Flowering of Contrasting White Clover Varieties in Relation to Temperature in Controlled Environments

Effects of temperature on floral initiation of ten white clovervarieties growing in controlled environments are described.Plants grown under long days (16 h) were subjected to constanttemperatures of 26, 18 and 10 ^°C. Relationships betweenmorphological and physiological traits and flowering were examined. Most plants flowered at the two higher temperatures but only10 per cent of plants flowered at 10 ^°C. Larger leaved typestended to produce more reproductive buds per stolon at the highertemperatures than did smaller leaved varieties. Of the floral characters studied, floret number was least affectedby temperature. Ovule number and peduncle length were greatestat 18 ^°C. Variation in, and absolute level of nectar secretionwas greatest at the highest temperature. Trifolium repens, white clover, flowering, temperature     

Influence of Temperature and Ageing of Ovules and Pollen on Reproductive Success in Trifolium repens L.

Four clones of Trifolium repens L., originating from Iceland,Denmark and Israel, were cultivated in controlled environmentchambers at 10, 14, 18 and 22°C. Seed set at 10°C was,on average, 45% lower than at 18°C. Generally, seed frequencieswere higher in the stylar ovule positions than in the basal.This polarization became more pronounced at low temperatures.At least 30% of the decrease in seed set was caused by inadequatepollen tube growth. This was further supported by in vitro studieswhich showed that pollen developed significantly shorter pollentubes at low temperatures and a strong positive correlationbetween the capacity of male parents to induce seed set in basalovule positions and the length of pollen tubes in vitro. Femaleage, determined by days relative to anthesis, was also foundto have a significant influence on seed set, i.e. when pollinationwas delayed until the fifth day of flowering, seed set decreasedby an average of 60% when compared to florets pollinated onthe first day of anthesis. Pollen age did not influence seedset. Ageing of pollen decreased in vitro germinability, tubeelongation and the percentage of pollen tubes with sperm cells.Seed set, pollen germinability, pollen tube lengths, and abilityto form sperm cells in vitro varied significantly among clones.Copyright1994, 1999 Academic Press Trifolium repens, white clover, temperature, pollen tube, sperm cells, ovule fertility, seed set     

Floral Initiation of White Clover (Trifolium repens L.) in Response to Daylength and Temperature

The response to different combinations of daylength (14, 15and 16 h) and day/night temperature (20/10°C, 30/15°C)of four white clover (Trifolium repens) varieties in terms offloral initiation and development was studied. More plants flowered under long days (16 h) at 30°C thanin any other combination. Varietal differences in response todaylength were large but temperature was the factor controllinginitiation. Daylength was more important in regulating peduncle length,floret and ovule numbers than was temperature. The amount ofnectar produced was lowest at the higher temperature althoughnectar concentration was unaffected by temperature. Nectar concentrationwas lowest in the shortest daylength (14 h). White clover, floral initiation, temperature, daylength     

Characteristics of Nectar Secretion by the Extrafloral Nectaries of Ricinus communis

The characteristics of nectar secretion by excised extrafloralnectaries of Ricinus have been examined. Secreted nectar wasfound to contain three sugars: sucrose, glucose and fructose,with glucose and fructose occurring in a 1: 1 ratio. All threesugars supported secretion when used in the culture medium andthe yield of nectar sugar was found to be concentration-dependent.Other sugar sources failed to support secretion. Experimentsusing ¹⁴C-sugars and ¹⁴CO₂ fed to intact plants allowed themovement of sugars through the nectary to be examined. Sucrosesynthesis occurs when excised glands are fed glucose and thisoccurs very early in the transport through the nectary. Themain sugar transported was sucrose, with little hydrolysis occurringuntil the final step of secretion. There was no evidence thatsucrose hydrolysis occurs either by invertase in the nectaror by a microbial flora. Inhibitors of respiration were foundto inhibit secretion as did anaerobiosis. Temperature also hada marked effect, with a temperature coefficient of 1.8. However,secretion of sucrose was not affected by anaerobic conditions,low temperatures or inhibitors of respiration as markedly asthat of glucose and fructose. Electron microscopy revealed the presence of a thickened andheavily stained wall at the inner border of the secretory epidermallayer. This wall contained numerous plasmodesmata at a frequencyof 14 per µm² and may represent an apoplastic barrier.Light microscope cytochemistry revealed that acid phosphataseis primarily located in the nectiferous tissue, while ATPaseis concentrated in the epidermis. The possibility that the nectarycontains two pathways for sucrose secretion, both apoplasticand symplastic, is discussed. Key words: Invertase, nectary, plasmodesmata, Ricinus communis, sucrose     

Effect of Temperature and Nitrogen Supply on the Growth of Perennial Ryegrass and White Clover. 2. A Comparison of Monocultures and Mixed Swards

White clover (Trifolium repens L.) and Perennial ryegrass (Loliumperenne L.) plants were grown, in Perlite, in simulated swardsas either monocultures or mixtures of equal plant numbers. Theywere supplied with a nutrient solution either high (220 µgg^–1) or low (40 µg g^–1) in ¹⁵N-labelled nitrateand grown to ceiling yield at either high (20°C day/15°Cnight) or low (10°C day/8°C night) temperature. Temperature had little effect on the maximum rates of grosscanopy photosynthesis which were similar in High-N grass andHigh-N and Low-N clover monocultures. However these maxima werereached more slowly in clover than grass, and more slowly atlow rather than high temperature. Nitrogen supply increasedphotosynthesis in grass but not in clover. Clover had higherN contents than grass in all four treatments, although in anygiven treatment its N content was lower, and contribution ofN₂-fixation relative to nitrate uptake higher, in mixture thanin monoculture. Conversely, grass had higher N contents in mixturethan monoculture, because more nitrate was available per plantand not because of transfer of biologically fixed N from clover. Under Low-N, clover outyielded grass in mixture, particularlyat high temperature. The grass plants in the Low-N mixtureshad higher N contents and higher SLA, LAR and shoot: root ratiosthan those in monoculture. It is proposed that competition forlight is the cause of the low relative yield and negative aggressivityof grass in these swards. Under High-N, grass outyielded cloverin monoculture and mixture, at both temperatures but particularlyat low temperature when grass had a high aggressivity. Nitrogenand yield component analyses shed no light on clover's apparentlylow competitive ability and evidence is drawn from the previouspaper to demonstrate that grass grew faster than clover onlyas spaced individuals during non-com petitive growth. The relativemerits of measures of competitive ability based on final harvestdata and physiological data taken over a growth period are discussed. Trifolium repens L., white clover, Lolium perenne, perennial ryegrass, competition, temperature, nitrogen     

Supercooling ability of Rhododendron flower buds in relation to cooling rate and cold hardiness

The freezing process and supercooling ability in flower budsof 11 native Rhododendron species were examined with referenceto the cooling rate and cold hardiness by differential thermalanalysis. The freezing patterns of the excised whole buds variedwith the season: in autumn, buds froze as whole units, whilein winter, freezing was initiated in the scales and propagatedto each floret. The supercooling ability of florets was enhancedduring winter. The freezing patterns in winter buds were stronglyinfluenced by the cooling rate (1 to 30°C/hr). Althoughthe first exotherm in scales occurred at –5 to –10°Gand was rate-independent, the occurrence of several floret exothermsshifted considerably to lower subzero temperatures at slowerrates. The most reliable cooling rate for testing maximum supercoolingability was l°C/hr. The exotherm in florets of hardier speciesoccurred at –20 to –25°C and at –7 to–20°C for less hardy ones, and were well correlatedwith their killing temperatures. Water relations within budtissues in response to freezing are briefly discussed. (Received June 26, 1980; )     

Supercooling ability of Rhododendron flower buds in relation to cooling rate and cold hardiness

The freezing process and supercooling ability in flower budsof 11 native Rhododendron species were examined with referenceto the cooling rate and cold hardiness by differential thermalanalysis. The freezing patterns of the excised whole buds variedwith the season: in autumn, buds froze as whole units, whilein winter, freezing was initiated in the scales and propagatedto each floret. The supercooling ability of florets was enhancedduring winter. The freezing patterns in winter buds were stronglyinfluenced by the cooling rate (1 to 30°C/hr). Althoughthe first exotherm in scales occurred at –5 to –10°Gand was rate-independent, the occurrence of several floret exothermsshifted considerably to lower subzero temperatures at slowerrates. The most reliable cooling rate for testing maximum supercoolingability was l°C/hr. The exotherm in florets of hardier speciesoccurred at –20 to –25°C and at –7 to–20°C for less hardy ones, and were well correlatedwith their killing temperatures. Water relations within budtissues in response to freezing are briefly discussed. (Received June 26, 1980; )     

Temporal changes in floral nectar production, reabsorption, and composition associated with dichogamy in annual caraway (Carum carvi; Apiaceae)

The dynamics of nectar production were studied in perfect florets of two varieties (Karzo, Moran) of annual caraway (Carum carvi L., Apiaceae). Florets were protandrous and strongly dichogamous, lasting 7-15 d but producing nectar from the stylopodia for 4-12 d, in an interrupted fashion. Nectar secretion began during a floret's phase of stamen elongation and anther dehiscence. After reabsorption of uncollected nectar, at which point nectary surfaces were completely dry, the two styles elongated and a second bout of secretion commenced during the female phase, up to 5 d later, when a floret became receptive to pollination. During the male and female phases, respectively, 0.392 ± 0.064 μL and 1.083 ± 0.261 μL of nectar of similar solute concentration (844 mg/mL) was produced per ten florets. On a daily basis, florets yielded 1.5-fold more nectar in the female than during the male phase. First-time nectar removal throughout the female phase did not match the sum of nectar quantities from male and female phases combined, suggesting that under natural conditions, any uncollected male-phase nectar, once reabsorbed, is not made available to visitors of the same florets when in the female phase. Nectar-sugar composition differed between bouts of secretion; it was hexose-rich (59.6% fructose, 26.9% glucose, 13.6% sucrose) initially, but hexose-dominant (70.2, 26.8, 3.1) during the female phase. A 5.7-fold difference in mean nectar production per floret occurred among plants.     

The Growth and Photosynthesis of Seedling Plants of White Clover at Low Temperature

The growth of white clover (Trifolium repens L.) in conditionstypical of April in Southern England (8 °C day/4 °Cnight, 12 h photoperiod of 90 J m^–2 s^–1 visibleradiation) was extremely slow, whether the plants were dependentfor nitrogen on fixation by their root nodules or were suppliedwith abundant nitrate; although growth was slower in the nodulatedplants. The reasons for slow growth were a large root: shootratio and a small leaf area, particularly in the nodulated plants,and a low photosynthetic rate in all plants. The probable effectsof these characteristics on the growth of white clover withgrasses in mixed pastures are discussed. Trifolium repens L, white clover, low temperature, leaf area, photosynthetic rate, nitrogen supply, growth     

Concurrent Rates of N2 Fixation, Nitrate and Ammonium Uptake by White Clover in Response to Growth at Different Root Temperatures

Nodulated white clover plants (Trifolium repens L. cv. Huia)were grown for 71 d in flowing nutrient solutions containingN as 10 mmol m_–3 NH₄NO₃, under artificial illumination,with shoots at 20/15°C day/night temperatures and root temperaturereduced decrementally from 20 to 5°C. Root temperatureswere then changed to 3, 7, 9, 11, 13, 17 or 25°C, and theacquisition of N by N₂ fixation, NH₄+ and NO₃^– uptakewas measured over 14 d. Shoot specific growth rates (d. wt)doubled with increasing temperature between 7 and 17°C,whilst root specific growth rates showed little response; shoot:root ratios increased with root temperature, and over time at11°C. Net uptake of total N per plant (N₂ fixation + NH₄++ NO₃^–) over 14 d increased three-fold between 3 and 17°C.The proportion contributed by N₂ fixation decreased with increasingtemperature from 51% at 5°C to 18% at 25°C. Uptake ofNH₄+ as a proportion of NH₄+ + NO₃^– uptake over 14 d variedlittle (55–62%) with root temperature between 3 and 25°C,although it increased with time at most temperatures. Mean ratesof total N uptake per unit shoot f. wt over 14 d changed littlebetween 9 and 25°C, but decreased progressively with temperaturebelow 9°C, due to the decline in the rates of NH₄+ and NO₃^–uptake, even though N₂ fixation increased. The results suggestthat N₂ fixation in the presence of sustained low concentrationsof NH₄+ and NO₄^– is less sensitive to low root temperaturethan are either NH₄+ or NO₃^– uptake systems. White clover, Trifolium repens L. cv. Huia, root temperature, nitrogen fixation, ammonium, nitrate     

The Effect of High Temperature at Different Stages of Ripening on Grain Set, Grain Weight and Grain Dimensions in the Semi-dwarf Wheat 'Banks'

Grain number in the wheat cultivar Banks was reduced by up to11 % with a rise in temperature from 21/16 °C to 30/25 °Cover a 10-d period immediately following first anthesis in general,the upper ‘d’ and ‘c’ florets were moreaffected by high temperature than the basal ‘a’and ‘b’ florets within a spikelet and florets fromthe upper spikelets were more sensitive than those lower onthe ear Grain weight and grain length at maturity were most affectedby a 10 d period of high temperature commencing 7–10 dafter anthesis However, if dry-matter accumulation between thestart of a treatment and grain maturity was used as a base forcomparison, the response was more uniform throughout development,with a peak in sensitivity 25 d after anthesis Although grainposition within an ear did not have a large effect on the responseto temperature, grains from the basal spikelets were more sensitivethan those from the apex, and the upper floret grains of a spikeletwere more sensitive to high temperature than those at the base There is a need to obtain, for a range of cultivars, more comprehensivedata on the effect of the timing and degree of temperature stressfollowing anthesis, for use in interpreting the response torising temperatures late in the development of the crop in thefield Triticum aestivum L, wheat, temperature, grain development     

Requirements for Floral Induction in Contrasting White Clover (Trifolium repens) Populations

Norm, I. B. 1987. Requirements for floral induction in contrastingwhite clover (Trifolium repens) populations.—J. exp. Bot.38: 900–907. Floral initiation and development of four contrasting whiteclover (Trifolium repens) populations was examined after differentinduction treatments (16 h, 5 ?C and 8 h, 5 ?C. The number of reproductive stolons and of reproductive budsper stolon was increased after cold induction. Varietal differencesin response to daylength were large; some varieties respondingbetter to a long day cold period, others to a short day coldperiod while one variety required no induction at all. Whetherthe daylength effect was due to photoperiod, irradiance or totheir interaction was not known. The induction periods had a subsequent effect upon pedunclelength, floret and ovule number. Short days and chilling reducedpeduncle length but increased ovule number, whereas long daysand chilling tended to increase floret number. Nectar concentrationwas highest after short day induction. Key words: White clover, floral initiation, floral induction     

Photoperiod and Temperature Effects on Late Developmental Stages of Stylosanthes guianensis

Seedlings of Stylosanthes guianensis var. guianensis cv. Cookand S. guianensis var. pauciflora cv. Bandeirante were defoliatedand placed in a naturally lit glasshouse at 23/18 °C, 28/23°C or 33/28 °C (day/night). After exposure to 14 h daysand after floral induction with 30 cycles of 11 h, plants wereallocated to 11, 12, 13 or 14 h during flowering and seed formation. Floral initiation occurred after 10–15 short-day cycles.Flower appearance was hastened by warm temperatures and spikenumber per plant at 20 d after flower appearance was negativelyrelated to temperature and greater in Cook than in Bandeirante.Exposure to 13- and 14-h days reduced the continued differentiationof inflorescences in Bandeirante, and in Cook in warm temperatures.Floret number per spike was greatest at 23/18 °C and a higherproportion of florets aborted in Bandeirante at 33/ 28 °C.Variations in seed setting of the bi-articulate loment of Bandeiranteare described. Highest potential seed yield occurred if afterfloral induction 11 or 12 h days were maintained with 23/18°C or 28/23 °C temperatures. Photoperiod, temperature, development, Stylosanthes guianensis, flowering     

Freezing Avoidance Mechanisms by Supercooling in Some Rhododendron flower Buds with Reference to Water Relations

Excised florets of some hardy Rhododendron species did not toleratefreezing at –5°C when ice-inoculated due to intracellularfreezing. Florets in intact December buds, however, could besupercooled to about –30°C. When flower buds of R.japonicum were slowly cooled with daily decrements of 5°Cto temperatures ranging from 0 to –20°C, the exothermtemperatures of the florets drastically decreased. This wasaccompanied by a decrease in water content of florets and peduncleand an increase in that of scales. The water in florets andthe peduncle is thought to migrate to scales and other tissuesduring the early stages of freezing; the dehydrated floret hasa lower freezing point which enhances its supercooling abilityand the dehydrated peduncle helps to maintain the supercooledstate of the florets. This hypothesis would explain the dependenceon the cooling rate of supercooling in Rhododendron flower buds.Water migration within flower buds was observed in other hardyRhododendron species with some variation in ice formation siteand the quantity of migrated water. The exotherm temperatureof excised florets was inversely proportional to their watercontent. Dehydration of flower buds by wind at 0°C alsoenhanced their supercooling ability. Mechanisms of freezingavoidance by supercooling in Rhododendron flower buds and therelationship of supercooling to freezing tolerance are discussed. ¹ Contribution No. 2254 from the Institute of Low TemperatureScience ² This is a revised form of the master's thesis of the seniorauthor (M.I.) which is cited in the present and previous papers(Sakai 1979a, b, etc.). (Received August 11, 1980; Accepted June 1, 1981)     

STRUCTURE OF FLORAL NECTARIES OF ALFALFA (MEDICAGO SATIVA L.) IN RELATION TO NECTAR PRODUCTION

Alfalfa (Medicago sativa L.) plants differ in nectar volume production per floret. These differences are heritable, but no information is available on what type of structural variation may be responsible for these differences. One high, one intermediate, and one low nectar-volume-producing clone was selected from each of two alfalfa cultivars. Results from light and scanning electron microscopy indicated that alfalfa has an annular nectary located on the staminal column and primarily on the receptacle. It is composed of several cell layers subtended by vascular bundles containing both xylem and phloem. Vascular tissue does not extend into nectariferous tissue. Permanently open stomata are present in the epidermal layer and are thought to function in nectar secretion. These stomata did not respond to stimuli known to affect leaf stomata. Number of stomata per nectary among the six clones ranged from 24.7 ± 1.9 to 6.8 ± 0.5. Nectar-reservoir diameters among clones ranged from 1.07 ± 0.09 mm to 0.70 ± 0.01 mm. Clones producing the most nectar were characterized by a large nectar reservoir and moderate to high numbers of stomata.     

Photosynthetic 14CO2 fixation products in altitudinal ecotypes of Trifolium repens L. with different temperature requirements

Summary Two Trifolium repens clones from natural meadows at 600 m and 2030 m above sea level, and with differing dependence on temperature of their rate of apparent photosynthesis, were grown under controlled environments. Radioactive products in detached leaves were examined after 20 and 40 s periods of steady state photosynthesis in ¹⁴CO₂ at 3° C and 24° C. Glycine and serine were hardly labeled at 3° C. At 24° C, the leaves of the alpine clone showed significantly, (P<0.025) more activity in these amino acids than those from the low altitude clone. It is suggested that the alpine clone has a higher photorespiration. This is supported by the labeling pattern of glucose, fructose, sucrose, and glucose-6-phosphate.The high altitude clone requires lower temperatures for photosynthesis than the low altitude clone. It is suggested, that this is caused by its higher photorespiration, which reduces net photosynthesis at high temperatures. The lower photorespiration activity of the low altitude clone can be interpreted as an adaptation to its warmer habitat.     

Supercooling Ability, Water Content and Hardiness of Rhododendron Flower Buds during Cold Acclimation and Deacclimation

The relationship between supercooling ability and water contentand killing temperature of flower buds during cold acclimationand deacclimation were studied using R. kiusianum and R. x akebono.The occurrence of multiple floret exotherms and their shiftto a narrow range at lower subzero temperatures, as well asthe marked decrease of florets water content, were observedas the symptoms of cold acclimation occuring in flower budsfrom fall to winter, and vice versa in spring buds during deacclimation.In R. kiusianum, the fully acclimated period was from Novemberto March and two months longer than that of R. x akebono. Thesupercooling ability of the former was about –25°Cand about –20°C in the latter. Although the watermigration within bud tissues during the freezing process wasdetermined in the acclimated and deacclimated buds for R. xakebono, no significant water changes could be observed, evenin the acclimated buds. Thus, it is conceivable that deep supercoolingin florets may result not necessarily from water migration fromflorets and bud axes to scales in response to freezing, butfrom low water content in situ of cold-acclimated or artificiallydehydrated flower buds. (Received July 29, 1981; Accepted October 12, 1981)     

The Effect of Temperature on Seed Set and Seed Development in Detached Spikelets of Perennial Ryegrass (Lolium perenne L.)

In perennial ryegrass seed yield is low. Pollination, fertilizationand early stages of seed development are critical periods forthe realization of the yield potential. As intact plants aredifficult to handle, we used detached spikelets to study effectsof temperature and genotype on seed set and seed development. Plants of four cultivars were grown in pots. Just prior to flowering,spikelets were detached from the ears and put on water. Afterpollination, various spikelets of each plant were placed atdifferent temperatures (14/14, 17/12 or 20/15°C). Sevendays after pollination the percentage seed set and the lengthof the developing seeds (caryopses) were assessed. Only floretsin the four basal positions were used. For each cultivar an analysis of variance for unbalanced datawas carried out. There were highly significant difference amonggenotypes for seed set and caryopsis length. There was no evidencefor genotypic variation for tolerance to low temperatures. Neithertemperature nor floret position had an effect on seed set. Thecaryopsis length, however, increased with increasing temperatureand basal florets produced longer caryopses than distal florets.The detached spikelets proved to be a useful experimental tool.Copyright1993, 1999 Academic Press Lolium perenne L., perennial ryegrass, seed yield, seed set, detached spikelets culture, temperature, floret position     

Ultrastructural Aspects of the Nectary Spur of Limodorum abortivum (L) Sw. (Orchidaceae)

The ultrastructure of the nectary spur of Limodorum abortivum(L) Sw. was examined before and after anthesis. In cross sectionthe nectary spur shows an internal epidermal layer of thin-walledcells bordering the secretory cavity and 10–12 layersof parenchyma cells. The ultrastructure of the secretory cellssuggests the involvement of ER, Golgi and plastids in nectarsecretion. The nectar accumulated in the sub-cuticular spaceis released into the nectariferous cavity by rupture of theouter layer of the cuticle. Limodorum abortivum (L) Sw., Orchidaceae, nectary spur, nectar secretion, ultrastructure, anthesis, endoplasmic reticulum, dictyosomes, plastids     

Effect of Temperature on Nitrogenase Activity in White Clover

Single, clonal plants of white clover were grown without inorganicnitrogen in four contrasting day/night temperature regimes,with a 12 h photoperiod, in controlled environments. Root andnodule respiration and acetylene reduction activity were measuredin a flow-through system during both day and night for plantsacclimated to day/night regimes of 23/18, 15/10 and 10/5 ?C.Similar measurements were made on plants acclimated to 20/15?C and stepwise at temperatures from 4 to 33 ?C. Peak rate of ethylene production, nitrogenase-linked respirationand basal root + nodule respiration increased approximatelylinearly from 5 to 23 ?C both in temperature-acclimated plantsand in plants exposed to varying measurement temperatures. Themeasured attributes did not vary significantly between day andnight. Temperatures above 23–25 ?C did not further enhancethe rate of ethylene production, which remained essentiallythe same up to the maximum measured temperature of 33 ?C. The measurements of nitrogenase-linked respiration between 5and 23 ?C, during both day and night, demonstrated a constant‘energetic cost’ of acetylene reduction of 2.9 µmolCO₂ µmol C₂H₄^–1,. Over the same temperature range,the approximate activation energy of acetylene reduction was60 kJ mol^–1. The integrated day plus night nitrogenase-linkedrespiration accounted for 13.4–16% of the plant‘snet shoot photosynthesis in a single diurnal period: there wasno significant effect of temperature between 5 and 23 ?C. Key words: Trifolium repens, white clover, temperature, N₂ fixation, respiration