Understanding the effects of isolation on seed and pollen flow, spatial genetic structure and effective population size of the dioecious tropical tree species Myracrodruon urundeuva

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Contemporary pollen flow, mating patterns and effective population size inferred from paternity analysis in a small fragmented population of the Neotropical tree Copaifera langsdorffii Desf. (Leguminosae-Caesalpinioideae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Genetic analyses reveal high levels of seed and pollen flow in hawthorn (Crataegus monogyna Jacq.), a key component of hedgerows

Hedgerows represent important components of agri-environment landscapes that are increasingly coming under threat from climate change, emergent diseases, invasive species and land use change. Given that population genetic data can be used to inform best-practice management strategies for woodland and hedgerow tree species, we carried out a study on hawthorn (Crataegus monogyna Jacq.), a key component of hedgerows, on a regional basis using a combination of nuclear and chloroplast microsatellite markers. We found that levels of genetic diversity were high and comparable to, or slightly higher than, other tree species from the same region. Levels of population differentiation for both sets of markers, however, were extremely low, suggesting extensive gene flow via both seed and pollen. These findings suggest that a holistic approach to woodland management, one which does not necessarily rely on the concept of “seed zones” previously suggested, but which also takes into account populations with high and/or rare chloroplast (i.e. seed-specific) genetic variation, might be the best approach to restocking and replanting.

The risk of pollen-mediated gene flow from exotic Corymbia plantations into native Corymbia populations in Australia

Sectors of the forest plantation industry in Australia are set to expand in the near future using species or hybrids of the spotted gums (Corymbia, Section Politaria). Plantations of these taxa have already been introduced across temperate and subtropical Australia, representing locally exotic introductions from native stands in Queensland and New South Wales. A literature review was undertaken to provide insights into the potential for pollen-mediated gene flow from these plantations into native populations. Three factors suggest that such gene flow is likely; (1) interspecific hybridisation within the genus has frequently been recorded, including between distantly related species from different sections, (2) apparent high levels of vertebrate pollinator activity may result in plantation pollen being moved over hundreds of kilometres, (3) much of the plantation estate is being established among closely related taxa and therefore few barriers to gene flow are expected. Across Australia, 20 of the 100 native Corymbia taxa were found to have regional level co-occurrence with plantations. These were located most notably within regions of north-east New South Wales and south-east Queensland, however, co-occurrence was also found in south-west Western Australia and eastern Victoria. The native species found to have co-occurrence were then assessed for the presence of reproductive barriers at each step in the process of gene flow that may reduce the number of species at risk even further. The available data suggest three risk categories exist for Corymbia. The highest risk was for gene flow from plantations of spotted gums to native populations of spotted gums. This was based on the expected limited existence of pre- and post-zygotic barriers, substantial long-distance pollen dispersal and an apparent broad period of flowering in Corymbia citriodora subsp. variegata plantations. The following risk category focussed on gene flow from Corymbia torelliana × C. c. variegata hybrid plantations into native C. c. variegata, as the barriers associated with the production and establishment of F1 hybrids have been circumvented. For the lowest risk category, Corymbia plantations may present a risk to other non-spotted gum species, however, further investigation of the particular cross-combinations is required. A list of research directions is provided to better quantify these risks. Empirical data will need to be combined within a risk assessment framework that will not only estimate the likelihood of exotic gene flow, but also consider the conservation status/value of the native populations. In addition, the potential impacts of pollen flow from plantations will need to be weighed up against their various economic and environmental benefits.

To self, or not to self ... A review of outcrossing and pollen-mediated gene flow in neotropical trees

Despite the typically low population densities and animal-mediated pollination of tropical forest trees, outcrossing and long-distance pollen dispersal are the norm. We reviewed the genetic literature on mating systems and pollen dispersal for neotropical trees to identify the ecological and phylogenetic correlates. The 36 studies surveyed found >90% outcrossed mating for 45 hermaphroditic or monoecious species. Self-fertilization rates varied inversely with population density and showed phylogenetic and geographic trends. The few direct measures of pollen flow (N = 11 studies) suggest that pollen dispersal is widespread among low-density tropical trees, ranging from a mean of 200 m to over 19 km for species pollinated by small insects or bats. Future research needs to examine (1) the effect of inbreeding depression on observed outcrossing rates, (2) pollen dispersal in a wide range of pollination syndromes and ecological classes, (3) and the range of variation of mating system expression at different hierarchical levels, including individual, seasonal, population, ecological, landscape and range wide.

The role of pollen in the changing environmental conditions of Scots pine

Rapid change in climate is challenge for the adaptation of forest trees in the future. In wind pollinated tree species pollen mediated long distance gene flow may provide alleles that are (pre)adapted to a future climate. In order to examine the long distance pollen flow in Scots pine (Pinus sylvestris L.), we measured the amount and viability of airborne pollen and flowering phenology in central, northern, and northernmost Finland during four years. Viable airborne pollen grains were detected during female flowering and before local pollen shedding in all study sites. The situation when there was nonlocal pollen in the air lasted from one to four days depending on the year and study site. The amount of nonlocal airborne pollen varied also between years and study sites, the total amount of nonlocal viable pollen in the air was 2.3% from all detected viable pollen grains. The effect of pollen origin on seeds siring ability was studied with artificial pollination experiments. Pollen genotypes originating from southern Finland sired 76% and 48 % of the analysed seeds in competition studies where both pollen origin were introduced simultaneously into the female strobili. We examined the importance of arrival order of pollen grains in to the strobili in a study where pollen genotypes of different origin were introduced in two hours interval. Northern genotypes sired 76% of the analysed seeds when it was injected first, but in the "southern first" experiment both pollen types sired equal amount of seeds. The first pollen grain in the pollen chamber do not always fertilizes the ovum, instead there likely is more complex way of competition between pollen grains. To examine chemically mediated pollen-pollen interactions we conducted in vitro germination experiment where different pollen genotypes had chemical but not physical contact. Both positive and negative effects of interactions were found. We found highly negative effects in germinability of northern pollen grains when they were germinating with southern pollen, and increase in the germinability of southern pollen. There were no variation in the size of the dry pollen grains between pollen origins, and minor variation between different genotypes. After hydration and germination northern pollen grains were larger than southern pollen. Pollen genotypes having high hydration rates had low germinability and tube growth rate, however, germinated pollen grains were larger in size than nongerminated. This supports the suggestion that the early germination and growth of pollen tube is dependent on pollen storage materialsand less dependent on water intake and hydration. Long distance pollen movements and good competition ability of southern pollen makes gene flow possible, although rising temperature and timing of pollen movements may affect pollen competition and the amount of gene flow.

独花兰和扇脉杓兰的传粉生态学

兰科植物精巧的花部结构及其独特的传粉机制为自然选择理论和异花受粉优势学说提供了强有力的证据。开展兰科植物的传粉生态学研究对进一步探讨植物进化中的一些关键问题（如生殖隔离、物种形成、适应和繁育系统进化等）具有重要意义。本文通过对独花兰 (Changnienia amoena) 和扇脉杓兰 (Cypripedium japonicum) 两种兰科植物进行植物与传粉者之间关系、种内形态变异、花粉散布等方面的研究，探讨其适应进化方式、传粉系统的进化趋势，同时为开发适用的分子标记，在独花兰中初步摸索和探讨了微卫星标记的分离。主要研究结果如下： 1. 濒危植物独花兰的传粉生态学 在神农架2个地点进行了连续2年的野外观测和实验。结果表明，独花兰是一种自交亲和、需要昆虫传粉的欺骗性植物。在2个地点独花兰的传粉者种类不同，在龙门河三条熊蜂 (Bombus (Diversobombus) trifasciatus) 是主要传粉者，在关门山仿熊蜂 (B. (Tricornibombus) imitator) 是唯一的传粉者。尽管它们出现的丰度和觅食行为不同，但其传粉行为和携粉部位非常相同，因此，可以看作是一个功能群（functional group）。从种群水平上看，开花个体的分布式样不是显著的偏斜曲线图，传粉者的有效访问主要集中在开花的前、中期。由于没有花蜜，传粉者在花内停留的时间很短，花粉块输出和传粉发生在熊蜂劳而无获的访问、退出花时。自然条件下，独花兰的结实率很低，只有6%-12%，并呈现明显的年份和地点变化。传粉者限制是结实率低的主要原因。与同亚族近缘种布袋兰相比，分布于中国中部的独花兰受冰期的影响很小，其传粉环境相对稳定，在进化历史中，形成了一种稳定的传粉系统。本研究结果纠正了前人对独花兰繁殖方式的错误认识，并为其保护策略的制订提供了重要资料。 2. 独花兰种群大小、传粉者访问和花粉流 为了检验独花兰种群大小、空间格局与传粉者访问之间的关系，连续2年对10个独花兰种群的花粉块输出和输入进行了统计分析，同时对花粉块进行标记研究其散布式样。结果显示，花粉块输出比例与种群大小之间关系在2003年呈显著负相关，但在2004年这种负相关关系不显著。尽管独花兰的空间分布格局在不同种群不同，但传粉者的访问除了在有蜂巢的种群为聚集式访问外，其余种群内均为随机访问式样。花粉块的散布式样呈尖峰态分布，多数花粉块散布很短的距离，少数散布较远。花粉块的平均散布距离在龙门河为7.3 m，在关门山为10.6 m。由于各种群之间相隔很远，种群之间花粉块交流受到限制，很少有种群外的花粉块输入。花粉块传递只在种群内近缘个体中进行，有可能导致种内遗传或形态分化。 3. 独花兰种内形态变异及其适应意义 对庐山、新宁和神农架3个地点15个独花兰自然种群的形态变异进行了研究，探讨了形态多样性水平和地理变异式样及其可能的适应机制。结果表明，在物种水平上独花兰形态性状存在丰富的变异。单因素方差分析显示3个地区间多个形态性状存在极显著差异（P<0.01），UPGMA聚类分析也表明这3个地区分别形成明显不同的分支，说明3个地区种群植物形态已经出现分化。在神农架地区，龙门河和关门山两个地点间出现明显的形态分化，而这两个地点的传粉者在形体大小表现出显著差异。适合度与形态性状之间的相关性分析显示，独花兰种内形态分化是以传粉者为媒介的自然选择作用的结果。移栽实验显示，本地传粉者对本地和移栽种群花的访问表现出一定的选择性。 4. 扇脉杓兰的传粉生态学研究 与独花兰同域分布、花期不遇的扇脉杓兰是杓兰属的一个特殊类群，具有典型“Japonicum”型唇瓣类型。对神农架6个种群连续两年野外观测和实验结果表明，扇脉杓兰是一种自交亲和、需要昆虫传粉的欺骗性植物。在种群水平上开花个体的分布式样呈显著的偏斜曲线图，在短时间内迅速到达盛花期。扇脉杓兰的传粉者是三种熊蜂，其访问频次很低，访问时间很短，有效访问主要集中在开花的前、中期。自然结实率只有4.3％－8.5％，人工授粉实验证明，传粉者限制是结实率低的主要原因，且传粉者限制程度在花期不同阶段和不同地点存在差异。对欺骗性植物聚集生长、开花有利于吸引传粉者这一假说进行检验，结果显示，克隆群丛大小与传粉者有效访问（花粉块输出比例）呈弱负相关关系。聚集生长的结实率与分散生长的没有显著差异，说明繁殖成功与开花个体的密度无关。扇脉杓兰的花粉集结为未蜡质化的花粉块，是杓兰属中粘质花粉和蜡质花粉块的中间进化类型。传粉生态学和形态学证据显示C. acaule和扇脉杓兰是东亚-北美间断分布的姐妹种对。 5. 独花兰微卫星位点的分离 为了深入研究独花兰种群的基因流和交配系统，用Dynabeads和选择性杂交法分离微卫星位点、筛选引物。首先将基因组DNA用合适的内切酶消化为400-1000 bp大小的片段，然后用生物素标记的简单重复寡核苷酸序列做探针与其杂交，杂交复合物用抗生链霉素蛋白包裹的磁珠吸附，经过一系列洗脱、沉淀，得到富含重复片段的DNA，然后在进行克隆、测序，利用SSR两侧翼区设计引物，经过多态性分析可得到微卫星标记。结果显示，35个克隆序列中18个（51.4％）含有重复片段（SSRs），说明用Dynabeads富集效率明显高于传统方法。到目前为止所得引物共4对。这一实验方法的探索为以后分离杓兰属等兰科植物微卫星位点、进而进行群体遗传学研究奠定了一定的基础。

Distância e padrões de dispersão contemporânea de pólen e sistema de reprodução em pequeno fragmento isolado de Copaifera Langsdorfii Desf. (Leguminosae – Caesalpinoideae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudo hierárquico do sistema de reprodução entre e dentro de frutos, fluxo de pólen e estrutura genética espacial em um fragmento e em árvores isoladas na pastagem de hymenaea stigonocarpa mart. ex hayne na região de cerrado

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Genetic effects of chronic habitat fragmentation on tree species: the case of Sorbus aucuparia in a deforested Scottish landscape

Sustainable forest restoration and management practices require a thorough understanding of the influence that habitat fragmentation has on the processes shaping genetic variation and its distribution in tree populations. We quantified genetic variation at isozyme markers and chloroplast DNA (cpDNA), analysed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in severely fragmented populations of Sorbus aucuparia (Rosaceae) in a single catchment (Moffat) in southern Scotland. Remnants maintain surprisingly high levels of gene diversity (H-E) for isozymes (H-E = 0.195) and cpDNA markers (H-E = 0.490). Estimates are very similar to those from non-fragmented populations in continental Europe, even though the latter were sampled over a much larger spatial scale. Overall, no genetic bottleneck or departures from random mating were detected in the Moffat fragments. However, genetic differentiation among remnants was detected for both types of marker (isozymes Theta(n) = 0.043, cpDNA Theta(c) = 0.131; G-test, P-value < 0.001). In this self-incompatible, insect-pollinated, bird-dispersed tree species, the estimated ratio of pollen flow to seed flow between fragments is close to 1 (r = 1.36). Reduced pollen-mediated gene flow is a likely consequence of habitat fragmentation, but effective seed dispersal by birds is probably helping to maintain high levels of genetic diversity within remnants and reduce genetic differentiation between them.

High nuclear genetic diversity, high levels of outcrossing and low differentiation among remnant populations of Quercus petraea at the margin of its range in Ireland

Background and Aims Quercus petraea colonized Ireland after the last glaciation from refugia on mainland Europe. Deforestation. however. beginning in Neolithic times, has resulted in small, scattered forest fragments, now covering less than 12 000 ha. Methods Plastid (three fragments) and microsatellite variation (13 loci) were characterized in seven Irish populations sampled along a north-south gradient. Using Bayesian approaches and Wright's F-statistics, the effects of colonization and fragmentation on the genetic structure and mating patterns of extant oak populations were investigated. Key-Results All Populations possessed cytotypes common to the Iberian Peninsula. Despite the distance from the refugial core and the extensive deforestation in Ireland, nuclear genetic variation was high and comparable to mainland Europe. Low population differentiation was observed within Ireland and populations showed no evidence for isolation by distance. As expected of a marker with an effective Population size of one-quarter relative to the nuclear genome, plastid variation indicated higher differentiation. Individual inbreeding coefficients indicated high levels of outcrossing. Conclusions Consistent with a large effective Population size in the historical migrant gene pool and/or with high gene flow among populations, high within-population diversity and low population differentiation was observred within Ireland. It is proposed that native Q. petraea populations in Ireland share a common phylogeographic history and that the present genetic structure does not reflect founder effects. (C) 2004 Annals of Botany Company.

Genetic resource impacts of habitat loss and degradation; reconciling empirical evidence and predicted theory for neotropical trees

The theoretical impacts of anthropogenic habitat degradation on genetic resources have been well articulated. Here we use a simulation approach to assess the magnitude of expected genetic change, and review 31 studies of 23 neotropical tree species to assess whether empirical case studies conform to theory. Major differences in the sensitivity of measures to detect the genetic health of degraded populations were obvious. Most studies employing genetic diversity (nine out of 13) found no significant consequences, yet most that assessed progeny inbreeding (six out of eight), reproductive output (seven out of 10) and fitness (all six) highlighted significant impacts. These observations are in line with theory, where inbreeding is observed immediately following impact, but genetic diversity is lost slowly over subsequent generations, which for trees may take decades. Studies also highlight the ecological, not just genetic, consequences of habitat degradation that can cause reduced seed set and progeny fitness. Unexpectedly, two studies examining pollen flow using paternity analysis highlight an extensive network of gene flow at smaller spatial scales (less than 10 km). Gene flow can thus mitigate against loss of genetic diversity and assist in long-term population viability, even in degraded landscapes. Unfortunately, the surveyed studies were too few and heterogeneous to examine concepts of population size thresholds and genetic resilience in relation to life history. Future suggested research priorities include undertaking integrated studies on a range of species in the same landscapes; better documentation of the extent and duration of impact; and most importantly, combining neutral marker, pollination dynamics, ecological consequences, and progeny fitness assessment within single studies.

Low levels of realized seed and pollen gene flow and strong spatial genetic structure in a small, isolated and fragmented population of the tropical tree Copaifera langsdorffii Desf

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Pollen and seed flow patterns of Carapa guianensis Aublet. (Meliaceae) in two types of Amazonian forest

Various factors affect spatial genetic structure in plant populations, including adult density and primary and secondary seed dispersal mechanisms. We evaluated pollen and seed dispersal distances and spatial genetic structure of Carapa guianensis Aublet. (Meliaceae) in occasionally inundated and terra firme forest environments that differed in tree densities and secondary seed dispersal agents. We used parentage analysis to obtain contemporary gene flow estimates and assessed the spatial genetic structure of adults and juveniles. Despite the higher density of adults (diameter at breast height >= 25 cm) and spatial aggregation in occasionally inundated forest, the average pollen dispersal distance was similar in both types of forest (195 +/- 106 m in terra firme and 175 +/- 87 m in occasionally inundated plots). Higher seed flow rates (36.7% of juveniles were from outside the plot) and distances (155 +/- 84 m) were found in terra firme compared to the occasionally inundated plot (25.4% and 114 +/- 69 m). There was a weak spatial genetic structure in juveniles and in terra firme adults. These results indicate that inundation may not have had a significant role in seed dispersal in the occasionally inundated plot, probably because of the higher levels of seedling mortality.