黑河上游灌丛建群种中国沙棘自由授粉子代父本分析和花粉流

利用SSR分子标记对中国沙棘(Hippophae rhamnoides ssp. sinensis Rousi)自由授粉的种子进行父本分析,研究其子代的父本来源和花粉散布模式。结果显示:在80%的置信水平上,193个子代中有104个个体可以确定花粉来源;在20个确定的父本中,16个为中国沙棘,4个为肋果沙棘(H. neurocarpa S. W. Liu et T.N. He)。传粉格局分析结果显示,中国沙棘有效花粉散布范围为3～71 m,平均距离为20.4 m,2株母本分别有87.23%和78.95%的有效花粉来自距其30 m的范围之内,研究结果表明中国沙棘自然种群以近距离传粉为主。此外,在黑河上游沙棘杂交带中,中国沙棘子代中的肋果沙棘花粉平均贡献率达到14.84%,表明中国沙棘与肋果沙棘存在较高的种间当代花粉流。     

利用SSR分子标记对鹅掌楸自由授粉子代的父本分析

利用自行开发的12个EST-SSR分子标记,采用最大似然法对鹅掌楸（L.chinense （Hemsl.）Sarg.）实验群体3个半同胞家系的180个子代进行父本分析。结果表明,每个SSR位点的等位基因数为3-7,平均为4.67;其平均观测杂合度（Ho）、平均期望杂合度（He）及平均多态信息含量（PIC）分别为0.458、0.635和0.580。利用12个SSR标记可在95%的可信度确定114个子代的父本,占子代群体的63.3%,其累积排除概率为98.52%。自由授粉状态下,鹅掌楸的自交率为11.6%,而北美鹅掌楸自交率为0,且种内交配比例大于种间交配。鹅掌楸平均有效花粉散布距离为20-30m,最大散布距离为70m。     

利用SSR 分子标记对鹅掌楸自由授粉子代的父本分析

利用自行开发的12个EST-SSR分子标记, 采用最大似然法对鹅掌楸(L.chinense (Hemsl.) Sarg.)实验群体3个半同胞家系的180个子代进行父本分析。结果表明, 每个SSR位点的等位基因数为3-7, 平均为4.67; 其平均观测杂合度(Ho)、平均期望杂合度(He)及平均多态信息含量(PIC)分别为0.458、0.635和0.580。利用12个SSR标记可在95%的可信度确定114个子代的父本, 占子代群体的63.3%, 其累积排除概率为98.52%。自由授粉状态下, 鹅掌楸的自交率为1.6%, 而北美鹅掌楸自交率为0, 且种内交配比例大于种间交配。鹅掌楸平均有效花粉散布距离为20-30 m, 最大散布距离为70 m。     

马尾松二代无性系种子园子代父本分析及花粉散布

目前国内较早建立的马尾松(Pinus massoniana)二代种子园正陆续进入正常开花结实期。研究马尾松二代种子园花粉散布和自由授粉子代的父本组成, 可为生产上指导马尾松高世代种子园的规划设计和遗传管理提供理论依据。该文利用筛选的11对SSR引物, 对马尾松二代无性系种子园内8个无性系单株的320个自由授粉子代和48个候选父本进行了扩增, 并采用最大似然法对子代进行父本分析。结果表明: 11个位点共检测到61个等位基因, 每个位点的等位基因数在2-11之间, 平均为5.55个。试验亲本和子代群体的总平均观测杂合度(H_o)、期望杂合度(H_e)及多态信息含量(PIC)分别为0.428、0.433和0.387。在80%的可信度水平下可为232 (72.50%)个子代确定其父本。平均每个采种母树与19个父本产生子代。在自由授粉状态下, 马尾松二代种子园自交率为1.72%, 自交现象很弱, 其交配方式以异交为主。绝大多数亲本无性系的雄性繁殖适合度在1.00%-4.00%之间, 候选父本平均繁殖适合度为2.17%, 平均形成5个后代。马尾松有效花粉的散布距离和固定交配距离的父本繁殖适合度均符合正态分布, 两者呈极显著负相关, 其主要散布距离集中在0-100 m, 而检测到的最大散布距离为192 m。种子园花粉污染率较低, 仅为4.06%。总体看来, 树冠南面子代亲本交配距离较北面有增加的趋势, 但树冠南、北面子代父本组成数并未表现明显的规律。     

中国沙棘(Hippophae rhamnoides L.ssp. sinensis Rousi)的开花特性及风媒传粉距离的检测

连续两年对雌雄异株的中国沙棘(Hippophae rhamnoides L.ssp. sinensis Rousi)的花期物候、开花式样进行了观察,检测了柱头可授性、花粉活力和花粉-胚珠比,并进行了人工授粉、套袋实验,检测结实率.结果显示中国沙棘风媒传粉发生在4月下旬或5月上旬,单花花期约7d;传粉盛期在第3天到第5天,柱头的可授性从第2天到第4天或第5天.中国沙棘花先叶开放、花小、无花冠、花药成熟时萼片从两个侧面的纵缝中裂开,形成有利于花粉散布的对流风洞.花粉活力和花粉-胚珠比都很高,具有典型的适应风媒传粉的花部特征和性状.中国沙棘花粉的传播距离受外界环境影响较大,通过重力玻片法检测可以看出,在风速小于3m/s时,主要集中在15～25m之间,当风速大于3m/s时,顺风向可超过85m以外.传粉效率对结实率的影响较大.     

中国沙棘开花生物学的初步研究

对雌雄异株的中国沙棘的花期物候、开花式样进行了观察,检测了柱头可授性、花粉活力和花粉-胚珠比(P/O).中国沙棘风媒传粉发生在四月下旬或五月上旬,单花花期约7天;传粉盛期在第3天到第5天,柱头的可授性从第2天到第4天或第5天.中国沙棘花先叶开放、花小、无花冠、花药成熟时萼片从两个侧面的纵缝中裂开,形成有利于花粉散布的对流风洞,具有典型的适应风媒传粉的花部特征.开花期内花粉的散发与温度、相对湿度、日照强度有关,受风向、风速作用显著;不同方位的花粉散布距离不等,当风速大于3 m/s时,主风方向花粉可散布到85 m以外.     

肋果沙棘(Hippophae neurocarpa S.W.Liu et T.N.He)自然种群的风媒传粉特征

野外调查了青藏高原特有植物肋果沙棘的花期物候、开花式样以及风媒传粉特征,对其花粉生活力、柱头可授性及单花花粉量进行了检测.结果表明,肋果沙棘有许多适应风媒传粉的特征:雌雄异株,花小,无花冠,先叶开放,雄花萼片纵裂形成对流的"风洞",花粉近球形,单花花粉量大;雌花柱头细长捻状,伸出萼筒,授粉面大,密布乳突.花粉生活力可以持续16.5 d,柱头可授性为10～20d.肋果沙棘传粉多发生在昼夜温差大的5月中下旬,在多变的气候条件下,种群花期可持续约12～25d,但在晴朗的天气条件下具有爆发性的集中开花机制,是对恶劣条件的适应.重力玻片法检测表明,离花粉源15m左右远的地点花粉密度最大,135m仍能接收到花粉,夜间接收到的花粉数量也很可观,这些研究结果表明,该植物集中在有利的晴朗天气昼夜进行长距离的风媒散粉,能较好地适应青藏高原的复杂环境.     

杭州石荠苧的传粉、小坚果散布和邻群大小

作者在杭州葛岭对杭州石荠苧自然居群的传粉和小坚果散布进行了详细的观察和实验。结果显示 ,以 2 6 0个传粉昆虫的飞行距离推算的花粉绝对散布方差为 0 .0 82 m2 ,798个小坚果的亲子散布方差为0 .1587m2。在上述散布特点和异交率为 0 .92 5条件下的邻群面积为 2 .2 33m 2 ,在有效密度为 4 7.7株 / m 2的有效密度下的邻群大小为 10 6 .5株。杭州石荠以异交为主 ,非常有限的基因散布导致邻群面积很小 ,居群容易发生遗传分化。     

油松种子园自由授粉与控制授粉种子父本分析

利用“微卫星”标记对油松(Pinus tabulaeformis)种子园控制授粉和自由授粉的子代进行了父本分析, 探讨自由授粉状态下种子园子代遗传组成与花粉传播的关系, 以及人工控制授粉状态下花粉形态等指标与子代遗传组成的相关性。其中,利用SSR对281粒油松种子进行父本分析, 在80%的置信水平下确定了其中126粒种子的花粉来源; 对2株母树自由授粉种子的父本分析表明, 种子园内花粉传播距离最大为85 m, 有效距离在30 m以内; 对11#和24＃母树分别用21个无性系的等体积混合花粉进行控制授粉发现, 子代由少数无性系的花粉受精产生, 其中 50＃、7＃、25＃、33#无性系对2株母树子代的花粉贡献率分别达到50.62%和61.54%; 花粉粒大小、花粉等体积重和花粉活力与选择性受精相关不显著, 而花粉活力偏低的无性系也有授粉成功的可能性。     

二倍体杂交种棱果沙棘双向杂交起源及其母本主要来源于中国沙棘的分子证据简

棱果沙棘为同域分布的中国沙棘和肋果沙棘同倍化自然杂交形成的。本文利用母系遗传的cpDNA trnS-G序列检测青海祁连棱果沙棘及其亲本中国沙棘和肋果沙棘同域分布的两个地区（拱北湾、八宝河滩）共93个个体的遗传关系。结果表明棱果沙棘及其亲本在拱北湾和八宝河滩分别有12个和7个单倍型,两地区的棱果沙棘都与其亲本共享单倍型,其中拱北湾棱果沙棘共36个个体中有28个与中国沙棘共享3个单倍型(H2,H4,H5),有2个个体与肋果沙棘共享单倍型(H11),八宝河滩的棱果沙棘共10个个体中有7个与中国沙棘共享一个单倍型(H4),3个与肋果沙棘共享单倍型(H7)。应用最大简约法（MP）分别对两地区的棱果沙棘及其亲本trnS-G序列构建的系统发育树中棱果沙棘的大部分个体都与中国沙棘聚在一起,另外,棱果沙棘4种特有单倍型(H3、H7、H8、H9)的6个个体在系统树上也与中国沙棘聚为一支。以上结果进一步证明了二倍体自然杂交种棱果沙棘为双向杂交起源,但其主要母本来源应为中国沙棘。     

祁连山两侧中国沙棘不同居群的遗传多样性研究

应用RAPD技术研究了祁连山中段中国沙棘(Hippophae rhamnoidoes ssp．sinensis)4个居群和1个对照居群的遗传多样性水平,探讨中国沙棘边缘居群的遗传变异以及天然屏障祁连山对中国沙棘居群遗传结构的影响。结果显示,祁连山地区中国沙棘居群水平的Nei’基因多样度(h)和Shannon多态信息指数(I)分别为0．2070和 0．297 4,基因分化系数(Gst)为0．319 3,均高于中国沙棘整个分布区的平均值,表明该地中国沙棘边缘居群遗传多样性水平及居群间的遗传分化都有增加的趋势;居群间每代迁移数(Nm=1．065 9)显著低于异交风媒植物(Nm= 5．380),表明祁连山的隔离对中国沙棘居群间的基因交流有限制作用,造成了该地中国沙棘边缘居群间较高的遗传分化。     

Clarification of the systematic position of Hippophae goniocarpa (Elaeagnaceae)

New information on (1) Hippophae goniocarpa Y.S. Lian et al . ex Swenson et Bartish and (2) H. goniocarpa ssp. litangensis (Lian et X.L. Chen ex Swenson et Bartish) Lian et K. Sun from China is provided and illustrated. Based on morphological characters and distribution, H. litangensis Y.S. Lian et X.L. Chen ex Swenson et Bartish is re-described as a subspecies. H. goniocarpa (including ssp. litangensis ) is a distinct species and should be placed in Section Gyantsensis of Hippophae , although it has been suggested that sspp. litangensis and goniocarpa are the result of hybridization between H. rhamnoides ssp. sinensis and H. neurocarpa ssp. neurocarpa , and H. rhamnoides ssp. yunnanensis and H. neurocarpa ssp. stellatopilosa , respectively. The type subspecies of H. goniocarpa is distributed in Songpan and Hongyuan Counties of Sichuan Province and Qilian County of Qinghai Province, at 3500–3750 m, whereas ssp. litangensis is found only in Jiawa, Litang County, Sichuan Province at c . 3700 m. A key including the four taxa of sect. Gyantsensis is provided.  © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society , 2003, 142 , 425−430.     

Karyotype Analysis of Hippophae L. in China

The present paper reports karyotypes in 4 species and 5 subspecies of Hippophae L. in China. It is found that all of the species and subspecies are diploid, with 2n=24, their complements are made of m and sm chromosomes and of them only H. thibetana has a pair of satellites. All of these karyotypes are symmetrical and primitive. It can be simplied as follows: Hippophae neurocarpa 2n = 2x = 24 = 18m+6sm, H. thibetana 2n = 2x = 24 = 14m (2sat)+ 8sm, H. rhamnoides L. ssp. gyantsensis 2n=2x=24= 18m+ 6sm, H. salicifolia 2n = 2x = 24 = 10m + 14sm, H. rhamnoides L. ssp. sinensis 2n=2x= 24 = 18m + 6sm, H. rhamnoides L. ssp. turkestanica 2n = 2x = 24 = 20m + 4sm; Hi. rhamnoides L. ssp. yunnanensis 2n = 2x = 24 = 14 + 10sm, H. rhamnoides L. ssp. mongolica 2n = 2x = 24 = 16m + 8sm.     

On phylogeny of Hippophae (Elaeagnaceae)

A cladistic analysis of the genus Hippophae (Elaeagnaceae) was undertaken. based on 37 characters. The five species of the genera Shepherdia and Elaeagnus were included in the analysis as the outgroup taxa. The operational taxonomic units of Hippophae were the species H. gyantsensis, H. neurocarpa, Hsalicifolia and H. tibetana plus eight subspecies of H. rhamnoides . The analysis revealed 117 or 14 equally parsimonious trees depending on whether the multistate characters were treated as ordered whenever possible, or all characters were treated as unordered. The results are not directly in accordance with any of the formerly proposed taxonomies. After successive weighting only two trees were retained both with ordered and unordered characters and the combinable component (semistrict) consensus trees give support to recognition of the two species in the genus, H. rhamnoides and Hsalicifolia . Three new combinations H. rhamnoides ssp. neurocarpa, H. salicifolia ssp. sinensis and H. salicifolia ssp. yunnanensis (Rousi) Hyvönen are proposed.     

Molecular Genetic Variation in Chinese Populations of Three Subspecies of Hippophae rhamnoides

To provide a population-level genetic profile for investigation and conservation of genetic diversity of the sea buckthorn (Hippophae rhamnoides L.), 300 individuals of fifteen natural populations of sea buckthorn in China were analyzed by using ISSR (inter-simple sequence repeats) markers. On the basis of Shannon's index and Nei's genetic diversity, the mean genetic diversity detected in the natural populations of Hippophae rhamnoides ssp. yunnanensis, ssp. sinensis, and ssp. gyantsensis was 0.1944, 0.2169, and 0.1372, respectively. The coefficient of gene differentiation (Gst) of seven ssp. yunnanensis populations is 0.2790, and that of seven ssp. sinensis populations is 0.4184. This means that 28% of the total molecular variance of seven ssp. yunnanensis populations existed among populations, and 42% for seven ssp. sinensis populations, suggesting that the subspecies have different genetic structures. No significant correlation between genetic and geographic distances of the populations was found using ISSR markers.     

Patterns of pollen dispersal in a small population of Pinus sylvestris L. revealed by total-exclusion paternity analysis

Patterns of pollen dispersal were investigated in a small, isolated, relict population of Pinus sylvestris L., consisting of 36 trees. A total-exclusion battery comprising four chloroplast and two nuclear microsatellites (theoretical paternity exclusion probability EP=0.996) was used to assign paternity to 813 seeds, collected from 34 trees in the stand. Long-distance pollen immigration accounted for 4.3% of observed matings. Self-fertilization rate was very high (0.25), compared with typical values in more widespread populations of the species. The average effective pollen dispersal distance within the stand was 48 m (or 83 m excluding selfs). Half of effective pollen was dispersed within 11 m, and 7% beyond 200 m. A strong correlation was found between the distance to the closest tree and the mean mating-distance calculated for single-tree progenies. The effective pollen dispersal distribution showed a leptokurtic shape, with a large and significant departure from that expected under uniform dispersal. A maximum-likelihood procedure was used to fit an individual pollen dispersal distance probability density function (dispersal kernel). The estimated kernel indicated fairly leptokurtic dispersal (shape parameter b=0.67), with an average pollen dispersal distance of 135 m, and 50% of pollen dispersed beyond 30 m. A marked directionality pattern of pollen dispersal was found, mainly caused by the uneven distribution of trees, coupled with restricted dispersal and unequal male success. Overall, results show that the number and distribution of potential pollen donors in small populations may strongly influence the patterns of effective pollen dispersal.     

中国特有植物卧龙沙棘自然群体的RAPD分析

应用12个随机引物对卧龙沙棘全部2居群共28个个体进行了RAPD分析。结果表明,卧龙沙棘具有较丰富的遗传多样性,多态位点百分率为78.05%,Nei’s基因多样性h=0.2553,Shannon’s多态性信息指数Ι=0.3841。分布范围狭窄的卧龙沙棘在亚种水平的遗传多样性明显高于分布较广的中国沙棘。在居群水平上,卧龙沙棘同样具有很高的遗传多样性,平均多态位点百分率为63.42%,Nei’s基因多样性h=0.2193,Shannon’s多态性信息指数I=0.3287。卧龙沙棘的基因分化系数Gst=0.1425,表明遗传变异主要存在于居群内。AMOVA的结果也表明,在全部的遗传变异中,19.18%的遗传变异存在于居群之间,与Gst值基本一致。这一结果符合卧龙沙棘风媒、异交的繁育系统特点。     

Patterns of pollen flow in a dense population of the insect-pollinated canopy tree species Castanopsis sieboldii

Insect pollinations of tree species with high-density populations have rarely been studied. Since the density of adults can affect effective pollen dispersal, short-distance pollination, even by insects, may frequently occur in high-density populations. To test this prediction, we investigated pollination patterns in a high-density population of the insect-pollinated canopy tree species Castanopsis sieboldii by paternity analysis using genotypes at 8 microsatellite loci of 145 adult trees and 439 seeds from 11 seed parents in a 4-ha plot. We then explored their genetic effects on the population by calculating other population genetics parameters. Although C. sieboldii has high potential for long-distance dispersal of pollen (as indicated by a fat-tailed dispersal kernel), the cumulative pollination at the local scale was spatially limited and strongly dependent on the distance between parents due to the high density of adults. Genetic diversity estimates for pollen pools accepted by each seed parent converged on a maximum as the effective number of pollen parents increased. The genetic diversity of pollen pool bulked over all the seed parents from inside the plot did not differ from that of the total pollen pools. Therefore, although pollen flow from distant pollen parents may help to maintain the genetic diversity of offspring, pollen parents neighboring seed parents may be the main contributors to the genetic diversity of the offspring at the seed stage.     

Outcrossing between an agroforestry plantation and remnant native populations of Eucalyptus loxophleba

Gene dispersal among populations of a species is an important force influencing their genetic structure. Dispersal may also occur between taxa that would normally be isolated when nonendemic, domesticated or transgenic species are planted within the natural range of interfertile taxa. Such a mosaic of populations is typical of many agricultural landscapes, and investigations are needed to assess the risks of genetic contamination of the endemic populations but a combination of approaches may be necessary because of the limitations of research in this landscape. This study used microsatellite markers and a range of analyses (mating system, paternity exclusion, Bayesian assignment) to examine gene dispersal between remnants of the endemic Eucalyptus loxophleba ssp. supralaevis and a plantation of a nonendemic subspecies. Our results indicate that remnant populations are connected by significant dispersal to pollen sources up to 1.94 km away including the plantation. The combined analyses showed that the pollen pool and outcrossing rates of individuals within remnants varied significantly probably because of asynchronous flowering and that the likelihood of paternity was not correlated with spatial proximity. More than half of all progeny had male parents from outside their stand with the largest proportions estimated to come from the plantation by exclusion (42.4%) or Bayesian analyses (18.8–76%). Fragmentation may not be associated with decreased gene dispersal between populations of tree species, natural or planted, so that the distances required to buffer endemic trees in fragmented rural landscapes are likely to be large.     

Extensive long-distance pollen dispersal in a fragmented landscape maintains genetic diversity in white spruce

Conifers are among the most genetically diverse plants but show the lowest levels of genetic differentiation, even among geographically distant populations. High gene flow among populations may be one of the most important factors in maintaining these genetic patterns. Here, we provide empirical evidence for extensive pollen-mediated gene dispersal between natural stands of a widespread northern temperate/boreal conifer, Picea glauca. We used 6 polymorphic allozyme loci to quantify the proportion of seeds sired by pollen originating from different sources in a landscape fragmented by agriculture in North Central Ontario, Canada. In 7 stands, a small proportion of seeds were sired by self-pollen or neighboring trees but 87.1% (+/-1.7% standard error [SE]) of seeds were sired by pollen from at least 250 to 3000 m away. In 4 single isolated trees, self-fertilization rates were low and more than 96% (+/-1.3% SE) of seeds were sired by immigrant pollen. The average minimum pollen dispersal distance in outcrossed matings was 619 m. These results provide strong evidence that extensive long-distance pollen dispersal plays a primary role in maintaining low genetic differentiation among natural populations of P. glauca and helps maintain genetic diversity and minimize inbreeding in small stands in a fragmented landscape.