Phylogeny of chromosome races ofFestuca arundinacea andF. mairei (Poaceae) as indicated by seed protein electrophoresis

Seed protein electrophoresis of four chromosomes races ofFestuca arundinacea, F. mairei and their progenitors showed variation in banding patterns. High protein similarities betweenF. arundinacea, F. mairei, F. scariosa, andF. pratensis indicate close phylogenetic relationships of these species. The ancestry ofF. arundinacea cytotypes could be narrowed to three diploid species:F. scariosa, F. pratensis, andF. rubra or to their close relatives.     

Variable effects of endophytic fungus on seedling establishment of fine fescues

Seedborne systemic endophytic fungi of grasses are thought to be plant mutualists, because they have been shown to improve their host’s resistance against biotic and abiotic stresses. The interactions in plant–endophyte associations vary from mutualistic to parasitic with environmental conditions and the genotypes of interacting species. The possible pros and cons of endophytic fungi are expected to be most evident during the seedling establishment, where host fitness is most directly affected. If this holds true, endophytes may play a focal role in local adaptation of hosts to different environments. We examined if endophyte-infected and uninfected seeds and seedlings of two native grass species, Festuca rubra and F. ovina, differ in seed germination and seedling growth rates under greenhouse conditions. The germination of F. rubra seeds was also studied in the field. This is the first time that the effects of Epichloë endophyte on seedling establishment of fine fescues from natural populations have been experimentally evaluated. Mother plant (seed family) had a marked effect on many response variables in both grass species. Length and mean biomass of tillers of endophyte-infected (E+) F. ovina seedlings were lower, but root:shoot ratios were higher than in endophyte-free (E?) seedlings. In F. rubra, the effects of the endophyte were dependent on the habitat where the seeds were collected. The E+ seeds from river banks germinated faster than E+ seeds from meadows, and E+ seedlings from the river banks produced fewer but taller and heavier tillers than the other seedlings. Our data suggest that the effects of the endophyte infection on the seedling stage of fine fescues are dependent the species of grass, host genetic background and mother plant habitat. The germination strategy and growth form of E+ red fescue seedlings from river banks may be beneficial to surviving in the harsh conditions of that habitat.     

Phylogeny of tall fescue and related species using RFLPs

The wild species of tall fescue (Festuca arundinacea var.genuina Schreb.) represent a wide range of genetic variation and constitute potential germplasm for tall fescue improvement. Our objective was to evaluate genome specificity of the previously-identified DNA probes and to examine the phylogenetic relationship of tall fescue with six related species by using RFLP data. A total of 29 DNA probes from aPstI-genomic library of tall fescue were hybridized toEcoRI-orHindIII-digested DNA of 32 plants from sixFestuca species and fromLolium perenne L. Fifteen probes hybridized to all seven species. The remaining 14 probes showed differential hybridization patterns (i.e., ±), especially at the diploid and tetraploid levels. This hybridization pattern reflected genome divergence in these species. The DNA probes will be useful markers in breeding programs involving interspecific and intergeneric hybridization. Cluster analyses were performed using the average genetic distances calculated with the RFLP data from 53 probe-enzyme combinations. Generally, genotypes from the same species were grouped in the same cluster. These data indicated that tall fescue has a close relationship withF. pratensis Huds. (diploid),F. arundinacea var.glaucescens Boiss. (tetraploid), andL. perenne L. (diploid) and thatFestuca pratensis andL. perenne had the closest degree of relationship.This paper is a contribution of the Missouri Agricultural Experimental Station, Journal Series no. 11,798     

Effect of bryophytes on survival of conifer seedlings in subalpine forests of central Japan

Bryophytes form the major seedbed for coniferous trees in the subalpine forests of central Japan. Field experiments were conducted on the floor of a closed stand dominated byTsuga on Mt Fuji to examine the seedling survival ofAbies veitchii andTsuga diversifolia in seven substrate types in relation to the morphological characteristics of the seeds and seedlings. NeitherAbies norTsuga seedlings could survive on dwarf-bamboo litter beyond 2 years after the seed rain.Abies seedlings survived not only in all types of bryophyte communities but also in larch litter. In contrast, the survival ofTsuga seedlings was restricted to communities of smaller-statured bryophytes found on logs. The seeds and 1st year seedlings ofA. veitchii were larger than those ofT. diversifolia. Abies seedlings produced hypocotyls taller than any bryophyte community and radicles which were long enough to penetrate into the moist humus layer, whereasTsuga seedling radicles presumably penetrate into humus only in bryophyte communities forming a thin mat on logs. The effect of bryophytes on survival of tree seedlings therefore depends on the relationships between the morphology of seeds and seedlings and the structure of bryophyte communities.     

Fertile plant regeneration from protoplasts of meadow fescue (Festuca pratensis Huds.)

Suspension cultures from mature embryo-derived compact callus were initiated in seven meadow fescue (Festuca pratensis Huds.) cultivars. Four to six months after initiation, embryogenic suspension cultures with a moderate growth rate were established from three of them (cvs. Barmondo, Belimo and Leopard). These suspension cultures showed the capacity, maintained over six months, to regenerate green plants which could be grown to maturity under greenhouse conditions.Morphogenic suspension cultures from single genotypes of three F. pratensis cultivars (cvs. Barmondo, Belimo and Leopard) yielded large numbers of protoplasts, which upon culture in agarose beads using nurse cells formed microcalli with an overall plating efficiency in the range of 10^-3 to 10^-4. Mature plants were reproducibly regenerated and established in soil, from such protoplasts during a period of six months. The regeneration of fertile plants from protoplasts derived from suspension cultures of meadow fescue and its implications on gene transfer technology for this species are discussed.Abbreviations 2,4-D 2,4-dichlorophenoxy-acetic acid.     

Phylogeographical history of the widespread meadow fescue (Festuca pratensis Huds.) inferred from chloroplast DNA sequences

Aim Here we explore the variation in chloroplast DNA (cpDNA) in a widespread Eurasian diploid forage grass, meadow fescue (Festuca pratensis Huds.), to address its phylogeographical history. In particular, we aim to answer whether the post‐glacial migration routes of meadow fescue are associated with the spread of agriculture or concurrent with well‐documented natural migration pathways from glacial refugia. Location A total of 56 Eurasian accessions of F. pratensis were analysed, representing the entire native distribution area as well as non‐native areas in northernmost Europe. Methods Based on initial sequencing of 10 non‐coding cpDNA regions, three regions were sequenced for all F. pratensis accessions. For reference, three closely related species [the diploid Lolium perenne L. and the polyploids Festuca arundinacea Schreb. and Festuca gigantea (L.) Vill.] were also sequenced, as well as the more distantly related Festuca ovina L. Divergence times were estimated assuming a simple molecular clock, calibrated using a previously published estimate of 9 Myr for the divergence between fine‐leaved (F. ovina) and broad‐leaved fescues (F. pratensis, F. arundinacea and F. gigantea). Results Limited, but geographically structured, cpDNA variation was observed in F. pratensis. Three haplotypes, estimated to have diverged 0.16 Ma, were identified: one western European (A), one with a wide eastern distribution from central‐eastern Europe into Asia (B) and one Caucasian (C). The haplotypes of the polyploids and L. perenne were estimated to have diverged from haplotype A in F. pratensis 0.8–1.3 Ma. Main conclusions We found no definite evidence for migration of the diploid F. pratensis associated with the spread of agriculture from the Fertile Crescent after the last glaciation. The distinct geographical structuring of the present‐day variation in cpDNA can rather be explained by northwards expansion of the western haplotype from an Iberian refugium, expansion of the eastern haplotype from an unlocated (south‐)eastern refugium and glacial survival without subsequent expansion from a Caucasian refugium. The high level of cpDNA divergence observed between this diploid and the polyploids which have probably been derived from it may suggest that the very low level of cpDNA variation in the diploid is caused by a recent bottleneck. Today, F. pratensis is widespread in the open agricultural landscape but appears otherwise confined to naturally open habitats such as river banks, and its populations may have been decimated when dense forests dominated in the previous interglacial.     

Relationships between three species ofFestuca sect.Bovinae (Poaceae)

Seed protein electrophoresis confirms the existence of polymorphism among hexaploid populations ofFestuca arundinacea. Both protein and morphological results suggest thatF. pratensis andF. arundinacea should retain independent specific status. High protein homology of these two species withF. gigantea points towards phylogenetic links between these taxa.     

Species relationships inFestuca (sect.Ovinae,Poaceae)

Morphological and seed protein studies of selected species ofFestuca were performed to elucidate the relationships between species of sect.Ovinae and their affinity with other fescues.Festuca rubra andF. heterophylla (extravaginale group) had higher phenetic affinity with taxa of sect.Scariosae, Montanae andBovinae than with members of their own section. 5 species of the intravaginale group (F. ovina, F. capillata, F. valesiaca, F. sulcata, F. trachyphylla) were fairly uniform and well defined by protein and morphological data. This supports current systematic thinking and does not agree withHackel's (1882) classification of this group.     

Drei neue Arten desFestuca ovina-Formenkreises (Poaceae) aus dem Osten Österreichs

WithinFestuca ovina agg. two hexaploid taxa:F. brevipila andF. carnuntina, and one tetraploid:F. eggleri, are described as new species.F. pseudovina Hack. exWiesb. is better treated asF. valesiaca Schleich. exGaudin subsp.parviflora (Hack.), comb. nova.

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Drei neue Arten desFestuca ovina-Formenkreises (Poaceae) aus dem Osten Österreichs

     

DasFestuca vivipara-Problem in den Alpen

The differences betweenFestuca vivipara (tetraploid) andF. ovina subsp.supina (diploid), two often confused taxa, are demonstrated in regard to morphology, leaf anatomy, ecology and distribution. New maps illustrate the distribution ofF. vivipara in the Alps and the Northern Hemisphere. The development of different polyploidy levels withinF. vivipara as a consequence of occasional sexual processes is discussed. (English Summary on p. 39.)

     

Effect of wet and dry cycles in calcareous soil on mineral nutrient uptake of two grasses,Agrostis stolonifera L. and Festuca ovina L.

Shoot uptake of mineral nutrients (Ca, Cu, Fe, K, mg, Mn, P, S, Zn) by Agrostis stolonifera L. was compared with Festuca ovina L. under wet and dry cycles. Such conditions are typical for A. stolonifera sites, whereas F. ovina is growing mostly on consistently drier and better-drained soils. Plants were grown in a glasshouse, at controlled temperature and light conditions, using two moisture regimes, one constant at 60% WHC (water holding capacity), one wet/dry fluctuating between 35 and 100% WHC. Above ground and total biomass production was lower under wet/dry treatment than at constant water regime in F. ovina, but did not differ between regimes in A. stolonifera. Shoot uptake of most elements was severely reduced in F. ovina at the wet/dry regime. Shoot uptake and concentrations of most elements studied (Cu, K, Mn, P, S, Zn) were lower (p<0.05) under wet/dry treatment than at constant regime in A. stolonifera and tended to be lower also of Fe and Mg. Differences in biomass production observed are consistent with field evidence that A. stolonifera grows in sites which are periodically flooded but may become quite dry during other periods, and that F. ovina is limited to sites which are consistently drier and better drained. Evidence from the present study, however, does not support any view that alternating wet and dry cycles, as typical of A. stolonifera field sites, would be beneficial to nutrient acquisition of this species but that biomass production may develop normally at the lower uptake of most mineral nutrients measured under the wet/dry regime. Such regimes are decidedly unfavourable to both growth and nutrient acquisition of F. ovina.     

Assemblages of native bryophytes in secondary forests with introduced Quercus rubra

We have studied the diversity of bryophytes in planted Polish post‐agricultural forests dominated by the native Scots pine Pinus sylvestris and the alien (North American) red oak Quercus rubra. The planted sites would be suitable for a mesic coniferous forest (abbreviation: CFS) or mesic broadleaved forest (abbreviation: BFS). We analysed the structure and composition of the bryophyte assemblages in relation to forest site and substrate availability. Special attention was paid to the introduced Q. rubra as a host species for native bryophytes. A total of 54 bryophyte species (9 liverworts and 45 mosses) were found in the 90 plots ( = phytosociological relevés, 10 × 10 m in area; 45 at each forest site) studied. DCA analysis showed that the bryophyte assemblages of the P. sylvestris–Q. rubra secondary forest community differed significantly between CFS and BFS sites; the similarity of the composition of bryophyte species was 36.8%. The substrate preferences (epigeic, epixylic, epiphytic), as well as the growth form and life form of the recorded bryophytes, also differed between CFS and BFS, while the proportion of bryophytes that had a particular life strategy was very similar. The introduced Q. rubra was inhabited by 28 bryophyte species, including two liverworts. This tree hosted 64% of the CFS and 47% of BFS bryophyte flora and as a host for epiphytes the species successfully fulfilled the functional role of the native oaks (Q. robur and Q. petraea). Thus, the introduction of Q. rubra may contribute to the restoration of post‐agricultural forests and to the conservation of epiphytic bryophyte species. On the other hand, the negative impact of Q. rubra observed on the ground flora (including bryophytes) puts the benefits of Q. rubra for the conservation of native biodiversity in general in question.     

Structural and numerical chromosomal polymorphism in natural populations ofAlopecurus (Poaceae)

Considerable structural and numerical chromosomal variation has been found in natural populations ofAlopecurus. Interchange heterozygotes, identified by multivalent formation during meiosis, have been recovered in four out of six species studied and are reported for the first time in the diploidsA. bulbosus, andA. myosuroides, and the tetraploidA. pratensis. B chromosomes have been found in two species,A. pratensis andA. myosuroides and also in inter-specific hybrids betweenA. pratensis andA. arundinaceus. The characteristics, distribution and meiotic behaviour of both interchange heterozygotes and B chromosomes are described.     

Infection with the fungal endophyte Epichloë festucae may alter the allelopathic potential of red fescue

Red fescue (Festuca rubra) is a perennial grass used as both forage and turfgrass. Asymptomatic plants of this species are systemically infected by the fungal endophyte Epichloë festucae, which has a beneficial effect on the infected plants. The aim of this study was to determine the effect of the endophyte Epichloë festucae on the allelopathic potential of F. rubra against four associated pasture species that are also considered as weeds in lawns, Trifolium pratense, Trifolium repens, Lotus corniculatus and Plantago lanceolata. Two experiments were designed to evaluate the allelopathic effect of extracts from the roots and leaves of endophyte‐infected (E+) and non‐infected (E?) plants on the germination and seedling growth of the four target species. Regardless of the endophyte status of the host plant, leaf extracts elicited a stronger reduction in germination and seedling growth than root extracts. Extracts from E+ plants reduced the speed of germination index of Trifolium spp. to a greater extent than those from E? plants. Radicle length of the target species was the parameter most affected by the presence of the endophyte in F. rubra. Root extracts from E+ plants had a greater inhibitory effect on the radicle growth of the target species than did root extracts from E? plants. A greater concentration in total phenolic compounds was found in the roots of E+ plants than of E?; however, this difference was not observed in the leaves. Thus, the allelopathic potential of F. rubra is altered in infected plants.     

Alder-Frankia interaction and alder-poplar association for biomass production

Summary Alders have an important role to play in biomass producing stands because of their N₂-fixing ability and their capacity to withstand soils having an excess of moisture. The objectives of preliminary trials were (1) to find if there is any alder-genotype xFrankia-strain interaction when the effect of inoculating the bacteria was compared to no inoculation in seed beds of different species and provenances of alder, (2) to measure the possible effect of black alders interplanted in poplars compared to pure poplar plots. Two trials were laid out to study the alder-Frankia interaction. Both produced interaction. In the first one the inoculation had a favorable effect onAlnus glutinosa at age 2 years andA. cordata at age 1 and 2 and no effect onA. rubra. In the second one the inoculation had a depressive effect at age 1 on 2 of 3 provenances ofA. rubra and no effect on 1A. rubra, 3A.glutinosa and 3A. cordata provenances.A closely spaced field trial associating one black alder provenance and the poplar clone UNAL gives no superiority of mixed plots compared to pure plots. The results suggest that the N₂-fixation of alders is not profitable to poplars at age 3 with a 1.5×2 m spacing.     

Intergeneric relationships ofLolium,Festuca, andVulpia (Poaceae) and their phylogeny

Morphological and seed protein analyses of 26 species of the generaLolium, Festuca andVulpia confirmed their close systematic affinities. Six inflorescence characters readily differentiatedFestuca fromLolium. Protein similarities betweenFestuca of sect.Bovinae and cross-pollinated species ofLolium, coupled with cytogenetic and crossability data, substantiate that they should be united into one genus.Vulpia had phenetic similarities with sect.Scariosae, Montanae andOvinae ofFestuca. Lolium, Festuca, andVulpia are most likely derived from a common ancestral form which was close toFestuca pratensis andLolium perenne.     

A novel method for characterisation and quantification of plant root exudates

A microcosm unit is described which readily allows manipulation of experimental conditions to enable the subsequent impact on root exudation release to be monitored with time. Festuca ovina and Plantago lanceolata seedlings were grown in this microcosm unit over a 34 day experimental period under conditions of high (3.75 mol m^–3 N) or low (1.25 mol m^–3 N) nitrate-nitrogen treatment. At the end of the experimental period the seedlings in the microcosms were labelled with [¹⁴C]-CO₂ and the fate of the label within the plant and its release by the roots monitored. Total organic carbon (TOC) content of the collected exudate material was measured throughout the experimental period as well as during the ¹⁴C-chase period and comparison of plant C budgets using these two measurements is discussed. Nitrogen treatment as found to have a greater effect on exudate release by F. ovina than by P. lanceolata seedlings as indicated by both the total organic carbon and ¹⁴C results. The use and applications of the microcosm unit are discussed.     

A linkage map of meadow fescue (<Emphasis Type="Italic">Festuca pratensis</Emphasis> Huds.) and comparative mapping with other Poaceae species

A genetic linkage map has been constructed for meadow fescue (Festuca pratensis Huds.) (2n=2x=14) using a full-sib family of a cross between a genotype from a Norwegian population (HF2) and a genotype from a Yugoslavian cultivar (B14). The two-way pseudo-testcross procedure has been used to develop separate maps for each parent, as well as a combined map. A total number of 550 loci have been mapped using homologous and heterologous RFLPs, AFLPs, isozymes and SSRs. The combined map consists of 466 markers, has a total length of 658.8 cM with an average marker density of 1.4 cM/marker. A high degree of orthology and colinearity was observed between meadow fescue and the Triticeae genome(s) for all linkage groups, and the individual linkage groups were designated 1F–7F in accordance with the orthologous Triticeae chromosomes. As expected, the meadow fescue linkage groups were highly orthologous and co-linear with Lolium, and with oat, maize and sorghum, generally in the same manner as the Triticeae chromosomes. It was shown that the evolutionary 4AL/5AL translocation, which characterises some of the Triticeae species, is not present in the meadow fescue genome. A putative insertion of a segment orthologous to Triticeae 2 at the top of 6F, similar to the rearrangement found in the wheat B and the rye R genome, was also observed. In addition, chromosome 4F is completely orthologous to rice chromosome 3 in contrast to the Triticeae where this rice chromosome is distributed over homoeologous group 4 and 5 chromosomes. The meadow fescue genome thus has a more ancestral configuration than any of the Triticeae genomes. The extended meadow fescue map reported here provides the opportunity for beneficial cross-species transfer of genetic knowledge, particularly from the complete genome sequence of rice.Communicated by P. Langridge     

Seedling establishment ofPolygonum cuspidatum andPolygonum weyrichii var.alpinum at high altitudes of Mt Fuji

Seedling establishment ofPolygonum cuspidatum (Seib. et Zucc.) andPolygonum weyrichii (F. Schmit) var.alpinum (Maxim.), dominant early-successional species in a volcanic gravel area on Mt Fuji, Japan, was compared for current-year seedlings at 2500 m a.s.l. The rate of survival in the first winter after germination at 2500 m was 2% and 24% forP. cuspidatum andP. weyrichii seedlings, respectively, and correlated with the upper distribution limit of each (2500–2600 m forP. cuspidatum; 3200–3300 m forP. weyrichii). The critical size for survival was 12 mg dry weight for overwintering current-year seedlings of both species. At 2500 m, the percentage of seedlings that attained the critical size for survival at the end of the first growing season was 19% and 85% forP. cuspidatum andP. weyrichii, respectively. Growth curves indicated that the larger-seeded characteristic ofP. weyrichii contributed to larger end-of-season size and resulted in higher rate of seedling survival at 2500 m, compared with smaller-seededP. cuspidatum. Current-year seedlings ofP. weyrichii, at least up to 3250 m, attained the first year annual growth critical for overwintering and therefore for successful seedling establishment, due to the large-seeded characteristic. At the top (3770 m) beyond the upper distribution limit ofP. weyrichii, most seedlings raised artificially failed to attain the critical size for survival in the first growing period due to the reduced growing period. Failure of seedling establishment would thus appear to restrict the distribution of the species at altitudes higher than 3300 m.     

Low allelopathic potential of an invasive forage grass on native grassland plants: a cause for encouragement?

Tall fescue (Festuca arundinacea Schreb.), a highly competitive European grass that invades US grasslands, is reportedly allelopathic to many agronomic plants, but its ability to inhibit the germination or growth of native grassland plants is unknown. In three factorial glasshouse experiments, we tested the potential allelopathic effects of endophyte-infected (E+) and uninfected (E−) tall fescue on native grasses and forbs from Midwestern tallgrass prairies. Relative to a water control, at least one extract made from ground seed, or ground whole plant tissue of E+ or E− tall fescue reduced the germination of 10 of 11 species in petri dishes. In addition, the emergence of two native grasses in potting soil was lower when sown with E+ and E− tall fescue seedlings than when sown with seeds of conspecifics or tall fescue. However, when seeds of 13 prairie species were sown in sterilized, field-collected soil and given water or one of the four tall fescue extracts daily, seedling emergence was lower in one extract relative to water for only one species, and subsequent height growth did not differ among treatments for any species. We conclude that if tall fescue is allelopathic, its inhibitory effects on the germination and seedling growth of native prairie plants are limited, irrespective of endophyte infection. On the other hand, the apparent inability of these plants to detect tall fescue in field soil could hinder prairie restoration efforts if germination near this strong competitor confers fitness consequences. We propose that lack of chemical recognition may be common among resident and recently introduced non-indigenous plants because of temporally limited ecological interactions, and offer a view that challenges the existing allelopathy paradigm. Lastly, we suggest that tall fescue removal will have immediate benefits to the establishment of native grassland plants.