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PubMed2003

Changes in the high-mountain vegetation of the Central Iberian Peninsula as a probable sign of global warming.

Aerial images of the high summits of the Spanish Central Range reveal significant changes in vegetation over the period 1957 to 1991. These changes include the replacement of high-mountain grassland communities dominated by Festuca aragonensis, typical of the Cryoro-Mediterranean belt, by shrub patches of Juniperus communis ssp. alpina and Cytisus oromediterraneus from lower altitudes (Oro-Mediterranean belt). Climatic data indicate a shift towards warmer conditions in this mountainous region since the 1940s, with the shift being particularly marked from 1960. Changes include significantly higher minimum and maximum temperatures, fewer days with snow cover and a redistribution of monthly rainfall. Total yearly precipitation showed no significant variation. There were no marked changes in land use during the time frame considered, although there were minor changes in grazing species in the 19th century. It is hypothesized that the advance of woody species into higher altitudes is probably related to climate change, which could have acted in conjunction with discrete variations in landscape management. The pronounced changes observed in the plant communities of the area reflect the susceptibility of high-mountain Mediterranean species to environmental change.

Ecosystem↗
PubMed2006

Comparison of four nuclear isolation buffers for plant DNA flow cytometry.

BACKGROUND AND AIMS: DNA flow cytometry requires preparation of suspensions of intact nuclei, which are stained using a DNA-specific fluorochrome prior to analysis. Various buffer formulas were developed to preserve nuclear integrity, protect DNA from degradation and facilitate its stoichiometric staining. Although nuclear isolation buffers differ considerably in chemical composition, no systematic comparison of their performance has been made until now. This knowledge is required to select the appropriate buffer for a given species and tissue. METHODS: Four common lysis buffers (Galbraith's, LB01, Otto's and Tris.MgCl2) were used to prepare samples from leaf tissues of seven plant species (Sedum burrito, Oxalis pes-caprae, Lycopersicon esculentum, Celtis australis, Pisum sativum, Festuca rothmaleri and Vicia faba). The species were selected to cover a wide range of genome sizes (1.30-26.90 pg per 2C DNA) and a variety of leaf tissue types. The following parameters were assessed: forward (FS) and side (SS) light scatters, fluorescence of propidium iodide-stained nuclei, coefficient of variation of DNA peaks, presence of debris background and the number of nuclei released from sample tissue. The experiments were performed independently by two operators and repeated on three different days. KEY RESULTS: Clear differences among buffers were observed. With the exception of O. pes-caprae, any buffer provided acceptable results for all species. LB01 and Otto's were generally the best buffers, with Otto's buffer providing better results in species with low DNA content. Galbraith's buffer led to satisfactory results and Tris.MgCl2 was generally the worst, although it yielded the best histograms in C. australis. A combined analysis of FS and SS provided a 'fingerprint' for each buffer. The variation between days was more significant than the variation between operators. CONCLUSIONS: Each lysis buffer tested responded to a specific problem differently and none of the buffers worked best with all species. These results expand our knowledge on nuclear isolation buffers and will facilitate selection of the most appropriate buffer depending on species, tissue type and the presence of cytosolic compounds interfering with DNA staining.

Buffers↗
PubMed2006

Evaluation of cool- and warm-season grasses for resistance to multiple chinch bug (Hemiptera: Blissidae) species.

Chinch bugs are common pests of many agronomic and horticulturally important crops and turfgrasses. The extensive overlap of plant hosts and geographic distribution of Blissus leucopterus leucopterus (Say), Blissus leucopterus hirtus Montandon, Blissus insularis Barber, and Blissus occiduus Barber underscores the importance of identifying resistant germplasm. Cool- and warm-season turfgrasses and sorghum, Sorghum bicolor (L.) Moench, were evaluated for resistance to chinch bugs in the Blissus complex, and the presence of multiple resistance was documented. Greenhouse studies established that B. occiduus-resistant ('Prestige', formerly NE91-118) and -susceptible ('378') buffalograsses,, Buchloë dactyloides (Nuttall) Engelmann, were susceptible to all other chinch bug species. KS94 sorghum exhibited resistance to both B. occiduus and B. l. leucopterus, whereas B. insularis-resistant St. Augustinegrass, Stenotaphrum secundatum (Walter) Kuntze ('Floratam'), was also resistant to B. occiduus. B. l. leucopterus-susceptible sorghum ('Wheatland') and B. insularis-susceptible St. Augustinegrasses ('Raleigh' and 'Amerishade') were highly resistant to B. occiduus. Endophyte-free and -enhanced fine fescues (Festuca spp.) were moderately to highly susceptible to B. l. hirtus but moderately to highly resistant to B. occiduus. The results of this research showed the buffalograsses evaluated, including B. occiduus-resistant Prestige, are moderately to highly susceptible to the three other chinch bug species. In contrast, B. occiduus did not cause considerable damage to any of the turfgrasses or sorghum cultivars evaluated, other than buffalograss, irrespective of whether or not they are resistant to another chinch bug species. This information is increasingly important as various grasses become adapted to regions that may possess chinch bug species other than those with which they are typically associated. These levels of Blissus resistance should be included when selecting resistant germplasm for managing Blissus species pests.

Animals↗
PubMed2000

Annuality, perenniality and cell death.

This essay considers annuality and perenniality as quantitative traits and discusses the application of established and new genetic tools to the analysis of plant life histories. Annual/perennial status is a function of meristem determinacy in combination with the processes of cell death and disposal employed by plants to generate well-adapted anatomies and morphologies. Creeping perennials, like clover or bracken, seem to move around in the environment. They do this by extending into unoccupied space while the oldest tissues behind the growing and mature regions senesce, die and decompose. Trees do essentially the same thing, except that they develop vertically and the old dead tissue does not disappear but instead persists as wood. A root system is a kind of upended vertical perennial. The balance between exploratory growth and the wave of tissue death that succeeds it is a major determinant of perenniality. So although perenniality and annuality may appear to be dramatically different traits, extremes of behaviour can arise by a relatively minor change in the relationship between growth and death. This conclusion is supported by evidence from genome dosage studies, from the practical experiences of breeding perennial-type traits into annual backgrounds and from molecular cladistics. Applications of methods for the genetic analysis of quantitative characters are described, including the exploitation of introgression mapping in Lolium-Festuca and quantitative trait locus mapping in cereals and other species.

Apoptosis↗
PubMed2001

Differential expression of XET-related genes in the leaf elongation zone of F. pratensis.

Festuca pratensis Huds. is a forage grass with the ability to withstand harsh climatic conditions. However, its potential agronomic use is limited by its poor competitive ability, which can be traced to limitations in leaf growth. In order to characterize this process and to identify genes which might function as markers for leaf growth, three XET-related genes in the leaf elongation zone (LEZ) of F. pratensis are reported. A detailed expression analysis is presented of the three genes in two F. pratensis genotypes with contrasting leaf growth characteristics grown under two nitrogen levels. By means of a detailed spatial analysis of growth and XET encoding transcript pattern along the LEZ, a specific correlation is shown between FpXET1 expression and tissue elongation that is maintained under the different growth conditions, while the two other XETs expressed in the LEZ show different transcript dynamics. Tissue localization of FpXET1 and FpXET2 transcripts indicate an accumulation throughout young tissue, which is consistent with the encoded proteins playing roles in cell wall modification processes during growth. It is proposed that FpXET1 is a potential marker for tissue elongation and leaf growth in F. pratensis.

Amino Acid Sequence↗
PubMed1998

Mortality of horn fly (Diptera: Muscidae) larvae in bovine dung supplemented with loline alkaloids from tall fescue.

Larvae of arthropod ectoparasites of livestock, such as the horn fly, Haematobia irritans (L.), may be exposed to acyl-loline alkaloids in dung of ruminant livestock ingesting herbage of the tall fescue (Festuca arundinacea Schreb.)-endophyte association [Neotyphodium coenophialum (Morgan-Jones & W. Gams) Glenn, Bacon & Hanlin comb. nov.]. Biological activity of alkaloid-supplemented bovine dung was assayed by growth, development, and survival of 1st instars of horn fly. An extract from tall fescue seed, containing N-formyl loline (NFL), N-acetyl loline (NAL), and loline (59:21:20 by mass, respectively) caused 100% mortality of horn fly larvae when dung was supplemented at > or = 100 micrograms/g. Probit analysis of data corrected for natural mortality indicated a LD50 of 30 micrograms/g (95% fidicial limits: 20-49 micrograms/g). When horn fly larvae were introduced to dung supplemented with up to 50 microM of acyl-loline derivatives, mortality of larvae varied significantly between alkaloids (P < 0.0001). Probit analysis indicated that NFL [LD50: 34 microM (95% fidicial limits: 3-53 microM)] was more toxic than NAL [LD50: 46 microM (0-83 microM)], and that loline hydrochloride was not toxic.

Alkaloids↗
PubMed1999

Mortality of horn fly larvae (Diptera: Muscidae) in bovine dung supplemented with ergotamine and N-formyl loline.

Dung-dwelling larvae of ectoparasites of livestock such as the horn fly, Hematobia irritans (L.), may be exposed to > or = 1 different alkaloid species in dung from animals ingesting herbage of the tall fescue (Festuca arundinacea Schreb.)--endophyte association (Neotyphodium coenophialum (Morgan-Jones & W. Gams) Glenn, Bacon & Hanlin comb. nov.). First-instar horn flies were exposed to bovine dung supplemented with up to 50 microM each of N-formyl loline and ergotamine tartrate in factorial combination. In the absence of ergotamine tartrate, N-formyl loline caused a linear decline in the number of pupae recovered, and probit analysis indicated an LC50 of 36 microM. In the absence of N-formyl loline, significant quadratic responses of larvae to ergotamine tartrate were established, and probit analysis indicated a LC50 of 34 microM. An interaction (P < 0.001) was found between the 2 alkaloids for larval survival. This interaction showed that ergotamine tartrate moderated the toxicity of N-formyl loline and indicates that a membrane-bound receptor may be involved. There was no evidence of carryover of effects of alkaloids on subsequent stages of development or expressed as abnormalities of pupae or adults. Interactions between alkaloids probably are involved in other plant-herbivore relationships of endophyte-infected grasses.

Alkaloids↗
PubMed2002

The contribution of roots and shoots to whole plant nitrate reduction in fast- and slow-growing grass species.

The hypothesis was tested that slow-growing grass species perform a greater proportion of total plant NO3- reduction in their roots than do fast-growing grasses. Eight grass species were selected that differed in maximum relative growth rate (RGR) and net NO3- uptake rate (NNUR). Plants were grown with free access to nutrients in hydroponics under controlled-environment conditions. The site of in vivo NO3- reduction was assessed by combining in vivo NO3- reductase activity (NRA) assays with biomass allocation data, and by analysing the NO3- to amino acid ratio of xylem sap. In vivo NRA of roots and shoots increased significantly with increasing NNUR and RGR. The proportion of total plant NO3- reduction that occurs in roots was found to be independent of RGR and NNUR, with the shoot being the predominant site of NO3- reduction in all species. The theoretical maximum proportion of whole plant nitrogen assimilation that could take place in the roots was calculated using information on root respiration rates, RGR, NNUR, and specific respiratory costs associated with growth, maintenance and ion uptake. The calculated maximum proportion that the roots can contribute to total plant NO3- reduction was 0.37 and 0.23 for the fast-growing Dactylis glomerata L. and the slow-growing Festuca ovina L., respectively. These results indicate that slow-growing grass species perform a similar proportion of total plant NO3- reduction in their roots to that exhibited by fast-growing grasses. Shoots appear to be the predominant site of whole plant NO3- reduction in both fast- and slow-growing grasses when plants are grown with free access to nutrients.

Algorithms↗
PubMed2006

Endophytic fungus decreases plant virus infections in meadow ryegrass (Lolium pratense).

We studied the effects of fungal endophyte infection of meadow ryegrass (Lolium pratense=Festuca pratensis) on the frequency of the barley yellow dwarf virus (BYDV). The virus is transferred by aphids, which may be deterred by endophyte-origin alkaloids within the plant. In our experiment, we released viruliferous aphid vectors on endophyte-infected and endophyte-free plants in a common garden. The number of aphids and the percentage of BYDV infections were lower in endophyte-infected plants compared to endophyte-free plants, indicating that endophyte infection may protect meadow ryegrass from BYDV infections.

Animals↗
PubMed1976

Numerical taxonomy of some yellow-pigmented bacteria isolated from plants.

Phenetic data on over 60 heterotrophic, Gram-negative, yellow chromogenic bacteria from plant material were collected and analysed using numerical taxonomic methods. Marker strains representing 42 taxa were included in the analyses. At similarity levels of 80% or above, eight distinct clusters were obtained, the first four of which included yellow chromogens. Custer I contained isolates from green healthy leaves of Agrostis tenuis, Festuca rubra, Holcus lanata, Lolium perenne and Poa pratensis, and clusters 2 and 3 consisted of isolates from Holcus lanata seeds and leaves of P. pratensis respectively. Cluster 4 contained seven subgroups and was equated with the family Enterobacteriaceae. Erwinia herbicola strains from a variety of sources formed a homogeneous subgroup, readily distinguishable from authentic strains of E. amylovora, E. carotovora, other representative erwiniae, and from all other enterobacteria studied. These data emphasize the heterogeneous nature of yellow-pigmented bacteria from plants, and support the inclusion of E. herbicola and other Erwinia species in the Enterobacteriaceae.

Bacteria↗
PubMed1993

A Novel Fungal Protease Expressed in Endophytic Infection of Poa Species.

The fungus Acremonium typhinum produces a novel endoprotease during symbiotic endophytic infection of the grass, Poa ampla. This protease is unusual because it is highly active in the presence of sodium dodecyl sulfate. The enzyme is a thiol-containing serine protease and is localized to a crude membrane fraction. Similar protease activity has been detected in endophyte-infected Poa autumnalis and Poa sylvestris plants. Expression of this protease may be important in endophytic infection of Poa spp., because similar activity has not been detected in endophyte-infected Festuca arundinacea or Lolium perenne.

Journal Article↗
PubMed1993

Novel O-D-galacturonoyl esters in the pectic polysaccharides of suspension-cultured plant cells.

Driselase digestion of uronate-6-14C-labeled primary walls of cultured spinach (Spinacia oleracea L.) cells yielded about 18 novel uronate-containing compounds, most of which could be hydrolyzed by cold dilute alkali to yield oligo-[14C]galacturonides. One typical Driselase digestion product (compound 17) yielded alpha-(1-->4)-D-[14C]galacturonotriose(GalA3) upon very mild treatment with alkali (50% yield of GalA3 in 7.2 min at pH 11 and 25 degrees C). One of the three galacturonate residues in compound 17 was reducible to a galactose residue with sodium borohydride, indicating that that GalA residue was esterified, via its--COOH group, to a putative alcohol. Compound 17 had a higher mobility than GalA3 on paper chromatography, indicating that the putative alcohol was relatively nonpolar. The putative alcohol could not have been methanol because Driselase readily hydrolyzed mono-, di-, and trimethyl esters of GalA3 to yield free galacturonic acid. Another Driselase digestion product (compound 12) was a derivative of GalA3 that apparently possessed two nonpolar esterified substituents: one about as labile as in compound 17, and the other approximately 10 times more stable. Compounds 12 and 17 could not labeled by in vivo feeding of [U-14C]cinnamate, suggesting that they were not phenolic conjugates. Similar but chromatographically distinguishable uronate-14C-labeled esters were obtained by Driselase digestion of walls of cultured carrot (Daucus carota L.), Paul's Scarlet rose (Rosa sp.), and tall fescue (Festuca arundinacea Schreber) cells. In spinach, the novel compounds constituted about 5% of the total galacturonate residues of the cell wall. The observations suggest that pectic polysaccharides are linked, via O-D-galacturonoyl ester bonds, to relatively hydrophobic constituents of the primary cell wall. Their possible role in wall architecture is discussed.

Carbohydrate Conformation↗
PubMed1994

Nitrogen Use within the Growing Leaf Blade of Tall Fescue.

Leaf elongation rate (LER) of grasses depends on N supply and is expressed mostly through cell production, whereas most N in mature leaf tissues is chloroplastic. Our objective was to evaluate a possible competition for N between cell production and chloroplast development processes, utilizing the gradient of cell development along the leaf growth zone of tall fescue (Festuca arundinacea Schreb.). Under the two contrasting N regimes, total N content was highest in the cell production zone, declined sharply as cells elongated, and remained relatively constant in more distal positions, at values close to those measured in mature tissues. A similar pattern was found for N in proteins and nucleic acids that were not soluble in 80% ethanol. Content of N compounds soluble in 80% ethanol was higher in the cell production and elongation zones than in mature parts of the leaf. NO3- N content was low in the cell production zone and increased in the cell elongation zone for high-N plants. The deposition rate of total N in the growth zone was much higher with plants in high N than in those shifted to no N. For both N regimes, most N was deposited during cell production and early cell elongation. Little N was deposited during cell maturation where ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) was being actively synthesized. This suggests that synthesis of Rubisco, and probably other chloroplastic proteins, occurs largely from recycling of N that was previously incorporated into proteins during cell production. Thus, Rubisco content in mature tissues is more closely associated with N deposited during cell production than with N deposited during its biosynthesis.

Journal Article↗
PubMed1994

Response of Fructan to Water Deficit in Growing Leaves of Tall Fescue.

Changes in dry matter and water-soluble carbohydrate components, especially fructan, were examined in the basal 25 mm of expanding leaf blades of tall fescue (Festuca arundinacea Schreb.) to assess their roles in plant response to water deficit. Water was withheld from vegetative plants grown in soil in controlled-environment chambers. As stress progressed, leaf elongation rate decreased sooner in the light period than it did in the dark period. The decrease in growth rate in the dark period was associated with a decrease in local relative elongation rates and a shortening of the elongation zone from about 25 mm (control) to 15 mm. Dry matter content of the leaf base increased 23% during stress, due mainly to increased water-soluble carbohydrate near the ligule and to increased water-soluble, carbohydrate-free dry matter at distal positions. Sucrose content increased 258% in the leaf base, but especially (over 4-fold) within 10 mm of the ligule. Hexose content increased 187% in the leaf base. Content of total fructan decreased to 69% of control, mostly in regions farther from the ligule. Fructan hydrolysis could account for the hexose accumulated. Stress caused the osmotic potential to decrease throughout the leaf base, but more toward the ligule. With stress there was 70% less direct contribution of low-degree-of-polymerization fructan to osmotic potential in the leaf base, but that for sucrose and hexose increased 96 and 67%, respectively. Thus, fructan metabolism is involved but fructan itself contributes only indirectly to osmotic adjustment.

Journal Article↗
PubMed1995

Fructosyltransferase Activities in the Leaf Growth Zone of Tall Fescue.

High concentrations of water-soluble carbohydrates, mainly fructan, accumulate in the growth zone of tall fescue (Festuca arundinacea Schreb.) leaf blades. We studied sucrose-hydrolyzing activities in the leaf growth zone because of their importance in carbohydrate partitioning. Sucrose hydrolysis in the basal 1.5 cm was largely due to fructosyltransferases, which had activities up to 10 times higher than in fully developed leaf tissue. Three fructosyltransferases (F1, F2, and F3) were purified from the leaf growth zone. Each synthesized, from either sucrose or 1-kestose, a mixture of trisaccharides and higher-order oligofructans identical with the low-degree of polymerization fructan extracted from similar plant tissue. The highly purified fructosyltransferases retained ability (13%) to transfer fructose from sucrose to water. Time-dependent and substrate-dependent studies, using sucrose as the substrate, showed proportional production of fructose and glucose, indicating that both products are from the same enzyme. Fructosyltransferase was calculated to contribute about half the total transfer of fructose to water in the basal 1.5 cm. Invertase activity increased to near 2.0 cm when fructosyl transfer to sucrose and other oligofructans decreased. Invertase was the major activity for sucrose hydrolysis at positions distal to 3.0 cm.

Journal Article↗
PubMed1995

Partitioning of Electrons between the Cytochrome and Alternative Pathways in Intact Roots.

To test the hypothesis that the cytochrome pathway is not invariably saturated when the alternative pathway is engaged, we titrated root respiration of several species with KCN (an inhibitor of the cytochrome pathway), both in the absence and presence of an inhibitor of the alternative pathway (salicylhydroxamic acid, SHAM). The slopes of the resultant KCN [rho] plots ([rho]cyt) were then used to determine whether the cytochrome pathway was saturated in each species. The species used were Festuca ovina ssp. ovina L., Phaseolus vulgaris L., and six Poa species (Poa pratensis L., Poa compressa L., Poa trivialis L., Poa alpina L., Poa costiniana Vick., and Poa fawcettiae Vick.). Although the cytochrome pathway was saturated in a number of species (i.e. [rho]cyt values were 1.0), several others exhibited [rho]cyt values of less than 0.5. Alternative pathway capacity correlated negatively with [rho]cyt, with [rho]cyt values of less than 1.0 occurring in tissues in which the alternative pathway capacity was greater than 25 to 30% of total respiration. The species that did not show full engagement of the cytochrome pathway rarely exhibited SHAM inhibition in the absence of KCN. We conclude that this lack of SHAM inhibition is not due to a lack of alternative pathway engagement but rather to the diversion of electrons from the alternative pathway to the unsaturated cytochrome path following the addition of SHAM.

Journal Article↗
PubMed2000

Cloning and functional analysis of sucrose:sucrose 1-fructosyltransferase from tall fescue.

Enzymes of grasses involved in fructan synthesis are of interest since they play a major role in assimilate partitioning and allocation, for instance in the leaf growth zone. Several fructosyltransferases from tall fescue (Festuca arundinacea) have previously been purified (Lüscher and Nelson, 1995). It is surprising that all of these enzyme preparations appeared to act both as sucrose (Suc):Suc 1-fructosyl transferases (1-SST) and as fructan:fructan 6(G)-fructosyl transferases. Here we report the cloning of a cDNA corresponding to the predominant protein in one of the fructosyl transferase preparations, its transient expression in tobacco protoplasts, and its functional analysis in the methylotrophic yeast, Pichia pastoris. When the cDNA was transiently expressed in tobacco protoplasts, the corresponding enzyme preparations produced 1-kestose from Suc, showing that the cDNA encodes a 1-SST. When the cDNA was expressed in P. pastoris, the recombinant protein had all the properties of known 1-SSTs, namely 1-kestose production, moderate nystose production, lack of 6-kestose production, and fructan exohydrolase activity with 1-kestose as the substrate. The physical properties were similar to those of the previously purified enzyme, except for its apparent lack of fructan:fructan 6(G)-fructosyl transferase activity. The expression pattern of the corresponding mRNA was studied in different zones of the growing leaves, and it was shown that transcript levels matched the 1-SST activity and fructan content.

Amino Acid Sequence↗
PubMed1971

Photosynthetic CO(2) Fixation Products and Activities of Enzymes Related to Photosynthesis in Bermudagrass and Other Plants.

After a 5-second exposure of illuminated bermudagrass (Cynodon dactylon L. var. ;Coastal') leaves to (14)CO(2), 84% of the incorporated (14)C was recovered as aspartate and malate. After transfer from (14)CO(2)-air to (12)CO(2)-air under continuous illumination, total radioactivity decreased in aspartate, increased in 3-phosphoglyceric acid and alanine, and remained relatively constant in malate. Carbon atom 1 of alanine was labeled predominantly, which was interpreted to indicate that alanine was derived from 3-phosphoglyceric acid. The activity of phosphoenolpyruvate carboxylase, alkaline pyrophosphatase, adenylate kinase, pyruvate-phosphate dikinase, and malic enzyme in bermudagrass leaf extracts was distinctly higher than those in fescue (Festuca arundinacea Schreb.), a reductive pentose phosphate cycle plant. Assays of malic enzyme activity indicated that the decarboxylation of malate was favored. Both malic enzyme and NADP(+)-specific malic dehydrogenase activity were low in bermudagrass compared to sugarcane (Saccharum officinarum L.). The activities of NAD(+)-specific malic dehydrogenase and acidic pyrophosphatase in leaf extracts were similar among the plant species examined, irrespective of the predominant cycle of photosynthesis. Ribulose-1, 5-diphosphate carboxylase in C(4)-dicarboxylic acid cycle plant leaf extracts was about 60%, on a chlorophyll basis, of that in reductive pentose phosphate cycle plants.We conclude from the enzyme and (14)C-labeling studies that bermudagrass contains the C(4)-dicarboxylic acid cycle and that pyruvate-phosphate dikinase does not exist exclusively in C(4)-dicarboxylic acid cycle plants, and we propose that in C(4)-dicarboxylic acid cycle plants the transfer of carbon from a dicarboxylic acid to 3-phosphoglyceric acid involves a decarboxylation reaction and then a refixation of carbon dioxide by ribulose-1, 5-diphosphate carboxylase.

Journal Article↗