Population genetics and breeding system of Tupistra pingbianensis (Liliaceae), a naturally rare plant endemic to SW China

The levels and partitioning of genetic diversity and inbreeding depression were investigated in Tupistra pingbianensis, a narrow endemic of South-east Yunnan, China, characterized by a naturally fragmented distribution due to extreme specialization on a rare habitat type. Here genetic diversity and patterns of genetic variation within and among 11 populations were analyzed using AFLP markers with 97 individuals across its whole geographical range. High levels of genetic variation were revealed both at the species level (P₉₉ = 96.012%; H_t = 0.302) and at the population level (P₉₉ = 51.41%; H_s = 0.224). Strong genetic differentiation among populations was also detected (F_ST = 0.2961; ⍬^嘦= 0.281), which corresponded to results reported for typical animal-pollinated, mixed selfing and outcrossing plant species. This result was consistent with mating patterns detected by our pollination experiments. The indirect estimate of gene flow based on ⍬^嘦 was low (N_m = 0.64). Special habitat and its life history traits may play an important role in shaping the genetic diversity and the genetic structure of this species. A pollination experiment also fail to detect significant inbreeding depression upon F₁ fruit set, seed weight and germinate rate fitness-traits. Since naturally rare species T. pingbianensis is not seriously genetically impoverished and likely to have adapted to tolerating a high level of inbreeding early in its history, we propose this species need only periodic monitoring to ensure their continued persistence but not intervention to remain viable.     

Population genetics and breeding system of Tupistra pingbianensis (Liliaceae),a naturally rare plant endemic to SW China

The levels and partitioning of genetic diversity and inbreeding depression were investigated in Tupistra pingbianensis, a narrow endemic of southeast Yunnan, China, characterized by a naturally fragmented distribution due to extreme specialization on a rare habitat type. Here genetic diversity and patterns of genetic variation within and among 11 populations were analyzed using amplified fragment length polymorphism markers with 97 individuals across its whole geographical range. High levels of genetic variation were revealed both at the species level (P_(99)=96.012%; H_t=0.302) and at the population level (P_(99)=51.41%; H_8= 0.224). Strong genetic differentiation among populations was also detected (F_(ST)= 0.2961; θ~Ⅱ= 0.281), which corresponded to results reported for typical animal-pollinated, mixed selfing, and outcrossing plant species. This result was consistent with mating patterns detected by our pollination experiments. The indirect estimate of gene flow based on θ~Ⅱwas low (N_m=0.64). Special habitat and its life history traits might play an important role in shaping the genetic diversity and the genetic structure of this species. A pollination experiment also failed to detect significant inbreeding depression upon F_1 fruit set, seed weight, and germinate rate fitness-traits. As a naturally rare species, T. pingbianensis is not seriously genetically impoverished and likely to have adapted to tolerating a high level of inbreeding early in its history, we propose this species need only periodic monitoring to ensure their continued persistence, but not intervention to remain viable.     

Genetic diversity in Isoetes yunguiensis, a rare and endangered endemic fern in China

Isoetes yunguiensis is an endangered and endemic fern in China.Field survey indicated that only one population and no more than 50 individuals occur in the wild.The genetic variation of 46 individuals from the population remaining at Pingha (Guizhou Province,China)was assessed by Random Amplified Polymorphic DNA (RAPD)fingerprinting.Twelve primers were screened from sixty ten-bp arbitrary primers,and a total of 95 DNA fragments were scored.Of these,62.1%were polymorphic loci,which indicated that high level genetic variation existed in the natural population.The accumulation of genetic variation in the history of the taxon and the apparent minimal reduction effect on genetic diversity following destruction of habitat might be responsible for the high level genetic diversity presently remaining in the I.yunguiensis population.However,with the continuing decrease of population size,the genetic diversity will gradually be lost.We suggest that the materials from the extant population should be used for re-establishment of the populations.     

Genetic diversity and population structure of the serpentine endemic Calystegia collina (Convolvulaceae) in northern California

We used enzyme electrophoresis to evaluate genetic diversity in 32 populations of Calystegia collina, a clonal plant species endemic to serpentine outcrops in northern California (USA). Of 34 loci examined 56% were polymorphic, but on average only 17% were polymorphic within local populations. Neither the total number of alleles nor the number of multilocus genotypes differed significantly between populations in small vs. large serpentine outcrops. Genetic and geographic distances between populations were positively correlated, but this relationship was not significantly affected by the isolation of serpentine outcrops. Populations were highly differentiated (F(st) = 0.417) and little genetic variation was explained by geographic region or serpentine outcrop.Observed heterozygosity within populations almost always exceeded Hardy-Weinberg expectations. In many populations, all 30 sample ramets were uniformly heterozygous at one or more loci yet were genetically variable at other loci. These results imply that many C. collina populations originate from one or a few genetic founders, with little recruitment from seeds. Genetic variation within uniformly heterozygous populations must be the product of multiple, closely related founders or somatic mutations within the population. We conclude that vegetative reproduction, perhaps coupled with somatic mutation, helps maintain genetic diversity in these isolated but long-lived populations.     

Genetic diversity in Isoetes yunguiensis, a rare and endangered endemic fern in China

Isoetes yunguiensis is an endangered and endemic fern in China. Field survey indicated that only one population and no more than 50 individuals occur in the wild. The genetic variation of 46 individuals from the population remaining at Pingba (Guizhou Province, China) was assessed by Random Amplified Polymorphic DNA (RAPD) finger-printing. Twelve primers were screened from sixty ten-bp arbitrary primers, and a total of 95 DNA fragments were scored. Of these, 62.1% were polymorphic loci, which indicated that high level genetic variation existed in the natural population. The accumulation of genetic variation in the history of the taxon and the apparent minimal reduction effect on genetic diversity following destruction of habitat might be responsible for the high level genetic diversity presently remaining in the I. yunguiensis population. However, with the continuing decrease of population size, the genetic diversity will gradually be lost. We suggest that the materials from the extant population should be used for re-establishment of the populations. Translated from Journal of Wuhan University (Natural Sciences Edition), 2005, 51(6): 767–770 [译自: 武汉大学学报 (理学版)]     

Fine-scale spatial genetic structure and gene flow in a small, fragmented population of Sinojackia rehderiana (Styracaceae), an endangered tree species endemic to China

Populations of Sinojackia rehderiana are highly threatened and have small and scattered distribution due to habitat fragmentation and human activities. Understanding changes in genetic diversity, the fine-scale spatial genetic structure (SGS) at different life stages and gene flow of S. rehderiana is critical for developing successful conservation strategies for fragmented populations of this endangered species. In this study, 208 adults, 114 juveniles and 136 seedlings in a 50 × 100-m transect within an old-growth forest were mapped and genotyped using eight microsatellite makers to investigate the genetic diversity and SGS of this species. No significant differences in genetic diversity among different life-history stages were found. However, a significant heterozygote deficiency in adults and seedlings may result from substantial biparental inbreeding. Significant fine-scale spatial structure was found in different life-history stages within 19 m, suggesting that seed dispersal mainly occurred near a mother tree. Both historical and contemporary estimates of gene flow (13.06 and 16.77 m) indicated short-distance gene dispersal in isolated populations of S. rehderiana. The consistent spatial structure revealed in different life stages is most likely the result of limited gene flow. Our results have important implications for conservation of extant populations of S. rehderiana. Measures for promoting pollen flow should be taken for in situ conservation. The presence of a SGS in fragmented populations implies that seeds for ex situ conservation should be collected from trees at least 19-m apart to reduce genetic similarity between neighbouring individuals.     

Intersimple sequence repeat (ISSR) variation in Lactoris fernandeziana (Lactoridaceae), a rare endemic of the Juan Fernández Archipelago, Chile

Sixteen populations and 89 individuals of Lactoris fernandeziana were examined for variation in intersimple sequence repeat (ISSR) banding patterns. The species is a rare endemic of Masatierra Island in the Juan Fernández Archipelago, and is the only member of the endemic family Lactoridaceae. Five populations showed a single genotype whereas the other 11 populations had from two to 16 multilocus genotypes. Over 73% of the ISSR diversity occurred across populations, with only about 27% within populations. Diversity among populations results from the presence of different subsets of loci within each population rather than unique loci within populations; only two populations displayed novel loci, with one and three in each. Levels of differentiation at ISSR loci among populations are not correlated with geographic distance on Masatierra; rather, the pattern of variation is mosaic. The presence of differentiated local populations is concordant with the geitonogamous breeding system of the species and suggests low levels of long distance pollen or seed dispersal. The mosaic pattern of ISSR variation on Masatierra may result, in part, from drift and inbreeding in small populations following fragmentation of a once more continuous distribution of Lactoris with the formation of canyons by erosion. Also, the generation of new ISSR loci by mutation could occur with rare, sporadic gene flow among populations accounting for the mosaic pattern of variation and the paucity of unique alleles within populations. The ISSR results for Lactoris suggest that studies of morphological, ecological and physiological features may elucidate differentiation among populations of L. fernandeziana . Field studies have demonstrated that plants occur both in the dense forest understory and in the full sunlight in forest openings.     

Genetic structure and AFLP variation of remnant populations in the rare plant Pedicularis palustris (Scrophulariaceae) and its relation to population size and reproductive components

We investigated plant reproduction in relation to genetic structure, population size, and habitat quality in 13 populations of the rare biennial plant Pedicularis palustris with 3-28500 flowering individuals. We used AFLP (amplified fragment length polymorphism) profiles to analyze genetic similarities among 129 individuals (3-15 per population). In a cluster analysis of genetic similarities most individuals (67%) were arranged in population-specific clusters. Analysis of molecular variance indicated significant genetic differentiation among populations and among and within subpopulations (P < 0.001). Gene flow (N(e) m) was low (0.298). On average, plants produced 55 capsules, 17 seeds per fruit, and 42 seedlings in the following growing season. The number of seeds per capsule was independent of population size and of genetic variability. In contrast, the number of capsules per plant (P < 0.05) and the number of seedlings per plant (P < 0.05) were positively correlated with population size. The relation between population size and the number of seeds per plant was not significant (P = 0.075). The number of capsules and of seeds and seedlings per plant (P < 0.01) were positively correlated with genetic variability. Genetic variability was independent of actual population size, suggesting that historical population processes have to be taken into account, too. Stepwise multiple regressions revealed additional significant relationships of habitat parameters (soil pH, C:N ratio), vegetation composition, and standing crop on reproductive components. We conclude that populations of P. palustris are genetically isolated and that reproductive success most likely is influenced by population size, genetic variability, and habitat quality. Management strategies such as moderate grazing, mowing, and artificial gene flow should endeavor to increase population size as well as genetic variation.     

鹅掌楸贵州烂木山居群的微卫星遗传多样性及空间遗传结构

濒危植物鹅掌楸(Liriodendron chinense)目前仅零散分布于我国亚热带及越南北部地区, 残存居群生境片断化较为严重。研究濒危植物片断化居群的遗传多样性及小尺度空间遗传结构(spatial genetic structure)有助于了解物种的生态进化过程以及制定相关的保育策略。本研究采用13对微卫星引物, 对鹅掌楸的1个片断化居群进行了遗传多样性及空间遗传结构的研究, 旨在揭示生境片断化条件下鹅掌楸的遗传多样性及基因流状况。研究结果表明: 鹅掌楸烂木山居群内不同生境斑块及不同年龄阶段植株的遗传多样性水平差异不显著(P>0.05), 居群内存在寨内和山林2个遗传分化明显的亚居群。烂木山居群个体在200 m以内呈现显著的空间遗传结构, 而2个亚居群内的个体仅在20 m的距离范围内存在微弱或不显著的空间遗传结构。鹅掌楸的空间遗传结构强度较低(Sp = 0.0090), 且寨内亚居群的空间遗传结构强度(Sp = 0.0067)要高于山林亚居群(Sp = 0.0053)。鹅掌楸以异交为主, 种子较轻且具翅, 借助风力传播, 在一定程度上降低了空间遗传结构的强度。此外, 居群内个体密度及生境特征也对鹅掌楸的空间遗传结构产生了一定影响。该居群出现显著的杂合子缺失, 近交系数(F_IS)为0.099 (P < 0.01), 表明生境片断化的遗传效应正逐渐显现。因此, 对鹅掌楸的就地保护应注意维护与强化生境的连续性, 促进基因交流。迁地保护时, 取样距离应不小于20 m, 以涵盖足够多的遗传变异。     

Genetic diversity and hierarchical population structure of a rare autotetraploid plant,Aster kantoensis (Asteraceae)

We sampled 17 populations of a rare autotetraploid Aster kantoensis (Asteraceae) from three river systems located in central Japan, and studied them for allelic variation at 22 enzyme loci. There was no significant correlation between the actual population size and three genetic diversity parameters, suggesting that the effective population size was very small even for the large populations, i.e., even large populations may still have a high probability of being of recent origin and remain influenced by the founder effect. Compared to other autotetraploid species, the total genetic variation of A. kantoensis is small. The number of alleles and gene diversity of a population were not significantly different among the river systems, although the percentage of polymorphic loci was different. Genetic differentiation among river systems was larger than between populations within the river systems, thereby indicating that gene flow between river systems is small, especially between the Kinu River system and Tama or Sagami River systems.     

Genetic impoverishment and cross-incompatibility in remnant genotypes of Ziziphus celata (Rhamnaceae), a rare shrub endemic to the Lake Wales Ridge, Florida

The loss of genetic diversity in fragmented populations ofself-incompatible plant species may result in sexual reproductive failure andlocal extinctions. Florida ziziphus (Ziziphus celata) is aself-incompatible clonal shrub known only from five genetically depauperatepopulations on the Lake Wales Ridge, Florida, USA. Recovery of this speciesrequires identification of cross-compatible genotypes that can be used to createviable (i.e., sexually reproducing) populations. To further development of arecovery program for this highly imperiled species, we investigated its geneticstructure and sexual reproductive viability. We used random amplifiedpolymorphic DNAs (RAPDs) to investigate genetic variability within remnantpopulations and we conducted experimental compatibility trials to determine thecross-compatibility of remnant genotypes. One hundred and ninety-nine uniquestem samples collected from one ex situ and fivein situ populations were assayed for the presence orabsence of a band for 32 RAPD markers. Based on unweighted pair-group meancluster analysis (UPGMA), only 11 multi-locus genotypes (MLGs) were identified.Eight of these MLGs correspond to MLGs identified in an earlier allozyme study.In addition, we identified three new RAPD-based MLGs. Three of the five naturalpopulations consisted of only one MLG, while the largest and most geneticallydiverse population comprised only four MLGs. Coefficients of similarity rangedfrom 96.6% for the most closely related MLGs to 20.7% for the most distantlyrelated. The compatibility trials demonstrated that most MLGs arecross-incompatible. With 69% of all possible one-way crosses tested (38/55), wehave identified only eight compatible crosses via germination trials. Based onthe results of the compatibility trials, we assigned MLGs toself-incompatibility (SI) mating types. On present evidence, the currentbreeding population of Florida ziziphus may comprise as few as two SI matingtypes. These SI mating types will be used to guide future breeding efforts andan experimental introduction.     

Understanding the genetic effects of recent habitat fragmentation in the context of evolutionary history: phylogeography and landscape genetics of a southern California endemic Jerusalem cricket (Orthoptera: Stenopelmatidae: Stenopelmatus)

Habitat loss and fragmentation due to urbanization are the most pervasive threats to biodiversity in southern California. Loss of habitat and fragmentation can lower migration rates and genetic connectivity among remaining populations of native species, reducing genetic variability and increasing extinction risk. However, it may be difficult to separate the effects of recent anthropogenic fragmentation from the genetic signature of prehistoric fragmentation due to previous natural geological and climatic changes. To address these challenges, we examined the phylogenetic and population genetic structure of a flightless insect endemic to cismontane southern California, Stenopelmatus'mahogani' (Orthoptera: Stenopelmatidae). Analyses of mitochondrial DNA sequence data suggest that diversification across southern California began during the Pleistocene, with most haplotypes currently restricted to a single population. Patterns of genetic divergence correlate with contemporary urbanization, even after correcting for (geographical information system) GIS-based reconstructions of fragmentation during the Pleistocene. Theoretical simulations confirm that contemporary patterns of genetic structure could be produced by recent urban fragmentation using biologically reasonable assumptions about model parameters. Diversity within populations was positively correlated with current fragment size, but not prehistoric fragment size, suggesting that the effects of increased drift following anthropogenic fragmentation are already being seen. Loss of genetic connectivity and diversity can hinder a population's ability to adapt to ecological perturbations commonly associated with urbanization, such as habitat degradation, climatic changes and introduced species. Consequently, our results underscore the importance of preserving and restoring landscape connectivity for long-term persistence of low vagility native species.     

Genetic diversity, population structure, and conservation of Sophora moorcroftiana (Fabaceae), a shrub endemic to the Tibetan Plateau

Sophora moorcroftiana is a perennial leguminous low shrub endemic to the middle reaches of Yarlung Zangbo River in Tibet. It is an important species to fix sand dunes and to avoid the formation of shifting sands; therefore, its progressive over-exploitation may enhance land desertification. The levels and distribution of genetic variability of this species were evaluated from 10 natural populations at 24 loci encoding 13 enzymes, using allozyme analysis by starch gel electrophoresis. Data obtained revealed moderate levels of genetic variation within populations (Pp=27.5%, Ap=1.5, Hep=0.122) and a considerable divergence among populations (FST=0.199). Significant positive correlations (r2=0.49, p<0.05; r2=0.46, p<0.05) were found between elevation and both mean number of alleles per locus (A) and gene diversity (He) in the studied populations of S. moorcroftiana. Lower genetic diversity in lower elevation populations might be due to the negative effects of human pressures and habitat fragmentation, to adaptation to high altitudes as a consequence of a peripatric speciation process, or to directional gene flow along the river basin from the source populations located in the west at higher altitudes. The evaluation of the degree of threat has led to the inclusion of this species in the category of EN ("endangered"), and conservation strategies for this endemic species are discussed on the basis of these findings.     

Utilizing a multifaceted approach to assess the current distribution and conservation status of an uncommon species: the golden mouse (Ochrotomys nuttalli) in Florida

Aim

Our goal was to assess the conservation status of the understudied and naturally uncommon habitat specialist, the golden mouse (Ochrotomys nuttalli), at the edge of its range where its historically fragmented habitat has been subjected to severe loss.

Location

Peninsular Florida, north of approximately 27° latitude, USA.

Methods

We used data gathered from museum collections, regional biologists, geographic information systems (GIS) layers, field surveys and DNA sequencing to determine the habitats that best explain the distribution of the species, examine changes in the geographic extent of both the species and its habitats, and compare genetic differentiation between populations occupying disjunct regions. The results from these multiple analyses were combined to assess the conservation status of the species.

Results

Golden mouse occurrence records align well with the distribution of hardwood habitats in Florida. These habitats occur naturally as ‘islands’, but have become increasingly fragmented by anthropogenic land use. Despite habitat loss, the location of the southern range periphery has remained relatively unchanged in location over the past century. Genetic analysis reveals a history of limited dispersal of females among habitat ‘islands’ that likely predates anthropogenic landscape fragmentation. This pattern suggests that isolated populations that are extirpated will have little to no chance of successful recolonization.

Main conclusions

The combined results from multiple analyses produced a more complete picture of the threats faced by this previously data‐deficient species than any single analysis would have. Although the species' southern range limit cannot be shown to have retracted in the face of human expansion, habitat fragmentation clearly has put the species at increased risk. Conservation and management of hardwood habitats are critical to the persistence of the golden mouse at the edge of its range.     

RAPD variation in relation to population size and plant fitness in the rare Gentianella germanica (Gentianaceae)

We investigated the distribution of genetic variation and the relationship between population size and genetic variation in the rare plant Gentianella germanica using RAPD (random amplified polymorphic DNA) profiles. Plants for the analysis were grown from seeds sampled from 72 parent plants in 11 G. germanica populations of different size (40-5000 fruiting individuals). In large populations, seeds were sampled from parents in two spatially distinct subpopulations comparable in area to the total area covered by small populations. Analysis of molecular variance revealed significant genetic variation among populations (P <0.001), while genetic variation among subpopulations was marginally significant (P <0.06). Average molecular variance within subpopulations in large populations did not differ significantly from whole-population values. There was a positive correlation between genetic variation and population size (P <0.01). Genetic variation was also positively correlated with the number of seeds per plant in the field (P <0.02) and the number of flowers per planted seed in a common garden experiment (P <0.051). We conclude that gene flow among natural populations is very limited and that reduced plant fitness in small populations of G. germanica most likely has genetic causes. Management should aim to increase the size of small populations to minimize further loss of genetic variation. Because a large proportion of genetic variation is among populations, even small populations are worth preserving.     

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.     

Phenology,pollination, and breeding system of the threatened tree Caesalpinia echinata Lam. (Fabaceae), and a review of studies on the reproductive biology in the genus

Caesalpinia echinata (brazilwood) is a threatened tree endemic to the Brazilian Atlantic forest with a global importance as it is worldwide used to manufacture the most desirable bows for violins, violas, and cellos. Although there is an international initiative for its conservation, there is a lack of studies on the reproductive biology of the species, as for the genus, a fundamental understanding for the conservation of any species. In this paper, we investigated the phenology, pollination, and breeding system of C. echinata, presenting some conservation guidelines for the species, together with a review of studies on the reproductive biology in Caesalpinia s.l. The genus has a complex taxonomic history with recent attempts to reassign its limits. Pending a final resolution of the placing of some species (including C. echinata) our revision covers the traditional Caesalpinia s.l. and includes annotations indicating to which segregate genus each species is now considered to belong, as well as the appropriate combination where this has already been published. Field work was undertaken from October/2004 to December/2006 at the Tapacurá Ecological Station, northeastern Brazil. Flowering occurred mainly in the dry season and the peak of seed dispersal was at the beginning of the wet season. Anthesis is diurnal, lasting 1 day. The flowers are zygomorphic, yellow, sweet-scented, and the effective pollinators were mainly medium-sized to large bees of the genera Centris and Xylocopa, together with the introduced Apis mellifera. Nectar volume and sugar concentration averaged 2.9±1.0 μL and 29.5±9.4%, respectively. The ovary has 1–4 ovules (2.35±0.58) and the pollen/ovule ratio was 5631.2; pollen viability was high (95.9±4.8%). Natural fruit set was low (9.2%) with mature fruits averaging 1.7±0.9 seeds. Results of the controlled pollinations and analysis of pollen tube growth revealed C. echinata presents late-acting self-incompatibility. The pollination biology and breeding system of C. echinata are discussed, together with available data on its genetics and physiology, in terms of best conservation practice for this endangered species. Data on the reproductive biology of the genus are scarce, revealing the predominance of bee pollination and SI system, with the occurrence of late-acting self-incompatibility mechanisms in some species.     

Genetic Diversity of a Tropical Tree Species,Shorea leprosula Miq. (Dipterocarpaceae), in Malaysia: Implications for Conservation of Genetic Resources and Tree Improvement1

Genetic diversity and population genetic structure of Shorea leprosula was investigated using seven natural populations distributed throughout Peninsular Malaysia and one natural population from Borneo. The mean population and species level genetic diversity were exceptionally high (H_e= 0.369 ± 0.025 and 0.406 ± 0.070, respectively). Heterozygosity varied among populations, ranging from 0.326 to 0.400, with the highest values found in the populations from central Peninsular Malaysia. Correlations among ecological factors (longitude, latitude, and annual rainfall) were not significant (P > 0.05), indicating that these ecological variables were not responsible for the observed genetic differences among populations. The Bangi adult population exhibited a higher level of observed heterozygosity but lower fixation indices in comparison to its seedling population. All other seedling populations also showed positive fixation indices (f), indicating a general excess of homozygotes. This also may suggest selection against homozygotes between the seedling and adult stages. A low level of population differentiation was detected (G_ST= 0.117 with the Lambir population and GST= 0.085 without the Lambir population). Furthermore, gene flow (N_m) between populations was not significantly correlated with geographical distances for the populations within Peninsular Malaysia. Cluster analysis also did not reflect geographical proximity and gave little insight into the genetic relatedness of the populations. This may indicate that the populations sampled are part of a continuous population with fragmentation having occurred in the recent past.     

Unmanaged sexual reproduction and the dynamics of genetic diversity of a vegetatively propagated crop plant, cassava (Manihot esculenta Crantz), in a traditional farming system

Occurrence of intervarietal or interspecific natural crosses has been reported for many crop plants in traditional farming systems, underlining the potential importance of this source of genetic exchange for the dynamics of genetic diversity of crop plants. In this study, we use microsatellite loci to investigate the role of volunteer seedlings (plants originating from unmanaged sexual reproduction) in the dynamics of genetic diversity of cassava (Manihot esculenta Crantz), a vegetatively propagated crop, in a traditional farming system in Guyana. A previous field study showed that farmers incorporate such plants into the germplasm for vegetative propagation, and that many of them are likely to be assigned by farmers to recognized varieties. Under strict vegetative propagation clonality of varieties is expected. The high proportion of polyclonal varieties observed suggests that incorporation of seedlings into the germplasm for propagation is a frequent event. The molecular variability assessed with microsatellite markers shows that there is high differentiation among heterozygous varieties, whereas populations of seedlings do not depart from the proportions expected under Hardy-Weinberg assumptions. Assignment of seedlings to a recognized variety on the basis of morphological similarity greatly increases genetic diversity within the variety. We argue that recombination and gene flow play a major role in the dynamics of genetic diversity of cassava in traditional farming systems. Documenting unmanaged sexual reproduction and its genetic consequences is a prerequisite for defining strategies of in situ conservation of crop plant genetic resources.     

Clonal and spatial genetic structure in natural populations of Luohanguo (Siraitia grosvenorii), an economic species endemic to South China,as revealed by RAPD markers

Clonal growth is generally expected to have significant effects on the spatial genetic structure within populations. In this study, random amplified polymorphic DNA (RAPD) markers were used to reveal clonal and spatial genetic structure of four natural populations of Luohanguo (Siraitia grosvenorii), an economic vine species endemic to South China. A total of 351 ramets were assigned to 76 distinct multi-locus genotypes (i.e. genets), with the G/N varying from 0.121 to 0.350. No widespread genet was found across different populations. The clonal diversity (D) and evenness (E) ranged from 0.333 to 0.828 and from 0 to 0.741, respectively. While most genets consisted of fewer than five ramets, we observed some dominant genets that had much more (up to 69) ramets and spread over large areas. Spatial autocorrelation analyses revealed a spatial genetic structure (i.e. significant positive autocorrelation within 20 m and negative autocorrelation beyond 40 m) in one population, but not in other three populations with smaller population size. This study highlights the importance of clonal growth in shaping the spatial genetic structure in Luohanguo, which may have complex effects on the dynamics and evolution of its declining populations.