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Poisoning of livestock in oregon in the 1940s to 1960s attributed to corynetoxins produced by Rathayibacter in nematode galls in chewings fescue (Festuca nigrescens).

Tunicaminyluracil antibiotics, similar to the corynetoxins produced by Rathayibacter toxicus in Australia and South Africa, were found in old nematode seed-galls from Festuca nigrescens from New Jersey (USA) and New Zealand (NZ). The toxin profiles from the NZ and USA galls were similar to each other, but differed from those produced by R toxicus from Australia and South Africa, suggesting that a geographical variant of R toxicus or closely related species may be involved. The NZ galls gave a positive response to a R toxicus-specific monoclonal antibody assay, albeit a considerably weaker response than that seen with Australian R toxicus galls, but the older USA galls were negative, possibly due to deterioration of the antigen. From these findings, it is postulated that livestock deaths associated with the feeding of nematode and bacterial infected screenings of F nigrescens in Oregon, USA, in the 1940s to 1960s were caused by corynetoxin-like toxins produced by the bacterium.

Actinomycetales↗

Intergeneric somatic hybridization in Gramineae: somatic hybrid plants between tall fescue (Festuca arundinacea Schreb.) and Italian ryegrass (Lolium multiflorum Lam.).

Tall fescue (Festuca arundinacea Schreb.) protoplasts, inactivated by iodoacetamide, and non-morphogenic Italian ryegrass (Lolium multiflorum Lam.) protoplasts, both derived from suspension cultures, were electrofused and putative somatic hybrid plants were recovered. Two different genotypic fusion combinations were carried out and several green plants were regenerated in one of them. With respect to plant habitus, leaf and inflorescence morphology, the regenerants had phenotypes intermediate between those of the parents. Southern hybridization analysis using a rice ribosomal DNA probe revealed that the regenerants contained both tall fescue- and Italian ryegrass-specific-DNA fragments. A cloned Italian ryegrass-specific interspersed DNA probe hybridized to total genomic DNA from Italian ryegrass and from the green regenerated somatic hybrid plants but not to tall fescue. Chromosome counts and zymograms of leaf esterases suggested nuclear genome instability of the somatic hybrid plants analyzed. Four mitochondrial probes and one chloroplast DNA probe were used in Southern hybridization experiments to analyze the organellar composition of the somatic hybrids obtained. The somatic hybrid plants analyzed showed tall fescue, additive or novel mtDNA patterns when hybridized with different mitochondrial gene-specific probes, while corresponding analysis using a chloroplast gene-specific probe revealed in all cases the tall fescue hybridization profile. Independently regenerated F. arundinacea (+) L. multiflorum somatic hybrid plants were successfully transferred to soil and grown to maturity, representing the first flowering intergeneric somatic hybrids recovered in Gramineae.

Cells, Cultured↗

Transformation of Acremonium coenophialum, a protective fungal symbiont of the grass Festuca arundinacea.

Acremonium coenophialum is a mutualistic mycosymbiont and natural agent of biological protection of the widely distributed grass Festuca arundinacea (tall fescue). An electroporative transformation system was developed for A. coenophialum. Segments of DNA 5' to the beta-tubulin gene (tub2) of the closely related ascomycete Epichloë typhina, fused to the Escherichia coli hph gene encoding hygromycin B phosphotransferase, conferred hygromycin resistance when introduced into A. coenophialum by electroporation. The incorporation of the Emericella nidulans trpC terminator greatly increased protoplast germination on selective medium and improved transformation efficiencies 30-200% depending on the plasmid construct. Plasmid pCSN43, which incorporates the trpC controlling elements for hph expression, was also used to transform A. coenophialum. Southern blot analysis of ten pCSN43 transformants indicated the possibility of random integration of this vector into the genome.

Acremonium↗

The chromosomes of Festuca pratensis Huds. (Poaceae): fluorochrome banding, heterochromatin and condensation.

The karyotype of meadow fescue Festuca pratensis Huds. (2n = 14) has been studied by means of fluorescence banding techniques. AT-enriched heterochromatic bands were revealed in the metaphase chromosomes 1-6. A single GC-rich band was associated with the chromosome 2 secondary constriction. Delay of condensation of the heterochromatic regions was discovered. The heterochromatic bands reported are markers which allow identification of all meadow fescue chromosomes.

Chromosome Banding↗

Genome plasticity in Festuca arundinacea: direct response to temperature changes by redundancy modulation of interspersed DNA repeats.

The response of the genome of Festuca arundinacea seedlings to changes in the temperature at which they were grown was investigated. Fifteen repeated sequences in the nuclear DNA were isolated and hybridized to the genomic DNA of seedlings grown at 10 degrees C or 30 degrees C. The redundancies of sequences recognized by four probes ( FaA5, FaH8, FaH13 and FaH14), were found to differ significantly in the two DNAs. DNA sequences recognized by FaH8, FaH13 and FaH14 were more represented in the genome of the 30 degrees C-raised seedlings than in the genome of the 10 degrees C-raised seedlings (76.5 x 10(3), 1.9 x 10(3), and 111.8 x 10(3) copies per haploid, 1C genome vs 62.7 x 10(3), 1.3 x 10(3), and 80.8 x 10(3) copies, respectively). In contrast, FaA5-related sequences were more represented in the genome of seedlings grown at the lower temperature (15.5 x 10(3) vs 10.2 x 10(3) copies, respectively). Southern-blot hybridization of these repeats to digested genomic DNA produced patterns which indicated that the probe sequences were part of longer repeated sequences having a limited degree of structural heterogeneity. These patterns were partly different when the probes were hybridized to the DNA from seedlings grown at 10 degrees C or 30 degrees C. In situ hybridization showed that the DNA sequences recognized by each probe were scattered along the length of all the chromosomes, with preferential location of FaA5- and FaH13-related sequences at given, mainly centromeric, regions of certain chromosomes. These findings suggest that redundancy modulations of interspersed repeated sequences allow direct responses of the genome of F. arundinacea to changes in environmental temperature.

Journal Article↗

Effects of water stress on selenium accumulation in tall fescue (Festuca arundinacea Schreb) from a selenium-contaminated soil.

A greenhouse experiment was conducted evaluating the effects of water stress on uptake and accumulation of selenium (Se) as well as on uptake of calcium (Ca), potassium (K), and magnesium (Mg) in tall fescue (Festuca arundinacea Schreb). Also, this study has documented changes in soil Se inventory of Se-contaminated field soil. The findings of this experiment suggest that low soil moisture and plant water stress had a positive effect on plant tissue Se concentration. However, the negative effect of water stress on plant growth superimposed the increase of plant tissue Se concentration. Therefore, Se accumulation and soil Se removal by tall fescue became largely dependent on plant dry weight production. Strategies utilizing for remediation of Se-contaminated soils should maximizing the growth rates of tall fescue by maintaining soil moisture levels above water-stress conditions (greater than -1.0 MPa). The results of this study also suggest that the relative sensitivity of net uptake of Se and the three macronutrients to reduced soil moisture is: Ca uptake < Se uptake < Mg uptake < K uptake.

Calcium↗

Genetic and physical analysis of a single Festuca pratensis chromosome segment substitution in Lolium perenne.

Molecular marker analysis and genomic in situ hybridisation (GISH) were used to examine the process of chromosome segment introgression in BC2 diploid hybrids (2n=2x=14) between Lolium perenne and Festuca pratensis. Two genotypes having what appeared to be the same, single, introgressed chromosome segment of F. pratensis in the L. perenne background were crossed with diploid L. perenne to produce a recombinant series for the introgressed region. Physical and genetic analysis of this series showed that, while recombination seemed to be possible at all points along the chromosome arm, the rate of recombination varied depending on relative position: more recombination was detected in the interstitial region as compared with the centromeric or telomeric regions. The implications of these results for the use of GISH and molecular marker analysis in the measurement of linkage drag in backcross breeding programmes is discussed.

Alleles↗

Linear, steroidal, and triterpene esters, and steryl glycosides from Festuca argentina.

Ester waxes and steryl glycosides of the grass Festuca argentina were studied. Saponification of the waxes from the petroleum ether extract led to n-hexacosanol as the major single linear alcohol, along with pentacyclic triterpenols, such as beta-amyrin, germanicol, isobaurenol, lupeol, hopenol-a and hopeol, and low amounts of sterols, such as cholesterol, campesterol, stigmasterol, sitosterol and dihydrositosterol, identified by gas chromatography/mass spectrometry (GC/MS). Fatty acids were identified as methyl esters as C12:0, C14:0, C16:0, C18:0, C18:2, and C20:0. The occurrence of a wide chainlength range of fatty acids and a single linear alcohol closely matched for other reports on the tribe Festuceae. On the contrary, pentacyclic triterpenols with a variety of skeletons, especially isobauerenol, are not usual as esters of fatty acids in the Gramineae. Low amounts of steryl glycosides were also obtained from the methylene chloride percolate of the methanol extract. Upon acetylation followed by hydrolysis, aglycones were identified by capillary gas-liquid chromatography (GLC) and GC/MS. As delta 7-cholesterol, campesterol, stigmasterol, sitosterol, dihydrositosterol, and the sugars as glucose, xylose, and arabinose by GLC of the respective alditol acetates. This is the first report on the linear, steryl, and triterpenyl esters of F. argentina. It is noteworthy that delta 7-steryl glycosides are rare, and steryl monoarabinosides have not been previously reported on the family Gramineae.

Alcohols↗

The effects of zinc on cell viability and on mitochondrial structure in contrasting cultivars of Festuca rubra L. - a rapid test for zinc tolerance.

A 20-min exposure to 5.0 microg Zn cm(-3) reduced the percentage of viable root meristematic cells in three cultivars of Festuca rubra L.: Merlin (Zn-tolerant), Hawk (salt-tolerant but with a degree of Zn tolerance) and S59 (Zn-sensitive). The Zn-induced cell mortality in S59 was approximately twice that of the tolerant cultivars. The mean area of mitochondrial profiles in root meristematic cells of Zn-untreated roots was similar in S59 and Merlin but that of Hawk was smaller. A 4-day exposure to 0.2 microg Zn cm(-3) resulted in mitochondrial swelling in the Zn-sensitive cultivar; there was a 25% increase in the mean area of mitochondrial profiles in this cultivar, but no significant increase occurred in Hawk or Merlin. Zn treatment caused a collapse of the cristae and a localized condensation of the mitochondrial matrix in S59, but not in Hawk or Merlin. The marked increase in cell mortality after only a 20-min Zn exposure and the relative simplicity of the technique, indicates that this procedure could be used as a rapid and independent measure of Zn tolerance.

Journal Article↗

Volatile compounds of endophyte-free and infected tall fescue (Festuca arundinacea Schreb.).

Volatile compounds produced by intact plants and ground leaf tissue from endophyte-infected (E+) and endophyte-free (E-) tall fescue (Festuca arundinacea Schreb.) were collected by a purge-and-trap procedure and analyzed by gas chromatography/mass spectrometry The volatile compound profile from ground leaf tissue was similar between E+ and E- clonal plants; however, the sheaths of E+ clonal plants produced higher levels of 1-octen-3-ol, a characteristic volatile compound derived from lipid peroxidation in fungi, which was absent in E- clonal plants. Intact plants produced fewer volatiles than macerated leaves. At 25 degrees C, (Z)-3-hexen-1-ol acetate was the most abundant compound, accounting for 77 and 89% of the total volatile emission from E+ and E- plants, respectively. Higher temperature (32 degrees C) significantly reduced the production of (Z)-3-hexen-1-ol acetate. Nonanal was the most abundant compound at 32 degrees C accounting for 52 and 45% of the total volatile emission from E+ and E- plants. Treatment of E+ and E- plants with jasmonic acid (JA) dramatically altered the volatile compound profile. The levels of (E)-beta-ocimene increased more than 200-fold and accounted for at least 43% of the total volatile emission. Although the presence of endophyte resulted in some qualitative and quantitative differences in the production of volatile compounds, they are unlikely to account for the differences in insect resistance between E+ and E- plants. Nevertheless, the production of a unique spectrum of volatiles after JA treatment may represent a significant plant-based defense response in tall fescue that is independent of endophyte.

Claviceps↗

Alkaloids from endophyte-infected Festuca argentina.

Five loline-type alkaloids, the known lolinine, N-formylloline, loline, N-methylloline and the new 5,6-dehydro-N-acetylloline, were characterized from the pooid-grass endophyte-infected Festuca argentina. These compounds are not responsible for the toxicity of the plant, as shown upon i.p. injection into mice. Their probable phytoalexine role and chemotaxonomical significance are discussed.

Acremonium↗

Effects of elevated ultraviolet radiation and endophytic fungi on plant growth and insect feeding in Lolium perenne, Festuca rubra, F. arundinacea and F. pratensis.

Plants of perennial ryegrass (Lolium perenne L.), red fescue (Festuca rubra L.), tall fescue (F. arundinacea Schreb.) and meadow fescue (F. pratensis Huds) were exposed at an outdoor facility located in Edinburgh, UK to modulated levels of UV-B radiation (280-315 nm) using banks of cellulose diacetate filtered UV-B fluorescent lamps that also produce UV-A radiation (315-400 nm). The plants were derived from a single clone of each species and were grown both with and without colonization by naturally-occurring fungal endophytes. The UV-B treatment was a 30% elevation above the ambient erythemally-weighted level of UV-B during July to October. Growth of treated plants was compared with plants grown under elevated UV-A radiation alone produced by banks of polyester filtered lamps and with plants grown at ambient levels of solar radiation under banks of unenergized lamps. At the end of the treatment period, sample leaves were collected for feeding trials with the desert locust Schistocerca gregaria (Forsk). The UV-B treatment produced no effects on the aboveground biomass of any of the four grasses. The UV-B treatment and the UV-A control exposure both increased plant height and the number of daughter plants formed by rhizome growth in F. rubra. There were significant effects of endophyte presence on the total fresh and dry weights of F. arundinacea and F. rubra, on fresh weight only in F. pratensis, and on the fresh and dry weights of inflorescence in F. arundinacea and L. perenne. There were no effects of UV treatments on the absolute amounts of leaf consumed or on the feeding preferences of locusts for leaves with or without endophyte in three species: F. rubra, F. arundinacea and L. perenne. In F. pratensis there was no effect of UV treatment on the weight of leaves consumed but a significant UV x endophyte interaction caused by a marked change in feeding preference between leaves with and without endophyte that differed between the UV-B treatment and UV-A control exposures. The alkaloid compounds known as lolines were analysed in leaves of F. pratensis and were only found in plants grown with endophyte. However, there was no significant relationship between total loline content and insect feeding preference. These effects illustrate the potential complexities of species interactions under increasing levels of UV-B. The experiment also demonstrates the importance of appropriate controls in UV lamp supplementation experiments for interpretation of both plant growth and insect feeding effects.

Animals↗

A DNA marker for the duplicated cytosolic PGI genes in sheep's fescue (Festuca ovina L.).

Active duplicate PgiC genes in sheep's fescue, Festuca ovina, are associated with a PCR marker of specific length (about 370 bp, of which 231 are in an intron). Using this marker, the frequency of plants with duplicate genes is estimated to be about 10% in a population from southern Sweden. The close molecular similarity between the electrophoretically different duplicated genes is in accordance with the conclusion reached earlier that they are indeed alleles at the same locus.

Alleles↗

Effect of phenolic structures on the degradability of cell walls isolated from newly extended apical internode of tall fescue (Festuca arundinacea Schreb.).

Apical internodes of tall fescue (Festuca arundinacea Schreb. var. Clarine) harvested at flowering were sectioned into 5 or 10 equal parts to study in situ degradability and cell wall composition, respectively. The basal (youngest) section had the greatest primary wall content. Cell walls in the upper (older) sections had the highest xylose/arabinose ratio and lignin content and a lignin rich in syringyl units, all typical of extensive secondary wall development. Almost all of the p-coumaric (p-CA) and about half of the ferulic acid (FA) were released by 1 M NaOH and presumed to be ester-linked. The total FA content was approximately double that of p-CA in all sections other than the youngest with a distribution similar to that of total p-CA. However, the ratio of esterified to ether and ether plus ester linked (Et & Et+Es) FA differed with age. Whereas the esterified form remained essentially constant ( approximately 4.5 g/kg of cell wall), Et & Et+Es ferulate increased with increasing age of the tissue and was significantly related to lignin deposition (r = 0.79, P < 0.01). The extent of cell wall degradation after 48 h of incubation in the rumen was inversely related to maturity, falling from 835 g/kg of dry matter in the youngest section to 396 g/kg in the oldest. Both the rate and extent of cell wall degradation were significantly negatively related to the ratio of xylose to arabinose, lignin content, proportion of syringyl units present in lignin, and concentration of Et & Et+Es FA present. A positive relationship between Et & Et+Es FA was also found, with the rate (P < 0.01) being better correlated than the extent (P < 0.05) of cell wall degradation. Application of the newly extended internode model to fescue produced results consistent with the view that both the lignin content and the extent to which lignin was covalently bound to the other wall polymers crucially influenced the rate and extent of degradation.

Animal Feed↗

Differential expression of alpha- and beta-expansin genes in the elongating leaf of Festuca pratensis.

Grasses contain a number of genes encoding both alpha- and beta-expansins. These cell wall proteins are predicted to play a role in cell wall modifications, particularly during tissue elongation. We report here on the characterisation of five alpha- and three vegetative beta-expansins expressed in the leaf elongation zone (LEZ) of the forage grass, Festuca pratensis Huds. The expression of the predominant alpha-expansin (FpExp2) was localised to the vascular tissue, as was the beta-expansin FpExpB3. Expression of another beta-expansin (FpExpB2) was not localised to vascular tissue but was highly expressed in roots and initiating tillers. This is the first description of vegetative beta-expansin gene expression at the organ and tissue level and also the first evidence of differential expression between members of this gene family. In addition, an analysis of both alpha- and beta-expansin expression along the LEZ revealed no correlation with growth rate distribution, whereas we were able to identify a novel xyloglucan endotransglycosylase (FpXET1) whose expression profile closely mimicked leaf growth rate. These data suggest that alpha- and beta-expansin activities in the grass leaf are associated with tissue differentiation, that expansins involved in leaf growth may represent more minor components of the spectrum of expansin genes expressed in this tissue, and that XETs may be useful markers for the analysis of grass leaf growth.

Amino Acid Sequence↗

Reduced atmospheric CO2 inhibits nitrogen mobilization in Festuca rubra.

In defoliated grasses, where photosynthesis is reduced due to removal of leaf material, it is well established that remobilization of nitrogen occurs from both older remaining leaves and roots towards the younger growing leaves. In contrast, little is known about the movement of nitrogen within intact grass plants experiencing prolonged inhibition of photosynthesis. We tested the following hypotheses in Festuca rubra L. ssp. rubra cv. Boreal: that both reduction of the atmospheric CO2 concentration and defoliation (1) induce mobilization of nitrogen from roots and older leaves towards growing leaves and (2) elicit similar directional change in the abundance of proteins in roots and older leaves relevant to the process of nitrogen mobilization including, glutamine synthetase (GS), EC 6.3.1.2; papain, EC 3.4.22.2; chymopapain, EC 3.4.22.6; ribulose bisphosphate carboxylase/oxygenase (Rubisco), EC 4.1.1.39; and the light harvesting complex of photosystem II (LHCPII). After growth at ambient atmospheric CO2 concentration, plants of F. rubra were subject to atmospheres containing either ambient (350 micro l l-1) or deplete (< 20 micro l l-1) CO2. Concurrently, plants were either left intact or defoliated on one occasion. Steady state 15N labelling coupled with a series of destructive harvests over a 7-day period enabled changes in the nitrogen dynamics of the plants to be established. Proteins pertinent to the process of nitrogen mobilization were quantified by immunoblotting. Irrespective of defoliation, plants in ambient CO2 mobilized nitrogen from older to growing leaves. This mobilization was inhibited by deplete CO2. Greater concentration of Rubisco and reduced chymopapain abundance in older remaining leaves of intact plants, in deplete compared with ambient CO2, suggested the inhibition of mobilization was due to inhibition of protein degradation, rather than to the export of degradation products. Both deplete CO2 and defoliation induced nitrogen mobilization from roots to growing leaves. In plants at ambient CO2, defoliation did not affect nitrogen uptake or its allocation. Therefore in F. rubra nitrogen mobilization can occur independently of any downregulation of nitrogen uptake. This suggests either different signal compounds may act to downregulate uptake and upregulate mobilization, or if one particular signalling compound is used its concentration threshold differs for induction of mobilization and downregulation of uptake. The abundance of the cysteine proteases papain and chymopapain was low in roots suggesting that they were not involved in protein degradation in this tissue.

Journal Article↗

Transgenic plants of tall fescue (Festuca arundinacea Schreb.) obtained by direct gene transfer to protoplasts.

Chimeric hygromycin phosphotransferase (hph) and phosphinothricin acetyltransferase (bar) genes were introduced, using polyethylene glycol treatment, into protoplasts isolated from embryogenic cell suspension cultures of tall fescue (Festuca arundinacea Schreb.), a graminaceous plant that is an important forage crop in temperate pastures. Colonies resistant to either 200 mg/l hygromycin or 100 mg/l phosphinothricin, respectively, were recovered upon selection using bead-type culture systems. Stable integration of the transgenes in the genomes of plants regenerated from resistant callus clones was shown by Southern hybridization analysis. In situ hybridization of a labeled transgene-probe to metaphase chromosomes is shown for one transgenic primary regenerant. Expression of the transgenes in mature plants was demonstrated by HPH enzyme assay or by phosphinothricin-herbicide spraying.

Acetyltransferases↗