Policy plans and management measures to restore eelgrass (Zostera marina L.) in the Dutch Wadden Sea

The Dutch Wadden Sea has been changed dramatically over the last centuries by human activities like land reclamation and different forms of fishery. This has, amongst other things, led to changes in the number of biological communities. One of the changes was the near extinction of eelgrass (Zostera marina L.) in the Dutch Wadden Sea. The deterioration of the area led to policy plans in the late 1980s that aimed at restoring the original natural communities of which the eelgrass community was one. This paper presents a restoration strategy which contains a selection procedure for suitable transplantation sites. The selection procedure is based on factors such as sediment composition, exposure time, current velocity and wave action. These were combined in a GIS-based map integrating these factors. One important action in the restoration process is to increase the number of freshwater discharge points to meet the requirements of the brackish water community in general and the growing conditions for eelgrass in particular. Received: 27 October 1999 / Received in revised form: 3 February 2000 / Accepted: 3 February 2000     

Zostera marina population genetics in Barnegat Bay, New Jersey, and implications for grass bed restoration

Within Barnegat Bay, New Jersey, Zostera marina populations have declined by 62% over the last 20 years, and restoration efforts have met with mixed success. We have completed a microsatellite-based genetic investigation of eight populations of Z. marina within Barnegat Bay to determine whether the genetic stock origins of the plants used in management projects may affect restoration success. Additionally, we assessed the genetic diversity of Z. marina in Barnegat Bay to better understand its population structure. Clonal diversity ranged from 0.70 to 0.95 for the populations studied. Individually, Barnegat Bay populations are not genetically diverse, and there is also little divergence among populations. The Atlantic populations had mean Hobs values (0.20–0.34) that were far lower than the Hexp values (0.69–0.83). Also, the F _IS values in all of the eastern populations indicate a surfeit of homozygotes over heterozygotes, suggesting a low degree of outcrossing in the Barnegat Bay populations. Six of the ten populations studied (Ham Island, Manahawkin Bay, Shelter Island, Marsh Elder, Harvey Cedar Sedge, and Long Island) show evidence of historical bottlenecks. Mean estimated F _ST values would suggest that most alleles are undergoing moderate genetic differentiation, with values that range from 0.06 to 0.13. Oyster Creek and Sedge Island demonstrate the largest estimated effective population sizes and may be the most appropriate populations for use in future eelgrass restoration projects.     

Effects of herbivorous birds on intertidal seagrass beds in the northern Wadden Sea

During autumn migration (September to December), brent geese (Branta b. bernicla) and wigeon (Anas penelope) feed on the seagrass Zostera noltii in the nearshore, upper tidal zone leeward of the island of Sylt (eastern North Sea). To graze on leaves and shoots above the sediment and on rhizomes and roots below, these birds reworked the entire upper 1 cm layer of sediment eight times within this 3-month period. In addition, brent geese excavated pits 3–10 cm deep by trampling in order to feed on below-ground phytomass. About 12% of the seagrass beds became pitted to an average depth of 4.5 cm. Using net exclosures, it was estimated that birds removed 34 g dry weight m^–2 of above-ground and 28 g of below-ground phytomass. This corresponds to 45% of the phytomass in September. Of the overall loss of phytomass from September to December, 63% was caused by birds. Roughly half of the leaves fell off anyway until December and the other half were taken by the birds. Below the ground, phytomass remained almost constant where birds were excluded, while with birds phytomass of rhizomes and roots was halved. In spite of this strong effect, in the next vegetation period the blade density was lower at former exclosure sites compared to the ambient seagrass bed. The underlying process seems to be a self-inhibition of dense overwintering seagrass by mud accretion. Assuming our experimental results can be scaled up to the entire seagrass bed, we hypothesize that in the sheltered upper intertidal zone, seasonal erosion caused by herbivorous geese and ducks is necessary for the persistence of Z. noltii. Received: 7 January 1999 / Received in revised form: 23 August 1999 / Accepted: 25 August 1999     

Effects of pesticides on seagrass beds

In the German Wadden Sea there has been a remarkable decline in seagrass beds. It was the aim of this study to test whether herbicide contamination could be a reason for this. Concentrations of triazine herbicides such as atrazine, simazine and terbutylazine as well as phenylurea herbicides were measured in Wadden Sea sediments within or in the neighbourhood of seagrass meadows. Sediments were thus used as a marker for medium-term contamination of the Wadden Sea. The respective concentrations were examined in relation to the density and status of the seagrass meadows. Preliminary results show that there may be a connection between seagrass decline and herbicide contamination in the parts of the Wadden Sea sampled. A comparison with other contamination is also given. Received: 11 January 1999 / Received in revised form: 1 December 1999 / Accepted: 8 December 1999     

Tidal impact on planktonic primary and bacterial production in the German Wadden Sea

 Tidal variation of biological parameters was studied at three anchor stations in selected inlet channels of the northern German Wadden Sea in May and July 1994. Concentrations of bacteria, chlorophyll a and suspended matter as well as primary and bacterial production were assessed over a period of 25 h in the surface and in the bottom water. Diurnal variation in primary production was found both under in situ light conditions and under constant illumination. Tidal turbulence caused the introduction of detritus, bacteria and pigments from the sediment into the water column. The impact of sediment resuspension was most evident in the bottom water, leading to tidally oscillating bacterial production rates which were high during high stream velocity and low during the slack times. Estimations of the areal daily phytoplankton production and corresponding bacterial carbon demands were unbalanced. Primary production accounted for only 25–45% of the total bacterial carbon requirement. This discrepancy is due to the shallow euphotic depth in the Wadden Sea, allowing net primary production only in the upper 2–3 m of the water column, while the relatively high levels of bacterial activity do not show a vertical decline. Assuming that the specific biological activities in the water columns over the tidal flats are similar to those found in the inlet channels, it was found that production processes dominate in shallow areas whereas decomposition processes dominate in the deep channels. Moreover, the predominance of heterotrophic processes in the inlet channels means that additional organic carbon sources must contribute to the heterotrophic metabolism in the deep parts of the Wadden Sea, and that the horizontal flux of material is important in this turbid mesotidal ecosystem. Received: 18 April 1998 / Accepted: 12 November 1998     

Distribution patterns of bacterioplankton and chlorophyll-a in the German Wadden Sea

 In a first synoptic evaluation, the temporal and spatial distribution of bacterioplankton and chlorophyll-a were determined in the German Wadden Sea. Three surveys were undertaken in winter, spring, and summer of 1994 using up to eight ships simultaneously between the river Ems and Sylt island. Despite intensive hydrodynamic mixing of the Wadden Sea water, spatial gradients were obvious. The abundance of bacterioplankton ranged from 0.4 to 26×10⁵ ml^–1 and chlorophyll-a varied between <0.1 and 79 μg l^–1. In winter, relatively homogeneous distribution patterns of both parameters with small gradients were found. Highest chlorophyll-a values connected with a highly patchy structure were observed in spring, while in summer both total chlorophyll-a values and the complexity of the distribution pattern had decreased. In contrast, bacterial numbers increased steadily from January to July with the highest bacterial densities and greatest patchiness observed in summer. Moreover, in some regions of the Wadden Sea, a trophic succession of algae as carbon producers and bacteria as consumers was evident. Correlation analysis verified the relationship between bacteria and chlorophyll a, indicating bottom-up control of bacterial abundance in the northern part of the German Wadden Sea. Since the observed regression slope is remarkably low (0.12–0.46) compared to literature values (0.5–0.8), we suggest that the link between phytoplankton and bacteria found here is a special characteristic of the Wadden Sea as a transition zone between the coastal region and the outer North Sea. Received: 26 May 1998 / Accepted:12 November 1998     

Was total primary production in the western Wadden Sea stimulated by nitrogen loading?

Borum and Sand-Jensen (1996) described empirical relationships between nitrogen (N) loadings from land and total (benthic + pelagic) primary production rates in shallow coastal marine waters. We applied these relationships to N loadings of the western Wadden Sea system, and compared the production estimates with actually observed primary production rates of autotrophic components (phytoplankton, microphytobenthos, macroalgae and seagrasses) for those years for which field data were available. During the 1980s and early 1990s, primary production values appear in good agreement with those derived from the empirical relationships. During the 1960s and early 1970s, however, these relationships substantially overestimated the total primary production in the western Wadden Sea. Based on ambient nutrient concentrations and the Redfield ratio, production in that period was considered not to be limited by N but by phosphorus (P) during most of the time. It is concluded that primary production is not invariably stimulated by N loading from land. If other factors (i.e. additional nutrient sources, N:P ratios, internal nutrient dynamics and co-limiting effects of nutrients and light) are not taken into account, management regulations that are targeted at diminishing the effects of eutrophication hold the risk of seriously under- or overestimating nutrient reductions that are thought necessary to achieve their goals. Received: 30 November 1998 / Received in revised form: 12 July 1999 / Accepted: 15 July 1999     

An analysis of the population genetics of restored Zostera marina plantings in Barnegat Bay, New Jersey

Within Barnegat Bay, New Jersey, eelgrass (Zostera marina) populations have declined by 62 % over the last 20 years. To better understand the consequences of this devastation, we have previously employed microsatellite DNA polymorphisms to analyze the population structure of Z. marina within Barnegat Bay, as well as along the eastern United States seaboard. We have restored populations of Z. marina in Barnegat Bay over the last 10 years to help assess the best planting conditions and ecotypes that might be used in long-term restoration strategies. In this study, we examined the genetic health of the restored populations compared to that of the donor eelgrass populations within the bay. Using microsatellites, we can identify which parental founding ecotypes survived the restoration process over multiple generations. The frequency of observed heterozygotes, although higher than in the natural populations, still indicates reduced levels of diversity and connectivity. The inbreeding frequency is high in the restored populations, but lower than what is seen in the native populations. All restored populations have effective population values >50, suggesting a high probability of survival in the short term.     

大叶藻对氮磷营养盐的吸收动力学特征

营养盐是影响海草生长的关键因子, 目前有关海草不同组织对不同形式氮和磷的吸收特征尚不明确。该研究通过利用海草地上和地下组织分隔培养装置, 设置不同的氨态氮、硝态氮和磷酸盐浓度, 探究了大叶藻(Zostera marina)植株及其地上和地下组织对氮磷营养盐的吸收动力学特征。结果显示: (1)大叶藻对氮磷的吸收符合饱和吸收动力学特征, 吸收速率和水体氮磷浓度可用米式方程描述; (2)大叶藻植株对NH₄⁺-N的最大吸收速率(V_max, 52 μmol·g^-1·h^-1)显著高于其对NO₃^--N的V_max (39 μmol·g^-1·h^-1); (3)大叶藻地上组织和地下组织均可吸收氮磷, 但地上组织对氨态氮、硝态氮、磷酸盐的V_max分别为43.1、30.5和15.6 μmol·g^-1·h^-1, 为地下组织的2.6、1.2和6倍。结果表明, 大叶藻对氨态氮的吸收能力高于硝态氮, 且对氮磷的吸收可能主要依赖地上组织(叶片)。结果为查明大叶藻对氮磷的吸收利用机制及评估大叶藻的海洋生态效应提供了理论依据。     

An Effective Method for Collecting and Storing Seeds from Zostera marina (Eelgrass) in the Yellow Sea,China

The coast of the Yellow Sea in China, like many other temperate coastal zones, has been experiencing a dramatic decline in the abundance of seagrass. Intensive efforts have been made to restore seagrass communities along the coast to restore the function of the coastal ecosystem. Transplanting adult Zostera marina shoots is labor‐intensive, time‐consuming, expensive, and detrimental to donor beds; thus, restoring seagrass communities through the use of seeds is highly valued in current, large‐scale restoration trials. In this study, an effective method for collecting, processing, and storing Z. marina seeds was developed. From 2009 to 2013, respectively, 122,000, 421,000, 364,000, 1,041,000, and 1,091,000 seeds were successfully collected. Two‐way analysis of variance (ANOVA) showed the interaction between salinity and temperature significantly affected the cumulative germination rate (CGR) (p < 0.01) during the storage period and the viability (p < 0.01) of seeds after storage. The germination rate after storage was significantly affected by salinity and temperature (p < 0.01). The highest viability (89.8 ± 1.0%) and germination rate (75.6 ± 4.5%) were found among seeds stored at 4°C and a salinity of 44.5 psu for 7 months. The cost for planting 1 ha of sea bottom with Z. marina seeds ranged from $2,613 to $80,900 depending on the seeding density and seed loss during storage. The average cost per Z. marina seed in this study was $0.00586.     

Habitat selection of wintering passerines in salt marshes of the German Wadden Sea

The salt marshes of the Wadden Sea are important wintering areas for some species of granivorous passerines, which have declined considerably since the 1960s. We investigated the habitat choice of all wintering passerines in eight study areas in German salt marshes with special consideration of human impact on these habitats. Granivorous species that almost exclusively winter in salt marshes, Shorelark (Eremophila alpestris), Snow Bunting (Plectrophenax nivalis) and Twite (Carduelis flavirostris) were concentrated in the lower salt marsh vegetation and in the driftlines, while all other species preferred the high upper salt marsh communities, although Rock Pipits (Anthus petrosus littoralis) fed in muddy areas along ditches. Shorelarks switched habitat in conditions where seeds were scarce to feed instead on arthropods in upper salt marshes. Intensively sheep-grazed upper salt marshes resemble lower salt marshes in their vegetation and were therefore mainly visited by Shorelarks, Snow Buntings and Twites. In winter, the driftline is preferred by the two former species, while in autumn and spring more birds foraged in the salt marshes. Twites prefer to feed mainly on seeds of Salicornia. Areas with S. europaea are visited mainly in late autumn and early winter, while areas with S. stricta are used throughout the winter because of a steady supply of seeds. Several years after embankment, polders are hardly used any more by the lower salt marsh species as the habitat changes into freshwater marshes. Large embankment projects since the early 1960s have included salt marshes and intertidal flats, and the resultant loss of habitat is responsible for the decline of lower salt marsh species. For other passerine species the effects of reclamation are unknown. The effects of intensified grazing on the wintering populations of Shorelark, Snow Bunting and Twite are still unresolved. Although grazing supports lower salt marsh vegetation, the seed production per plant is much lower there and some important seed producers hardly occur. Since grazing was reduced and embankment projects have been stopped, the salt marsh areas (especially lower salt marshes) have increased and so have the wintering populations of Shorelark, Snow Bunting and Twite. For the other species, the consequences of habitat changes are unknown, although it is suggested that reduced grazing will support them. Reducing the human impact on salt marshes will, in the long run, probably lead to a natural salt marsh with much variety in elevation and in its corresponding vegetation and bird communities. Meanwhile, management by grazing might be required in parts of the salt marshes.     

Assessing establishment success and suitability analysis of Zostera marina transplants using staple method in northern lagoons

2009年利用植株枚订移植法在我国北方典型澙湖——山东荣成天鹅湖逐月进行大叶藻(Zostera marina)植株移植,并于当年逐月对移植植株的存活率、定居时间和生长进行监测,分析该方法在我国北方澙湖的有效性和适宜性。结果显示:(1)4–6月移植植株的存活率为76.5%–90.4%,其中4月移植植株的存活率最低,7–9月移植植株的存活率达到100%;(2)6–9月移植植株的定居时间均为1个月,5月移植植株的定居时间为2个月,而4月移植植株的定居时间长达4个月;(3)除个别监测月份外,移植植株的叶长和叶鞘长均显著小于天然植株,而茎节直径和根长均与天然植株无明显差异;(4)我国北方澙湖较适宜大叶藻植株移植的区域为海水透明度高、水深不超过1 m的潮下带,且底质为泥含量较高的泥砂底质海区,9月份是适宜的移植时间。     

北方澙湖大叶藻植株枚订移植法的效果评估与适宜性分析

2009年利用植株枚订移植法在我国北方典型澙湖——山东荣成天鹅湖逐月进行大叶藻(Zostera marina)植株移植, 并于当年逐月对移植植株的存活率、定居时间和生长进行监测, 分析该方法在我国北方澙湖的有效性和适宜性。结果显示: (1) 4-6月移植植株的存活率为76.5%-90.4%, 其中4月移植植株的存活率最低, 7-9月移植植株的存活率达到100%; (2) 6-9月移植植株的定居时间均为1个月, 5月移植植株的定居时间为2个月, 而4月移植植株的定居时间长达4个月; (3)除个别监测月份外, 移植植株的叶长和叶鞘长均显著小于天然植株, 而茎节直径和根长均与天然植株无明显差异; (4)我国北方澙湖较适宜大叶藻植株移植的区域为海水透明度高、水深不超过1 m的潮下带, 且底质为泥含量较高的泥砂底质海区, 9月份是适宜的移植时间。     

Seed- versus transplant-based eelgrass (Zostera marina L.) restoration success in a temperate marine lake

Despite active seagrass restoration gaining traction as a tool to halt and reverse worldwide seagrass losses, overall success remains limited. Restoration strategies, through seeding or transplantation, face different environmental bottlenecks that limit success. Choosing the most appropriate strategy of the two for a specific location, however, is hampered by lack of direct practical comparisons between strategies within a single system. To investigate potential life stage dependent bottlenecks, we compared seed-based and transplant-based restoration of Zostera marina in the subtidal saltwater Lake Grevelingen. Our results demonstrate that seedling recruitment was negatively impacted by bioturbation from the lugworm Arenicola marina and sediment movement due to hydrodynamic exposure. Transplant-based restoration was clearly more successful but surprisingly best predicted by leaf gluing by the ragworm Platynereis dumerilii. This previously undescribed interaction caused seagrass leaves to clump and reduce effective photosynthetic surface and leaf movement. We suggest that the observed behavior of these worms may result from a lack of foodweb interactions, illustrating the importance of  trophic control for seagrass restoration. Thus, in addition to recognizing life stage dependent environmental bottlenecks for restoration strategy selection, seagrass restoration may also require the active recovery of their associated food webs.     

Density-dependent recruitment after winter disturbance on tidal flats by the lugworm Arenicola marina

The polychaete Arenicola marina is abundant and widespread on intertidal sand flats in K?nigshafen (island of Sylt, North Sea). Juveniles overwinter in subtidal channels and then colonize the upper tidal zone above the range of the adults. In the summers of 1995 and 1997, after a mild and a moderate winter, a distinct nursery belt fringing the shoreline was apparent. The severe winter of 1995/96 changed this pattern. Population size of the adults was halved, and in summer 1996 the juveniles were no longer restricted to an upper intertidal belt and settled over a wide range of tidal flats where adults were diminished. This spatial expansion of recruits is assumed to be a density-dependent response. An overcompensation by one-third of the previous population size did not carry over into the next year. Compared to some other species of the tidal-flat fauna, the disturbance effect by the severe winter on the lugworm population was small and brief. Received in revised form: 7 May 2001 Electronic Publication     

Desiccation,Moisture Content and Germination of Zostera marina L. Seed

Zostera marina is the only seagrass species whose seeds have been successfully used in large‐scale restoration. Although progress has been made in refining Z. marina restoration protocols, additional information on Z. marina seed physiology is necessary as the science of seagrass restoration evolves. We tested the germination rates of Z. marina seeds under different relative humidities and temperatures for different periods of time. Z. marina seed moisture content (MC) and germination rates were also tested when seeds were exposed to a temperature of 25°C and relative humidity of 50%. Z. marina seeds suffered higher mortality when exposed to lower relative humidity and higher temperature for longer period of exposure time. A significant negative correlation was detected between seed germination rate and MC. Z. marina seeds are sensitive to desiccation exposure and long periods of exposure to air should be prevented to minimize seed mortality when seeds are used in restoration projects.     

North Atlantic phylogeography and large-scale population differentiation of the seagrass Zostera marina L

As the most widespread seagrass in temperate waters of the Northern Hemisphere, Zostera marina provides a unique opportunity to investigate the extent to which the historical legacy of the last glacial maximum (LGM18 000-10 000 years bp) is detectable in modern population genetic structure. We used sequences from the nuclear rDNA-internal transcribed spacer (ITS) and chloroplast matK-intron, and nine microsatellite loci to survey 49 populations (> 2000 individuals) from throughout the species' range. Minimal sequence variation between Pacific and Atlantic populations combined with biogeographical groupings derived from the microsatellite data, suggest that the trans-Arctic connection is currently open. The east Pacific and west Atlantic are more connected than either is to the east Atlantic. Allelic richness was almost two-fold higher in the Pacific. Populations from putative Atlantic refugia now represent the southern edges of the distribution and are not genetically diverse. Unexpectedly, the highest allelic diversity was observed in the North Sea-Wadden Sea-southwest Baltic region. Except for the Mediterranean and Black Seas, significant isolation-by-distance was found from ~150 to 5000 km. A transition from weak to strong isolation-by-distance occurred at ~150 km among northern European populations suggesting this scale as the natural limit for dispersal within the metapopulation. Links between historical and contemporary processes are discussed in terms of the projected effects of climate change on coastal marine plants. The identification of a high genetic diversity hotspot in Northern Europe provides a basis for restoration decisions.     

Variable responses of native eelgrass Zostera marina to a non-indigenous bivalve Musculista senhousia

The transport and establishment of non-indigenous species in coastal marine environments are increasing worldwide, yet few studies have experimentally addressed the interactions between potentially dominant non-native species and native organisms. We studied the effects of the introduced mussel Musculista senhousia on leaf and rhizome growth and shoot density of eelgrass Zostera marina in San Diego Bay, California. We added M. senhousia over a natural range in biomass (0–1200 g dry mass/m²) to eelgrass in transplanted and established beds. The effects of the non-indigenous mussel varied from facilitation to interference depending on time, the abundance of M. senhousia, and the response variable considered. Consistent results were that mussel additions linearly inhibited eelgrass rhizome elongation rates. With 800 g dry mass/m² of M. senhousia, eelgrass rhizomes grew 40% less than controls in two eelgrass transplantations and in one established eelgrass bed. These results indicate that M. senhousia, could both impair the success of transplantations of eelgrass, which spread vegetatively by rhizomes, and the spread of established Z. marina beds to areas inhabited by M. senhousia. Although effects on leaf growth were not always significant, in August in both eelgrass transplantations and established meadows leaf growth was fertilized by mussels, and showed a saturation-type relationship to sediment ammonium concentrations. Ammonium concentrations and sediment organic content were linear functions of mussel biomass. We found only small, non-consistent effects of M. senhousia on shoot density of eelgrass over 6-month periods. In established eelgrass beds, but not in transplanted eelgrass patches (≈0.8 m in diameter), added mussels suffered large declines. Hence, eelgrass is likely to be affected by M. senhousia primarily where Z. marina beds are patchy and sparse. Our study has management and conservation implications for eelgrass because many beds are already seriously degraded and limited in southern California where the mussel is very abundant. Received: 31 May 1997 / Accepted: 4 September 1997     

Characterization of single nucleotide polymorphism markers for eelgrass (Zostera marina)

We characterized 37 single nucleotide polymorphism (SNP) makers for eelgrass Zostera marina. SNP markers were developed using existing EST (expressed sequence tag)-libraries to locate polymorphic loci and develop primers from the functional expressed genes that are deposited in The ZOSTERA database (V1.2.1). SNP loci were genotyped using a single-base-extension approach which facilitated high-throughput genotyping with minimal optimization time. These markers show a wide range of variability among 25 eelgrass populations and will be useful for population genetic studies including evaluation of population structure, historical demography, and phylogeography. Potential applications include haplotype inference of physically linked SNPs and identification of genes under selection for temperature and desiccation stress.     

An evaluation of small-scale genetic diversity and the mating system in Zostera noltii on an intertidal sandflat in the Wadden Sea

Background and Aims

The dwarf eelgrass, Zostera noltii, is a predominant inhabitant of soft-bottom intertidal regions along the coasts of northern Europe. It is a monoecious, protogynous angiosperm in which the potential for self-fertilization and inbreeding are high, especially if clone sizes exceed pollen dispersal distances. The aim of the present study was to determine the relationship between mating system and clonal structure, examine the relative roles of geitonogamous selfing and biparental inbreeding, measure pollen availability (multiple paternities) and estimate pollen dispersal.

Methods

A 100-m² plot was established in a large, intertidal Z. noltii meadow on the island of Sylt in the German Wadden Sea. A total of 256 adult shoots was sampled: one from the centre of 100 fixed 1-m² quadrats (large scale resolution) and an additional 156 from within eight randomly selected 1-m² sub-quadrats (small-scale resolution). DNA was extracted from seeds and leaf tissue of all samples and genotyped with nine microsatellite loci.

Key Results

Mating system analysis revealed high multilocus and single locus outcrossing rates. Average pollen dispersal distance was nearly the same as the average genet (clone) size. Multiple paternity was common and 20–30 % of mature seeds originated from matings within the plot. Among inbred seeds, most resulted from geitonogamy rather than biparental inbreeding.

Conclusions

Moderate disturbances intrinsic to the intertidal habitat appear to facilitate seed recruitment by gap formation. Pollen dispersal distances are sufficient to maintain outcrossing and high clonal diversity.