Effects of temperature and photoperiod on vegetative growth of white clover (Trifolium repens) ecotypes

Effects of photoperiod and temperature on vegetative growth of seedling populations and clones of white clover ( Trifolium repens L.) originating from various latitudes (58°48'–69°54'N) and altitudes (up to 1100 m above sea level) were studied in a phytotron. Dry matter production, stolon elongation, petiole elongation and leaf lamina size were enhanced by long photoperiod. The requirement for long photoperiod increased with decreasing temperature. At 6°C the maximum growth was recorded under 24-h daylength. At 18°C already an increase in photoperiod from 12 to 15 h significantly enhanced growth, and maximum growth was obtained at 21-h photoperiod. The studied populations responded similarly to daylength, and the results did not indicate photoperiodic ecotypes in the material. The southernmost clones and populations generally had the highest dry matter production at all temperature treatments (6–18°C). Variation between clones within one location was, however, significant, and rapidly growing clones were found also in high-latitude locations. Dry matter production was poorly correlated with the morphological characters observed, but in some cases significant correlation with leaf lamina size was found.     

Studies on the feeding of Sitona lineatus L. (Coleoptera: Curculionidae) on white clover (Trifolium repens L.) seedlings

Measurements of feeding damage by sitona weevil (Sitona lineatus L.) adults on differing numbers of seedlings of white clover (Trifolium repens L.) at the first and fourth trifoliate leaf stage were made in the glasshouse at 20°C. S. lineatus consumed more of the trifoliate component of the seedling. Sitona adults caused significant yield reduction at all levels of plant population. Total clover consumption increased with increasing size of sitona population, but consumption per adult weevil was reduced.     

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.     

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.     

Symbiotic growth of indigenons white clover (Trifolium repens) with local Rhizobium leguminosarum biovar trifolii

To study a possible adaptation of the symbiosis between white clover (Trifolium repens L.) and Rhizobium leguminosarum biovar trifolii with regard to light and temperature at northern latitudes, local seed populations of white clover and isolates of R. leguminosarum biovar trifolii from 3 different latitudes in Norway, 58°48'N, 67°20'N and 69°22'N, were used. The commercial cultivar Undrom was used as a reference plant. The experiments were done at 18 and 9°C under controlled conditions in a phytotron during the natural growing season at 69° 39'N. Growth of the plants was evaluated by number and size of leaves, dry matter production and total N-content. At 18°C the white clover plants were harvested twice while at 9°C there was only one growth period. The results from first harvest at 18°C and total growth at 9°C, showed that white clover populations from northern Norway had a lower growth potential than the population from the south and cv. Undrom. This difference was not apparent in the second growth period at 18°C. Growth of the plants from seeds to first harvest was enhanced by mineral nitrogen compared to plants dependent on Rhizobium only. However, after a second growth period dry weight and total nitrogen content of the plants with nitrogen fixation were comparable to the plants receiving mineral nitrogen. Statistical analysis showed that the most important factor for the variation in dry matter production was the plant population. Within the populations at 9°C and at first harvest at 18°C, there were no significant differences in dry matter production with different Rhizobium inoculum. In the second growth period at 18°C, different inoculum gave significantly different amount of dry matter within a population. The results showed a significant interaction between plant population and Rhizobium inoculum, and the results indicated that plants from the north gave higher yield when nodulated by Rhizobium from the north than from the south.     

Incidence and severity of pest and disease damage to white clover foliage at 16 sites in England and Wales

From 1985 to 1987, leaves of white clover ( Trifolium repens ) cv. Grasslands Huia were examined for damage by pests and fungal diseases on seven occasions at up to 16 sites in England and Wales. Pest damage was recorded at all sites on all sampling dates. Over all sampling dates, the mean number of leaves damaged by slugs ranged from 23% to 67%, and that by weevils ( Sitona spp.) ranged from 3% to 62%. Also, up to 30% of leaves were damaged by other, unidentified pests. At individual sites, total pest damage frequently exceeded 90% of leaves. The area of leaf damaged by pests ranged from 2 - 12%. Fungal diseases were recorded in July and September but not in May, and were more prevalent in September 1985 than in September 1986 or 1987. Black blotch ( Cymadothea trifolii ) was the most frequently recorded disease, and the mean number of leaves damaged ranged from 4% to 21%. The mean area of leaves covered by lesions was 1–2%. Infections by viruses were assessed on two occasions, using indicator plants and electron microscopy, and only a very low incidence of arabis mosaic virus and red clover necrotic mosaic virus was recorded.     

Sensitivity of white clover to UV-B radiation depends on water availability, plant productivity and duration of stress

Abstract Nine populations of white clover ( Trifolium repens L.) were grown for 12 weeks with supplemental application of 13.3 kJ m⁻² d⁻¹ ultraviolet-B (UV-B) radiation under controlled environmental conditions. Drought was applied during the last four weeks of the experiment. Under well-watered conditions, UV-B decreased white clover growth on average by 20%. Cultivars bred for agricultural performance were sensitive to UV-B, while slow-growing ecotypes were UV-B-tolerant. After four weeks of water stress, there were no significant population differences in UV-B responsiveness. UV-B sensitivity decreased with increasing exposure to drought and with longer duration of UV-B irradiation, suggesting that the direction and extent of the UV-B 3× drought interaction depends on the duration of stress. The population comparisons indicate that low constitutive growth rate and adaptation to other forms of stress may be related to UV-B tolerance under well-watered conditions, but not during extended periods of drought.     

Photoinhibition of white clover seed germination at low water potential

Photosensitivity of germination of white clover ( Trifolium repens L. cv. Podkowa) seeds was studied under water deficit (low water potential) conditions at 25°C. The seeds showed negative photoblastism, which was most pronounced at -0.03 MPa polyethylene glycol solution. Inhibition was observed at two different wavelength bands with maxima at 660 nm (R) and around 730 nm (FR). Red light acted identically to white light (maximum inhibition ca 50%). The effect of far-red illumination was less inhibitory (20–30%). The photoresponse required long illuminations (3 h exposures); saturation level was at 0.1 W m⁻², independently of the light quality. White clover seed germination showed no reversibility of the effects of R and FR light. Prolonged illumination with R and FR increased the inhibition, and intermittent illumination had a higher effect than a continuous one. It was concluded that the photoinhibition of germination of seeds of Trifolium repens involves a reaction dependent on the rate of phytochrome interconversion, a property that is characteristic for the high irradiance reaction.     

The influence of ectomycorrhiza on nitrogen nutrition and growth of Pinus sylvestris seedlings

Ectomycorrhizal seedlings of Scots pine ( Pinus sylvestris L. cv.), inoculated with the fungus Suillus bovinus (L. ex Fr.) O. Kuntze, and non-mycorrhizal controls were grown in growth units with a circulating culture solution. Steady-state nutrition and constant relative growth rates were achieved by means of varied relative nutrient addition rates and free access of nutrients. Typical mycorrhizas always formed within a short period of time after inoculation. The nutrition/growth relationships were in principle similar to previous studies under steady-state conditions: there were close linear relationships between relative addition rate, relative growth rate and internal nitrogen concentration, i.e. an equilibrium established between nutrients added and taken up. This occurred when infected and uninfected seedlings were grown separately. When grown together in the same growth unit, there are indications that the fungus influenced the exudation pattern of the uninfected seedlings. More carbon was thus provided to the unspecified microflora in the cultivation system, and it was able to grow and withhold nitrogen from the seedlings. The mycorrhizal infection did not increase the specific uptake capacity of the roots, and the fungus constituted a sink for carbon. However, the nitrogen productivity (growth rate per unit of nitrogen per unit of time) was similar for mycorrhizal and non-mycorrhizal seedlings, so that there might be mechanisms which compensate for the carbon cost.     

Effect of pollen competition and stigmatic receptivity on seed set in white clover (Trifolium repens L.)

The effect of competition between incompatible and compatible pollen grains on the seed production of white clover was studied. Stigmatic receptivity was also studied. A selection line of white clover that has red leaves was used as a pollen donor and as a genetic marker to allow determination of the pollen donor responsible for ovule fertilisation. Results show that incompatible pollen did not inhibit compatible pollen grains from fertilising ovules and producing seeds, although it did slightly impair seed yield. At temperatures of 20/10°C (day/night) stigmas of white clover remained receptive to additional pollination up to 32 h after an initial compatible pollination and 40 h after an incompatible pollination. The results imply that factors other than inadequate pollination and the self-incompatibility system are responsible for failure of white clover to attain its potential seed yield. Received: 31 August 1999 / Revision accepted: 28 February 2000     

Photosynthesis and carbon export in white clover plants grown at various levels of phosphorus supply

Net photosynthesis, concurrent carbon export and starch, sucrose and inorganic phosphorus concentrations were measured in leaves of white clover ( Trifolium repens L. cv. Grasslands Huia) grown at four levels of phosphorus supply in the presence or absence of mineral nitrogen. The nitrogen treatments had no effect on growth, photosyntheis or carbon export. At the three higher levels of phosphorus supply, the amount of carbon exported was about 77% of net fixation. Photosynthesis and export per leaf decreased with phosphorus supply, primarily through the effect of phosphorus supply on leaf area. The rate of photosynthesis was reduced only at the lowest level of phosphorus supply.
Inorganic phosphorus rose with phosphorus supply but starch concentration was unaffected. Sucrose was reduced at the lowest level of phosphorus supply but not significantly affected at higher levels. The ratio between starch and sucrose concentration was also unaffected at the higher levels, but was increased at the lowest level of supply. There thus appeared to be direct effects of phosphorus supply on photosynthesis, partitioning of carbon to carbohydrates and, by implication, export, only at the lowest level of phosphorus supply. As leaf area and plant growth were affected over the whole range of phosphorus supply, factors other than photosynthesis per se must have determined the response of growth to phosphorus supply.     

Phosphorus concentrations in the leaves of defoliated white clover affect abscisic acid formation and transpiration in drying soil

Increased leaf phosphorus (P) concentration improved the water-use efficiency (WUE) and drought tolerance of regularly defoliated white clover plants by decreasing the rate of daily transpiration per unit leaf area in dry soil. Night transpiration was around 17% of the total daily transpiration. The improved control of transpiration in the high-P plants was associated with an increased individual leaf area and WUE that apparently resulted from net photosynthetic assimilation rate being reduced less than the reductions in the transpiration (27% vs 58%). On the other hand, greater transpiration from low-P plants was associated with poor stomatal control of transpirational loss of water, less ABA in the leaves when exposed to dry soil, and thicker and smaller leaf size compared with high-P leaves. The leaf P concentration was positively related with leaf ABA, and negatively with transpiration rates, under dry conditions ( P < 0.001). However, leaf ABA was not closely related to the transpiration rate, suggesting that leaf P concentration has a greater influence than ABA on the transpiration rates.     

Involvement of a high irradiance response in photoinhibition of white clover seed germination at low water potential

Seeds of white clover ( Trifolium repens L., cv. Podkowa) were germinated at water potential ψ=−0.3 MPa in darkness, at 25°C. A short exposure to blue light (B) inhibited germination in a manner similar to that described earlier for red (R) and far‐red (FR) light (Niedźwiedź‐Siegień and Lewak 1989). No reversibility of B, R and FR effects was observed. Saturation irradiance and energy was the lowest for R and the highest for B. The reciprocity of irradiance versus time of exposure was observed only for non‐saturating irradiances of B, R and FR.     

The influence of phosphorus deficiency on growth and nitrogen fixation of white clover plants

The effects of P deficiency on growth, N(2)-fixation and photosynthesis in white clover (Trifolium repens L.) plants were investigated using three contrasting relative addition rates of P, or following abrupt withdrawal of the P supply. Responses to a constant below-optimum P supply rate consisted of a decline in N(2)-fixation per unit root weight and a small reduction in the efficiency with which electrons were allocated to the reduction of N(2) in nodules. Abrupt removal of P arrested nodule growth and caused a substantial decline in nitrogenase activity per unit root weight, but not per unit nodule mass. Similarly, the rate of photosynthesis per unit leaf area was unaffected by abrupt P removal, whereas CO(2) acquisition for the plant as a whole decreased due to a decline in total leaf area, leaf area per unit leaf weight and utilization of incoming radiation. These changes followed the decline in tissue P concentrations. The ratio between CO(2)-fixation and N(2)-fixation was maintained under short-term P deprivation but increased under long-term low P supply, indicating a regulatory inhibition of nodule activity following morphological and growth adjustments. It is concluded that N(2)-fixation did not limit the growth of clover plants experiencing P deficiency. A low P status induced changes in the relative growth of roots, nodules and shoots rather than changes in N and/or C uptake rates per unit mass or area of these organs.