Ribosomal RNA genes in species of theCynareae tribe (Compositae). II

Summary The structure of the ribosomal DNA has been analyzed in three species of theCynareae tribe using Southern blot hybridization.Silybum marianum possesses two types of ribosomal genes 12.3 and 13.4 kb long;Cirsium vulgare has at least four types of rDNA repeats 10.6, 10.5, 11.7 and 11.9 kb long;Carlina acaulis only one type of ribosomal genes 9.7 kb long. The rRNA genes of the three species studied showed an identical restriction mapping in the 18 S and 25 S regions. However species differentiation in length and/or nucleotide sequences are present in the external spacer and very probably in the internal transcribed spacer.By cytophotometric studies and byin vitro rRNA/DNA filter hybridization, the DNA amount/4 C nucleus and the rDNA percentage were calculated in nine species of theCynareae tribe:Cynara cardunculus subsp.scolymus (artichoke),Cynara cardunculus subsp.cardunculus (wild artichoke),Onopordon acanthium, O. illyricum, Galactites tomentosa, Carduus nutans, Silybum marianum, Cirsium vulgare andCarlina acaulis. The DNA amount/4 C nucleus in eight species are similar, ranging from 4.24 pg inGalactites tomentosa to 5.96 pg inCirsium vulgare, whileCarlina acaulis has a DNA amount/ 4C nucleus of 11.8 pg. The rDNA percentages range from 0.192% inOnopordon acanthium to 1.022% inSilybum marianum, whileCarlina acaulis has 0.038% of rDNA. This latter species is clearly distinct within theCynareae tribe.     

Shift from facilitative to neutral interactions by the cushion plant Silene acaulis along a primary succession gradient

Background

The stress‐gradient hypothesis predicts a shift from facilitative to competitive plant interactions with decreasing abiotic stress. This has been supported by studies along elevation and temperature gradients, but also challenged by the hypothesis of a facilitation collapse at extremely harsh sites. Although facilitation is known to be important in primary succession, few studies have examined these hypotheses along primary succession gradients.

Aim

To examine whether there is a relationship between the presence of the circumpolar cushion plant Silene acaulis and other species, and if so, whether there is a shift between positive and negative interactions along a primary succession gradient in a glacier foreland.

Location

Finse, southern Norway.

Methods

We examined the performance of the common alpine forb Bistorta vivipara, species richness of vascular plants, bryophytes and lichens, and the number of seedlings and fertile vascular plants in S. acaulis cushions, and control plots without S. acaulis, along a succession gradient with increasing distance from a glacier front, and thus decreasing abiotic stress. To examine if S. acaulis cushions modify the abiotic environment, we recorded soil temperature, moisture, organic content and pH in cushions and control plots.

Results

Bistorta vivipara performed better, as shown by bigger leaves in S. acaulis cushions compared to control plots in the harshest part of the gradient close to the glacier. There were few differences in B. vivipara performance between cushion and control plots in the more benign environment further away from the glacier. This suggests a shift from facilitative to mainly neutral interactions by S. acaulis on the performance of B. vivipara with decreasing abiotic stress. A trend, although not significant, of higher vascular species richness and fertility inside S. acaulis cushions along the whole gradient, suggests that S. acaulis also facilitates community‐level species richness. The causal mechanism of this facilitation is likely that the cushions buffer extreme temperatures.

Conclusions

Our results support the stress‐gradient hypothesis for the relationship between the cushion plant S. acaulis and the performance of a single species along a primary succession gradient in a glacier foreland. S. acaulis also tended to increase vascular plant species richness and fertility regardless of stress level along the gradient, suggesting facilitation at the community level. We found no collapse of facilitation at the most stressful end of the gradient in this alpine glacier foreland.     

GENETIC VARIATION IN ALETES ACAULIS AND ITS RELATIVE,THE NARROW ENDEMIC A. HUMILIS (APIACEAE)

We compared the organization of genetic variability in Aletes humilis, a species restricted to seven populations in north-central Colorado, with A. acaulis, a closely related widespread species. Genetic variability was scored at 11 electrophoretically detectable protein loci. Nine of these loci were polymorphic (i.e., P > 0.95) in both taxa, and were used for interpopulation and interspecific comparisons. Levels of genetic variability and patterns of organization of this variability are very comparable in both species. In contrast to many other narrow endemics, A. humilis is not genetically depauperate when compared to its presumed progenitor. Both biochemical and morphological evidence suggest that A. humilis is derived from A. acaulis; the comparable levels of variability in the two species suggest that A. humilis was derived from a source containing a substantial portion of the original A. acaulis genome, because there is no evidence of a genetic bottleneck or appreciable loss of allelic variability.     

Small‐scale spatial associations between Artemisia frigida and Potentilla acaulis at different intensities of sheep grazing

Questions: The formerly overgrazed Inner Mongolia steppe was subject to retrogressive succession. Today, Artemisia frigida and Potentilla acaulis are two dominant species in different phases of successive degradation. To investigate the impact of grazing intensity on spatial community structure, we investigated the small‐scale spatial association between A. frigida and P. acaulis at zero, light, medium and heavy sheep grazing, and proposed factors involved in the spatial associations between these two species along a grazing intensity gradient. Location: The Inner Mongolia Grassland Ecosystem Research Station of the Chinese Academy of Sciences. Methods: Four grazing intensities were selected: zero, light (1.33 sheep/ha), medium (4.0 sheep/ha) and heavy (6.7 sheep/ha). After 13 years of grazing three 2 m × 2 m quadrats with 100 × 100 cells of size 2 cm × 2 cm were randomly selected in each treatment in July and August 2002. The presence of A. frigida and P. acaulis in each cell was recorded and the positions of the individuals were mapped using Cartesian coordinates in each quadrat. The small‐scale spatial associations between A. frigida or P. acaulis were quantified with the L₁₂(d), J₁₂(d) functions (both derived from K₁₂(d), the former indicating the type of the spatial association, the latter indicating the strength of the spatial association), using Monte Carlo simulations. Results: A. frigida was negatively associated with P. acaulis at short distances (0–100 cm) under zero and light grazing, and negatively or independently under medium and heavy grazing. Increasing grazing intensities suppressed the peak negative associations. More intense grazing enhanced the tendency towards independent distribution of these two species. Conclusions: The small‐scale spatial associations between A. frigida and P. acaulis were significantly different at four different intensities of sheep grazing. Grazing disturbance, clonal growth habit of species, and interspecific competition are the main factors leading to a difference of spatial associations between these two species at different grazing intensities.     

Cuticular studies in some Nelsonioideae (Acanthaceae)*

The foliar epidermis and cuticle of Staurogyne longifolia (Nees) Kuntze, Elytraria acaulis (L.f.) Lindau var. acaulis, E. acaulis var. lyrata (Nees) Bremek. and Nelsonia campestris R.Br, have been investigated, revealing broad similarities with those of the rest of the Acanthaceae; the presence of diacytic stomata in the Nelsonioideae is evidence of its affinity with the Acanthaceae in general-, while the presence of panduriform glandular hairs and the absence of the cystoliths in Nelsonioideae indicate its particular affinity with the Thunbergioideae. Substantial evidence is provided to support the retention of Nelsonioideae as a subfamily of the Acanthaceae, rather than its transfer to the Scrophulariaceae.     

Isozyme variation and species relationships in the genus Lolium L. (ryegrasses,Graminaceae)

Thirty-two natural populations belonging to the eight species of the genus Lolium (ryegrass) or to Festuca pratensis (meadow fescue) were recorded for allelic frequencies at 13 isozyme loci. Cultivated ryegrass (L. perenne and L. multiflorum), meadow fescue, and the annual L. rigidum, are true outbreeders. The other species are true inbreeders, except for L. canariense, which shows a moderate level of cross fertilisation (20%). Hierarchical clustering from Nei's unbiased distance leads to four groups. The three self-pollinating, weed species, L. temulentum, L. remotum and L. persicum, belong to the first cluster, which is the most differentiated one. The second cluster comprises L. multiflorum, L. subulatum and most populations of L. rigidum. All L. perenne populations belong to the third cluster, as do two of L. rigidum. The average genetic distance within the L. perenne group is very low. Surprisingly, the fourth cluster groups together L. canariense and Festuca pratensis. The data suggest that L. rigidum is the species with the greatest diversity, and could be a common ancestor of the genus. Knowledge of historical processes of domestication could help to calibrate the molecular clock.     

Cytochrome P450 CYP81A10v7 in Lolium rigidum confers metabolic resistance to herbicides across at least five modes of action

Rapid and widespread evolution of multiple herbicide resistance in global weed species endowed by increased capacity to metabolize (degrade) herbicides (metabolic resistance) is a great threat to herbicide sustainability and global food production. Metabolic resistance in the economically damaging crop weed species Lolium rigidum is well known but a molecular understanding has been lacking. We purified a metabolic resistant (R) subset from a field evolved R L. rigidum population. The R, the herbicide susceptible (S) and derived F₂ populations were used for candidate herbicide resistance gene discovery by RNA sequencing. A P450 gene CYP81A10v7 was identified with higher expression in R vs. S plants. Transgenic rice overexpressing this Lolium CYP81A10v7 gene became highly resistant to acetyl-coenzyme A carboxylase- and acetolactate synthase-inhibiting herbicides (diclofop-methyl, tralkoxydim, chlorsulfuron) and moderately resistant to hydroxyphenylpyruvate dioxygenase-inhibiting herbicide (mesotrione), photosystem II-inhibiting herbicides (atrazine and chlorotoluron) and the tubulin-inhibiting herbicide trifluralin. This wide cross-resistance profile to many dissimilar herbicides in CYP81A10v7 transgenic rice generally reflects what is evident in the R L. rigidum. This report clearly showed that a single P450 gene in a cross-pollinated weed species L. rigidum confers resistance to herbicides of at least five modes of action across seven herbicide chemistries.     

Molecular diversity and derivations of populations of Silene acaulis and Saxifraga oppositifolia from the high Arctic and more southerly latitudes

A survey of allozyme diversity within and between populations of Silene acaulis from Spitsbergen, Norway, Iceland and Scotland, showed that populations from the high Arctic (Spitsbergen, > 76°N) contained high levels of diversity and were genetically similar to populations from more southern locations. Indirect measures of gene flow (Nm), calculated from Wrigh's F indicated that there had been extensive gene flow between Spitsbergen and some Norwegian populations. A restriction site analysis of chloroplast DNA (cpDNA) in S. acaulis revealed that all populations contained a single identical cpDNA haplotype, except one population from Norway which also contained a second haplotype. In contrast, five different cpDNA haplotypes were distinguished in a more limited survey of cpDNA variation in Saxifraga oppositifolia, with all five haplotypes present in one of two Spitsbergen populations surveyed. The contrasting cpDNA results for the two species suggest that whereas high-Arctic populations of Silene acaulis have most likely been derived from immigrants which arrived from the south after the last glacial period, high-Arctic populations of Saxifraga oppositifolia may be derived, in part, from ancient northern stocks which survived the last glaciation in high-Arctic refugia.     

The dicotyledonous species Erodium moschatum (L) L'Hér. ex Aiton is sensitive to haloxyfop herbicide due to herbicide-sensitive acetyl-coenzyme A carboxylase

Most plants are resistant to herbicides which inhibit acetyl-coenzyme A carboxylase (ACCase) because they have both eukaryotic ACCase and herbicide-insensitive, prokaryotic ACCase. Members of the Gramineae are killed because they have only herbicide-sensitive, eukaryotic ACCase. Here we report that a dicot, Erodium moschatum, is sensitive to the ACCase-inhibiting herbicide haloxyfop because it has herbicide-sensitive ACCase. Erodium moschatum was controlled by haloxyfop application at rates which controlled the gramineous species Digitaria ciliaris and a susceptible Lolium rigidum biotype but did not control the dicot Nicotiana tabacum or a haloxyfop-resistant L. rigidum biotype WLR96. Similarly, the haloxyfop acid concentration required to inhibit activity by 50% in E. moschatum ACCase assays (1.0 μM) was similar to that required for D. ciliaris (2.3 μM) and susceptible L. rigidum (0.4 μM) but much less than that for the resistant L. rigidum biotype WLR96 (353 μM) or the dicots  N. tabacum (182 μM) and Pisum sativum (150 μM). Leaf protein extracts from  N. tabacum and P. sativum contained both eukaryotic ACCase and prokaryotic subunits of ACCase, but E. moschatum, D. ciliaris and both L. rigidum biotypes exhibited only the eukaryotic ACCase. Thus, the dicot  E. moschatum is sensitive to haloxyfop because it lacks the herbicide-insensitive prokaryotic ACCase, a protein that has been considered ubiquitous in dicot species. Received: 12 May 1998 / Accepted: 27 June 1998     

Variations in <Emphasis Type="Italic">Anarthrophyllum rigidum</Emphasis> radial growth,NDVI and ecosystem productivity in the Patagonian shrubby steppes

The lack of long-term records of productivity is a critical limitation to the study of ecosystem dynamics. Annual rings, a measure of growth in woody species, are a useful tool to document ecosystem dynamics. Time series of the Normalized Difference Vegetation Index (NDVI) provide estimates of ecosystem productivity through satellite-derived data on the fraction of photosynthetic active radiation absorbed by vegetation. In the Patagonian steppes, we relate changes in NDVI to interannual variations in the radial growth of the shrub Anarthrophyllum rigidum. A widely distributed network of 15 ring-width chronologies of A. rigidum was used to estimate changes in NDVI across the Patagonia steppe (35°–50°S). In most sites, interannual variations in shrub growth and NDVI are regulated by winter precipitation. The water accumulated in the soil during winter is used by A. rigidum during the growing season, concurrent with the maximum NDVI values. At 10 from the 15 selected sites, variations in the radial growth of A. rigidum explained between 23 and 62% of the total variance in seasonal NDVI, suggesting that the A. rigidum growth at some sites provides good estimates of productivity in the Patagonian shrubby steppes during the growing season. However, we were unable to determine clear relationships between radial growth and NDVI at high-elevation mountainous sites or where intensive grazing by sheep masked the effect of climate variability on shrub growth. We conclude that dendrochronological methods can complement other estimates to reconstruct variations of productivity, supplementing and extending the few short records available in the Patagonian steppe.     

Pollination in Geonoma macrostachys and Three Congeners,G. acaulis,G. gracilis,and G. interrupta

A continuous 15 month study of the floral ecology of four syntopic understorey palm species of Genoma was conducted in Amazonian Peru lowland rainforest. The spicate inflorescences of G. macrostachys, G. acaulis and G. gracilis are strictly protandrous and the plants are functionally dioecious. Data suggest that in G. macrostachys and G. acaulis pollination is based on a mimicry system, the pistillate flowers mimicking the staminate ones in colour, shape and scent. Pollen-collecting meliponine bees (Hymenoptera, Apidae, Meliponinae) and pollen-feeding syrphid flies (Diptera, Syrphidae) which visit inflorescences during both sexual stages are the pollinators of G. macrostachys. Geonoma acaulis is pollinated by small pollen-feeding weevils (Coleoptera, Curculionidae, Derelomini) that visit male and female spikes. Additionally, in G. macrostachys another pollinator type, viz. euglossine bees (Hymenoptera, Apidae, Euglossinae), which are attracted and rewarded by both types of flowers may account for long-distance pollination. The palm G. gracilis shows a very distinct pollination system. Although opportunistic insect visitors are attracted to the inflorescences of this species it seems to be mainly anemophilous because pollen becomes powdery during an thesis. The branched inflorescences of G. interrupta are also protandrous, but unlike the other species of Geonoma observed, staminate and pistillate anthesis of individual flowers are, for the most, overlapping. A broad spectrum of visitors is attracted (bees, wasps, flies, and beetles), which all may act as pollinators. Outcrossing is especially encouraged during the purely female phase at the end of the flowering cycle when there are no more staminate flowers in the inflorescence. Effects on the reproductive biology and population structure of different pollination systems and breeding system are discussed.     

Distribution and population structure of the anther smut Microbotryum silenes‐acaulis parasitizing an arctic–alpine plant

Cold‐adapted organisms with current arctic–alpine distributions have persisted during the last glaciation in multiple ice‐free refugia, leaving footprints in their population structure that contrast with temperate plants and animals. However, pathogens that live within hosts having arctic–alpine distributions have been little studied. Here, we therefore investigated the geographical range and population structure of a fungus parasitizing an arctic–alpine plant. A total of 1437 herbarium specimens of the plant Silene acaulis were examined, and the anther smut pathogen Microbotryum silenes‐acaulis was present throughout the host's geographical range. There was significantly greater incidence of anther smut disease in more northern latitudes and where the host locations were less dense, indicating a major influence of environmental factors and/or host demographic structure on the pathogen distribution. Genetic analyses with seven microsatellite markers on recent collections of 195 M. silenes‐acaulis individuals revealed three main genetic clusters, in North America, northern Europe and southern Europe, likely corresponding to differentiation in distinct refugia during the last glaciation. The lower genetic diversity in northern Europe indicates postglacial recolonization northwards from southern refugia. This study combining herbarium surveys and population genetics thus uniquely reveals the effects of climate and environmental factors on a plant pathogen species with an arctic–alpine distribution.     

Long distance pollen-mediated flow of herbicide resistance genes in <Emphasis Type="Italic">Lolium rigidum</Emphasis>

Gene flow promotes genetic exchange among plant populations mediating evolutionary dynamics; yet, the importance of gene flow at distance via pollen movement is poorly understood. A field experiment at the landscape level was conducted with Lolium rigidum herbicide-susceptible individuals (population VLR1) placed into an otherwise Lolium-free bushland environment at increasing distances from adjacent large commercial crop fields infested with herbicide-resistant L. rigidum. Herbicide resistance was used as a marker to quantify the distance and the rate of pollen-mediated gene flow. About 21,245 seeds were produced on the isolated, susceptible mother plants of which 3,303 seedlings were tested for herbicide resistance and 664 seedlings were found to be resistant. Pollen-mediated gene flow occurred at 3,000 m (maximum tested distance). Both Mendelian and molecular analyses (sequencing and CAPS markers) confirmed the introgression of herbicide resistance genes. This is the first documented case of long-distance gene flow in L. rigidum. The results are important for future modeling simulations of herbicide resistance evolution and subsequent mobility. The adoption of integrated agronomic strategies, the control of potential receptor plants on fields’ margins and conservative use of herbicides can be realistic options to minimize herbicide resistance spread. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Trichomanes resinosum (Hymenophyllaceae), a new species from southern Venezuela and adjacent Guyana

A new species,Trichomanes resinosum, is described and compared to two similar species,T. elegans Rich. andT. rigidum Sw., with which it has been confused.     

The phylogeny and biogeography of Gentiana L. sect. Ciminalis (Adans.) Dumort.: A historical interpretation of distribution ranges in the European high mountains

Gentiana sect. Ciminalis consists of seven mostly ecologically or geographically vicariant and closely related species which are distributed throughout the South and Central European high mountains. The analysis of a RAPD data set and trn L-intron and ITS sequences resulted in slightly different phylogenetic hypotheses. In the preferred hypothesis the group consists of two completely resolved main lineages: 1) G. clusii and G. alpina. 2) G. dinarica, G. acaulis, G. ligustica, G. angustifolia and G. occidentalis. The most important conclusions we have drawn from this phylogenetic hypothesis and from the observed patterns of molecular variation are: 1) The calcifuge ecology of G. acaulis and G. alpina evolved independently from calcicole ancestors. 2) Among the calcicole taxa speciation proceeded from East to West in a simple vicariant pattern. 3) The application of a provisional molecular clock indicates that speciation events in sect. Ciminalis probably occurred in the Quaternary. 4) Differences in infraspecific genetic variation among the widespread species suggest that G. alpina probably experienced more recent dispersal or gene flow than G. clusii and G. acaulis. 5) The large number of mutations in the lineage leading to G. angustifolia, compared to the few mutations in the lineage leading to G. dinarica, may be a result of their different population histories. While the extant range of G. angustifolia was strongly affected by Quaternary climatic fluctuations, that of G. dinarica has had a more stable climatic history. 6) The low number of mutations and the basal position in one clade of the preferred cladogram leads to the conclusion that G. dinarica is the species most similar to the last common ancestor of sect. Ciminalis.     

Enhanced rates of herbicide metabolism in low herbicide‐dose selected resistant Lolium rigidum

Lolium rigidum is an obligately cross‐pollinated, genetically diverse species and an economically important herbicide resistance‐prone weed. Our previous work has demonstrated that recurrent selection of initially susceptible L. rigidum populations with low herbicide rates results in rapid herbicide resistance evolution. Here we report on the mechanisms endowing low‐dose‐selected diclofop‐methyl resistance in L. rigidum. Results showed that resistance was not due to target‐site ACCase mutations or overproduction, or differential herbicide leaf uptake and translocation. The in vivo de‐esterification of diclofop‐methyl into phytotoxic diclofop acid was rapid and similar in resistant versus susceptible populations. However, further metabolism of diclofop acid into non‐toxic metabolites was always faster in resistant plants than susceptible plants, resulting in up to 2.6‐fold lower level of diclofop acid in resistant plants. This corresponded well with up to twofold higher level of diclofop acid metabolites in resistant plants. The major polar metabolites of diclofop acid chromatographically resembled those of wheat, a naturally tolerant species. Clearly, recurrent selection at reduced herbicide rates selected for non‐target‐site‐based enhanced rates of herbicide metabolism, likely involving cytochrome P450 monooxygenases.     

Mycorrhizae from Denali National Park and Preserve,Alaska

 Roots of 40 taxa of higher plants (Pteridophyta, Spermatophyta) from two alpine study sites in Denali National Park and Preserve in central Alaska were examined for their mycorrhizal colonization. We observed ectomycorrhizae on six species: Betula nana, Salix reticulata, Salix polaris, Salix arctica, Polygonum viviparum, and Dryas octopetala. Seven taxa, Cassiope tetragona, Empetrum nigrum, Ledum palustre subsp. decumbens, Ledum palustre subsp. groenlandicum, Loiseleuria procumbens, Vaccinium uliginosum and Vaccinium vitis–idaea (all Ericales), had ericoid mycorrhizae. One species, Arctostaphylos alpina, formed a typical arbutoid mycorrhiza. Two species (Sibbaldia procumbens and Aconitum delphinifolium) showed well-developed VA mycorrhizae, whereas three species of plants (Lycopodium clavatum, Silene acaulis and Oxytropis scammaniana) had vesicles, but no arbuscules. The roots of 11 other plants (Lycopodium clavatum, Lycopodium selago, Silene acaulis, Gentiana algida, Lupinus arcticus, Oxytropis scammaniana, Pedicularis langsdorffii, Pedicularis capitata, Pedicularis verticillata, Artemisia sp. and Carex bigelowii) had a variety of intracellular colonizations which are referred to as dark septate fungi. No mycorrhizae were found on 12 other plants: Equisetum arvense, Equisetum variegatum, Lycopodium alpinum, Polygonum bistorta, Saxifraga hieracifolia, Saxifraga hirculus, Astragalus alpinus, Pedicularis kanei, Petasites frigidus, Carex podocarpa, Carex microchaeta and Poa arctica. A possible ecological role of dark septate fungi is discussed. Accepted: 4 August 1995     

The effect of B chromosomes on homoeologous pairing in species hybrids

Six diploid populations of Lolium rigidum were used in crosses with a standard homozygous line of Lolium temulentum in order to analyse the variation in the effect of B chromosomes on homoeologous pairing in the interspecific hybrid. — Large differences in chiasma frequency were found, both between the progeny of the different populations and, in one case, between the progeny of different plants from the same population. That this variation was due to differences in B chromosome genotype was ruled out by the use of appropriate 1B Lolium rigidum parents. It is concluded therefore that the effect of B's on homoeologous pairing in this interspecific hybrid can be modified quite substantially by the A chromosome genotype of the parents.