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PubMed2006

Chromosome pairing in triploid intergeneric hybrids of Festuca pratensis with Lolium multiflorum, revealed by GISH.

Genomic in situ hybridisation (GISH) was used to reveal chromosome pairing in two partly fertile, triploid (2n = 3x = 21) hybrids obtained by crossing the diploid (2n = 2x = 14) Festuca pratensis Huds. (designated FpFp), used as a female parent, with the autotetraploid (2n = 4x = 28) Lolium multiflorum Lam. (designated LmLmLmLm), used as a male parent. The pattern of chromosome pairing calculated on the basis of the mean values of chromosome configurations identified in all 100 PMCs analysed, was: 0.71I Lm + 2.24I Fp + 2.18II Lm/Lm + 0.54II Lm/Fp + 4.18III Lm/Lm/Fp. A relatively high number of Lm/Lm bivalents and Fp univalents, and a low number of Lm/Fp bivalents and Lm univalents indicated that the pairing was preferential between L. multiflorum chromosomes. Other observations regarding chromosome pairing within the Lm/Lm/Fp trivalents also confirmed this preferential pairing in the analysed triploids, as the Fp chromosome was not randomly located in the chain- and frying-pan-shaped trivalents. The similarities and differences in chromosome pairing at metaphase I and the level of preferential pairing between Lolium chromosomes in the different triploid Lolium-Festuca hybrids are discussed.

Chromosome Pairing↗
PubMed2004

An SSR- and AFLP-based genetic linkage map of tall fescue (Festuca arundinacea Schreb.).

Tall fescue (Festuca arundinacea Schreb.) is commonly grown as forage and turf grass in the temperate regions of the world. Here, we report the first genetic map of tall fescue constructed with PCR-based markers. A combination of amplified fragment length polymorphisms (AFLPs) and expressed sequence tag-simple sequence repeats (EST-SSRs) of both tall fescue and those conserved in grass species was used for map construction. Genomic SSRs developed from Festuca x Lolium hybrids were also mapped. Two parental maps were initially constructed using a two-way pseudo-testcross mapping strategy. The female (HD28-56) map included 558 loci placed in 22 linkage groups (LGs) and covered 2,013 cM of the genome. In the male (R43-64) map, 579 loci were grouped in 22 LGs with a total map length of 1,722 cM. The marker density in the two maps varied from 3.61 cM (female parent) to 2.97 (male parent) cM per marker. These differences in map length indicated a reduced level of recombination in the male parent. Markers that revealed polymorphism within both parents and showed 3:1 segregation ratios were used as bridging loci to integrate the two parental maps as a bi-parental consensus. The integrated map covers 1,841 cM on 17 LGs, with an average of 54 loci per LG, and has an average marker density of 2.0 cM per marker. Homoeologous relationships among linkage groups of six of the seven predicted homeologous groups were identified. Three small groups from the HD28-56 map and four from the R43-64 map are yet to be integrated. Homoeologues of four of those groups were detected. Except for a few gaps, markers are well distributed throughout the genome. Clustering of those markers showing significant segregation distortion (23% of total) was observed in four of the LGs of the integrated map.

Chromosome Mapping↗
PubMed2005

A decade of "chromosome painting" in Lolium and Festuca.

GISH has been a particularly useful technique for studying the Lolium-Festuca species complex of forage grasses. The reason for this utility is two-fold: (i) the complex is unique amongst crop plants in which fertile hybrids, and backcross progenies, can be produced which recombine genomes and promiscuously exchange their genes through homoeologous recombination; (ii) dispersed repetitive DNAs differ between species, and this allows tracking of the identity of chromosomes and chromosome segments. This tracking property has enabled several fruitful lines of research to produce a harvest of new information for both fundamental and practical purposes. We review this first decade of GISH (genomic in situ hybridization) in Lolium-Festuca, and discuss and summarize the achievements which have accrued.

Chromosome Painting↗
PubMed2005

[Research on Festuca arundinacea transformation mediated by Agrobacterium tumefaciens].

The embryo-derived calli from four types of tall fescues (Festuca arundinacea Schreb) were transformed with two Agrobactrium tumefaciens strains AGL1 and GV3101. AGL1 harbors an intron-AtNHX1 expression vector pROK2U containing ubiqutin promoter and npt II marker gene. GV3101 harbors an intron-AtNHX1 expression vector pROK2 containing 35S promoter and npt II gene. After infection and co-culture with AGL1 or GV3101, the embyogenic calli were selected with 50-150 mg/L paromomycine and 1126 resistant plants regenerated from the resistant calli. All plants were selected further with 10-20 mg/L Kanamycin and 525 of them remained green. Genome DNA of the resistant plants was checked with specific primers and probe from AtNHX1 gene. The results of PCR assay and Southern blot analysis indicted that exogenous target gene (AtNHX1 gene) had been transferred into different cultivars of Festuca arundinacea. Different transformation frequencies among the four cultivars were obtained.

Agrobacterium tumefaciens↗
PubMed1996

Susceptibilities of Different Test Systems from Maize (Zea mays), Poa annua, and Festuca rubra to Herbicides That Inhibit the Enzyme Acetyl-Coenzyme A Carboxylase

The susceptibilities of maize (Zea mays cv. Champ) and two graminicide-resistant grass species, Poa annua (annual meadow grass) and Festuca rubra (red fescue), to two aryloxyphenoxypropionates (quizalofop and fluazifop) and a cyclohexanedione (sethoxydim) graminicide were evaluated in leaf blades and isolated chloroplasts, and by assaying acetyl-coenzyme A carboxylase (ACCase) in desalted leaf homogenates. The graminicide resistance of P. annua and F. rubra appeared to be at the level of ACCase. Festuca rubra ACCase was highly insensitive and P. annua ACCase was partially insensitive to the graminicides that were tested. Fatty acid synthesis in isolated maize chloroplasts was more susceptible to inhibition than was ACCase activity from whole leaves. There was a smaller difference in graminicide sensitivity between these two test systems in P. annua. The developmental pattern of ACCase specific activity and its inhibition by quizalofop was measured in maize and P. annua leaf blades. There was an age-dependent increase in the sensitivity of maize leaf ACCase activity to inhibition by quizalofop. Together with the greater susceptibility of chloroplasts compared with leaf homogenates this could imply that a graminicide-insensitive (extrachloroplastic) ACCase isoform is less highly expressed in older leaves. Poa annua ACCase did not significantly alter in sensitivity as leaves aged, consistent with the smaller difference in the level of inhibition between chloroplasts and leaf homogenates in this species. A small pyruvate carboxylase activity was detected in maize leaves after 9 days. By 38 days, when leaves were senescing, pyruvate carboxylase activity predominated over ACCase.

Journal Article↗
PubMed2001

Molecules and morphology in concert. II. The Festuca brachyphylla complex (Poaceae) in Svalbard.

We used a combined molecular and morphological approach to unravel variation in the autogamous Festuca brachyphylla polyploid complex in the arctic archipelago of Svalbard. Forty populations were analyzed for random amplified polymorphic DNAs (RAPDs) and 46 morphological characters. Eighteen RAPD multilocus phenotypes were observed in the 86 plants analyzed, based on 30 polymorphic markers. Multivariate analyses of the RAPD data revealed four distinct groups of multilocus phenotypes; in contrast, the variation was more or less continuous in multivariate analyses of the morphological data. However, we identified several individual morphological characters that unambiguously discriminated among the four groups of RAPD multilocus phenotypes. Analysis of type material suggests that the four groups in Svalbard can be referred to Festuca baffinensis, F. brachyphylla, F. hyperborea, and F. edlundiae. This study shows that concerted analysis of molecules and morphology is a powerful tool in low-level taxonomy.

Journal Article↗
PubMed2002

[Quantitative characteristics of degenerative succession in Festuca sinensis sowing grassland in the alpine pastoral area].

Quantitative characteristics of degenerative succession in Festuca sinensis sowing grassland in the alpine pastoral area were studied. The results showed that with the succession developing, number of species, richness index, diversity index, and evenness index were all increased, but ecological dominance was decreased. Analysis on important values, niche width, and niche overlaps proved that some herb species such as Ajania tenuifolia and Polygomum sibiricum were pioneer plants invading sowing grassland in the succession process. The pioneer plants had wider niche widths than other species, and their important values increased with the succession developing. Niche overlaps between Festuca sinensis and other species were smaller than that between two latifoliate herbs species in most case. The invasion of latifoliate herbs was one of the most important factors, which made the sowing grassland more degenerative.

Cold Climate↗
PubMed2001

Meiosis and fertility of reciprocal triploid hybrids of Lolium multiflorum with Festuca pratensis.

Diploid and tetraploid forms of Lolium multiflorum and Festuca pratensis were crossed under controlled conditions and after embryo rescue all four combinations of autoallotriploid hybrids were obtained. Male and female fertility and chromosome pairing at metaphase I of meiosis were studied in several plants from each hybrid combination. The hybrids with two genomes of L. multiflorum and one of F. pratensis (genomic formulae LmLmFp and FpLmLm) were male and female fertile while the hybrids with two genomes of F. pratensis and one of L. multiflorum had a reduced fertility (FpFpLm) or were completely sterile (LmFpFp). Chromosome pairing at metaphase I varied among hybrid combinations depending on their genomic composition. LmLmFp and FpLmLm hybrids had similar patterns of pairing (1.83I + 5.29II + 2.85III and 2.22I + 5.22II + 2.75III, respectively) but they differed from those of FpFpLm (3.65I + 4.65II + 2.68III) and especially from LmFpFp (4.78I + 5.87II + 1.49III). Conventional analysis of meiosis failed to explain the differences in chromosome behaviour and fertility/sterility levels between the autoallotriploid hybrids with two Lolium or two Festuca genomes.

Journal Article↗
PubMed2003

Field performance of transgenic tall fescue (Festuca arundinacea Schreb.) plants and their progenies.

Tall fescue (Festuca arundinacea Schreb.) is a hexaploid, outcrossing grass species widely used for forage and turf purposes. Transgenic tall fescue plants were generated by biolistic transformation of embryogenic cell suspension cultures that were derived from single genotypes of widely used cultivar Kentucky-31. Primary transgenics from two genotypes, their corresponding regenerants from the same genotypes and control seed-derived plants were transferred to the field and evaluated for 2 years. Progenies of these three classes of plants were obtained and evaluated together with seed-derived plants in a second field experiment. The agronomic characteristics evaluated were: heading date, anthesis date, height, growth habit, number of reproductive tillers, seed yield and biomass. The agronomic performance of the primary transgenics and regenerants was generally inferior to that of the seed-derived plants, with primary transgenics having fewer tillers and a lower seed yield. However, no major differences between the progenies of transgenics and the progenies of seed-derived plants were found for the agronomic traits evaluated. Primary transgenics and regenerants from the same genotype were more uniform than plants from seeds. Progenies of transgenics performed similarly to progenies of the regenerants. The addition of a selectable marker gene in the plant genome seems to have had little effect on the agronomic performance of the regenerated plants. No indication of weediness of the transgenic tall fescue plants was observed. Our results indicate that outcrossing grass plants generated through transgenic approaches can be incorporated into forage breeding programs.

Biolistics↗
PubMed2003

A linkage map of meadow fescue ( Festuca pratensis Huds.) and comparative mapping with other Poaceae species.

A genetic linkage map has been constructed for meadow fescue ( Festuca pratensis Huds.) (2n=2 x=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.

Chromosome Mapping↗
PubMed2005

Construction of a Festuca pratensis BAC library for map-based cloning in Festulolium substitution lines.

Introgression in Festulolium is a potentially powerful tool to isolate genes for a large number of traits which differ between Festuca pratensis Huds. and Lolium perenne L. Not only are hybrids between the two species fertile, but the two genomes can be distinguished by genomic in situ hybridisation and a high frequency of recombination occurs between homoeologous chromosomes and chromosome segments. By a programme of introgression and a series of backcrosses, L. perenne lines have been produced which contain small F. pratensis substitutions. This material is a rich source of polymorphic markers targeted towards any trait carried on the F. pratensis substitution not observed in the L. perenne background. We describe here the construction of an F. pratensis BAC library, which establishes the basis of a map-based cloning strategy in L. perenne. The library contains 49,152 clones, with an average insert size of 112 kbp, providing coverage of 2.5 haploid genome equivalents. We have screened the library for eight amplified fragment length polymorphism (AFLP) derived markers known to be linked to an F. pratensis gene introgressed into L. perenne and conferring a staygreen phenotype as a consequence of a mutation in primary chlorophyll catabolism. While for four of the markers it was possible to identify bacterial artificial chromosome (BAC) clones, the other four AFLPs were too repetitive to enable reliable identification of locus-specific BACs. Moreover, when the four BACs were partially sequenced, no obvious coding regions could be identified. This contrasted to BACs identified using cDNA sequences, when multiple genes were identified on the same BAC.

Blotting, Southern↗
PubMed2005

Quantitative trait loci controlling vernalisation requirement, heading time and number of panicles in meadow fescue (Festuca pratensis Huds.).

The "BF14/16xHF2/7" mapping population of meadow fescue (Festuca pratensis Huds.) was characterised for number of panicles produced by non-vernalised plants in the field, vernalisation requirement (number of weeks at 6 degrees C and 8 h photoperiod), as well as days to heading, number of panicles and proportion of shoots heading after a 12 weeks vernalisation treatment. Quantitative trait loci (QTLs) were identified and compared to QTLs and genes related to the induction of flowering in cereals and grasses. A region on chromosome 1F affected days to heading and the proportion of shoots heading. Chromosome 4F appeared to have several genes with a strong effect on vernalisation requirement. The strongest effects were located in the proximal end of 4F and may correspond to the earliness per se (eps) QTL eps6L.2 in barley and a heading time QTL in perennial ryegrass. A part of the meadow fescue orthologue of VRN1 was sequenced and mapped to another region of 4F that also had a strong effect on vernalisation requirement. The proximal end of chromosome 5F had QTLs for days to heading and proportion of heading shoots. Syntenic regions in wheat and barley contain eps-loci. A QTL for number of panicles in the field and a QTL for proportion of heading shoots were present on chromosome 6. A region on 7F affected the variation in number of panicles among plants without a vernalisation requirement, and is syntenic to regions in perennial ryegrass, barley and rice containing orthologues of Arabidopsis thaliana CO.

Arabidopsis Proteins↗
PubMed2005

Molecular diversity of local Norwegian meadow fescue (Festuca pratensis Huds.) populations and Nordic cultivars-consequences for management and utilisation.

Genetic diversity and relatedness were studied in 30 Norwegian local populations of meadow fescue (Festuca pratensis Huds.) using amplified fragment length polymorphism (AFLP) markers. The populations were also compared with 13 Nordic meadow fescue cultivars in order to analyse the distribution of variation in local populations and cultivars and to elucidate relationships between local populations and cultivars. Analysis of molecular variance (AMOVA: ) analysis showed that most of the variation was present within populations and that little variation was found between local populations and cultivars. Separate AMOVA: analyses of local populations and cultivars revealed a higher level of variation within registered cultivars than within local populations. A cluster analysis based on corrected average pairwise differences between populations showed that the populations could be divided into three clusters, of which one also contained the cultivars. These results were supported by principal coordinates analysis. The results indicate that the Norwegian meadow fescue has a narrow genetic basis and that the local populations in Norway can be divided into three groups following the most probable routes of introduction of the species into Norway. The inland populations are closely related to the cultivars and have most probably been established as a result of migration from sown meadows. The western and southern populations probably originate from human activity-for example, trade-to the coastal western and northern parts of the country and to the central parts of southern Norway.

Analysis of Variance↗
PubMed2006

Genome constitution and evolution in Lolium x Festuca hybrid cultivars (Festulolium).

Festulolium hybrids are being increasingly used worldwide as forage grasses. This is due to their superior agronomic characteristics, which combine yield performance of ryegrasses (Lolium multiflorum and L. perenne) and tolerance against abiotic stress of fescues (Festuca pratensis, F. arundinacea and F. arundinacea var. glaucescens). Despite the widespread use, only fragmentary information exists on their genomic constitution. We used genomic in situ hybridization (GISH) to analyze genomic constitution of over 600 plants from almost all commercially available cultivars of Festulolium. Our results revealed a surprisingly large range of variation in the proportions of parental genomes and in the extent of intergenomic recombination. Using fluorescence in situ hybridization (FISH) with probes for ribosomal DNA, we assessed the frequency of recombination and elimination of particular chromosomes and chromosome groups in three contrasting Festulolium cultivars. This study provides novel information that will aid in understanding the relationship between a genetic make-up and the phenotype of Festulolium hybrids. Our results indicate that GISH might be a useful tool to aid in Festulolium breeding and provide data for a more detailed description of registered cultivars.

Chromosomes, Plant↗
PubMed2005

Stabilization of metals in acidic mine spoil with amendments and red fescue (Festuca rubra L.) growth.

Stabilization of metals with amendments and red fescue (Festuca rubra, cv. Keszthelyi 2) growth was studied on an acidic and phytotoxic mine spoil (pH(KCl) 3.20-3.26; Cd 7.1 mg kg(-1), Cu 120 mg kg(-1), Pb 2154 mg kg(-1) and Zn 605 mg kg(-1)) from Gyöngyösoroszi, Hungary in a pot experiment. Raising the pH above 5.0 by lime (CaCO(3)), and supplementing with 40 mg kg(-1)nitrogen (NH(4)NO(3)) made this material suitable for plant growth. All cultures were limed with 0.5% (m/m) CaCO(3) (treatment 1), which was combined with 5% (m/m) municipal sewage sludge compost (treatment 2), 5% (m/m) peat (treatment 3), 7.5% (m/m) natural zeolite (clinoptilolite) (treatment 4), and 0.5 (m/m) KH(2)PO(4) (treatment 5). Treatments 1-5 were combined with each other (treatment 6). After 60 days of red fescue growth, pH of the limed mine spoil decreased in all cultures units. Application of peat caused the highest pH decrease (1.15), while decrease of pH was less than 0.23 in treatments 2, 5 or 6. Application of lime significantly reduced concentrations of metals in the 'plant available' fraction of mine spoil compared to non-limed mine spoil. Amendments added to limed mine spoil changed variously the ratio of Cd, Cu, Pb and Zn in exchangeable or 'plant available' fractions, differently influencing the phytoavailability of these metals. Most of the metals were captured in the roots of test plants. Treatment 2 caused the appearance of less Cd in shoots (<0.1 microg g(-1)) or roots (3.11 microg g(-1)), while treatment 5 resulted in the highest Cd concentration (2.13 microg g(-1)) in shoots. Treatments did not influence significantly the Cu accumulation in shoots. The Pb accumulation of roots (44.7 microg g(-1)) was most effectively inhibited by combined treatment, while the highest value (136 microg g(-1)) was found in the culture treated with potassium phosphate. Pb concentration in shoots was below the detection limit, except for treatments 5 and 6. Peat application resulted in higher Zn concentration (448 microg g(-1)) in shoots than other amendments, where these values were around 100 microg g(-1). All amendments influenced positively the dry matter yield of red fescue grown in limed mine spoil, however the application of 0.5 phosphate was less favourable. Liming, application of amendments and growth of red fescue can stabilize metals in acidic and phytotoxic mine spoil, and by phytostabilization they can reduce the risk of metal contamination of the food chain.

Biodegradation, Environmental↗
PubMed2005

Screening the wetland plant species Alisma plantago-aquatica, Carex rostrata and Phalaris arundinacea for innate tolerance to zinc and comparison with Eriophorum angustifolium and Festuca rubra Merlin.

Several wetland plant species appear to have constitutive metal tolerance. In previous studies, populations from contaminated and non-contaminated sites of the wetland plants Typha latifolia, Phragmites australis, Glyceria fluitans and Eriophorum angustifolium were found to be tolerant to high concentrations of metals. This study screened three other species of wetland plants: Alisma plantago-aquatica, Carex rostrata and Phalaris arundinacea for innate tolerance to zinc. The degree of tolerance was compared to known zinc-tolerant E. angustifolium and Festuca rubra Merlin. It was found that A. plantago-aquatica and P. arundinacea did not posses innate tolerance to zinc, but that C. rostrata was able to tolerate elevated levels of zinc, at levels comparable to those tolerated by E. angustifolium and F. rubra Merlin. The findings support the theory that some wetland angiosperm species tend to be tolerant to exposure to high levels of metals, regardless of their origin.

Alisma↗
PubMed2005

Mutational analysis of the active center of plant fructosyltransferases: Festuca 1-SST and barley 6-SFT.

The active center of the glycoside hydrolase family 32 contains the three characteristic motifs (N/S)DPNG, RDP, and EC. We replaced the N-terminal region including the (N/S)DPNG motif of barley 6-SFT (sucrose:fructan 6-fructosyltransferase) by the corresponding region of Festuca 1-SST (sucrose:sucrose 1-fructosyltransferase). The chimeric enzyme, expressed in Pichia, retained the specificity of 6-SFT. Attempts to replace a larger piece at the N-terminus including also the RDP motif failed. A point mutation introduced in the RDP motif of 1-SST abolished enzymatic activity. Interestingly, point mutations of the EC-motif resulted in an enzyme which had lost the capability to form 1-kestose and glucose from sucrose but still accepted 1-kestose, producing fructose and sucrose as well as nystose.

Binding Sites↗
PubMed2005

Agrobacterium-mediated high efficiency transformation of tall fescue (Festuca arundinacea).

Tall fescue (Festuca arundinacea) is the predominant cool-season pasture grass in the USA. Embryogenic calluses were induced from seeds/caryopsis of elite tall fescue cultivars Jesup and Kentucky-31, and were broken up into small pieces and used for Agrobacterium tumefaciens-mediated transformation. Agrobacterium strains LBA4404 and EHA105 harboring pCAMBIA vectors or the super-binary vector pTOK233 were used to infect the embryogenic callus pieces. The number of hygromycin resistant calluses obtained per dish of infected callus pieces was in the range of 2.0-5.8, and the number of transgenic plants recovered per dish of infected callus pieces was in the range of 0.4-1.7. When transformation efficiency was calculated based on the number of transgenic plants recovered and the number of original intact calluses used, the transformation frequency was in the range of 1.9-8.7%. The use of the easily available pCAMBIA vectors could produce equivalent results as the superbinary vector pTOK233. The transgenic nature of the regenerated plants was demonstrated by Southern hybridization analysis using undigested and digested genomic DNA samples. Expression of the transgenes was confirmed by northern hybridization analysis, GUS staining, and detection of GFP signals. Fertile transgenic plants were obtained after vernalization in the greenhouse. Progeny analysis revealed Mendelian inheritance of the transgenes.

Agrobacterium tumefaciens↗