Growth and development of soyabean cv. TK5 as affected by tropical daylengths, day/night temperatures and nitrogen nutrition

Two experiments were done in Saxcil growth cabinets in order to investigate the effects of climatic factors and nitrogen nutrition on the growth, reproductive development and seed yield of soyabean cv. TK5. In the first, plants were grown to maturity in eight environments comprising all combinations of two short daylengths (11 h 40 min and 13 h 20 min), two day (27 and 33^oC) and two night (19 and 24^oC) temperatures. In the second, day temperature was kept at 33^oC but the night temperature was varied (19 and 24^oC) as was the mineral nitrogen supply (20 and 197 ppm N) to plants which were either inoculated or not with an effective single strain of Rhizobium. Taller, more branched, later flowering plants were produced in the longer daylength but seed yield was hardly affected because the components of yield did not all respond similarly. In the higher day temperature treatments seed yield per plant was reduced by half because all yield components were adversely affected - pods per plant by 34 %, mean seed dry weight by 24 % and seeds per pod just slightly. There was a marked effect of the higher night temperature which promoted early vegetative growth, induced early flowering and although the number of pods per plant was, overall, reduced by 48 %, seed yield per plant was little affected as mean seed dry weight was increased by 37 % and the number of seeds per pod was also increased slightly. Prior to flowering, nodulated plants obtained about two thirds of their total nitrogen requirement via direct uptake and one third through the symbiotic system. Vegetative dry weight and plant nitrogen content were increased by the higher mineral nitrogen level and, although height was slightly diminished, more branches were produced. Seed yield, however, was only slightly increased. These experiments have shown that night temperature is an environmental factor of major importance for the growth of this soyabean cultivar. They have provided, also, a more rational basis for interpreting seasonal variations in growth and seed yield of soyabean in the tropics where, clearly, day and night temperature effects can override those of daylength and nitrogen nutrition.     

Effects of temperature, cultivar and isolate on the incubation period of white leaf spot (Mycosphaerella capsellae) on oilseed rape {Brassica napus)

When leaves of oilseed rape (cv. Cobra) were inoculated with conidial suspensions of Mycosphaerella capsellae (white leaf spot) and incubated in controlled environments, the lag period from inoculation to the appearance of the first lesions decreased, and the total number of lesions produced increased, as temperature increased from 5^oC to 20^oC, although differences between 15^oC and 20^oC were small. With incubation period estimated as the time from inoculation until 5%, 50% or 95% of the lesions were produced, there was a linear relationship between l/(incubation period in days) and temperature over the range 5^oC to 20^oC, from which values at intermediate temperatures could be estimated. Summed mean daily temperatures from inoculation to the production of 5% of the lesions were estimated as 115–130 degree-days in the controlled environment experiments at 5^oC to 20^oC. When pods or leaves of plants in oilseed rape crops (cv. Cobra or cv. Libravo) were inoculated with conidial suspensions of M. capsellae on five occasions from January to October, with variable temperatures during the incubation period, degree-days until the first appearance of lesions were in the range 115–230. The numbers of white leaf spot lesions cm^-2 which developed on inoculated leaves differed greatly between nine oilseed rape cultivars, with most on cv. Tapidor and fewest on cv. Libravo, but the incubation period differed little between cultivars. Similarly, the number of lesions which developed differed between four M. capsellae isolates from different regions but the incubation period did not.     

Seed Development in Phaseolus vulgaris L. cv Seminole: I. Developmental Independence of Seed Maturation

Phaseolus vulgaris L. cv Seminole pods removed from the plant continued their development when incubated in suitable conditions. Seeds continued to grow and develop and pods and seeds passed through an apparently normal developmental sequence to dryness. Seed growth was at the expense of pod dry weight (DW) reserves. Losses of pod DW paralleled DW gains by seeds in detached pods and in pod cylinders containing a seed. The transfer activity was apparent only within the period 10 to 30 days after anthesis (DAA) with maximal activity between 15 to 20 DAA. This period corresponds to maximum pod growth and the attainment of maximal DW. Seeds are in only the early phase of seed growth at this time. No DW transfer was observed at developmental stages beyond 30 to 35 DAA when normal senescence DW losses in pods became evident and seeds were in the later phase of seed fill. Pods or pod cylinders remained green and succulent over the transfer period, later passing through yellowing and drying phases characteristic of normal development. DW transfer was dependent on funicle integrity and was readily detectable in pod cylinders after 7 days incubation. The DW transfer activity may contribute to continuing nutrition of seeds under conditions where the normal assimilate supply to seeds becomes limiting. Defoliation and water stress treatments applied to Phaseolus plants reduced seed yields but allowed persistence of seed maturation processes such that all seeds developing to dryness were capable of germination.     

The effect of elevated temperatures and their duration on the incidence of hollow heart in pea seeds

Plants of Pisum sativum cv. Pania were exposed to high temperature (35 ^oC day/25 ^oC night) at three stages in development. These were: (i) at process crop maturity, when the seed moisture content (m.c.) was 80%, i.e. about 10 days prior to pod wrinkle, (ii) at a seed m.c. of 70 to 80%, about the onset of pod wrinkle, and (iii) when the seed m.c. was 50 to 60%, 10 days after the onset of pod wrinkle. At each stage of development the high temperature treatment was imposed for 2,4,6, 8 or 10 days. Plants exposed to high temperature at seed m.c. of 70 to 80% produced seed with the highest incidence of hollow heart and at all stages of development the incidence increased with the length of exposure to the high temperature. Where plants at the development stage of pod wrinkle (seed m.c. 70–80%) were exposed to a range of temperature regimes for 5 days, over 20% of the seeds had hollow heart when the mean day/night temperature was 25 ^oC. Above a mean temperature of 25 ^oC, the percentage of affected seed increased with increasing day or night temperature. Over 80% of the seeds had hollow heart symptoms after 5 days exposure to a daily mean temperature of 32-5 ^oC. The severity of symptom development was not related to the proportion of seeds with the disorder and, under laboratory conditions, seed germination was not affected.     

Effects of temperature on the flowering and seed production of Stylosanthes guianensis cultivars

Seedlings of Stylosanthes guianensis var. guianensis were grown for 75 days in 16 h photoperiods and transferred to controlled environment cabinets for a further 112 cycles set at 12 h days and 18, 21, 24 or 27 OC constant temperatures. Floral initiation (FI) occurred in cv. Cook after 21 short-day cycles of 12 h and was independent of temperature. Warm temperatures delayed FI in cv. Schofield and hastened it in cv. Endeavour. Rate of node appearance was positively and the duration of the phase FI to flower appearance negatively related to temperature. Spike production and floret number per spike were maximal for Cook and Endeavour at 24°C; seed setting was independent of temperature above 18°C in Cook. Growth, flowering, and seed production of Endeavour required warm temperatures, but Cook was more versatile in its adaptation to varying temperature regimens.     

Influence of Different Storage Conditions on the Mycotoxin Production and Quality of Fusarium-Infected Wheat Grain

Wheat seed samples with different initial infection levels of Fusarium culmorum were kept under different storage conditions for 36 weeks. Samples for analysis were drawn before storage and at intervals of 6‐8 weeks to determine the mycotoxin contents, seed health and seed quality. Zearalcnone (ZEA) accumulated to higher kernel contents towards the end of storage, when the seed was stored under warm and humid conditions [25°C/90% relative humidity (RH)], whereas the deoxynivalenol (DON) content of severely infected kernel samples (> 50%) remained unchanged under any of the conditions. On the other hand, DON contents increased in samples with a slight (4%) or moderate (15%) Fusarium infection level. when the seed was stored under Warm and humid conditions. Nivalenol (NIV) was not found in any samples immediately after harvest but later on in storage, and only under cool or warm but very humid conditions (15°C/84% RH and 25°C/90% RH). During storage, the mycotoxin contents of the samples did not reflect the percentage of Fusarium infected kernels. Under warm but dry conditions (25°C/62% RH) the seed germination rate showed a slight increase or remained nearly constant; at the same time the Fusarium infection level of the kernels decreased fairly fast. Cool and dry conditions (15°C/56% RH) maintained good seed quality but the Fusarium infection level of the kernels remained largely the same. Warm and humid conditions are not appropriate to maintaining quality of both seed and grain product.     

Interactive effect of cytokinin and potassium on sink-source relationships in Lupinus angustifolius

This study examined whether increased K supply in conjunction with BAPcould increase lupin seed yield and harvest index by enlarging sink volume (podnumber), increasing assimilate and improving assimilate partitioning to filltheadditional pods induced by BAP treatment. Narrow-leafed lupin(Lupinusangustifolius, cv. Danja abs^– mutant) was grown inaglasshouse, in pots containing sandy soil with four K treatments (0, 15, 60 and120 mg K/kg soil). BAP (2 mM) was applied daily toallmain stem flowers throughout the life of each flower from opening to senesced.BAP application did not affect assimilate production (as measured by totalabove-ground biomass), but changed assimilate partitioning. On BAP-treatedplants, there were greater proportions of seed to pod wall dry weight on themain stem but smaller proportions on the branches, and an increased weightratioof seed to pod wall overall which meant more assimilate was used for seedgrowthrather than pod wall growth. BAP increased the number of pods per plant by35% and this more than compensated for the decreases in seeds per podandseed weight. Therefore, there was an increased harvest index (+11%)and seed yield per plant (+13%) in BAP-treated plants. BAP alsoincreased the number of pods with filled seeds (146%) on the main stemand main stem seed K⁺ concentration (from 0.81% to0.87%). Added K increased biomass but only slightly affected assimilatepartitioning. As applied K increased, relatively more assimilate was used forpod wall growth rather than seed growth. Added K increased seed yield per plantby about 14% due to increases in seed weight and the number of pods onthe main stem. Moreover, K⁺ concentration in seeds and shootsincreased with increasing level of applied K. Seed yield was enhanced more byBAP when K was supplied at high levels. Increasing K supply interactedpositively with added BAP by increasing narrow-leaf lupin seed yield andharvestindex through increases in assimilate supply and its partitioning into seeds.     

Lengthening versus shortening dark periods and blossoming in sugar cane as affected by temperature

Sugar cane, an intermediate day plant, clearly received a stronger stimulus to flower during lengthening nights than during shortening nights. Flowering was vigorous under warm, lengthening nights (21°) but less so under cool, lengthening nights (16-17°). Warm or cool shortening nights either failed to induce flowering altogether or reduced it substantially. Under the warmer nights the inductive dark period was 10 hours 57 minutes to 11 hours 26 minutes whether the nights were lengthening or shortening. Under cooler conditions, it was longer by from 20 minutes to nearly 2 hours.     

Pre‐zygotic parental environment modulates seed longevity

The potential for the pre‐zygotic plant growth environment to play a role in determining seed longevity was investigated for a species that inhabits arid to semi‐arid Australia. Seed longevity is particularly important for wild populations in fluctuating environments because the longer a seed‐lot is able to survive in the soil seed bank the more likely it is to buffer the population from unpredictable environments. Thus Wahlenbergia tumidifructa plants received wet or dry soil moisture within a warm or cool glasshouse until flowering. Seeds subsequently produced by flowers that opened on the day that plants were moved to a common environment were collected at maturity and longevity assessed by controlled ageing at 60% relative humidity and 45°C. Mean seed longevity was similar for seeds produced by plants that grew in warm‐wet, warm‐dry and cool‐dry conditions (P₅₀ of about 20 days), but extended for plants in cool‐wet conditions (P₅₀ = 41.7 days). Cool temperatures resulted in seeds with a wider distribution of lifespans (σ = 20 days) than warm conditions (σ = 12 days); the large σ caused the extended P₅₀ for cool‐wet plants, but not cool‐dry as a result of a concomitant reduction in initial seed germination (K_i). After moving to the common environment, all plants generated new vegetative material, which went on to produce seeds with similar longevity (P₅₀ approx. 20 days) irrespective of original environment. Visible phenotypic responses of the parent to environmental conditions correlated with longevity and quality parameters of the progeny seeds, suggesting that a parental effect modified seed longevity. Our study provides novel empirical data showing that environmental conditions expected under climate change scenarios may potentially cause seed longevity to decline for a species that inhabits arid to semi‐arid Australia. These negative impacts on population buffering may weaken the storage effect mechanism of species coexistence in fluctuating environments.     

Intensity of pre‐dispersal seed predation in the invasive legume Leucaena leucocephala is limited by the duration of pod retention

Abstract The intensity of seed predation the invasive tropical legume Leucaena leucocephala by the bruchid Acanthoscelides macropthalmus was investigated in south‐eastern Queensland, Australia. The number of seeds damaged by A. macropthalmus as a proportion of total seeds available was found to increase the longer the pods remained on the tree. Seed predation ranged from a mean of 10.75% of seeds on pods that remained on the plant for 1 month and increased to 53.54% for pods that remained of the plant for 4 months. The low bruchid populations at high pod densities results in ‘predator satiation’. However, pods dehisce over time and the proportion of pods available over time to the bruchid correspondingly declines. By the time bruchid densities build up, most pods have dehisced and the seeds consequently escape predation. As a result the number of seeds lost to bruchid damage increases only marginally over time. Despite the levels of seed predation observed over the course of the study, the number of seeds in the soil seedbank almost doubled over time increasing from 8.5 seeds m^?3 to 15.5 seeds m^?3 over a 4‐month period. Levels of seed predation and addition of seeds to the soil seedbank were not correlated. The taxonomic (subspecies) status and apparency of host plants as measured by plant and patch traits (average plant height, density of podding plants and patch size) did not influence levels of seed predation. Pre‐dispersal seed predation studies need to take into account the pod/seed retention behaviour of the plant. The ability of the bruchid to regulate the invasiveness of Leucaena through influencing its demography is likely to be diminished if the insect populations cannot increase rapidly enough to use the seeds before pod dehiscence.     

High night temperatures during grain number determination reduce wheat and barley grain yield: a field study

Warm nights are a widespread predicted feature of climate change. This study investigated the impact of high night temperatures during the critical period for grain yield determination in wheat and barley crops under field conditions, assessing the effects on development, growth and partitioning crop‐level processes driving grain number per unit area (GN). Experiments combined: (i) two contrasting radiation and temperature environments: late sowing in 2011 and early sowing in 2013, (ii) two well‐adapted crops with similar phenology: bread wheat and two‐row malting barley and (iii) two temperature regimes: ambient and high night temperatures. The night temperature increase (ca. 3.9 °C in both crops and growing seasons) was achieved using purpose‐built heating chambers placed on the crop at 19:000 hours and removed at 7:00 hours every day from the third detectable stem node to 10 days post‐flowering. Across growing seasons and crops, the average minimum temperature during the critical period ranged from 11.2 to 17.2 °C. Wheat and barley grain yield were similarly reduced under warm nights (ca. 7% °C^?1), due to GN reductions (ca. 6% °C^?1) linked to a lower number of spikes per m². An accelerated development under high night temperatures led to a shorter critical period duration, reducing solar radiation capture with negative consequences for biomass production, GN and therefore, grain yield. The information generated could be used as a starting point to design management and/or breeding strategies to improve crop adaptation facing climate change.     

Effect of high soil temperatures on nodulation of cowpea, Vigna unguiculata

Cowpeas, inoculated with one of five effective strains of Rhizobium isolated from African soils, were grown at root temperatures of 30 ^oC continuously or at 36, 38, 40, 42 and 44 ^oC for 5 h/day and returned to glasshouse ambient for the intermediate period. Growth was best at 30 and 36 ^oC; above 40 ^oC growth was poor and no nodules formed. At 40 ^oC two strains failed to nodulate. Symbiotic performance was not dependent only on nodule production as nodule efficiency varied inversely with temperature. The number of nodules formed by strain R5000 after exposure of inoculated seed or seedlings to 40,42 or 44 ^oC for 5 h/day on each of 3 or 6 days depended on the age of plant and the duration of exposure to stress. When exposed to 42 or 44 ^oC during the first 3 days after sowing nodulation was reduced from 18 to 1–3 nodules/plant even after a further 40 days growth at ambient (30 day, 20 ^oC night). Nodulation was unaffected when 10–15-day-old seedlings were exposed to the same conditions. Numbers of strain R5000 on seed declined rapidly following three daily exposures of 5 h at 39 and 42 ^oC; at 45 ^oC less than three bacteria survived on each seed. Other rhizobia of the cowpea group varied greatly in their toleration of high temperatures, some survived well at 45 ^oC whereas others behaved like R5000.     

Pod photosynthesis and seed dark CO2 fixation support oil synthesis in developingBrassica seeds

Rate of photosynthesis and activities of photosynthetic carbon reduction cycle enzymes were determined in pods (siliqua), whereas rate of dark CO₂ fixation, oil content and activities of enzymes involved in dark CO₂ metabolism were measured in seeds ofBrassica campestris L. cv. Toria at different stages of pod/seed development. The period between 14 and 35 days after anthesis corresponded to active phase of seed development during which period, seed dry weight and oil content increased sharply. Rate of pod photosynthesis and activities of photosynthetic carbon reduction cycle enzymes were maximum in younger pods but sufficiently high levels were retained up to 40 days after anthesis. The rate of dark¹⁴CO₂ fixation in seeds increased up to 21 days after anthesis and declined thereafter but maintaining sufficiently high rates till 35 days after anthesis. Similarly various enzymes viz., phosphoenolpyruvate carboxylase, NAD⁺-malate dehydrogenase and NADP⁺-malic enzyme, involved in dark CO₂ metabolism retained sufficient activities during the above period. These enzyme activities were more than adequate to maintain the desired supply of malate which mainly arises from dark CO₂ fixation in seeds and further translocated to leucoplasts for onward synthesis of fatty acids. Enzyme localization experiments revealed phosphoenolpyruvate carboxylase and enzymes of sucrose metabolism to be present only in cytosol, whereas enzymes of glycolysis were present both in cytosolic and leucoplastic fractions. These results indicated that oil synthesis in developingBrassica seeds is supported by pod photosynthesis and dark CO₂ fixation in seeds as the former serves as the source of sucrose and the latter as a source of malate     

Effects of variation in ear temperature on growth and yield of spring wheat

Wheat plants were kept in a growth room at 15 ^oC from 7 days after an-thesis until maturity and their ears were warmed to 20 or 25 ^oC for various periods. Continuous warming initially increased the growth rate of the ear and decreased that of the stem, but hastened senescence of the ear and decreased final grain yield by decreasing dry weight per grain. Warming the ears increased the movement of ¹⁴C and nitrogen to them from the leaves and stem during the early stages of grain growth but decreased it later. Warming the ears for only the first 10 or 20 days also hastened ear senescence; grain yield decreased progressively with increase in duration of the warm period. All effects were greater at 25 than at 20 ^oC. The effects of changing ear temperature from 15 to 20 ^oC were independent of the temperature of the rest of the plant. Altering the humidity of the air around the ears by 4–7 mb at constant temperature had no effect on ear growth or senescence. Warming all of the plant except the ears from 15 to 20 ^oC increased ear growth slightly during early grain growth and decreased it later, irrespective of ear temperature: stem dry weight, leaf area and net photosynthetic rate of flag leaves and green stems were decreased and dark respiration rate of stems was increased.     

The effect of cold treatment during seed imbibition on lettuce radicle length and the uniformity of seedling emergence

Two experiments were conducted to determine how a period of exposure to 0 ^oC, between two periods of imbibition at 20 ^oC, affected radicle development and the uniformity of seedling emergence. In both experiments cold treatments, given 12 h after seed imbibition began, produced shorter radicles than those given at 0, 4, 8, 16 and 20 h. Increasing the duration of exposure to 0 ^oC gave longer radicles and reduced the spread of seedling emergence. It is concluded that lettuce seeds are particularly sensitive to a temperature change about 12 h after seed imbibition starts.     

Rate of Increase in Size and Dry Weight of Individual Pods of Field Grown Soya Bean Plants

The dry weight of the whole fruit, the pod wall and an enclosedseed of randomly harvested soya beans is estimated from theexternal dimensions of the attached pod. The relations betweendimensions and dry weight are independent of cultivar and growthcondition and can be used on pods from 1 cm in length untilthe seeds reach their maximum fresh weight. Dimensions of tagged pods of three cultivars of field grownsoya beans differing in time to reach maturity were measuredevery 2–3 days from initial pod elongation until maturation.Dry weights for each pod were estimated from the dimensions,and the dry weight accumulation with time was fitted to thelogistic function to find the growth rate that best characterizedthe data for each pod. The final weight, the specific growthrate and the maximum growth rate of the whole fruit, the podwall and a single seed were subjected to analysis of variance. The most significant difference between pods of these cultivarswas the specific growth rate of individual seeds, which decreasedwith increasing maturity group. There were no differences ingrowth of the pod wall. However, most of the variation was betweenindividual pods within a cultivar, where the rate of dry weightaccumulation of the whole fruit, governed largely by the seedgrowth rate times the number of seeds, was highly correlatedwith the earlier growth of the pod wall. This suggests thatthe growth of individual whole fruit was determined early inpod development and was slightly influenced by factors appliedduring the period of rapid seed growth. Glycine max (L.) Merrill, Soya bean, seed growth analysis, specific growth rate     

Significance of photosynthetic and respiratory exchanges in the carbon economy of the developing pea fruit

The nutritional economy of the developing fruit of Pisum sativum L. (cv. Greenfeast) was studied in terms of intake of translocate, incorporation of C and N into dry matter, transpiration, and CO₂ exchanges of the fruit with its external and internal atmospheres. The environmental conditions were 12-hr days (22 C, 850 μeinsteins m⁻² sec⁻¹ at fruit level); 12-hr nights of 15 C.     

Seed abortion in Pongamia pinnata (Fabaceae)

In Pongamia pinnata only one of the two ovules develops into a seed in most of the pods. Since pollen was not found to be limiting and reduced fertilization could not completely explain the observed frequency of seed abortion, it implied an effect of postfertilization factors. Aqueous extracts of developing seeds and maternal tissue (placenta) did not influence abortion in vitro, suggesting that abortion may not be mediated by a chemical. Experimental uptake of ¹⁴C sucrose in vitro indicated that both the stigmatic and the peduncular seed have similar inherent capacities of drawing resources, but the peduncular seed is deprived of resources in the presence of the stigmatic seed. This deprivation of the peduncular seed could be offset by supplying an excess of hormones leading to the subsequent formation of two seeds in a pod. The prevalence of single-seeded pods in P. pinnata seems therefore to be a result of competition between the two seeds for maternal resources. The evolutionary significance of single-seeded pods in P. pinnata is discussed with respect to possible dispersal advantage enjoyed by such pods.     

Pre- and post-harvest influences on seed dormancy status of an Australian Goodeniaceae species, Goodenia fascicularis

Background and Aims

The period during which seeds develop on the parent plant has been found to affect many seed characteristics, including dormancy, through interactions with the environment. Goodenia fascicularis (Goodeniaceae) seeds were used to investigate whether seeds of an Australian native forb, harvested from different environments and produced at different stages of the reproductive period, differ in dormancy status.

Methods

During the reproductive phase, plants were grown ex situ in warm (39/21 °C) or cool (26/13 °C) conditions, with adequate or limited water availability. The physiological dormancy of resulting seeds was measured in terms of the germination response to warm stratification (34/20 °C, 100 % RH, darkness).

Key Results

Plants in the cool environment were tall and had high above-ground biomass, yet yielded fewer seeds over a shorter, later harvest period when compared with plants in the warm environment. Seeds from the cool environment also had higher viability and greater mass, despite a significant proportion (7 % from the cool-wet environment) containing no obvious embryo. In the warm environment, the reproductive phase was accelerated and plants produced more seeds despite being shorter and having lower above-ground biomass than those in the cool environment. Ten weeks of warm stratification alleviated physiological dormancy in seeds from all treatments resulting in 80–100 % germination. Seeds that developed at warm temperatures were less dormant (i.e. germination percentages were higher) than seeds from the cool environment. Water availability had less effect on plant and seed traits than air temperature, although plants with reduced soil moisture were shorter, had lower biomass and produced fewer, less dormant seeds than plants watered regularly.

Conclusions

Goodenia fascicularis seeds are likely to exhibit physiological dormancy regardless of the maternal environment. However, seeds collected from warm, dry environments are likely to be more responsive to warm stratification than seeds from cooler, wetter environments.Key words: Goodenia fascicularis, Goodeniaceae, Australia, physiological dormancy, seeds, temperature, soil moisture, maternal influence, climate     

Thermal inactivation of cherry leaf roll virus in tissue cultures of Nicotiana rustica raised from seeds and meristem-tips

Nicotiana rustica tissue cultures derived from seeds or embryos infected with cherry leaf roll virus (CLRV), remained infected after culture at 22 ^oC. No infectivity was found in cultures held at 32 ^oC for 5 days but it was readily detected after such cultures were transferred to 25 ^oC for 8 days. Virus was permanently eradicated from most plants after 20 days incubation at 32 ^oC and from all plants after 7 days incubation at 40 ^oC. Partially purified preparations of CLRV lost infectivity after 9–12 days at 22^oC, 5 days at 32^oC and 3 days at 40^oC.