Growth, photosynthesis and assimilate partitioning in Lolium temulentum exposed to chilling temperatures

Vegetative plants of Lolium temulentum L, grown at 20°C with an 8 h photoperiod were transferred to either 5 or 2°C (8 h photoperiod) at 4th leaf maturity. Measurement of dry weight gain indicated a marked decline in relative growth rate below 5°C, with growth being reduced as much between 5 and 2°C as between 20 and 5°C. This reduction was not associated with increased mortality and was reversible if plants were returned to 20°C. Tissue explants from cold-treated plants retained the ability to extend if returned to 20°C Rates of extension in explants were less temperature sensitive than the relative growth rates observed in intact plants. Measurements of photosynthetic capacity, and of the patterns of accumulation of reserve carbohydrate in plants exposed to different temperatures, suggested that the inhibition of growth caused by chilling is not caused by an inability of the plants to fix sufficient carbon.     

Effects of Tiller Separation and Root Pruning on the Growth of Lolium perenne L.

Experiments have been carried out to assess the role of inter-tillerinteractions within plants of Lolium perenne L. cv. S24 as factorswhich could play a part in determining the growth-rate of thewhole plant. In order to do this, the effect of tiller separationon plant growth was studied as well as the influence of theremoval of different parts of the root and shoot system. Wheneverplants were supplied with nutrients by means of culture solutions,separation of tillers brought about a marked increase in theamount of dry matter produced. Root pruning and in particularthe removal of root apices brought about similar increases ingrowth. Separation of tillers in soil-grown plants did not bringabout increased growth. These results are interpreted as indicatingthat separation of tillers or the initial removal of root apicesin plants supplied with nutrients by means of culture solution,promotes root branching which brings about an increased netassimilation rate arising from an increase supply of a growth-promotingsubstance, probably a cytokinin, from the root apices. The resultsare not taken as indicative of an intertiller interaction whichrestricts the growth of intact plants.     

Asynapsis in Lolium perenne

Eighteen plants displaying varying degrees of asynapsis ranging from weak to very strong were found among four out of six populations of Lolium perenne L. (2n=14) which had been subjected to three cycles of directional phenotypic selection for productivity of green material. No plants were found displaying univalents in the original generation but the incidence increased with cycles of selection, indicating the genetic control and differential distribution of asynaptic genes among these populations. — The analysis of univalents and chiasma frequency of pollen mother cells (PMC) of six partially asynaptic plants chosen for detailed study revealed that univalents occurred throughout all PMC chiasma classes irrespective of chiasma frequency, but the higher the chiasma frequency of any PMC the less the likelihood of univalents occurring. The relationship between chiasma frequency and univalent frequency per PMC per plant was negative. — Mean chiasma frequency per bivalent increased for the asynaptic cells in comparison with the normal in both the weak and medium asynaptic groups which was explained by the availability of additional chiasmata for redistribution.     

Association of candidate genes with flowering time and water-soluble carbohydrate content in Lolium perenne (L.)

We describe a candidate gene approach for associating SNPs with variation in flowering time and water-soluble carbohydrate (WSC) content and other quality traits in the temperate forage grass species Lolium perenne. Three analysis methods were used, which took the significant population structure into account. First, a linear mixed model was used enabling a structured association analysis to be incorporated with the nine populations identified in the structure analysis as random variables. Second, a within-population analysis of variance was performed. Third, a tree-scanning method was used, in which haplotype trees were associated with phenotypes on the basis of inferred haplotypes. Analysis of variance within populations identified several associations between WSC, nitrogen (N), and dry matter digestibility with allelic variants within an alkaline invertase candidate gene LpcAI. These associations were only detected in material harvested in one of the two years. By contrast, consistent associations between the L. perenne homolog (LpHD1) of the rice photoperiod control gene HD1 and flowering time were identified. One SNP, in the immediate upstream region of the LpHD1 coding sequence (C-4443-A), was significant in the linear mixed model. Within-population analysis of variance and tree-scanning analysis confirmed and extended this result to the 2118 polymorphisms in some of the populations. The merits of the tree-scanning method are compared to the single SNP analysis. The potential usefulness of the 4443 SNP in marker-assisted selection is currently being evaluated in test crosses of genotypes from this work with turf-grass varieties.     

Cytomixis in Lolium perenne

Cytomixis is a spontaneous process occurring through the formation of cytoplasmic bridges between adjacent pollen mother cells. This phenomenon was observed in the pollen mother cells of 3 genotypes of Lolium perenne which had been subjected to directional selection for productivity of green material. — The process has led to the formation of up to 34.8% of PMCs with chromosome numbers deviating from the normal diploid number 2n=14. The abnormal PMCs contained chromosome numbers ranging from 2–56 as observed at first metaphase. — Abnormal PMCs were also observed with approximately the same frequency in Meiosis II. This phenomenon is under genetic control. The evolutionary significance of cytomixis and the cytological consequences are discussed.     

Photosynthesis and Photorespiration in Lolium multiflorum and Lolium perenne

The response of net photosynthesis to changing light-flux densityby leaves of Lolium multiflorum (S. 22) and L. perenne (S. 321)is more adequately described by current models when a term allowingfor photorespiration is included. The magnitude of this termwas determined from the changes in the slope of the light-responsecurves for net photosynthesis. A pseudo first-order rate-constantfor photorespiration, and a pseudo second-order rate-constantfor photosynthesis calculated by this technique for L. multiflorumwere found to be similar to corresponding parameters calculatedfrom light-compensation-point measurements using a simple modeldescribed by Brown (1969). The relative magnitudes of respirationand photosynthesis at light saturation for both Lolium specieswere similar to reported values for other temperate species(Lake, 1967). Two selection lines of L. perenne (S. 321) with contrastingdry-matter yields were found to have the same parameters forrespiration and photosynthesis.     

The dynamics of aneuploidy in an induced tetraploid population of Lolium multiflorum x Lolium perenne

Summary Populations of induced polyploids invariably contain a substantial proportion of aneuploid individuals. A model is described which can predict the level of aneuploidy in successive generations of a closed population of Lolium tetraploids. The results demonstrate clearly that the proportion of aneuploid individuals increases sharply for two to three generations and then stabilizes at a level determined by the gametic output of the euploid plants. A change in the gametic output of aneuploid individuals has a relatively small effect on the final level of aneuploidy reached.     

An association mapping approach to identify flowering time genes in natural populations of Lolium perenne (L.)

We describe an association mapping approach using natural populations of perennial ryegrass (Lolium perenne L.) to identify molecular markers associated with heading date, an important trait affecting seasonal production, tillering, digestibility and grassland management regimes. Twenty-three natural populations originating from throughout Europe, with heading date phenotypes ranging from very early to very late, as well as three synthetic populations (varieties) were used for molecular marker genotyping using AFLP. In total, 589 polymorphic markers were identified. Hierarchical clustering, principal coordinate and other statistical analyses identified four outlying populations forming a clearly distinct sub-group. Removal of those four populations from the subsequent analysis reduced population sub-structure twofold. However, this made relatively little difference to the result of the association analysis. Linear regression identified three markers whose frequency of occurrence correlated with the heading date phenotype. Moreover, these markers were shown to be closely linked to each other within a major QTL on Chromosome 7, explaining 70% of the total variation in heading date. Pairwise linkage disequilibrium among them was also significant. These results suggest that association mapping approaches may be feasible in L. perenne, and that the use of natural populations could provide a useful source of genetic variation in traits of importance in crop improvement.     

Temperature-induced alteration to root longevity in Lolium perenne

The production and mortality of roots is an important factor determining nutrient and carbon fluxes in terrestrial ecosystems. However, the effect of temperature on the longevity of roots is not understood. In this study the impact of changes in temperature on the longevity of Lolium perenne roots was determined.Plants were grown at 15 °C, 21 °C and 27 °C and their roots observed using a minirhizotron system. Major effects of temperature were measured with large reductions in root longevity occurring at higher temperatures. The implications of these data are discussed.     

Effects of photon flux density on carbon partitioning and rhizosphere carbon flow of Lolium perenne

The distribution and partitioning of dry matter and photoassimilateof Lolium perenne was investigated under two light regimes providingphotosynthetically active radiation of 350 µmol m^–2s^–1 (low light treatment) or 1000 µmol m^–2s^–1 (high light treatment). Plants were grown at specificgrowth conditions in either soil or sand microcosm units tofollow the subsequent release of carbon into the rhizosphereand its consequent incorporation into the microbial biomass(soil system) or recovery as exudates (sand system). The distributionof recent assimilate between the plant and root released carbonpools was determined using ¹⁴CO₂ pulse-chase methodology atboth light treatments and for both sand- and soil-grown seedlings.A significant (P     

Assessment of candidate reference genes for the expression studies with brassinosteroids in Lolium perenne and Triticum aestivum

Quantitative PCR studies need proper reference genes with expression stability exclusively validated under certain experimental conditions. The expression stability of several genes commonly used as references was tested under 24-epibrassinolide (EBR) and temperature treatment. Different statistical approaches (qBase^PLUS, BestKeeper, NormFinder) were used to prepare rankings of expression stability in two species of an economic importance: common wheat (Triticum aestivum) and perennial ryegrass (Lolium perenne). Candidate reference genes were shown to be regulated differentially in these two plant species. The maximum stability values indicated that the expression stability was higher in T. aestivum. Taking into account of all ranks it seems that TBP-1 and UBI in ryegrass and ACT, ADP and EF1A in wheat should be used as reference genes in the brassinosteroids and temperature involving studies.     

The Effect of New Tiller Growth on Carbohydrates, Nitrogen and Seed Yield per Ear in Lolium perenne L.

To clarify whether new vegetative tillers that develop aroundanthesis in aLolium perenne seed crop can depress seed yields,the possible competition for carbohydrates or nitrogen betweenthe seeds and new tillers that develop after the onset of anthesiswas investigated. In two greenhouse experiments the number oftillers per plant was varied by a combination of cutting, nitrogensupply, light quality and light intensity treatments. Two genotypeswith different tillering rates were used. Seed yield per earwas largely independent of the number of tillers and regrowthof cut tillers after the onset of anthesis. It increased inone genotype, but only under low light and a reduced nutrientavailability, and no new tillers were produced. The amount ofwater-soluble carbohydrates in the reproductive tillers increasedin both clones under these conditions. Under more favourableconditions the increased tillering rate and regrowth of tillersafter cutting did not adversely affect seed yield per ear ineither clone, although carbohydrate reserves in the floweringtillers were sharply reduced. Tiller removal increased the concentrationand amount of nitrogen in the remaining flowering tillers, irrespectiveof the amount of regrowth. It is concluded that competitionfor carbohydrates or nitrogen between the seeds and new vegetativetillers that develop after the onset of anthesis, is not a majorcause of the low and variable seed yields inL. perenne seedcrops. Processes within the ear itself are probably limiting. Lolium perenne L.; perennial ryegrass; seed yield; seed set; tillering; carbohydrates; nitrogen; competition.     

Carbon and nitrogen allocation in Lolium perenne in response to elevated atmospheric CO2 with emphasis on soil carbon dynamics

The effect of elevated CO2 on the carbon and nitrogen distribution within perennial ryegrass (L. perenne L.) and its influence on belowground processes were investigated. Plants were homogeneously 14C-labelled in two ESPAS growth chambers in a continuous 14C-CO2 atmosphere of 350 and 700 L L-1 CO2 and at two soil nitrogen regimes, in order to follow the carbon flow through all plant and soil compartments.After 79 days, elevated CO2 increased the total carbon uptake by 41 and 21% at low (LN) and high nitrogen (HN) fertilisation, respectively. Shoot growth remained unaffected, whereas CO2 enrichment stimulated root growth by 46% and the root/soil respiration by 111%, irrespective of the nitrogen concentration. The total 14C-soil content increased by 101 and 28% at LN and HN, respectively. The decomposition of the native soil organic matter was not affected either by CO2 or by the nitrogen treatment.Elevated CO2 did not change the total nitrogen uptake of the plant either at LN or at HN. Both at LN and HN elevated CO2 significantly increased the total amount of nitrogen taken up by the roots and decreased the absolute and relative amounts translocated to the shoots.The amount of soil nitrogen immobilised by micro-organisms and the size of the soil microbial biomass were not affected by elevated CO2, whereas both were significantly increased at the higher soil N content.Most striking was the 88% increase in net carbon input into the soil expressed as: 14C-roots plus total 14C-soil content minus the 12C-carbon released by decomposition of native soil organic matter. The net carbon input into the soil at ambient CO2 corresponded with 841 and 1662 kg ha-1 at LN and HN, respectively. Elevated CO2 increased these amounts with an extra carbon input of 950 and 1056 kg ha-1. Combined with a reduced decomposition rate of plant material grown at elevated CO2 this will probably lead to carbon storage in grassland soils resulting in a negative feed back on the increasing CO2 concentration of the atmosphere.     

Development and mapping of a public reference set of SSR markers in Lolium perenne L.

We report on the characterization and mapping of 76 simple sequence repeat (SSR) markers for Lolium perenne. These markers are publicly available or obtained either from genomic libraries enriched for SSR motifs or L. perenne expressed sequence tag (EST) clones. Four L. perenne mapping populations were used to map the SSR markers. A consensus linkage map of the four mapping populations containing 65 of the SSR markers is presented, together with primer information and a quality score indicating the usefulness of the SSR marker in different populations. The SSR markers identified all seven L. perenne linkage groups.     

Hardening and Dehardening of Lolium perenne in Response to Fluctuating Temperatures

Hardening and dehardening responses of two contrasting varietiesof Lolium perenne, measured as LT50 estimates, were followedin fluctuating temperature environments. Unhardened seedlingswere exposed to hardening environments for 7, 14, and 21 d inall combinations of 2, 4, 6, 8 and 10 C with either high dayand low night temperatures or low day and high night temperatures.Seedlings hardened for 28 d at 2 C were exposed to dehardeningenvironments in all combinations of 4, 6, 8, 10, and 12 C withhigh day and low night temperatures. A low day, or night, temperature of 2 C in combination withany other temperature increased hardening compared with theconstant higher temperature. For Premo, a hardy variety, thisincrease was 3 C when night temperature was reduced from 10to 2 C in combination with a day temperature of 10 C. Similarly,a low night temperature reduced the dehardening response ofPremo to higher day temperatures. At 12 C this effect on LT60was greater than 2 C. Much smaller responses to daily periodsof low temperature were found for the less hardy variety, GrasslandsRuanui. During each 24-h period, exposures to 2 C of longer than 4h were required to achieve greater hardening than that achievedin continuous 10 C treatments. Hardiness was not improved furtherby exposures longer than 8 h. Responses to diurnal temperature fluctuations were discussedin relation to possible mechanisms and to changes in hardinessduring the winter under different weather systems. Lolium perenne, cold hardening, cold dehardening, diurnal temperature fluctuations, varieties     

Environment-induced Modifications to Root Longevity in Lolium perenne and Trifolium repens

Understanding how environmental factors affect the longevityof roots is essential if root mortality linked nutrient cyclingprocesses within ecosystems are to be understood, and the impactof natural and anthropogenic climate change properly evaluated.In this study the longevity of roots at two geographically andclimatically distinct sites were compared to identify the scaleof change that can occur due to environmental differences. Minirhizotrontubes were inserted into swards sown with the same variety ofLolium perenne and Trifolium repens at sites in the UK and Italy.Roots were viewed using a video camera and digitized imagesof roots generated at intervals. From these images the lifehistory of individual roots was determined and compared. Therewere few differences in patterns of longevity between differentspecies at the same site. Major differences, however, were observedbetween roots of the same species at different sites. For example,73% of Trifolium repens roots failed to survive for 21 d inItaly compared to only 29% at the UK site. Similarly, over 84%of Lolium perenne roots failed to survive for more than 21 din Italy compared to 38% in the UK. These data suggest thatenvironmental factors can have a major impact on root longevity.Copyright 2000 Annals of Botany Company Climate change, Lolium perenne L., minirhizotron, perennial ryegrass, root longevity, Trifolium repens L., white clover     

Mineral nutrition and the reaction of Lolium perenne to defoliation

Summary Plants were grown in nutrient solutions, defoliated to remove all expanded leaf blades and the amount of regrowth measured a week later. Prior to defolation, plants in concentrated nutrient solutions grew more rapidly than those in dilute solutions. After defoliation, the plants in concentrated solutions did not produce, however, a greater quantity of regrowth relative to their size than those in dilute solutions. Other things being equal, in environments which produced plants with a low root: shoot ratio, the greatest amount of regrowth was produced by genotypes with a higher than average root: shoot ratio, and vice versa.These results are discussed in relation to repeated defoliation, size and age of the plants, and the role of roots and reserve substances in recovery from defoliation