色谱分析

采用薄层层析法对不同居群夏蜡梅(Sinocalycanthus chinensis)叶、同一居群夏蜡梅不同营养器官中的黄酮化合物进行分离,并以芦丁为标准品通过双波长锯齿扫描作定量分析。结果表明:不同居群夏蜡梅叶片中黄酮化合物含量以鸡窝坪居群最高,中坪居群次之,龙塘山居群最低,除双石边和白水坞两个居群差异不显著外,其它各居群间差异均达到显著水平;同一居群夏蜡梅黄酮化合物在不同营养器官中含量不同,叶片中含量显著高于其它营养器官,根、茎、一年生枝和二年生枝等4个营养器官中黄酮化合物含量差异不显著。薄层层析法分离黄酮化合物组分结果显示,除大明山居群叶片黄酮化合物为5种组分外,其它居群叶片均为7种组分;大明山居群夏蜡梅的根、茎、一年生枝和二年生枝等营养器官黄酮化合物均为6种组分,叶片中缺少Rf值为0.66的化合物。     

滇牡丹天然居群的遗传多样性分析

以云南中部及西北部的6个滇牡丹(Paeonia delavayi)天然居群为研究对象,进行株高、新枝长等9个表型性状的表型多样性分析和ISSR分析。结果表明:9个表型性状变异幅度为0.9%~39.8%,平均值达到了18.9%;居群间生殖器官的变异较大,居群内营养器官更容易产生变异。利用居群间欧式距离进行聚类分析,6个居群聚为4个类群,没有与实际地理位置相吻合,说明表型特征的性状与地理距离的相关性不大。遗传多样性分析结果表明:利用筛选得到的10条引物,在取自6个自然居群、180个个体中,检测到56个多态位点。在居群水平上,多态位点百分率(PPB)为60.2%,Nei's基因多样性指数(H)和Shannon's信息指数(I)分别为0.281和0.414。在物种水平上,Nei's基因多样性指数(H)和Shannon's信息指数(I)分别为0.409和0.596。居群间的遗传分化系数(Gst)达0.319。结果显示,表型性状在居群间和居群内均存在广泛变异。滇牡丹遗传多样性水平较高,居群间遗传分化较大,滇牡丹并不濒危。     

新疆野杏种质资源表型性状多样性研究

为了明确新疆野杏种质资源的遗传多样性和变异特点,以新疆伊犁地区3个野杏居群为研究对象,采集了135个单株样品,测定其植株35个形态特征相关指标,利用DPS 7.05软件对表型指标和主成分进行分析,采用UPGMA法对3个居群的欧氏距离进行聚类分析。结果显示:(1)数值性状和非数值性状的Simpson和Shannonweaver指数均为霍城居群最高(0.979 9、4.729 9;0.981 9、4.770 1),巩留居群居中(0.975 5、4.385 7;0.978 6、4.416 0),新源居群最小(0.944 7、3.241 9;0.945 2、3.277 1),说明霍城居群的表型最丰富,巩留居群居中,新源居群的表型多样性最差。(2)各居群间不同性状变异系数的变异幅度在6.16%~54.58%之间,大部分性状的变异系数都在10%以上;霍城居群和新源居群均为果形指数的变异系数最小,遗传较稳定,巩留居群叶片长宽比的变异系数最小;3个居群均以硬度的变异系数最大,另有单果重、鲜核重和壳厚的变异系数也普遍较大。(3)新源居群与巩留居群间的欧氏距离为20.445 3,其亲缘关系最远;霍城居群与新源居群间的欧氏遗传为19.218 6,其亲缘关系最近。(4)35个性状的主成分分析中前14个主成分的累计贡献率为80.64%,说明叶片长度、叶片宽度、叶片长宽比、叶基形态、叶尖形态、着色类型、单果重、鲜核重、鲜仁重、果实形状、可溶性固形物等是造成新疆野杏表型差异的主要因素。     

白檀自然居群遗传结构与遗传多样性研究

采用ISSR分子标记技术分析了湖南省5个地区6个居群白檀的遗传多样性水平和遗传结构。结果表明,9条ISSR引物对6个居群的149份白檀样品PCR扩增检测到122个扩增位点,其中多态性位点113个,多态性比率高达92.62%。Nei's基因多样性指数(He)为0.3264,Shannon多样性指数(I)为0.4873,居群之间产生较大的遗传变异(Gst=0.5215,Nm=0.4588),AMOVA分子差异分析表明白檀居群间遗传分化程度高,51.07%的变异存在于居群间,48.93%的变异存在于居群内;UPGMA聚类分析将6个白檀居群分为3大类:大围山和龙山居群为一类,岳阳和道县居群组成另一类;衡山居群单独成一类,聚类原则跟地理位置不一致,与海拔高度有一定的相关性。     

云南金钱槭果实、种子形态分化研究

对5个云南金钱槭(Dipteronia dyeriana Henry)天然居群的果实、种子形态进行研究.测量了果实和种子的长度、宽度、重量等12个表型性状指标.数据统计结果显示5个云南金钱槭居群按果实、种子大小进行排序为蒙自(MZ)>屏边(PB)>文山1(WSh1)>文山3(WSh3)>文山2(WSh2).表型性状在居群内和居群间均存在着一定程度的变异,其中居群间变异系数的平均值从0.064(果实整体形态,PL/PB)到0.197(种子重量,SW),相比之下果实整体形态最为稳定.居群间形态总体差异显著性配对t检验结果说明,多数居群间已产生较明显的形态分化.Ward聚类和相关分析结果都表明表型性状与生态因子、海拔高度之间存在着一定程度的相关性,多种生态因子的共同作用是导致果实、种子形态特征产生差异的主要原因.     

异质性生境对半红树植物海漆(Excoecaria agallocha)居群遗传结构的影响

选取半红树植物——海漆（Excoecaria agallocha）作为研究对象,采用简单重复序列（ISSR）分子标记技术,对不同地理位置、不同生境（潮间带和陆生）海漆居群的遗传多样性及遗传结构进行研究。结果表明,在同一地点,潮间带居群的遗传变异水平均高于其陆生居群。潮间带居群间的遗传分化水平（GST=0．191）略低于其陆生居群间的遗传分化（GST=0．218）．表明潮间带居群间通过漂浮的种子进行的基因交流较陆生居群间频繁。AMOVA分析显示,由异质性生境造成的分子变异为15．13％,而采样不同地区间（相距181～759km）造成的分子变异却只有11．63％．暗示环境胁迫造成的选择压力导致海漆居群适应性进化。同时地理隔离、南海北部的西南季风漂流及中国沿岸流和遗传漂变对海漆居群的遗传分化均有着重要影响。     

Effect of sire population on the genetic diversity and fitness of F1 progeny in the endangered Chinese endemic Sinocalycanthus chinensis

Sinocalycanthus chinensis Cheng et S. Y. Chang (Calycanthaceae), which has a unique systematic status, is listed as a national second‐class protected plant of China. In this study, the genetic diversity, performance, and fitness of F1 progeny from crosses between the Damingshan (DMS) population of S. chinensis and pollen parents from the Daleishan (DLS) and Longxushan (LXS) populations were examined. The DLS population has a relatively small population size, low genetic diversity, and considerable geographical and genetic distances from the DMS population relative to the LXS population. Compared with naturally occurring seeds, DLS‐sired seeds had the highest thousand‐seed weight, starch content, fat content, germination rate, germination index, and emergence rate, but the lowest protein content. Naturally occurring, open‐pollinated seeds had the lowest thousand‐seed weight, starch content, and fat content, but the highest protein content. Compared with natural F1 progeny, DMS × DLS seedlings had the highest genetic diversity, photosynthetic parameters, and growth characteristics, except for leaf mass ratio and stem mass ratio. Under strong light, DMS × DLS seedlings exhibited a F_v/F_m value of 0.75, while the other two seedling types exhibited F_v/F_m values of 0.65. DLS‐sired seeds had the most vigorous growth characteristics except for leaf mass ratio and stem mass ratio. These results suggest that genetic rescue by transplanting seedlings from the DLS population or hand pollination with pollen from the DLS population would be effective methods to reduce inbreeding depression and obtain strong offspring with high genetic diversity and fitness in the DMS population.     

Conservation genetics of Heritiera littoralis (Sterculiaceae), a threatened mangrove in China,based on AFLP and ISSR markers

AFLP and ISSR markers were used to determine the genetic variations in eight mangrove and non-mangrove populations of Heritiera littoralis (Sterculiaceae), a threatened species in China. Our results showed a moderate to high level of genetic variation in this species (P = 63.69%, H_T = 0.20 for AFLP; P = 76.07%, H_T = 0.22 for ISSR), and a relatively high level of genetic differentiation among populations (G_ST = 0.24 for AFLP and 0.27 for ISSR). Life history traits, long-distance dispersal of floating seeds, and local environments may play important roles in shaping the genetic diversity and genetic structure of this species. Investigation of the plant’s reproductive capacity showed that the natural seed production of H. littoralis was low, as was the germination rate and the transformation rate from juvenile to adult. H. littoralis is seriously threatened and is in urgent need of conservation in China.     

Highly differentiated populations of the narrow endemic and endangered species Primula cicutariifolia in China,revealed by ISSR and SSR

The perennial herb Primula cicutariifolia Pax is an endangered and endemic species with narrow distribution in eastern Anhui and Zhejiang provinces of China. In this study, the levels of genetic variation and the pattern of genetic structure in five natural populations of P. cicutariifolia were assessed by inter-simple sequence repeats (ISSR) and simple sequence repeat (SSR) markers. Both markers revealed that there was remarkably low genetic variation within populations (e.g., H_e = 0.19 and 0.18, for ISSR and SSR respectively) and high differentiation among populations (G_ST = 0.714 and 0.611; Φ_ST = 0.698 and 0.599, for ISSR and SSR respectively). The level of population genetic diversity was correlated to population size only detected by ISSR markers. The genetic structure of P. cicutariifolia may be explained by limited gene flow that was caused by habitat fragmentation and limited seeds and pollen dispersal ability, self breeding system and biennial life form. To protect and avoid disappearance of P. cicutariifolia, much more attentions should be paid to protect all the remnant populations and their habitats, and three management units, i.e. Tianmushan, Damodao, and Panan units, were proposed.     

Is the genetic diversity of small scattered forest tree populations at the southern limits of their range more prone to stochastic events? A wild cherry case study by microsatellite-based markers

Wild cherry (Prunus avium L.) is a widespread, partially asexual, noble hardwood European species characterized by a scattered distribution, small population sizes, and human exploitation for its valuable wood. These characteristics, especially at the southern limits of the species natural distribution where additional varying stresses may occur, render P. avium populations prone to potential stochastic, genetic, and demographic events. In this study, we used dominant inter simple sequence repeat (ISSR) and codominant simple sequence repeat (SSR) markers to infer the genetic structure of P. avium. Five populations from northern Greece were evaluated based on 46 ISSR and 11 SSR loci. Populations presented a relatively high level of genetic variation, with a mean genetic diversity of H _e?=?0.166 and H _e?=?0.740 regarding ISSR and SSR analysis, respectively. We observed moderate population differentiation for ISSR (G _ST?=?0.113) and SSR (F _ST?=?0.097) markers. AMOVA also detected significant differentiation among populations for ISSRs (?? _ST?=?0.338) and SRRs (?? _ST?=?0.162). According to linkage disequilibrium analysis, estimates of effective population size were generally sufficient for maintaining extant genetic variability and evolutionary potential. A possible bottleneck was detected for only one population. In general, it appears that despite the particular characteristics of the P. avium populations studied, genetic stochasticity events were not apparent. The studied populations, located at the rear edge of the species European distribution, reveal a wealth of genetic variation that is very valuable for the genetic conservation of local adaptive gene complexes, especially under contemporary climatic change scenarios.     

Pollen limitation in the endangered Chinese endemic species Sinocalycanthus chinensis

Pollen limitation negatively impacts endangered and endemic plants with small fragmented populations, such as Sinocalycanthus chinensis, an endangered plant endemic to China. In this study, we analyzed the pollen limitation of the S. chinensis Damingshan (DMS) population in 2006, 2009, and 2010, and crossed plants with mates separated by different distances, both within and between populations. The DMS population exhibited strong pollen limitation in fruit set, seed set, and seeds per fruit in 2006, 2009, and 2010. The average accumulated pollen limitation (for fruit set times seeds per fruit) was 0.510 ± 0.180. Progeny crossed with pollen from intermediate neighboring plants within the same population (separated by 30–50 m from pollen recipients) had the lowest fitness. No optimal outcrossing distance was found within the DMS population. Progeny from crosses with the Shunxiwu (SXW) and Daleishan (DLS) populations performed relatively better, while those from crosses with Qingliangfeng (QLF) and Longxushan (LXS) populations performed worse. Compared with average reproductive success, outbreeding depression was found in progeny from crosses with the LXS and QLF populations. Reproductive success from pure self‐pollination indicated S. chinensis is self‐compatible. Geitonogamous selfing increased reproductive success. Based on geitonogamous selfing, the proportion of selfed offspring was relatively high. These results provide basic references for the conservation of this species.     

Genetic diversity of Lilium tsingtauense in China and Korea revealed by ISSR markers and morphological characters

To study the genetic diversity and population structure of Lilium tsingtauense Gilg (Qingdao Lily), we collected 648 samples from 12 sites in China and Korea, and analyzed their Inter-Simple Sequence Repeat (ISSR) molecular markers and morphological characters. ISSR data revealed a relatively high genetic diversity at the species level, with 72.31% polymorphic loci, effective numbers of alleles of 1.437, average expected heterozygosity of 0.231 and Shannon’s information index of 0.369. Considerable genetic differentiation among the natural populations (G_ST = 0.144) and the gene flow (N_m = 1.487) were detected. AMOVA analysis and UPGMA-dendrogram suggested a hierarchical regional structure among populations, and spatial autocorrelation analysis showed a micro-scaled spatial structure. Furthermore, there was a high correlation between morphological characters and genetic parameters obtained from ISSR parameters. There was only a low genetic differentiation among the different morphological types of L. tsingtauence in China. Based on these findings, we recommend in situ and ex situ conservation strategies for the preservation of L. tsingtauense.     

茶条槭自然种群种子和果实表型多样性研究

为探析茶条槭(Acer ginnala)种子和果实表型差异程度及变异规律,该研究采用巢式方差分析、主成分分析、聚类分析等方法,对茶条槭主要分布区内7个种群种子和果实的12个表型性状进行比较研究,分析茶条槭种群间和种群内的表型多样性以及与地理生态因子的关系。结果表明:除种子长/宽(SLW)外,其它11个表型性状在茶条槭种群间和种群内均具有显著差异;各性状平均变异系数(CV)为13.90%,变异幅度为8.14%～32.08%;种群间翅果性状的平均变异系数(15.63%)高于种子性状的平均变异系数(8.71%),表明种子性状的稳定性高于果实性状。主成份分析中,果实形态特征对茶条槭种群表型的贡献率大于种子贡献率。种群间表型分化系数为35.47%,说明种群内变异(64.53%)大于种群间变异(35.47%),暗示变异主要来源于种群内变异。茶条槭种子和果实表型性状受地理生态因子影响较小,主要受自身遗传因素的影响。基于种群间欧式距离的聚类分析将茶条槭7个种群分为2大支,没有严格按照地理距离而聚类,表明茶条槭种群表型性状变异的不连续性。茶条槭不同种群种子和果实具有较高的表型多样性,与其分布范围和生物学特性有关。     

Genetic and Morphological Divergence in Three Strains of Brook Trout Salvelinus fontinalis Commonly Stocked in Lake Superior

Fitness related traits often show spatial variation across populations of widely distributed species. Comparisons of genetic variation among populations in putatively neutral DNA markers and in phenotypic traits susceptible to selection (Q_ST F_ST analysis) can be used to determine to what degree differentiation among populations can be attributed to selection or genetic drift. Traditionally, Q_ST F_ST analyses require a large number of populations to achieve sufficient statistical power; however, new methods have been developed that allow Q_ST F_ST comparisons to be conducted on as few as two populations if their pedigrees are informative. This study compared genetic and morphological divergence in three strains of brook trout Salvelinus fontinalis that were historically or currently used for stocking in the Lake Superior Basin. Herein we examined if morphological divergence among populations showed temporal variation, and if divergence could be attributed to selection or was indistinguishable from genetic drift. Multivariate Q_ST F_ST analysis showed evidence for divergent selection between populations. Univariate analyses suggests that the pattern observed in the multivariate analyses was largely driven by divergent selection for length and weight, and moreover by divergence between the Assinica strain and each of the Iron River and Siskiwit strains rather than divergent selection between each population pair. While it could not be determined if divergence was due to natural selection or inadvertent artificial selection in hatcheries, selected differences were consistent with patterns of domestication commonly found in salmonids.     

Genetic diversity within and among populations of the endangered and endemic species Primula merrilliana in China

Primula merrilliana Schltr. is an endangered and narrowly-distributed endemic species of southern Anhui Province in China. In this study, the level of genetic variation and the pattern of genetic structure in six natural populations of P. merrilliana were assessed by using ISSR (inter-simple sequence repeats) markers. Based on ten primers, 137 clear and reproducible DNA fragments were generated, of which 109 were polymorphic. The statistical results indicated that there was a relatively low genetic diversity within populations, and a high genetic differentiation among populations (G_ST = 0.53, Φ_ST = 0.49). The level of population genetic diversity was correlated to habitat type and the gene flow (N_m) was low with only 0.45. The unexpected genetic structure of P. merrilliana may be explained by limited gene flow that was caused by habitat fragmentation and limited seeds and pollen dispersal ability, self-compatible breeding system and biennial life form.