Senescence and decomposition of white clover stolons in grazed upland grass/clover swards

The fate of marked sections of stolons of white clover (Trifolium repens) over a 50-week period from May 1987 was followed in grazed grass/clover swards maintained at 5-cm sward surface height with and without N fertiliser. There was little effect of N treatment on the pattern of survival of stolon sections. The proportion of live stolons recovered decreased during the experiment, and in May 1988 on average only 29% of the marked sections remained alive. At all harvests only a small percentage of stolon sections showed signs of senescence; the maximum percentage, on average 20% of those marked, occurred in autumn, 15–20 weeks after marking. Following this period, i.e. in late autumn/winter, the most rapid increase in percentage of decomposed stolons was measured. Over 50% of stolon sections were buried within the 5-week period following marking and nearly all were buried after 20 weeks; generally a much smaller proportion of stolon tips was buried. Nutrient concentrations of N, P and K fell to their lowest levels in autumn, before increasing in the following spring. Results are discussed in relation to the cycling of nutrients via stolon senescence.     

Auxin-stimulated somatic embryogenesis from immature cotyledons of white clover

Cotyledons from immature embryos of white clover (Trifolium repens L.) cv. Osceola were exposed to 2,4-D or NAA to induce somatic embryogenesis. NAA at 10 or 20 mg 1^–1 was very inefficient at stimulating embryogenesis, while concentrations of 30 or 40 mg 1^–1 resulted in death of the explant tissue. Continuous exposure of cotyledons to 40 mg 1^–1 2,4-D resulted in somatic embryos which were arrested at the globular stage, or which underwent cycles of secondary embryogenesis, never proceeding beyond the globular stage. A 10 day exposure time to 2,4-D at the same concentration led to formation of somatic embryos, most of which had poorly developed cotyledons. Almost 10% of the somatic embryos converted into plants following transfer to medium devoid of growth regulators. Attempts to improve morphology of somatic embryos by using shorter exposure times to 2,4-D at 40 mg 1^–1, or by maintaining the 10 day exposure time while varying the concentration of 2,4-D, were not successful. Plants were obtained from all parents evaluated, although at different frequencies.     

The effects of low concentrations of aluminium on the growth and uptake of nitrate-N by white clover

Summary The effects of aluminium (Al³⁺) at concentrations of 0, 25, 50 and 100 μM on the growth of white clover, dependent upon N supplied as NO ₃ ⁻ , were examined in flowing solution culture. Plants were established with a normal nutrient supply for 7 weeks and then grown with carefully controlled pH (at 4.5) and P concentrations, and with 0, 25, 50 or 100 μM Al³⁺ for a further three weeks. There were rapid visual effects (i.e. symptoms of P deficiency and reduction in root extension) and the dry weights of shoots and roots were reduced at 50 and 100 μM. Less than 10% of Al absorbed from solution was transported to the shoots. The uptake of P, and its transport between roots and shoots, were reduced in plants grown with Al. The uptake of NO ₃ ⁻ stopped immediately after the introduction of 50 or 100 μM Al, and was significantly reduced at 25 μM after three weeks. During a second phase of the experiment, plants previously grown at 0, 25, 50 and 100 μM Al, were grown for a further 2 weeks either with NO ₃ ⁻ (with and without 50 μM Al³⁺) or without NO ₃ ⁻ but with inoculation by Rhizobia (and with or without 50 μM Al³⁺). The effects of the previous treatments with Al on N uptake were small during the second phase, but uptake by all plants was restricted when Al was present. Inoculation did not result in nodulation in the second phase when Al³⁺ was present in the solution, but Al already in the plant from the first phase did not prevent nodulation in the absence of Al during the second phase.     

Status of self-pollen in bee pollination efficiency of white clover (Trifolium repens L.)

Flowers of white clover (Trifolium repens L.) are hermaphrodite and self-incompatible; their cross-pollination depends entirely on insect visitors, mainly bees (Apoidea). Because self-pollination of white clover occurs before flower anthesis, we determined whether selfing affected the pollination efficiency of a honeybee visit. We compared pollen deposition in emasculated and intact flowers following (1) a single honeybee visit, (2) open-pollination for a day and (3) enclosure in a cloth bag to prevent insect visits. In emasculated flowers, open-pollination resulted in more pollen deposited than after one visit (+30%) which is consistent with flowers being visited more than once by pollinators during the course of a day. On intact flowers, saturation of the stigma was achieved after the first visit of a honeybee (near 280 grains) because of self-pollination. Additional visits did not increase pollen deposits, but they improved pollen efficiency in terms of numbers of pollen tubes reaching the ovules. In such a context of easily saturated stigmas, self-pollen does not inhibit cross-pollen activity, but represents a constraint for pollination which demands multiple bee visits to each flower to achieve maximum fertilization. Received: 20 May 1997 / Accepted: 25 October 1997     

Selenium uptake and its influence on the antioxidative system of white clover as affected by lime and phosphorus fertilization

Selenium (Se) is regarded as an antioxidant in animal and human nutrition, but its biological role in plants needs to be clarified. Plants vary considerably in their ability to tolerate Se, and their biochemical response to Se may be affected by liming or P fertilization. Two greenhouse experiments were conducted with white clover (Trifolium repens L.) to evaluate Se accumulation, tolerance, and the antioxidant response at increasing selenite supply levels (from 0 to 60 g Se ha⁻¹) and the effect of lime and P on both the Se uptake and the antioxidant activity of plants treated with 0, 20 and 40 g Se ha⁻¹. Selenium concentration in plant tissues was increased by Se applications, and plant growth was reduced at Se supply levels above 20 g ha⁻¹. At shoot concentration up to 200 μg kg⁻¹ DW, Se promoted antioxidant ability by increasing the free radical scavenging activity and by inhibiting lipid peroxidation (TBARS), whereas above this level TBARS accumulation increased. Significant changes in the activities of peroxidase (POD) and ascorbate peroxidase (APX) enzymes were also observed as a consequence of the increase in shoot Se concentration. The application of lime and P improved the plant nutrition, which increased the dry matter yield and enhanced the plant’s antioxidative system. Under different combinations of soil acidity and P fertilization a differential uptake of Se by the plant occurred. These factors appear to be responsible for beneficial or detrimental effects of Se in terms of lipid peroxidation of biological membranes and the activation of POD and APX in white clover.     

The measurement and analysis of competitive ability among populations of white clover and perennial ryegrass

Summary Analysis of the competitive interactions among a set of white clover and perennial ryegrass populations indicated that the intra-specific pressures exerted by the white clover plants upon themselves were significantly greater than the inter-specific pressures they exerted upon the perennial ryegrass. This partitioning of competitive effects could not be carried out on the ryegrass populations, however, because the data required the fitting of separate models to monocultures and duocultures. Although not significant at this stage of the experiment, trends were detected among some of the clover-ryegrass duocultures that could be related to their previous coexistence. The results are briefly discussed in relation to the problems surrounding the measurement of competitive effects among species that ideally require very different managements.     

The infectivity ofRhizobium trifolii into a minute excised root of white clover

The degree of root cell mass necessary for normal infection thread production and nodule formation by rhizobia was studied. Excised white clover root tissues of 5.0, 2.5 and 1.0 mm in length, obtained from two days seedlings, were cultured in the dark withRhizobium trifolii 4S. A culture period of seven days was separated into an initial period of three days and a later period of four days. Culture media of liquid on agar (0.8%) were used including Fåhraeus inorganic medium and an organic medium containing vitamins, sucrose, and an extrinsic substance isolated fromR. trifolii 4S cells (ES-6000). When ES-6000 was added in culture medium for the initial period and root segments had an apical meristem, infection threads and nodules were most numerous.     

Simple empirical intensity and buffering capacity measurements to predict potassium uptake by white clover

Summary This experiment examined the feasibility of predicting K uptake in white clover by the use of simple and relatively rapid tests that would dispense with the need of knowing the quantity of plant available potassium. Potassium uptake was found to correlate highly (R=>0.9) in linear bivariate regressions using K concentration in the soil solution displaced by centrifuging and an empirical estimate of potassium retention. There was no advantage in determining activity ratios because in at least some of the soils used the ratio law did not apply. Exchangeable potassium alone correlated rather poorly with uptake except at very low soil potassium status. This was not because nonexchangeable potassium was an important source of this nutrient to white clover but because of the large differences in the retention of K amongst the soils used.     

Influence of boron on the reproductive growth of white clover (Trifolium repens L.) cultivars

The effect of boron on the reproductive growth of four contrasting white clover cultivars was examined in a controlled environment and glasshouse experiment. Plants grown under a nutrient regime excluding boron and those receiving 0.5 mg/litre boron had fewer stolons, reproductive stolons and inflorescences than those receiving 1, 2 or 3 mg/litre. Inflorescences were also smaller and had fewer seeds per floret. A boron concentration of 1 mg/litre gave optimum reproductive growth, higher levels of boron giving no further increase. Peduncle length and nectar quantity, but not nectar quality, were also influenced by boron indicating a possible beneficial effect of this element on pollinator activity. Cultivars differed in both vegetative and reproductive growth. Cv. Gwenda, a small leaved cultivar, had more stolons but fewer inflorescences than cv. Menna and smaller inflorescences than cv. Olwen, a large leaved cultivar. Generally, these cultivars responded similarly to boron. The implications of these results for the reproductive growth of white clover are discussed.     

The response of white clover to different strains ofRhizobium trifolii in hill land reseeding: A second trial

Summary S. 184 white clover was surface seeded into natural molinia pasture on wet stagnogley soil containing no indigenousRhizobium trifolii. Seedlings were ‘spray inoculated’ after emergence with each of three strains ofR. trifolii. The best of these treatments produced an eight fold improvement in dry matter in the seeding year, followed by a 28% improvement in the following year. The results confirm the potential benefits which may be achieved by inoculating clover with suitable strains of rhizobia. The data are compared with a previously reported trial on an adjacent site where benefits were much greater in the first year. The difference is attributed to the overall advantages conferred in the present trial by much higher seedling populations and less severe competition from native species in the establishment phase.     

Some soil-plant and root-shoot relationships of copper,zinc and manganese in white clover and perennial ryegrass

Summary White clover and perennial ryegrass were grown separately, in pots maintained under controlled environment conditions, for a period of 7 months on ten soils. The proportion of the total soil content of each element taken up by the ryegrass, including that in roots, ranged from 0.88 to 2.18% for Cu, from 0.82 to 2.80% for Zn and from 0.25 to 3.15% for Mn. Uptake by the clover was within these ranges for Cu and Zn, but ranged from 0.10 to 1.71% for Mn.After adjustment for the effects of soil contamination, the ratio of root concentration: shoot concentration was always greater than 1 for both Cu and Zn, and for Cu, though not for Zn, it was considerably greater with ryegrass than with clover. For Mn, the ratio of root:shoot concentration was often greater than 1, and differences between clover and grass were not consistent.Concentrations of Cu and Zn in the shoots were always greater in the clover than in the grass, but concentrations of Mn were generally greater in the grass than the clover.     

Longevity of white clover (Trifolium repens) leaves, stolons and roots, and consequences for nitrogen dynamics under northern temperate climatic conditions

BACKGROUND AND AIMS: White clover (Trifolium repens) is, due to nitrogen (N) fixation, important to the N dynamics of several northern temperate agroecosystems. This study aimed at monitoring growth and death of major white clover plant organs to assess their potential contribution to within-season N input and risk of off-season N losses. METHODS: White clover ('Snowy') was studied in a plot and root window experiment in southeast Norway (60 degrees 42'N, 10 degrees 51'E). Leaves, stolons and roots were tagged for lifespan measurement in harvested and unharvested stands during two experimental years. The availability of soil inorganic N was measured by plant root simulator (PRS) probes. KEY RESULTS: The longevity of leaves and petioles ranged from 21 to 86 d (mean = 59 d), of main stolon sections from 111 to over 677 d (mean = 411 d) and of roots from 27 to 621 d (mean = 290 d). About 60 % of the leaves produced had turned over by the end of the growing season and another 30 % had died or disappeared by the subsequent spring. Harvesting reduced the longevity of stolons and increased plant fragmentation, but did not decrease leaf or root lifespan or increase soil N availability. From the plant organ turnover data, it was estimated that the gross N input to the soil-plant system from white clover in pure stand during two growing seasons corresponded to a 2.5-fold increase over the total N in harvestable shoots. CONCLUSIONS: The short lifespan and poor over-wintering of leaves showed their potential importance as a nitrogen source in the soil-plant ecosystem but also their potential contribution to the risk of off-season N losses.     

Coexistence and the productivity of white clover-perennial ryegrass mixtures

Summary Yield comparisons between five white cloverperennial ryegrass mixtures, whose individual components had previously coexisted, and a corresponding set of ten mixtures based on components that had not coexisted disclosed a yield advantage to the former group of 8.5% over a 4-year period. All five clover populations produced their highest yields when associated with their coexisting grass. The coexisting mixtures also yielded more grass in spring, probably due to the exotic origin of the majority of the companion grasses, reinforced by the nitrogen transfer process between clover and grass. The relative proportions of clover attained by the five populations was apparently unaffected by grass companion. These results are briefly discussed in the context of improving the productivity of white clover-perennial ryegrass mixtures.     

The growth and asymmetry of neighbouring plants of white clover (Trifolium repens L.)

Summary Transplants of white clover (Trifolium repens L.) were grown isolated from each other and in pairs placed at different distances apart. The paired plants developed asymmetrically and at the interface between paired clones both the density of nodes and of stolons appeared to reach ceiling values that were of the same order as those achieved in isolated clones. It is argued that the growth of plants of T. repens is controlled by the local conditions experienced by the plant parts and not by integrated growth of the whole. Transplants of three different genotypes of T. repens, which differed in growth form, were grown as neighbouring pairs and the calculated asymmetry of the plants was used to compare their mutual aggressivenes. The more compact (phalangeal) genotypes induced greater asymmetry in their neighbours than the more diffuse forms.     

Nodulation of white clover (Trifolium repens) in the absence ofRhizobium

Summary Spontaneous nodules developed on the roots of white clover (Trifolium repens cv. Ladino) in the absence ofRhizobium. A small subpopulation of uninoculated clover plants (0.2%) exhibited white, single-to-multilobed elongated structures on their root systems when grown without fixed nitrogen. Clonal propagation using aseptic stolons confirmed the genetic stability of the observation. Few if any viable bacteria of unknown origin were recovered from surfacesterilized structures. Nodule contents were incapable of eliciting nodulation. Histological observations showed that these structures possessed all the characteristic features of indeterminate nodules, such as active meristem, cortex, endodermal layer, vascular strands, and a central zone with parenchyma cells. Infection threads, intercellular or intracellular bacteria were absent. Instead, numerous starch grains were observed in the central zone, a feature absent in normal nitrogen-fixing nodules. Our observation broadens the concept of spontaneous nodulation, believed to be restricted to alfalfa (Medicago sativa), to other legumes, and suggests a degree of generality among indeterminately nodulated legumes displaying natural heterozygosity.     

Differential effects of low concentrations of aluminium on the growth of four genotypes of white clover

Summary The effects of aluminium (Al³⁺) on the growth of four cultivars of white clover dependent upon NO₃ ⁻−N were examined. Plants were grown in flowing solution culture with carefully maintained low concentrations (0, 12.5, 25 and 50 mmolm⁻³) of Al, and with P and pH (4.5) also held constant and appropriately low. A three-week treatment period resulted in major effects on the growth and elemental composition of shoots and roots at all concentrations of added Al. There were inherent differences between the cultivars in growth but the relative effects of Al were similar in each case. Examination by S.E.M. and x-ray microanalysis of one cultivar grown at 50 mmolm⁻³ Al, indicated that Al in the roots was associated with P, especially in old, outer epidermal cells. Aluminium reduced NO₃ ⁻ uptake and there were significant effects of Al on nitrate reductase activity (NRA). In contrast to the other characteristics, there were differential effects between the cultivars in NRA, both in the presence and absence of Al.