Conservation of genetic diversity in old‐growth forest communities of the southeastern United States

Question: How do studies of the distribution of genetic diversity of species with different life forms contribute to the development of conservation strategies? Location: Old‐growth forests of the southeastern United States. Methods: Reviews of the plant allozyme literature are used to identify differences in genetic diversity and structure among species with different life forms, distributions and breeding systems. The general results are illustrated by case studies of four plant species characteristic of two widespread old‐growth forest communities of the southeastern United States: the Pinus palustris – Aristida stricta (Longleaf pine – wiregrass) savanna of the Coastal Plain and the Quercus – Carya – Pinus (Oak‐hickory‐pine) forest of the Piedmont. Genetic variation patterns of single‐gene and quantitative traits are also reviewed. Results: Dominant forest trees, represented by Pinus palustris(longleaf pine) and Quercus rubra (Northern red oak), maintain most of their genetic diversity within their populations whereas a higher proportion of the genetic diversity of herbaceous understorey species such as Sarracenia leucophylla and Trillium reliquum is distributed among their populations. The herbaceous species also tend to have more population‐to‐population variation in genetic diversity. Higher genetic differentiation among populations is seen for quantitative traits than for allozyme traits, indicating that interpopulation variation in quantitative traits is influenced by natural selection. Conclusion: Developing effective conservation strategies for one or a few species may not prove adequate for species with other combinations of traits. Given suitable empirical studies, it should be possible to design efficient conservation programs that maintain natural levels of genetic diversity within species of conservation interest.     

Evaluation of Genetic Diversity and Development of a Core Collection of Wild Rice (Oryza rufipogon Griff.) Populations in China

Common wild rice (Oryza rufipogon Griff.), the progenitor of Asian cultivated rice (O. sativa L.), is endangered due to habitat loss. The objectives of this research were to evaluate the genetic diversity of wild rice species in isolated populations and to develop a core collection of representative genotypes for ex situ conservation. We collected 885 wild rice accessions from eight geographically distinct regions and transplanted these accessions in a protected conservation garden over a period of almost two decades. We evaluated these accessions for 13 morphological or phenological traits and genotyped them for 36 DNA markers evenly distributed on the 12 chromosomes. The coefficient of variation of quantitative traits was 0.56 and ranged from 0.37 to 1.06. SSR markers detected 206 different alleles with an average of 6 alleles per locus. The mean polymorphism information content (PIC) was 0.64 in all populations, indicating that the marker loci have a high level of polymorphism and genetic diversity in all populations. Phylogenetic analyses based on morphological and molecular data revealed remarkable differences in the genetic diversity of common wild rice populations. The results showed that the Zengcheng, Gaozhou, and Suixi populations possess higher levels of genetic diversity, whereas the Huilai and Boluo populations have lower levels of genetic diversity than do the other populations. Based on their genetic distance, 130 accessions were selected as a core collection that retained over 90% of the alleles at the 36 marker loci. This genetically diverse core collection will be a useful resource for genomic studies of rice and for initiatives aimed at developing rice with improved agronomic traits.     

Mapping genetic diversity of cherimoya (Annona cherimola Mill.): application of spatial analysis for conservation and use of plant genetic resources

There is a growing call for inventories that evaluate geographic patterns in diversity of plant genetic resources maintained on farm and in species' natural populations in order to enhance their use and conservation. Such evaluations are relevant for useful tropical and subtropical tree species, as many of these species are still undomesticated, or in incipient stages of domestication and local populations can offer yet-unknown traits of high value to further domestication. For many outcrossing species, such as most trees, inbreeding depression can be an issue, and genetic diversity is important to sustain local production. Diversity is also crucial for species to adapt to environmental changes. This paper explores the possibilities of incorporating molecular marker data into Geographic Information Systems (GIS) to allow visualization and better understanding of spatial patterns of genetic diversity as a key input to optimize conservation and use of plant genetic resources, based on a case study of cherimoya (Annona cherimola Mill.), a Neotropical fruit tree species. We present spatial analyses to (1) improve the understanding of spatial distribution of genetic diversity of cherimoya natural stands and cultivated trees in Ecuador, Bolivia and Peru based on microsatellite molecular markers (SSRs); and (2) formulate optimal conservation strategies by revealing priority areas for in situ conservation, and identifying existing diversity gaps in ex situ collections. We found high levels of allelic richness, locally common alleles and expected heterozygosity in cherimoya's putative centre of origin, southern Ecuador and northern Peru, whereas levels of diversity in southern Peru and especially in Bolivia were significantly lower. The application of GIS on a large microsatellite dataset allows a more detailed prioritization of areas for in situ conservation and targeted collection across the Andean distribution range of cherimoya than previous studies could do, i.e. at province and department level in Ecuador and Peru, respectively.     

Local adaptation across a climatic gradient despite small effective population size in the rare sapphire rockcress

When assigning conservation priorities in endangered species, two common management strategies seek to protect remnant populations that (i) are the most genetically divergent or (ii) possess the highest diversity at neutral genetic markers. These two approaches assume that variation in molecular markers reflects variation in ecologically important traits and ignore the possibility of local adaptation among populations that show little divergence or variation at marker loci. Using common garden experiments, we demonstrate that populations of the rare endemic plant Arabis fecunda are physiologically adapted to the local microclimate. Local adaptation occurs despite (i) the absence of divergence at almost all marker loci and (ii) very small effective population sizes, as evidenced by extremely low levels of allozyme and DNA sequence polymorphism. Our results provide empirical evidence that setting conservation priorities based exclusively on molecular marker diversity may lead to the loss of locally adapted populations.     

Biodiversity assessment using markers for ecologically important traits

Most studies of genetic variation within species to date are based on random markers. However, how well this correlates with quantitative variation is contentious. Yet, functional, or'ecotypic' variation in quantitative traits determines the ecological niche of a species, its future evolutionary potential, and, for livestock, crops and their wild relatives, their usefulness as a genetic resource for breeding. But nowadays we can also assess genetic diversity using markers directly targeted at specific genes or gene families. Such gene-targeted, multilocus profiles of markers can contribute to ex-situ management of genetic resources, ecological studies of diversity, and conservation of endangered species.     

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 (HE) for isozymes (HE = 0.195) and cpDNA markers (HE = 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.     

DNA marker technology for wildlife conservation

Use of molecular markers for identification of protected species offers a greater promise in the field of conservation biology. The information on genetic diversity of wildlife is necessary to ascertain the genetically deteriorated populations so that better management plans can be established for their conservation. Accurate classification of these threatened species allows understanding of the species biology and identification of distinct populations that should be managed with utmost care. Molecular markers are versatile tools for identification of populations with genetic crisis by comparing genetic diversities that in turn helps to resolve taxonomic uncertainties and to establish management units within species. The genetic marker analysis also provides sensitive and useful tools for prevention of illegal hunting and poaching and for more effective implementation of the laws for protection of the endangered species. This review summarizes various tools of DNA markers technology for application in molecular diversity analysis with special emphasis on wildlife conservation.     

Admixture mapping of quantitative traits in Populus hybrid zones: power and limitations

Uncovering the genetic architecture of species differences is of central importance for understanding the origin and maintenance of biological diversity. Admixture mapping can be used to identify the number and effect sizes of genes that contribute to the divergence of ecologically important traits, even in taxa that are not amenable to laboratory crosses because of their long generation time or other limitations. Here, we apply admixture mapping to naturally occurring hybrids between two ecologically divergent Populus species. We map quantitative trait loci for eight leaf morphological traits using 77 mapped microsatellite markers from all 19 chromosomes of Populus. We apply multivariate linear regression analysis allowing the modeling of additive and non-additive gene action and identify several candidate genomic regions associated with leaf morphology using an information-theoretic approach. We perform simulation studies to assess the power and limitations of admixture mapping of quantitative traits in natural hybrid populations for a variety of genetic architectures and modes of gene action. Our results indicate that (1) admixture mapping has considerable power to identify the genetic architecture of species differences if sample sizes and marker densities are sufficiently high, (2) modeling of non-additive gene action can help to elucidate the discrepancy between genotype and phenotype sometimes seen in interspecific hybrids, and (3) the genetic architecture of leaf morphological traits in the studied Populus species involves complementary and overdominant gene action, providing the basis for rapid adaptation of these ecologically important forest trees.     

湖北河岸带植物中华蚊母树遗传多样性的SRAP分析

利用SRAP分子标记技术,对湖北省河岸带植物中华蚊母树的4个自然居群和1个迁地居群的遗传多样性和遗传结构进行了分析。结果表明,中华蚊母树物种具有较高水平的遗传多样性,7对SRAP引物进行PCR扩增的多态性位点百分率(PPF)为80.43%,每个位点的等位基因数(A)为2,有效等位基因数(Ae)为1.34,总遗传多样性Nei’s基因多样性指数(Hp)为0.215 9,Shannon信息多样性指数(I)为0.350 9。在居群水平上,5个居群总的遗传变异Ht为0.218 8,居群内的遗传变异Hs为0.193 4,居群间的遗传分化系数Gst为0.116 1,表明在总的遗传变异中有88.39%变异存在于居群内,仅11.61%存在于居群间,居群间的基因流Nm为3.807 2,表明居群间有较大程度的基因交流。UPGMA聚类分析和主成分分析显示中华蚊母树主要分为两个居群组,在长江三峡沿岸香溪和乐天溪由于遗传距离比较近聚为一小类再与高家堰聚为一大类,而沿渡河和三峡植物园居群聚为另一大类,表明迁地居群三峡植物园的中华蚊母树与来自巴东沿渡河居群的样本亲缘关系最近,且三峡植物园迁地保护居群基本保育了其遗传多样性总水平。同时在分析讨论了中华蚊母树遗传多样性与其繁育系统、生境及其起源进化的关系的基础上,评价了中华蚊母树的保护策略,并在评价保护成果的基础上,提出了今后进一步保育的策略。结果还表明SRAP标记是分析中华蚊母树遗传多样性和遗传结构非常可靠的一种标记,而且这是使用SRAP标记研究中华蚊母树的首次报道。     

Genetic structure and diversity of Shorea obtusa (Dipterocarpaceae) in Thailand

Shorea obtusa is a keystone species of the dry deciduous dipterocarp forest in Thailand. In this study, the genetic structure and diversity of this species were evaluated by means of five microsatellite markers. A total of 146 trees were collected from five populations encompassing major forest regions of Thailand. High levels of genetic diversity were found among the five populations with the average He of 0.664. Genetic differentiations between populations, although significant, were low with approximately 3% of genetic variation partitioned among populations. This may indicate that the populations sampled were recently part of a continuous population. A tree constructed using the unweighted pair group method with arithmetic average, based on Nei's genetic distance, divided the populations into three groups. This separation was consistent with the altitudinal zonation of the populations, thus indicating that altitude might play a significant role in the genetic structure of S. obtusa. Areas of high genetic diversity were identified which could be considered priorities for conservation.     

No genetic diversity at molecular markers and strong phenotypic plasticity in populations of Ranunculus nodiflorus, an endangered plant species in France

BACKGROUND AND AIMS: Although conservation biology has long focused on population dynamics and genetics, phenotypic plasticity is likely to play a significant role in population viability. Here, an investigation is made into the relative contribution of genetic diversity and phenotypic plasticity to the phenotypic variation in natural populations of Ranunculus nodiflorus, a rare annual plant inhabiting temporary puddles in the Fontainebleau forest (Paris region, France) and exhibiting metapopulation dynamics. METHODS: The genetic diversity and phenotypic plasticity of quantitative traits (morphological and fitness components) were measured in five populations, using a combination of field measurements, common garden experiments and genotyping at microsatellite loci. KEY RESULTS: It is shown that populations exhibit almost undetectable genetic diversity at molecular markers, and that the variation in quantitative traits observed among populations is due to a high level of phenotypic plasticity. Despite the lack of genetic diversity, the natural population of R. nodiflorus exhibits large population sizes and does not appear threatened by extinction; this may be attributable to large phenotypic plasticity, enabling the production of numerous seeds under a wide range of environmental conditions. CONCLUSIONS: Efficient conservation of the populations can only be based on habitat management, to favour the maintenance of microenvironmental variation and the resulting strong phenotypic plasticity. In contrast, classical actions aiming to improve genetic diversity are useless in the present case.     

Population structure of an endangered species living in contrasted habitats: Parnassia palustris (Saxifragaceae)

In endangered species, it is critical to analyse the level at which populations interact (i.e. dispersal) as well as the levels of inbreeding and local adaptation to set up conservation policies. These parameters were investigated in the endangered species Parnassia palustris living in contrasted habitats. We analysed population structure in 14 populations of northern France for isozymes, cpDNA markers and phenotypic traits related to fitness. Within population genetic diversity and inbreeding coefficients were not correlated to population size. Populations seem not to have undergone severe recent bottleneck. Conversely to pollen migration, seed migration seems limited at a regional scale, which could prevent colonization of new sites even if suitable habitats appear. Finally, the habitat type affects neither within-population genetic diversity nor genetic and phenotypic differentiation among populations. Thus, even if unnoticed local adaptation to habitats exists, it does not influence gene flow between populations.     

Apportionment of global human genetic diversity based on craniometrics and skin color

A number of analyses of classical genetic markers and DNA polymorphisms have shown that the majority of human genetic diversity exists within local populations (approximately 85%), with much less among local populations (approximately 5%) or between major geographic regions or "races" (approximately 10%). Previous analysis of craniometric variation (Relethford [1994] Am J Phys Anthropol 95:53-62) found that between 11-14% of global diversity exists among geographic regions, with the remaining diversity existing within regions. The methods used in this earlier paper are extended to a hierarchical partitioning of genetic diversity in quantitative traits, allowing for assessment of diversity among regions, among local populations within regions, and within local populations. These methods are applied to global data on craniometric variation (57 traits) and skin color. Multivariate analysis of craniometric variation shows results similar to those obtained from genetic markers and DNA polymorphisms: roughly 13% of the total diversity is among regions, 6% among local populations within regions, and 81% within local populations. This distribution is concordant with neutral genetic markers. Skin color shows the opposite pattern, with 88% of total variation among regions, 3% among local populations within regions, and 9% within local populations, a pattern shaped by natural selection. The apportionment of genetic diversity in skin color is atypical, and cannot be used for purposes of classification. If racial groups are based on skin color, it appears unlikely that other genetic and quantitative traits will show the same patterns of variation.     

Development of robust genomic simple sequence repeat markers for estimation of genetic diversity within and among bulb onion (Allium cepa L.) populations

Improved understanding of genetic diversity in onion and shallot (Allium cepa L.) is required to inform breeding and genetic resource conservation, and to enable development of association genetics and seed quality assurance methods. To develop quantitative estimates of diversity we estimated within- and among-population heterozygosity in a set of onion populations using genomic simple sequence repeat (SSR) markers developed by genomic skim sequencing. Primer sets (166) designed to flank SSR motifs identified were evaluated in a diverse set of lines, with 80 (48?%) being polymorphic. The 20 most robust single copy markers were scored in 12 individuals from 24 populations representing short-day to long-day adapted material from diverse environments. The average genetic diversity estimate (H _e) per population was 0.3 (SD 0.08) and the average per marker was 0.49 (SD 0.2). The onion populations assessed in this survey were distinct with moderate to large population differentiation but also had high within-population variation (F _st?=?0.26). There was evidence of inbreeding (F _is?=?0.22) with observed heterozygosities lower than the expected. This marker resource will be applicable for DNA fingerprinting, measuring levels of inbreeding in breeding lines, assessing population structure for association mapping and expanding linkage maps that are principally based on expressed sequence tag-based markers. A Galaxy workflow was developed to facilitate bulk SSR marker design from next-generation sequence data. This study provides one of the first quantitative views of population genetic variation in onion and a practical toolset for further genetics.     

Strong spatial genetic structure in peripheral but not core populations of Sitka spruce [Picea sitchensis (Bong.) Carr.

We examined spatial genetic structure within eight populations of Sitka spruce classified as core or peripheral based on ecological niche, and continuous or disjunct based on species distribution. In each population, 200 trees were spatially mapped and genotyped for eight cDNA-based sequence tagged site (STS) codominant markers. Spatial autocorrelation was assessed by estimating p(ij), the average co-ancestry coefficient, between individuals within distance intervals. The distribution of alleles and genotypes within core populations was almost random, with nonsignificant co-ancestry values among trees as close as 50 m in core populations. In contrast, the distribution of alleles and genotypes within peripheral populations revealed an aggregation of similar multilocus genotypes, with co-ancestry values greater than 0.20 among trees up to 50 m apart and significant, positive values between trees up to 500 m. The relatively high density of reproductive adults in core populations may lead to highly overlapping seed shadows that limit development of spatial genetic structure. However, in peripheral populations with a lower density of adults, the distribution of alleles and genotypes was highly structured, likely due to offspring establishment near maternal trees and subsequent biparental inbreeding, as well as more recent population establishment at the leading edge of post-Pleistocene range expansion. Conserving genetic diversity in peripheral populations may require larger reserves for in situ conservation than required in core populations. These data on spatial genetic structure can be used to provide guidance for sampling strategies for both ex situ conservation and research collections.     

Isozyme studies in provenance research of forest trees

Summary The nature and origin of the isozymes and the techniques for their detection in forest trees are briefly reviewed. The theories used to interpret the isozyme variation are summarized. Recent isozyme variation studies in provenance research are discussed in relation to known variation pattern detected by classical nursery and field tests. The populations of a tree species can be sometimes, but not always, distinguished by their isozyme patterns. For a number of species, relationships between environment of origin of the provenances and some isozyme frequencies have been statistically established. In one case (Picea sitchensis) where direct comparison between the variation pattern detected by isozymes and the genetic variation of silviculturally important traits was possible, no meaningful relationships between both patterns could be detected. Nei's genetic distances and indices of gene diversity do not appear to be useful in provenance research. The concept of genetic distance based on gene frequencies is probably not very useful in provenance research either.     

Genetic structure and demographic history of the endangered tree species Dysoxylum malabaricum (Meliaceae) in Western Ghats,India: implications for conservation in a biodiversity hotspot

The impact of fragmentation by human activities on genetic diversity of forest trees is an important concern in forest conservation, especially in tropical forests. Dysoxylum malabaricum (white cedar) is an economically important tree species, endemic to the Western Ghats, India, one of the world's eight most important biodiversity hotspots. As D. malabaricum is under pressure of disturbance and fragmentation together with overharvesting, conservation efforts are required in this species. In this study, range‐wide genetic structure of twelve D. malabaricum populations was evaluated to assess the impact of human activities on genetic diversity and infer the species’ evolutionary history, using both nuclear and chloroplast (cp) DNA simple sequence repeats (SSR). As genetic diversity and population structure did not differ among seedling, juvenile and adult age classes, reproductive success among the old‐growth trees and long distance seed dispersal by hornbills were suggested to contribute to maintain genetic diversity. The fixation index (F_IS) was significantly correlated with latitude, with a higher level of inbreeding in the northern populations, possibly reflecting a more severe ecosystem disturbance in those populations. Both nuclear and cpSSRs revealed northern and southern genetic groups with some discordance of their distributions; however, they did not correlate with any of the two geographic gaps known as genetic barriers to animals. Approximate Bayesian computation‐based inference from nuclear SSRs suggested that population divergence occurred before the last glacial maximum. Finally we discussed the implications of these results, in particular the presence of a clear pattern of historical genetic subdivision, on conservation policies.     

岛屿植物舟山新木姜子居群遗传多样性的RAPD分析

基于随机扩增多态 DNA(RAPD)方法分析了舟山群岛濒危植物舟山新木姜子 (N eolitsea sericea) 6个居群的遗传多样性及分化程度。 10条随机引物扩增出 84个可分析位点 ,多态位点百分比 (PPL)为 3 8.10 %。经 POPOGENE分析发现 ,舟山新木姜子居群平均水平的多态位点百分比 (PPL )为 2 3 .18% ,Nei' s基因多样度 (HE)为 0 .0 793 ,Shannon信息指数 (H )为 0 .12 0 1,与其它岛屿植物比较具有中等偏低水平的遗传多样性 ;岛屿各居群间遗传分化程度较高 (Gst=0 .3 646) ;地理距离与遗传距离之间具有显著相关性 (r=0 .7697,P=96.62 % ) ,岛屿隔离效应是导致居群间遗传分化的重要因素。结合居群遗传多样性及UPGMA聚类分析 ,推测普陀山岛舟山新木姜子部分个体可能为大猫岛迁入的后裔 ,而朱家尖岛舟山新木姜子则由人为移植自普陀山岛。基于舟山新木姜子的物种保护及资源利用 ,建议加强现有自然居群的就地保护 ,促进居群自然更新 ;建立种质资源库 ,收集不同岛屿的种源进行混合繁殖 ,促进基因交流 ;选育优良品系用于海岛植被恢复及园林观赏     

Mammalian herbivores reveal marked genetic divergence among populations of an endangered plant species

Quantitative genetically based traits in dominant and keystone tree species can have extended effects on other biota and also on ecosystem processes. This has direct implications for managed plant systems, where choice of genetic stock in conservation or commercial plantings will affect the ecological and evolutionary trajectory of the associated biotic communities. Hence an understanding of genetic variation in quantitative traits, especially those that relate directly to fitness, should be incorporated into the management of species. In plants, quantitative traits such as foliar defences that mediate the complexity of biotic interactions (e.g. herbivory), may be key fitness traits to consider in the management of gene pools of species that are of high conservation value. In this paper we examine the interactions of an endangered eucalypt species, Eucalyptus morrisbyi and a marsupial herbivore, the common brushtail possum Trichosurus vulpecula. We investigate the genetic variability of resistance of plants sourced from two populations and genetic variability in foliage defences as key quantitative traits that may be essential for survival of this eucalypt species. Trichosurus vulpecula detect clear genetic divergence in the two E. morrisbyi populations as evidenced by their browsing preferences in the field. In addition, trees from the more susceptible population (Calverts Hill) suffered fitness consequences with lower flowering than trees from the more resistant population (Risdon Hills). Field feeding preferences were confirmed in captive feeding trials arguing differences were due to foliar attributes consistent with the genetic‐based differences observed in key chemical and physical foliage traits. Biotic interactions such as herbivory may affect populations of rare plant species. Results of this study highlight the need to understand the degree of genetic differentiation of resistance to herbivores and in the quantitative traits mediating these interactions in species of high conservation value, as these traits affect the adaptive potential of populations.     

Estimating inbreeding depression in natural plant populations using quantitative and molecular data

Inbreeding and inbreeding depression are important issues in the biology and conservation of natural plant and animal populations, primarily when subpopulation sizes are reduced due to habitat fragmentation. In this study, we propose a method for estimating inbreeding depression in progenies of natural plant populations, combining the estimation of the fixation index by codominant markers with the experimental evaluation of quantitative traits. Our technique estimates apparent inbreeding depression in structured natural populations using the linear regression of phenotypic means on the inbreeding coefficients estimated with codominant markers. This method was applied to data from 112 maternal progenies of 10 natural subpopulations of Eugenia dysenterica DC, a fruiting tree species from the Brazilian savanna (Cerrado). The results show that the proposed method was efficient at detecting the presence of inbreeding depression for seedling emergence and initial growth traits in the species. This corroborates the importance of maintaining high levels of heterozygosity for in situ conservation or genetic restoration of natural populations.