Sexual differences in physiological integration in the dioecious shrub Lindera triloba: a field experiment using girdling manipulation

Background and Aims

It is important to consider the modular level when verifying sexual dimorphism in dioecious plants. Nevertheless, between-sex differences in resource translocation among modules (i.e. physiological integration) have not been tested at the whole-plant level. In this study, sexual differences in physiological integration were examined among ramets, within a genet in the dioecious sprouting shrub Lindera triloba, by a field experiment with girdling manipulation.

Methods

Female and male genets were randomly assigned to girdled or intact groups. Girdling of the main ramets was conducted in May 2009 by removing a ring of bark and cambium approx. 1 cm wide at a height of 80–100 cm. The effects of treatment and sex on ramet dynamics (mortality, recruitment and diameter growth) and inflorescence production during 1 year after girdling were examined.

Key Results

The diameter growth rate of main ramets of both sexes was lower at ground level (D₀) but higher at breast height (dbh) in girdled than in intact groups. In sprouted ramets with a dbh of 0–2 cm, males in girdled groups had lower growth rates at D₀ than those of intact groups, whereas no girdling effect was found for females. The main ramets in girdled groups produced more inflorescences than intact groups, irrespective of sex, but male ramets showed a greater response to the treatment than females.

Conclusions

In L. triloba, physiological integration exists at the whole-plant level, and sprouted ramets are dependent on assimilates translocated from main ramets, but this dependence weakens as sprouted ramets get larger. Female sprouted ramets can grow in a physiologically independent manner from the main ramet earlier than those of males. This study highlights the importance of considering modular structures and physiological integration when evaluating sexual differences in demographic patterns of clonal plants.     

Sexual dimorphism in clonal growth forms and ramet distribution patterns in <Emphasis Type="Italic">Rumex acetosella</Emphasis> (Polygonaceae)

We investigated clonal traits in the dioecious herb Rumex acetosella to characterize sexual dimorphism in clonal forms and to correlate below-ground clonal patterns and above-ground ramet distributions. We recorded creeping root length, branching patterns, ramet and clump (caespitose ramets from the same position on the root) sprouting patterns, and biomass allocations in three females and males. We also estimated the patch size of flowering ramets within a quadrat. No sexual dimorphism was detected in the frequencies of branches and flowering ramets per root length. Male plants allocated proportionally more biomass to below-ground organs. Total root length did not differ between the sexes. Females sprouted more clumps with fewer flowering ramets per root length than males, which sprouted fewer clumps with more flowering ramets, which meant that clump sprouting patterns were phalanx-like in females and guerrilla-like in males. Flowering ramets were aggregately distributed in both females and males and patch sizes were similar between sexes, indicating that the spreader propagations were not found in the guerrilla-like males. We assumed that sexual dimorphism occurred in response to physiological integration for higher reproductive effort in females.     

Dynamics of distribution and performance of ramets constructing genets: a demographic�Cgenetic study in a clonal plant, Convallaria keiskei

Background and Aims

In clonal plants producing vegetative offspring, performance at the genet level as well as at the ramet level should be investigated in order to understand the entire picture of the population dynamics and the life history characteristics. In this study, demography, including reproduction and survival, the growth patterns and the spatial distributions of ramets within genets of the clonal herb Convallaria keiskei were explored.

Methods

Vegetative growth, flowering and survival of shoots whose genets were identified using microsatellite markers were monitored in four study plots for 3 years (2003–2005). The size structures of ramets in genets and their temporal shifts were then analysed. Their spatial distributions were also examined.

Key Results

During the census, 274 and 149 ramets were mapped in two 1 × 2 m plots, and 83 and 94 ramets in two 2 × 2 m quadrats. Thirty-eight genotypes were identified from 580 samples. Each plot included 5–18 genets, and most ramets belonged to the predominant genet(s) in each plot. Shoots foliated yearly for several years, but flowering ramets did not have an inflorescence the next year. A considerable number of new clonal offspring persistently appeared, forming a bell-shaped curve of the size structure of ramets in each genet. Comparing the structures modelled by the normal distributions suggested variation among ramets belonging to a single genet and variation among genets. Furthermore, spatial analyses revealed clumped and distant distributions of ramet pairs in a genet, in which the distant patterns corresponded to the linearly elongating clonal growth pattern of this species.

Conclusion

Characteristics of ramet performances such as flowering and recruitment of clonal offspring, in addition to growth, played a large part in the regulation of genet dynamics and distribution, which were different among the studied genets. These might be characteristics particularly relevant to clonal life histories.Key words: Clonal plant, Convallaria keiskei, demography, genet, genetic identification, growth pattern, life history, ramet, spatial distribution     

Patch structure and ramet demography of the clonal tree, Asimina triloba, under gap and closed-canopy

Clonal understory trees develop into patches of interconnected and genetically identical ramets that have the potential to persist for decades or centuries. These patches develop beneath forest canopies that are structurally heterogeneous in space and time. Canopy heterogeneity, in turn, is responsible for the highly variable understory light environment that is typically associated with deciduous forests. We investigated what aspects of patch structure (density, size structure, and reproductive frequency of ramets) of the clonal understory tree, Asimina triloba, were correlated with forest canopy conditions. Specifically, we compared A. triloba patches located beneath closed canopies and canopy gaps. We also conducted a three-year demographic study of individual ramets within patches distributed across a light gradient. The closed canopy-gap comparison demonstrated that the patches of A. triloba had a higher frequency of large and flowering ramets in gaps compared to closed-canopy stands, but total ramet density was lower in gaps than in closed canopy stands. In the demographic study, individual ramet growth was positively correlated with light availability, although the pattern was not consistent for all years. Neither ramet recruitment nor mortality was correlated with light conditions. Our results indicate that the structure of A. triloba patches was influenced by canopy condition, but does not necessarily depend on the responses of ramets to current light conditions. The lack of differences in ramet recruitment and mortality under varying canopy conditions is likely to be a primary reason for the long-term expansion and persistence of the patches. The primary benefit of a positive growth response to increasing light is the transition of relatively small ramets into flowering ramets within a short period of time.     

Partial mechanical stimulation facilitates the growth of the rhizomatous plant Leymus secalinus: modulation by clonal integration

Background and Aims

Mechanical stimulation (MS) often induces plants to undergo thigmomorphogenesis and to synthesize an array of signalling substances. In clonal plants, connected ramets often share resources and hormones. However, little is known about whether and how clonal integration influences the ability of clonal plants to withstand MS. We hypothesized that the effects of MS may be modulated by clonal integration.

Methods

We conducted an experiment in which ramet pairs of Leymus secalinus were subjected to three treatments: (1) connected ramet pairs under a homogeneous condition [i.e. the proximal (relatively old) and distal (relatively young) ramets were not mechanically stressed]; (2) connected ramet pairs under a heterogeneous condition (i.e. the proximal ramet was mechanically stressed but the distal ramet was not); and (3) disconnected ramet pairs under the same condition as in treatment 2. At the end of the experiment, we harvested all plants and determined their biomass and allocation.

Key Results

Clonal integration had no significant influence on measured traits of distal L. secalinus ramets without MS. However, under MS, plants with distal ramets that were connected to a mother ramet produced more total plant biomass, below-ground biomass, ramets and total rhizome length than those that were not connected. Partial MS exerted local effects on stimulated ramets and remote effects on connected unstimulated ramets. Partial MS increased total biomass, root/shoot ratio, number of ramets and total rhizome length of stimulated proximal ramets, and increased total biomass, root weight ratio, number of ramets and total rhizome length of connected unstimulated ramets due to clonal integration.

Conclusions

These findings suggest that thigmomorphogenesis may protect plants from the stresses caused by high winds or trampling and that thigmomorphogenesis can be strongly modulated by the degree of clonal integration.     

Buffalograss decreases ramet propagation in infertile patches to enhance interconnected ramet proliferation in fertile patches

Buffalograss (Buchloë dactyloides) is known for its low-nutrient tolerance. However, in natural habitats, nutrients are usually patchily distributed. For clonal plants like buffalograss, physiological integration is an important strategy to cope with adverse environmental conditions. In order to examine how integration helps buffalograss to survive in patchy conditions, a greenhouse experiment was conducted for 91 days. Interconnected ramet pairs of stoloniferous buffalograss were planted in two partitioned same-sized containers, and subjected to identical or contrasting nutrient supply. In contrast to normally perceived resource-sharing concepts, results showed that buffalograss genets reduced production of new ramets in nutrient-poor patches promoting at the same time propagation of interconnected ramets in nutrient-rich patches. Ramets in nutrient-rich patches gained significant benefit from heterogeneous treatments, whereas nutrient-poor ramets performed even worse than in uniform low-nutrient treatment. Younger ramets developed more biomass than elder ramets with the same amounts of nutrient supply under homogeneous treatment, while elder ramets were more tolerant when nutrients were scarce. Heterogeneity had a particular strong effect on stolons and new ramet production in nutrient-rich patches. Rooted ramets in nutrient-poor patches suffered from a by-pass of nutrients to interconnected ramets on nutrient-rich substrate that probably resulted from different transpiration rates. We conclude that this resource-sharing strategy is advantageous for buffalograss to concentrate more ramets in fertile patches, and facilitate the survivorship of more buffalograss ramets in adverse environments with uneven nutrient supply.     

Response of physiological integration in Trifolium repens to heterogeneity of UV-B radiation

Physiological integration has been documented in many clonal plants growing under resource heterogeneity. Little is still known about the response of physiological integration to heterogeneous ultraviolet-B radiation. In this paper, the changes in intensity of physiological integration and of physiological parameters under homogeneous and heterogeneous ultraviolet-B radiation (280-315 nm) were measured in order to test the hypothesis that in addition to resource integration a defensive integration in Trifolium repens might exist as well. For this purpose, homogeneous and heterogeneous ultraviolet-B radiation was applied to pairs of connected and severed ramets of the stoloniferous herb Trifolium repens. Changes in intensity of water and nutrient integration were followed with acid fuchsin dye and ¹⁵N-isotope labeling of the xylem water transport. In order to assess the patterns of physiological integration contents of chlorophyll, ultraviolet-B absorbing compounds, soluble sugar and protein were determined and activities of superoxide dismutase (SOD) and peroxidase (POD) measured. When ramets were connected and exposed to heterogeneous UV-B radiation, the velocity of water transportation from the UV-B treated ramet to its connected sister ramet was markedly lower and the percentage of ¹⁵N left in labelled ramets that suffered from enhanced UV-B radiation was higher and their transfer to unlabelled ramets lower. In comparison with clones under homogeneous ultraviolet-B radiation, the content of chlorophyll, ultraviolet-B absorbing compounds, soluble sugar and activities of SOD and POD increased notably if ultraviolet-B stressed ramets were connected to untreated ramets. Chlorophyll and UV-B absorbing compounds were shared between connected ramets under heterogeneous UV-B radiation. This indicated that physiological connection improved the performance of whole clonal plants under heterogeneous ultraviolet-B radiation. The intensity of physiological integration of T. repens for resources decreased under heterogeneous ultraviolet-B radiation in favor of the stressed ramets. Ultraviolet-B stressed ramets benefited from unstressed ramets by physiological integration, supporting the hypothesis that clonal plants are able to optimize the efficiency of their resistance maintaining their presence also in less favorable sites. The results could be helpful for further understanding of the function of heterogeneous UV-B radiation on growth regulation and microevolution in clonal plants.     

Shifting effects of physiological integration on performance of a clonal plant during submergence and de-submergence

Background and Aims

Submergence and de-submergence are common phenomena encountered by riparian plants due to water level fluctuations, but little is known about the role of physiological integration in clonal plants (resource sharing between interconnected ramets) in their adaptation to such events. Using Alternanthera philoxeroides (alligator weed) as an example, this study tested the hypotheses that physiological integration will improve growth and photosynthetic capacity of submerged ramets during submergence and will promote their recovery following de-submergence.

Methods

Connected clones of A. philoxeroides, each consisting of two ramet systems and a stolon internode connecting them, were grown under control (both ramet systems untreated), half-submerged (one ramet system submerged and the other not submerged), fully submerged (both ramet systems submerged), half-shaded (one ramet system shaded and the other not shaded) and full-shaded (both ramet systems shaded) conditions for 30 d and then de-submerged/de-shaded for 20 d. The submerged plants were also shaded to very low light intensities, mimicking typical conditions in turbid floodwater.

Key Results

After 30 d of submergence, connections between submerged and non-submerged ramets significantly increased growth and carbohydrate accumulation of the submerged ramets, but decreased the growth of the non-submerged ramets. After 20 d of de-submergence, connections did not significantly affect the growth of either de-submerged or non-submerged ramets, but de-submerged ramets had high soluble sugar concentrations, suggesting high metabolic activities. The shift from significant effects of integration on both submerged and non-submerged ramets during the submergence period to little effect during the de-submergence period was due to the quick recovery of growth and photosynthesis. The effects of physiological integration were not found to be any stronger under submergence/de-submergence than under shading/de-shading.

Conclusions

The results indicate that it is not just the beneficial effects of physiological integration that are crucial to the survival of riparian clonal plants during periods of submergence, but also the ability to recover growth and photosynthesis rapidly after de-submergence, which thus allows them to spread.     

美丽箬竹水分生理整合的分株比例效应——基于叶片抗氧化系统与光合色素

生理整合是克隆植物实现资源共享, 增强对异质生境适应能力的重要手段。其中, 水分生理整合是克隆植物最为重要的生理整合, 解析竹子水分生理整合特征对于竹林水分科学管理具有重要意义。该研究以分株地下茎相连的美丽箬竹(Indocalamus decorus)盆栽苗为试验材料, 设置2个盆栽基质相对含水率(高水势(90% ± 5%)和低水势(30% ± 5%))和5个分株比例(1:3、1:2、1:1、2:1、3:1, 高水势分株与低水势分株数量比值, 地下茎相连的分株总数12株)处理。处理后15、30、45、60天分别取不同处理的克隆分株成熟叶测定抗氧化酶活性、相对电导率和丙二醛含量、可溶性蛋白质含量、光合色素含量, 分析基于分株比例的美丽箬竹水分生理整合方向、强度和效率的变化规律。结果表明: 在异质水分条件下, 美丽箬竹分株间存在着从高水势供体分株向低水势受体分株进行水分转移的生理整合作用, 并随着分株比例的增大, 整合强度增强, 受体分株获益提高, 供体分株耗损增大。随着处理时间的延长, 处理前期分株间水分生理整合强度增强, 处理后期整合强度减弱, 反映出供体分株与受体分株间耗-益在时间序列上是有变化的, 处理前期耗-益更为明显。研究表明克隆系统分株比例对竹子水分生理整合有重要影响, 分株间水分梯度差是水分传导的潜在驱动力, 决定水分生理整合方向、强度和效率的是分株间水分供需关系。     

Clonal Diversity in Differently-Aged Populations of the Pseudo-Annual Clonal Plant Circaea lutetiana L.

Abstract: In many clonal plant species seedling recruitment is restricted to short colonization episodes early in the development of the population, and clonal diversity (i.e., genet diversity) in the population is expected to decrease with increasing population age. In established populations of the pseudo-annual Circaea lutetiana seedling recruitment has previously not been observed. Therefore, we expected established populations to have low clonal diversities. We analysed number and frequency of genets and spatial distribution of genets in six differently-aged C. lutetiana populations with the use of four informative RAPD primers. We found relatively low clonal diversities in young populations but very high clonal diversities in established populations. Therefore, the hypothesis was rejected that seedling recruitment does not occur in established populations. Moreover, we did not find large genet size asymmetries in established populations. Genet size differences can be caused by stochastic processes or by fitness related traits, such as differences in vegetative reproduction. Because vegetative propagation of ramets is dependent on ramet size, and the number of ramets and the size of each ramet determine genet size, we expected that large genets produced, on average, large ramets. However, this was not the case, suggesting that stochastic processes caused genet size differences. Genet size may also be bounded if spatial distribution of genets is affected by micro-habitat differences. For this we expected to find a clumped spatial distribution of ramets of the same genet. However, ramets of large genets were always found intermingled with ramets belonging to other genets.     

Clonal integration affects growth and sediment properties of the first ramet generation,but not later ramet generations under severe light stress

克隆整合影响严重光胁迫下第一分株世代的生长和沉积物特征但不影响 后续分株世代的生长和沉积物特征 克隆整合通过缓冲环境压力和提高资源获取效率使克隆植物受益。然而,在一个克隆系统中,受益于克隆整合的连接分株世代的数量很少受到关注。我们进行了一个盆栽实验来评估沉水植物苦草 (Vallisneria natans)克隆系统内的生理整合程度,该克隆系统由一个母株和3个依次连接的后代分株组成。 母株生长在正常光照下,而后代分株被严重遮荫。母株与后代分株间的匍匐茎被切断或保持连接,但3个后代分株之间的连接仍然存在。与遮荫的后代分株连接时,苦草未遮荫的母株的光合能力显著增强,但其生物量积累大大减少。克隆整合显著增加了第一分株世代(相邻分株)的生物量积累和土壤的碳氮可用性、胞外酶活性和微生物生物量,但没有增加后续分株世代的这些特征。我们的结果表明,在严重光胁迫下,来自苦草母株的支持可能仅限于克隆系统中相邻的后代分株,这暗示着一个分株世代的效应。我们的结果有助于更好地理解克隆植物的层次结构和分段化。这些发现表明克隆整合程度在分株种群的生态相互作用中起着至关重要的作用。     

Effects of soil heterogeneity and clonal integration on Alternanthera philoxeroides populations with a radial ramet aggregation

Some clonal plants can spread their ramet populations radially, and soil heterogeneity and clonal integration may greatly affect the establishment of these types of populations. We constructed Alternanthera philoxeroides populations with a radial ramet aggregation, allowing old ramets of clonal fragments to concentrate in central pots and younger ramets to root in peripheral pots. The peripheral pots were supplemented either with three different levels (high, medium and low) of soil nutrients to simulate a heterogeneous soil environment, or only one medium level of soil nutrients to simulate a homogeneous environment. Stolon connections between the central older ramets and the peripheral younger ramets were left intact or severed to test the effect of clonal integration. The maintenance of stolon connection could induce the division of labor between different‐aged ramets, by increasing the root investment in central ramets and the above‐ground growth in peripheral ramets. The maintenance of stolon connection could improve the growth of the central and peripheral ramets, clonal fragments and even the whole population. However, the positive consequence in peripheral ramets and whole fragments was only detected in the high‐nutrient patch of heterogeneous treatment. In sum, in the population with the radial ramet aggregation, clonal integration can play a key role in the rapid recruitment of young ramets of A. philoxeroides fragments, as well as the expansion of the whole population. The magnitude of clonal integration also became more obvious in the peripheral young ramets and whole fragments that experienced high‐nutrient patches.     

Influences of the genetic neighborhood on ramet reproductive success in a clonal desert cactus

Clonal structure in clonal plants can affect sexual reproduction. Individual ramets can decrease reproduction if their neighbors are ramets of the same genet due to inbreeding depression or self-incompatibility. We assessed ramet reproductive success in the partial self-incompatible Ferocactus robustus (Cactaceae) as a function of floral display size in focal ramets and floral display size and clonal structure of their reproductive neighborhoods. Ramets were labeled, sized in number of stems, mapped and genetically identified through RAPD markers in one population. A pollen dispersal area of 15-m radius was established for each ramet to determine the clonal diversity in the neighborhoods. Flower production and fruit set were counted on a monthly basis during one reproductive season as a surrogate of ramet fitness. We expected a decrease in individual ramet reproductive success as a function of the number of reproductive ramets of the same genet in the neighborhood. A total of 272 sampled ramets revealed 116 multilocus genotypes, showing high clonal diversity in the population (G/N = 0.43, D = 0.98). Clonal diversity of neighborhoods ranged from 0.06 to 1 and fruit set varied from 0 to 76.9%. Individual ramet reproductive success was influenced by (1) mate availability, (2) floral display size of a genet within the reproductive neighborhood, and (3) the proportion of distinguishable genotypes. Floral display size of genets and ramets coupled with the genetic diversity within the reproductive neighborhood determines the low sexual reproduction in F. robustus.     

The oldest known clones of Salix herbacea growing in the Northern Apennines,Italy are at least 2000 years old

Premise

Dominant in many ecosystems around the world, clonal plants can reach considerable ages and sizes. Due to their modular growth patterns, individual clonal plants (genets) can consist of many subunits (ramets). Since single ramets do not reflect the actual age of genets, the ratio between genet size (radius) and longitudinal annual growth rate (LAGR) of living ramets is often used to approximate the age of clonal plants. However, information on how the LAGR changes along ramets and how LAGR variability may affect age estimates of genets is still limited.

Methods

We assessed the variability of LAGR based on wood-section position along the ramets and on the duration of the growing season on three genetically distinct genets of Salix herbacea growing in the Northern Apennines (Italy). We compared genet ages estimated by dividing genet radius by the LAGRs of its ramets.

Results

LAGR increased significantly from the stem apex to the root collar; indicating that ramet growth rate decreased with time. Furthermore, a difference of ca. 2 weeks in the onset of the growing period did not impact LAGR. Considering the high LAGR variability, we estimated that the three genets started to grow between ~2100 and ~7000 years ago, which makes them the oldest known clones of S. herbacea even considering the most conservative age estimate.

Conclusions

Our findings indicate that analyzing ramets at the root collar provides an integrative measurement of their overall LAGR, which is crucial for estimating the age of genets.     

Demography of the clonal palm Geonoma brevispatha in a Neotropical swamp forest

Abstract We expected that in clonal plants with the phalanx growth form, prereproductive ramets function as a meristem bank that helps buffer ramet demography against environmental fluctuations. Specifically, we evaluated four expectations for the forest understorey clonal palm Geonoma brevispatha in a swamp forest in south‐eastern Brazil. Our expectations were are follows: (i) the vital rates of prereproductive ramets are more responsive to genet‐related characteristics (genet size, distance and size of the nearest genet neighbour, and reproductive ramet survivorship) than to environmental variables (canopy openness, basal area of small trees, flooding, and distance and size of the nearest large tree neighbour), whose effects are expected to affect ramet dynamics indirectly through physiological integration; (ii) the progress of prereproductive ramets to later stages is dependent on the death of reproductive ramets; (iii) the population dynamics of G. brevispatha is resistant to significant interannual changes in the vital rates of ramets, (i.e. the occurrence of good and bad years); and (iv) there are large negative covariances among demographic transitions within a given year, as well as between years, reflecting the flexibility of G. brevispatha to adjust to environmental variability. Expectations 1, 2 and 4 were confirmed, while expectation 3 was only partially confirmed. Matrix models predicted that the growth rate of the overall ramet population was significantly greater than unity in the first study year, but it was significantly smaller than 1.0 in the second study year. Ramet population growth rates at the level of individual genets were greater than unity in the majority of genets in the first year, but in the second year genets with growth rates smaller than unity prevailed. Ramet population dynamics seems to be resistant to the succession of good and bad years; in effect, the presence of a meristem bank of ramets helps stabilize λ.     

Arbuscular mycorrhizal fungi reduce effects of physiological integration in Trifolium repens

Background and Aims

One of the special properties of clonal plants is the capacity for physiological integration, which can increase plant performance through mechanisms such as resource sharing and co-ordinated phenotypic plasticity when plants grow in microsites with contrasting resource availabilities. However, many clonal plants are colonized by arbuscular mycorrhizal fungi (AMF). Since AMF are likely to reduce contrasts in effective resource levels, they could also reduce these effects of clonal integration on plasticity and performance in heterogeneous environments.

Methods

To test this hypothesis, pairs of connected and disconnected ramets of the stoloniferous herb Trifolium repens were grown. One ramet in a pair was given high light and low nutrients while the other ramet was given high nutrients and low light. The pairs were inoculated with zero, one or five species of AMF.

Key Results

Pairs of ramets grown without AMF developed division of labour and benefited from resource sharing, as indicated by effects of connection on allocation to roots, accumulation of mass, and ramet production. Inoculation with five species of AMF significantly reduced these effects of connection, both by inhibiting them in ramets given high nutrients and inducing them in ramets given high light. Inoculation with one species of AMF also reduced some effects of connection, but generally to a lesser degree.

Conclusions

The results show that AMF can significantly modify the effects of clonal integration on the plasticity and performance of clonal plants in heterogeneous environments. In particular, AMF may partly replace the effects and benefits of clonal integration in low-nutrient habitats, possibly more so where species richness of AMF is high. This provides the first test of interaction between colonization by AMF and physiological integration in a clonal plant, and a new example of how biotic and abiotic factors could interact to determine the ecological importance of clonal growth.Key words: Arbuscular mycorrhizal fungi, biomass allocation, clonal plant, division of labour, environmental heterogeneity, light availability, nutrients, white clover     

Spatial distribution pattern of a clonal species: effects of differential production of clonal and sexual offspring

The spatial distribution patterns of genets and ramets within populations are expected to change as a function of the frequency with which clonal species recruit different types of offspring (sexual and clonal). We used an integrated approach to study the spatial arrangement of clonal plants by combining molecular and ecological data using Opuntia microdasys as a study system. The species is able to produce two types of clonal (plantlets and cladodes) and one type of sexual (seeds) offspring. Additionally it is found in three habitats that cause differences in the ability of each type of offspring to establish. In 2007, all individuals in the three habitats (162 in BH = bajada, 264 in IDH = hill-piedmont, and 136 at HPH = interdunes) were tagged and mapped. Amplified inter-simple sequence repeats (ISSR’s) were used to determine the multilocus genotype and relatedness of each individual ramet using 120 polymorphic bands (104 in BH, 128 in HPH and 180 in IDH). The spatial distribution pattern of genets and ramets was analyzed with the Hopkins test and spatial autocorrelation analysis. For all habitats we found that O. microdasys displayed a spatial distribution characterized by clumps of aggregated ramets, but habitats differed in the number of genets present. As for other clonal species a strong positive spatial autocorrelation exists within 20 m, although all analyses suggest that adjacent ramets are genetically less related to each other or belong to different genets, that is, ramets of different genets are intermingled. The spatial arrangement of genets and ramets in O. microdasys between habitats closely matches the frequency of establishment of each type of offspring (e.g. the more clonal areas are clumped groups of related individuals). These results confirm that in two habitats (BH and IDH) clonal recruitment had been more common than in the other habitat (HPH).     

Clonal spread of invasive Ludwigia hexapetala and L. grandiflora in freshwater wetlands of California

Ludwigia hexapetala and L. grandiflora are recent, aggressive invaders of freshwater wetlands in California. To assess the relative role of sexual versus clonal reproduction in invasive spread, we used AFLP markers to genotype 794 ramets of L. hexapetala sampled from 27 populations in three watersheds of California, and 150 ramets of L. grandiflora from five populations in a fourth watershed. We then used two analytical methods, similarity thresholds and character incompatibilities, to distinguish genotypic variation within genets (clones) from variation between genets. Our analyses revealed extremely limited genotypic and genet variation in invasive L. hexapetala and L. grandiflora within California. Within L. hexapetala, 95% of the ramets analyzed represented a single genet. The genet was the only one detected in 20 populations. The remaining seven populations contained two to nine genets. Within L. grandiflora, all ramets were of only one genotype. Thus, invasive spread within and between populations, and across watersheds, appears to be almost exclusively clonal and brought about by the dispersal of vegetative propagules. The extremely low seedling recruitment indicates that management should target vegetative dispersal and growth.     

Effects of inoculation with native arbuscular mycorrhizal fungi on clonal growth of Potentilla reptans and Fragaria moschata (Rosaceae)

Many clonal plants live in symbiosis with ubiquitous arbuscular mycorrhizal (AM) fungi, however, little is known about their interaction with respect to clonal reproduction and resource acquisition. The effects of arbuscular mycorrhiza on the growth and intraclonal integration between ramets of two stoloniferous species were studied experimentally in a nutritionally homogenous soil environment. Two species coexisting at the same field site, Potentilla reptans and Fragaria moschata, were selected as model plants for the study. Pairs of their ramets were grown in neighbouring pots with each ramet rooted separately. Four inoculation treatments were established: (1) both mother and daughter ramets remained non-inoculated, (2) both ramets were inoculated with a mixture of three native AM fungi from the site of plant origin, (3) only mother or (4) daughter ramet was inoculated. The stolons connecting the ramets were either left intact or were disrupted. Despite the consistent increase in phosphorus concentrations in inoculated plants, a negative growth response of both plant species to inoculation with AM fungi was observed and inoculated ramets produced fewer stolons and fewer offspring ramets and had lower total shoot dry weights as compared to non-inoculated ones. A difference in the extent of the negative mycorrhizal growth response was recorded between mother and daughter ramets of P. reptans, with daughter ramets being more susceptible. Due to AM effect on ramet performance, and thereby on the source-sink relationship, inoculation also significantly influenced biomass allocation within clonal fragments. Physiological integration between mother and daughter ramets was observed when their root systems were heterogeneous in terms of AM colonization. These results hence indicate the potential of mycorrhizal fungi to impact clonal growth traits of stoloniferous plant species, with possible consequences for their population dynamics.     

Contrasting patterns of clonality and fine-scale genetic structure in two rare sedges with differing geographic distributions

For plants with mixed reproductive capabilities, asexual reproduction is more frequent in rare species and is considered a strategy for persistence when sexual recruitment is limited. We investigate whether asexual reproduction contributes to the persistence of two co-occurring, rare sedges that both experience irregular seed set and if their differing geographic distributions have a role in the relative contribution of clonality. Genotypic richness was high (R=0.889±0.02) across the clustered populations of Lepidosperma sp. Mt Caudan and, where detected, clonal patches were small, both in ramet numbers (⩽3 ramets/genet) and physical size (1.3±0.1 m). In contrast, genotypic richness was lower in the isolated L. sp. Parker Range populations, albeit more variable (R=0.437±0.13), with genets as large as 17 ramets and up to 5.8 m in size. Aggregated clonal growth generated significant fine-scale genetic structure in both species but to a greater spatial extent and with additional genet-level structure in L. sp. Parker Range that is likely due to restricted seed dispersal. Despite both species being rare, asexual reproduction clearly has a more important role in the persistence of L. sp. Parker Range than L. sp. Mt Caudan. This is consistent with our prediction that limitations to sexual reproduction, via geographic isolation to effective gene exchange, can lead to greater contributions of asexual reproduction. These results demonstrate the role of population isolation in affecting the balance of alternate reproductive modes and the contextual nature of asexual reproduction in rare species.