利用等位酶对油松无性系种子进行父本分析

利用水平淀粉凝胶电泳8种酶10个位点34个等位基因,对辽宁兴城油松无性系种子园中17个无性系20个单株的200粒种子作了父本分析。无性系雄花量的多少与授粉果率并不成正比;不同无性系对花粉的接受存在一定的选择性,分析试样单株与周围的无性系并没有构成固定的授粉组合,说明授粉期风向等气象因子,无性系的雌雄花器的构造,受精等对受粉的成败都有作用,种子园结实初期自交现象并不明显,在分析试样中只有16＃,17＃,19＃无性系存在自交现象;种子园内花偻的有效传播一般在半径30m范围内,同时,分析了种子园子代的等位酶变异和空间遗传结构,从另一侧面反映出采种母树周围花粉分布比较均匀,并说明散粉期间风向等气象因子对授粉成败影响较大。     

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。     

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。     

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。     

野菊与菊花杂交中花粉活力和柱头可授性及胚胎发育研究

应用石蜡制片、活体压片、光学显微镜及扫描电子显微镜观察等方法,研究了四倍体河南云台山野菊(Dendranthema indicum)与栽培菊花'钟山金山'(D.grandiflorum 'Zhongshanjinshan')种间杂交中父本花粉活力、花粉在柱头萌发、花粉管生长及胚胎发育情况等.结果发现,父本云台山野菊的花粉活力在授粉时为12%左右.人工授粉后的不同时间,在柱头上都观察到正常萌发的花粉粒,且花粉管都能进入柱头,其中,在授粉后0.5 h时,平均每柱头有5.9粒花粉萌发;12 h时,为59.9粒;而24和48 h时,则分别降为47.1和35.7粒.此外,在授粉后8、10、12和15 d时,分别在49.1%、40.8%、39.7%和38.5%子房内观察到正常发育的胚胎,最终杂交结实率为44.8%,而母本自然开放结实率为52.3%.研究表明,授粉前其多数母本雌蕊发育良好、授粉后多数花粉能在柱头正常萌发和花粉管正常生长,在受精后大部分胚胎发育正常是野菊与栽培菊种间杂交较高结实率的重要保证,而授粉前父本较低的花粉活力对杂交结实率影响不大.     

黄梅秤锤树孤立居群的遗传多样性及其小尺度空间遗传结构

研究濒危植物片断化孤立居群不同年龄阶段植株的遗传多样性和小尺度空间遗传结构有助于认识残存居群动态和制订保育策略。本研究选取黄梅秤锤树(Sinojackia huangmeiensis)的一个片断化孤立居群(面积160m×80m)为研究对象,对居群内60株成年个体、175株幼树和198株幼苗全部定位,采用8个微卫星位点检测了居群内不同生活史阶段植株的遗传多样性、空间遗传结构,并分析了花粉和种子传播距离和式样。结果表明,黄梅秤锤树居群3种不同年龄阶段植株的遗传多样性之间无显著差异;居群出现显著的杂合子缺失,可能是由近交造成的;在10m以内成年个体、幼树和幼苗植株均呈现出显著的空间遗传结构,说明种子扩散限制于成年母树周边;种子和花粉传播的平均距离分别为9.07±13.38和23.81±23.60m,且花粉和种子传播式样均呈"L"型分布;种子雨重叠少、有限的基因流、自疏以及近亲繁殖是造成各年龄阶段出现空间遗传结构的主要原因。本研究结果启示,在采集秤锤树迁地保护材料时个体间距离应超过10m,以降低采样个体的遗传相似性;同时在就地保护过程中需要人为促进基因流和加强幼苗管理,以降低近交风险。     

玉米胚珠离体受精成功

从未授粉的玉米植株的雌穗上,切除子房上部的三分之一,获得裸露的胚珠。在切口处授以经过消毒的玉米花粉,获得了14粒由胚珠发育的幼嫩种子。在授粉后20—22天幼嫩种子的胚已经成熟,并在果穗块上萌发和生长,现已获得两个植株。其中一株植株上有明显的父本标记的紫色素性状及类似父本的旗叶。根尖细胞染色体2n=20,F_2代有分离,因此证明是胚珠离体受精而来的杂种植株。另一株植株形态、色素、旗叶均与母本相似,染色体2n=20,生长健壮,雄穗很早抽出,但在抽雄后49天才长出弱小的雌穗,未获得种子。     

花粉数量对芝麻受精结实、胚胎发育和后代的作用

本文研究了芝麻授粉时的花粉数量对受精结实、胚胎发育和后代的作用。试验处理包括1、4、10、20、40粒花粉等各级限量授粉及大量授粉对照。结果表明:每一粒花粉理论上能使一粒胚珠受精结成一粒种子。花粉数量减少引起胚的早期发育延缓与不整齐,但后期发育渐趋正常,接近或赶上对照胚。限量授粉与大量授粉的后代(F_1、F_2)看不出显著差别。本文讨论了以下问题:(1)受精作用的基本形式是“一对一”还是“多对一”?(2)“多余”花粉起什么作用?(3)限量授粉是不是一种有效的育种途径?     

外来入侵植物粉花月见草的繁殖生物学特性

粉花月见草单花花期1d,整株花期50d左右;单花花粉量达8825个,花粉活力达53.2%,花粉萌发快,萌发率达44.6%,自花授粉;种子小,千粒重0.092g,种子发芽率达85%,萌发时间短,具有休眠机制。三年生株丛结果达1274个,单果种子高达206粒,单株丛种子量2.6×105。种子表面平滑,自然传播以重力为主和风力作用,传播距离在100cm范围内。结果表明:粉花月见草繁殖力、适应力强,易形成单一优势种群,种子小近圆形易形成种子库,人为活动使其远距离传播,具有较大的危害性。     

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。     

Effects of male fecundity, interindividual distance and anisotropic pollen dispersal on mating success in a Scots pine (Pinus sylvestris) seed orchard

Quantifying the effect of pollen dispersal and flowering traits on mating success is essential for understanding evolutionary responses to changing environments and establishing strategies for forest tree breeding. This study examined, quantitatively, the effects of male fecundity, interindividual distance and anisotropic pollen dispersal on the mating success of Scots pine (Pinus sylvestris), utilizing a well-mapped Scots pine seed orchard. Paternity analysis of 1021 seeds sampled from 87 trees representing 28 clones showed that 53% of the seeds had at least one potential pollen parent within the orchard. Pronounced variation in paternal contribution was observed among clones. Variations in pollen production explained up to 78% of the variation in mating success, which was 11.2 times greater for clones producing the largest amount of pollen than for clones producing the least pollen. Mating success also varied with intertree distance and direction, which explained up to 28% of the variance. Fertilization between neighboring trees 2.3 m apart was 2.4 times more frequent than between trees 4.6 m apart, and up to 12.4 times higher for trees downwind of the presumed prevailing wind direction than for upwind trees. The effective number of pollen donors recorded in the seed orchard (12.2) was smaller than the theoretical expectation (19.7). Based on the empirical observations, a mating model that best describes the gene dispersal pattern in clonal seed orchards was constructed.     

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.     

Patterns of pollen and seed dispersal in a small, fragmented population of the wind-pollinated tree Araucaria angustifolia in southern Brazil

Paternity analysis based on eight microsatellite loci was used to investigate pollen and seed dispersal patterns of the dioecious wind-pollinated tree, Araucaria angustifolia. The study sites were a 5.4 ha isolated forest fragment and a small tree group situated 1.7 km away, located in Paranalpha State, Brazil. In the forest fragment, 121 males, 99 females, 66 seedlings and 92 juveniles were mapped and genotyped, together with 210 seeds. In the tree group, nine male and two female adults were mapped and genotyped, together with 20 seeds. Paternity analysis within the forest fragment indicated that at least 4% of the seeds, 3% of the seedlings and 7% of the juveniles were fertilized by pollen from trees in the adjacent group, and 6% of the seeds were fertilized by pollen from trees outside these stands. The average pollination distance within the forest fragment was 83 m; when the tree group was included the pollination distance was 2006 m. The average number of effective pollen donors was estimated as 12.6. Mother-trees within the fragment could be assigned to all seedlings and juveniles, suggesting an absence of seed immigration. The distance of seedlings and juveniles from their assigned mother-trees ranged from 0.35 to 291 m (with an average of 83 m). Significant spatial genetic structure among adult trees, seedlings, and juveniles was detected up to 50 m, indicating seed dispersal over a short distance. The effective pollination neighborhood ranged from 0.4 to 3.3 ha. The results suggest that seed dispersal is restricted but that there is long-distance pollen dispersal between the forest fragment and the tree group; thus, the two stands of trees are not isolated.     

Paternity analysis of pollen-mediated gene flow for Fraxinus excelsior L. in a chronically fragmented landscape

Paternity analysis based on microsatellite marker genotyping was used to infer contemporary genetic connectivity by pollen of three population remnants of the wind-pollinated, wind-dispersed tree Fraxinus excelsior, in a deforested Scottish landscape. By deterministically accounting for genotyping error and comparing a range of assignment methods, individual-based paternity assignments were used to derive population-level estimates of gene flow. Pollen immigration into a 300 ha landscape represents between 43 and 68% of effective pollination, mostly depending on assignment method. Individual male reproductive success is unequal, with 31 of 48 trees fertilizing one seed or more, but only three trees fertilizing more than ten seeds. Spatial analysis suggests a fat-tailed pollen dispersal curve with 85% of detected pollination occurring within 100 m, and 15% spreading between 300 and 1900 m from the source. Identification of immigrating pollen sourced from two neighbouring remnants indicates further effective dispersal at 2900 m. Pollen exchange among remnants is driven by population size rather than geographic distance, with larger remnants acting predominantly as pollen donors, and smaller remnants as pollen recipients. Enhanced wind dispersal of pollen in a barren landscape ensures that the seed produced within the catchment includes genetic material from a wide geographic area. However, gene flow estimates based on analysis of non-dispersed seeds were shown to underestimate realized gene immigration into the remnants by a factor of two suggesting that predictive landscape conservation requires integrated estimates of post-recruitment gene flow occurring via both pollen and seed.     

Pollen Dispersal Between Isolated Trees in the Brazilian Savannah: A Case Study of the Neotropical Tree Hymenaea stigonocarpa

This case study examines the pollen dispersal distance, pollen dispersal patterns and intra‐family genetic structure for isolated trees in pastures of the bat‐pollinated Neotropical tree species Hymenaea stigonocarpa using six microsatellite loci and parentage analysis. The sampling included 28 grouped trees (referred to as the population) and six isolated trees in pastureland along a highway in Mato Grosso do Sul State, Brazil. From the population, we sampled 137 seeds from 12 seed‐trees, and from the isolated trees, we sampled 34 seeds from two seed‐trees. The results showed that pollen was dispersed over long distances (reaching 7353 m) and therefore the spatially isolated trees were not reproductively isolated. The pollen immigration rate in the population was also high (31%). Isolated trees presented a higher selfing rate (s=26%) than trees in the population (s=12%), suggesting that the spatial isolation of the trees increased selfing. However, selfing was responsible for only 30 percent of the inbreeding in offspring and mating among relatives was 70 percent. In the population, excluding selfing, ca 72 percent of the pollen was dispersed over distances <1000 m (average: 860 m). For the two isolated seed‐trees, excluding selfing, the average pollen dispersal distance was 5229 m. The results demonstrate that although pollen can be dispersed over long distances for H. stigonocarpa isolated trees, a high percentage of pollen comes from the same tree (selfing) and mating was correlated. Consequently, seeds must be collected from a large number of seed‐trees for conservation purposes.     

Short-distance pollen dispersal in a protogynous Annonaceae tree species from the Brazilian Cerrado

Annona crassiflora (Annonaceae) is a protogynous beetle-pollinated savannah tree species, widely distributed in the savannahs of the Cerrado biome. Studies on the mating system and pollen dispersal of protogynous species are very scarce. Here, we used six microsatellite loci to assess the mating system and pollen dispersal of A. crassiflora in a savannah remnant in Central Brazil. We mapped and sampled leaves of 112 adult trees and collected 74 fruits from 20 mother trees (1–4 fruits per plant) to obtain the seeds used (460) for mating system and parentage analyses. Annona crassiflora has predominantly allogamous mating systems, with a high multilocus outcrossing rate (t_m?=?0.974, SE?=?0.011) that did not differ among mother trees (F?=?1.32, p?=?0.165). However, t_m–t_s was variable among seed trees, indicating that some seeds were produced by mating among relatives. Our results also showed multiple paternity within fruits. Multilocus correlation of outcrossed paternity was high (r_p?=?0.302, SE?=?0.045), indicating that for each mother tree, the probability that the same pollen donor sired two random sibs was 30.2%, and the mean number of pollen donors per mother tree was high (6.3). We detected a maximum pollen dispersal distance of 360.7 m and an average of 124.3 m (SD?=?80 m), but most pollination events (73%) occurred at shorter distances (<?160 m), indicating short-distance pollen dispersal, most likely due to the pollinator behaviour.     

Contemporary pollen flow,mating patterns and effective population size inferred from paternity analysis in a small fragmented population of the Neotropical tree <Emphasis Type="Italic">Copaifera langsdorffii</Emphasis> Desf. (Leguminosae-Caesalpinioideae)

Pollen flow and dispersal patterns were investigated in a small, isolated forest fragment of the Neotropical insect pollinated tree Copaifera langsdorffii, using paternity analysis and eight microsatellite loci. We also investigated the coancestry and effective population size of progeny arrays for conservation and environmental restoration purposes. Open-pollinated seeds were collected from 15 seed trees within the forest fragment, in which all adult trees were mapped, measured and genotyped. Twenty seeds were also collected from the nearest neighbor tree located 1.2 km from the forest fragment. Our results show that levels of genetic diversity were significantly higher in adults than offspring and significant levels of inbreeding were detected in offspring (F = 0.226). From paternity analysis, we observed low levels of selfing (s = 8%) and pollen immigration (m = 8%) in the fragment, but very high levels were detected for the isolated tree (s = 20%; m = 75%), indicating that the population and the tree are not reproductively isolated and are connected by patterns of long distance pollen dispersal (maximum detected 1,420 m). Within the forest fragment, the pattern of pollen dispersal was a near neighbor pattern with 49% of the pollen being dispersed within 50 m. The effective population size of the progeny array was low, indicating the need to collect seeds from a large number of seed trees (at least 76) for conservation purposes. The results show that the spatial isolation of the population and isolated tree due to forest fragmentation has not disrupted genetic connectivity; however, spatial isolation does seem to increase selfing and correlated mating.     

FLOWER,FRUIT AND SEED ABORTION IN TROPICAL FOREST TREES: IMPLICATIONS FOR THE EVOLUTION OF PATERNAL AND MATERNAL REPRODUCTIVE PATTERNS

Flower, fruit and immature seed abortion was studied in seven self-incompatible species of trees in a tropical lowland semideciduous forest. The species showed considerable variability in fruit and seed set and the rate at which flowers and fruits were aborted. The amount of flower and fruit abortion also varied over time within species. Small samples of open-pollinated flowers in three species showed adequate amounts of pollen on the stigma, but it could not be determined whether the pollen was compatible or incompatible. In a species with multiseeded fruits, the aborted fruits contained significantly fewer seeds than those retained on the plant. Position of fruit within the inflorescence and of seed within the ovary also had a marked effect on abortion: fruits and seeds at certain positions had a higher probability of abortion than those at other positions. Experiments to test the effect of pollen source on abortion were inconclusive. The factors underlying abortions were evaluated in the context of three mutually non-exclusive hypotheses. It is concluded that selection for increased pollen dispersal and uncertainty in paternity of the zygotes are major factors underlying abortions.     

Increased selfing and correlated paternity in a small population of a predominantly outcrossing conifer, Pinus sylvestris

Outcrossing rate, the rates of ovule and seed abortion, and levels of correlated paternity were estimated in a small population of Pinus sylvestris, a predominantly outcrossing conifer, and were compared with estimates from two widely dispersed woodlands of the same species, showing a range of densities. On average, seed trees of the small population showed an eight-fold higher selfing rate (25 vs. 3%) and a 100-fold greater incidence of correlated paternity (19.6 vs. 0.2%) than did trees from the large populations. No evidence was found of pollen limitation within the remnant stand, as suggested by ovule abortion rates. Investigation of the mating patterns in the small population, based on the unambiguous genealogy of 778 open-pollinated seeds, showed a large departure from random mating. Only 8% of the possible mating pairs within the stand were observed. Correlated paternity rate within a maternal sibship was negatively associated (rs = -0.398, P < 0.050) with the distance to the nearest neighbour, and shared paternity among maternal sibships was negatively correlated (rs = -0.704, P < 0.001) with the distance between seed trees. Numerical simulations, based on the estimated individual pollen dispersal kernel, suggest that restricted dispersal might have been the key factor affecting mating patterns in the small population and, together with low population density, may account for the observed mating system variation between the small and the large populations. The results of this study show that a severe size reduction may substantially affect the mating system of a wind-pollinated, typically outcrossed plant species.