Role of lipids in theNeurospora crassa membrane. I. Influence of fatty acid composition on membrane lipid phase transitions

Summary The relationship between lipid composition and phase transition was investigated by differential scanning calorimetry for intact and membrane phospholipid extracts of wild-type (w/t) and thecel ^– (Tw 40) mutant ofNeurospora crassa. Thecel ^– (Tw 40) mutant (grown on minimal, sucrose medium supplemented with Tween 40 at 34 °C) had approximately twice the saturated fatty acid content ofw/t organisms grown at 22 °C. The gel-liquid crystal phase transitions of ergosterol-free extracts derived fromw/t andcel ^– (Tw 40) occur at –31 and –11 °C, respectively. The heats of transition (H) of these extracts were 1 and 13 cal/g, respectively. The addition of ergosterol (the predominant sterol inNeurospora) to the phospholipid extracts decreased the observed heats of transition, but did not alter the transition temperature. IntactNeurospora, whetherw/t orcel ^– (Tw 40) did not manifest similar gel-liquid crystal phase transitions in the differential scanning calorimeter. However, an endothermic peak at approximately 30 °C was observed in intact cells and extracted phospholipids of bothw/t andcel ^– (Tw 40) organisms. This peak was insensitive to the addition of ergosterol, had a low heat content (H1 cal/g), and was reversible.     

Cold-hardiness of Dermacentor marginatus (Acari: Ixodidae)

The cold-hardiness of Dermacentor marginatus using laboratory-reared offspring of ticks collected in Germany was characterized. Investigations of unfed stages revealed that adult ticks suffered 50% mortality at –10°C after 4–5 months, but larvae and nymphs suffered mortality within few days, whereas –15°C was lethal for all stages within a very short period. Larval hatch and moulting of engorged larvae and nymphs did not occur at 10°C. Embryonic development of eggs with larval hatch was considerably reduced by exposure of eggs to 10°C. Engorged females did not lay eggs at 10°C, the oviposition capability, however, persisted over 6 months at 10°C, 5 months at 5°C, 3 months at 0°C and 2 months at –10°C without substantial decrease of the oviposition capacity or reduction of viable eggs. These results present evidence that unfed adult ticks are the ecoepidemiologically most effective stages, which are capable to tolerate long and extremely cold winters without substantial impairment of the population density. It is also considered that engorged females interrupt their oviposition at low and subzero temperatures delaying it for months and so contribute in bypassing winter conditions. None of the stages survived supercooling indicating that D. marginatus is freeze intolerant. Mean supercooling point (SCP) ranged between –26°C in eggs and –12, 6°C in engorged females. Compared with eggs, the SCP of the other stages was significantly higher. In conclusion, the SCP is considered to have no predictive value in the context with cold-hardiness.     

Respiration rates of Chiloplacus sp. and other arctic nematodes at low and high temperatures

Summary Respiration of an undescribed species of soil nematode of the genus Chiloplacus from the Canadian High Arctic was measured at 2°, 5°, 10°, 15°, 20° and 25°C. The corresponding metabolic rates were 0.2697×10^-3 l, 0.3406×10^-3 l, 0.8408×10^-3 l, 0.8539×10^-3 l, 1.8420×10^-3 l and 2.9360×10^-3 l O₂ ind^-1 h^-1, respectively, for a nematode of 1.0 g dry weight. The relationship between respiration and dry weight for Chiloplacus sp. at 10°C is described by the function log R=-3.0693+0.8844 log W. Q₁₀ values for the 2°–5°, 5°–10°, 10°–15°, 15°–20° and 20°–25°C temperature intervals were 2.18, 6.09, 1.03, 4.65 and 2.54, respectively. Chiloplacus sp. showed raised metabolic rates at low tempetatures compared with species from warmer environments. Metabolic rates of representative samples of the soil, nematode fauna (dominated by individuals of the genus Plectus) from the same location were 0.1593×10^-3 l, 0.3603×10^-3 l and 0.5332×10^-3 l O₂ ind^-1 h^-1 at 5°, 10° and 15°C for an average nematode of 0.4297 g dry weight.     

Stimulation of potato tuber respiration by cold stress is associated with an increased capacity of both plant uncoupling mitochondrial protein (PUMP) and alternative oxidase

The CO₂ evolution of intact potato tubers (Solanum tuberosum, L., var. Bintje) was analyzed during a 10-day period of their warm (25 ± 2°C) or cold (5 ± 1°C) storage, to evaluate cold-stress effects on expression and activities of plant uncoupling mitochondrial protein (PUMP) and alternative oxidase (AOX). CO₂ evolution rates were analyzed at 20°C, to reflect their possible capacities. The 20°C CO₂ production declined from 13 to 8 mg kg^–1 h^–1 after 2 days of warm storage and then (after 3 to 7 days) decreased from 8 to 6.5 mg kg^–1 h^–1. In contrast, 20°C CO₂ evolution did not change after the first day of cold storage, increased up to 14.5 mg kg^–1 h^–1 after 2 days, and decreased to about 12 mg kg^–1 h^–1 after 3 to 7 days of cold storage. Cold storage increased PUMP expression as detected by Western blots and led to elevated capacities of both PUMP (44%) and CN-resistant AOX (10 times), but not the cytochrome pathway. Since we found that cold storage led to about the same mitochondrial respiration of 40 nmol O₂ min^–1 mg^–1 attributable to each of the respective proteins, we conclude that both AOX and PUMP equally contribute to adaptation of potato tubers to cold.     

Factors affecting oviposition and fecundity in the grain miteAcarus siro L. (Acarina:Acaridae), especially temperature and relative humidity

Oviposition and fecundity in the grain miteAcarus siro were studied at 5–30°C and 62.5–90% RH. At and above 20°C, 80% RH, mating and oviposition occurred soon after emergence, but at lower temperatures and humidities egg laying was progressively delayed from one to several days. Females needed to mate repeatedly in order to achieve maximum egg production, optimum conditions for which were 15°C, 90% RH, where total output per female averaged 435 with a maximum of 858. Oviposition rates were highest at higher temperatures, the mean daily rate at 20 and 25°C, 90% RH, rising to maximum levels of 28/29 eggs per female per day on day six.Oviposition followed clearly defined patterns, favourable conditions producing rapid increases in the mean daily oviposition rate to high peak levels reached at an early stage in the oviposition period. Less favourable conditions resulted in reduced outputs and lower, more uniform rates of egg laying. The mean oviposition period, varying with humidity, fell from 72–122 days at 5°C to 9–13 days at 30°C and the mean incubation period from 42–70 days at 5°C to 3–4 days at 30°C. Egg viability increased with increasing humidity but was little affected by temperature and unaffected by age of the female at time of oviposition.Males tended to live longer than females at most conditions; longevity—depending on humidity—averaging 13–15 days at 30°C and 129–175 days at 5°C. Adult life for females averaged 12–19 days at 30°C and 88–169 days at 5°C. An index of suitability, calculated from egg number, viability and duration of the egg stage and oviposition period, indicated that the most favourable conditions for oviposition and hatching were 20–25°C and 80–90% RH.     

Energy requirements for growth and maintenance of Scenedesmus protuberans Fritsch in light-limited continuous cultures

Scenedesmus protuberans Fritsch was grown in light-limited continuous cultures with a light-dark cycle, at temperatures of 20° and 28° C. At 20° irradiances of 12 and 38 W m^–2 were used, at 28° 38 W m^–2.The relationships between growth rate and light uptake rate were of diphasic linear character. With the lower growth rates the relationships were defined with the parameters _e , i.e. the specific maintenance rate constant, and c, the true efficiency of light energy conversion into biomass. The _e -value was dependent on temperature, the c on irradiance.In cultures, incubated in prolonged darkness, decrease rates of biomass were comparable to the derived _e -values.Both diphasic linear relationships between growth rate and light uptake rate and the same order of magnitude of _e -values could be derived from literature data on other green algae.     

Effect of acute pH depression on the survival of the freshwater amphipod Hyalella azteca at variable temperatures: field and laboratory studies

Field observations on temperature and pH of a small pond showed that a amphipod population of Hyalella azteca was exposed to variable seasonal pH between 5.10–5.85, and water temperatures between 2–21 °C. Laboratory experiments were designed to simulate seasonal temperatures and field pHs of a small pond habitat. Laboratory bioassay experiments were conducted to determine the survival of Hyalella azteca at pHs 4, 5, 6 and 7, and varying temperatures of 5°, 10°, 15°, 20° and 25 °C.The LT₁₀₀ at pH 4 and 25 °C was 5.7 ± 0.47 days, compared to 47.3 ± 2.49 days at 5 °C. An Analysis of Variance (ANOVA) showed temperature was a significant (p > 0.0001) source of variation in the acute lethality of pH to H. azteca. A Duncans Multiple Range Test (DMRT) further showed that in laboratory experiments at pH 4, there was a significant difference ( = 0.01) between the LT_100s at 5°, 10°, 15° and 20 °C, but not between temperatures 20° and 25 °C.     

Photosynthesis and growth rates of Laurencia brongniartii J. Agardh (Rhodophyta, Ceramiales) in preparation for cultivation

Laurencia brongniartii is usually found at depths below 4 m, but can be found in shallow subtidal areas in crevices and on the walls of a coral reef in Amami Oshima Island, Kagoshima Prefecture, Japan, where irradiances were significantly lower than those at similar depths in open water. In preparation for the possible cultivation of this species for its antibiotic compounds, the effects of temperature and irradiance on photosynthesis and growth were measured. Photosynthesis and growth rates of L. brongniartii explants were highest at 26 and 28 °C, which closely corresponded to temperatures found during August to late December when it was most abundant. The estimated maximum photosynthesis rate (P _max) was 4.41 mol photon m^–2 s^–1 at 26 °C and 4.07 mol photon m^–2 s^–1 at 28 °C. Saturating irradiance occurred at 95 mol photon m^–2 s^–1 at 26 °C and 65 mol photon m^–2 s^–1 at 28 °C. In contrast, growth experiments at 41.7 mol photon m^–2 s^–1 caused bleaching of explants and the maximum growth rate observed during the study was 3.02 ± 0.75% day^–1 at 28 °C and 25 mol photon m^–2 s^–1. The difference in the saturating irradiance for photosynthesis and the irradiance that caused bleaching in growth experiments suggests that long-term exposure to high irradiance was detrimental and should be addressed before the initiation of large scale cultivation.     

Distribution and biology of marine cladocerans in the coastal waters of southern China

Geographical and seasonal distributions of marine cladocerans in the coastal waters of southern China were studied. Penilia avirostris was the most common species, followed by Evadne tergestina and Podon schmackeri. P. avirostris and E. tergestina were most common during summer. P. schmackeri, found only in a small bay northeast of Hong Kong, showed no clear seasonal pattern of occurrence. P. avirostris and E. tergestina were found at temperatures ranging from 16–32°C and salinity ranging from 7.3–37.2. P. schmackeri was restricted to a temperature range of 17–29°C and a salinity range of 31.0–37.2. No significant relationships between marine cladoceran abundance and chlorophyll a concentration were found in samples taken from Tolo Harbour. Parthenogenetic brood size of P. avirostris and E. tergestina ranged from 1 to 14, while P. schmackeri was found to carry up to 19 embryos per brood. No geographical trend in fecundity patterns was observed. No correlation was found between body length and brood size. The occurrence of females with resting eggs was rare.     

The productivity of mesocyclops leuckrarti (Claus) in Lake Kinneret (Israel)

Specific rates of growth (Cw) of Mesocyclops leuckarti, young male and female instars varied between 0.03–0.26 and 0.03–0.17 g_(w.w.)/g_(w.w.)/day respectively at 15° and 22°C, whilst at IV–V copepodid stages females showed a higher Cw values. During 1969–1975 the averages of productivity and monthly P/B ratios were 44 (±23) g_(w.w.) (= 5g_(w.w.)/m²/month) and 3.1 respectively. P/B ratios were highly correlated (r(sup2)=0.98) with temperature changes. Metabolic parameters and P/B ratios were found to be similar to other water bodies in the world indicating an adaptation of M. leuckarti to different conditions.     

Light-temperature interactions on the growth of Antarctic diatoms

Summary The combined effect of various temperatures and light intensities on the growth of seven species of antarctic diatoms in culture has been studied. With the exception of Chaetoceros deflandrei whose thermal tolerance is fairly good, these obligatory psychrophils cannot survive in temperatures above 6° to 9° C. Their mean growth rate is relatively low, between 0.24 div d^–1 for Corethron criophilum and 0.63 div d^–1 for C. deflandrei. Regardless of light intensity, growth rate increased with the temperature to reach a maximum between 3° and 5° C. The highest rates were obtained between 115 and 220 mol m^–2 s^–1 with 0.38 div d^–1 for C. criophilum, 0.56 div d^–1 for Synedra sp. and between 0.71 and 0.88 div d^–1 for the other 5 species. A reduction in light intensity from 220 to 46 mol m^–2 s^–1 slowed growth by nearly 50%. These results suggest that the combined effect of temperature and light is one of the factors involved in the limitation of antarctic phytoplankton growth. The low temperatures of the environment do not permit rapid growth, which, even under optimal light conditions remains low. In addition, in the euphotic layer, the overall light energy available for algae is considerably reduced due to turbulence, a factor which exacerbates the reduced growth rate.     

The combined relationship of temperature and molybdenum concentration to nitrogen fixation byAnabaena cylindrica

The joint effects of growth temperature, incubation temperature, and molybdenum concentration on the nitrogen fixation rate ofAnabaena cylindrica were determined using the acetylene-reduction technique. The nitrogen-fixation response to increased molybdenum concentration varied among three growth temperatures (15°, 23°, and 30° C). The pattern of rate change was similar within a growth temperature but increased overall in magnitude with the three incubation temperatures (also 15°, 23°, and 30° C). The maximum rate of nitrogen fixation occurred at 30°C regardless of previous growth temperature. The minimum molybdenum concentration necessary to yield substantial acetylene reduction varied with growth temperature: at 15°C, 15g 1^–1 was effective; at 23°C, less than 5g 1^–1 was effective; and at 30°C, 50g 1^–1 was effective. At all three growth temperatures, increases in molybdenum concentration above the minimum effective concentration produced increases in acetylene reduction. However, at higher molybdenum concentrations inhibition of nitrogen fixation occurred.     

Über Vorzugstemperatur und Wasserhaushalt von Oncopeltus fasciatus Dall. (Heteroptera: Lygaeidae)

Zusammenfassung Die Temperaturpräferenz von Oncopeltus fasciatus Dall. wurde untersucht. Die Verteilung in einem Temperaturgradienten erfolgte über einen breiten Temperaturbereich von 12,5–35°C, der als Vorzugstemperaturbereich für Oncopeltus definiert werden kann. Innerhalb seiner Grenzen variiert die Vorzugstemperatur (V.T.) je nach physiologischem Zustand des Versuchstieres. Nach 48stündigem Wassermangel wählen die Wanzen 15–17,5°C als V.T. Bei Temperaturen unterhalb von 30°C bildet Oncopeltus fasciatus aufgrund ihres subsozialen Verhaltens Aggregationen, oberhalb von 30°C dagegen erfolgt keine Gruppenbildung. Die relative Luftfeuchtigkeit besitzt im Bereich von 35–100% keine Bedeutung für die Lebensdauer. Auch in Kombination mit Wassermangel zeigt sie keinen Effekt. Die aktive Wasseraufnahme jedoch dürfte eine dominierende Bedeutung für das Überleben der samensaugenden Wanze Oncopeltus fasciatus haben.

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On the Preferred Temperature and water economy of Oncopeltus fasciatus Dall. (Heteroptera: Lygaeidae)

Summary The Preferred Temperature of Oncopeltus fasciatus has been investigated. In a temperature gradient the bugs disperse within a broad range from 12,5 to 35°C (=Range of Preferred Temperature). Within this special range each individual selects its own smaller Preferred Temperature (P.T.), depending on its physiological condition. After water deficiency for 48 hrs. the bugs choose a P.T. of a range between 15–17,5°C. At temperatures below 30°C Oncopeltus fasciatus forms aggregations because the bug exhibits a subsocial behaviour. At temperatures above 30°C no aggregation is formed. At a range between 35–100% the Relative Humidity (R.H.) has no meaning for the duration of life, also in combination with water deficiency. But the active uptake of water seems to be the dominant factor for the survival of the seed-sucking bug Oncopeltus fasciatus.

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Durchgeführt mit Unterstützung der Deutschen Forschungsgemeinschaft im Rahmen einer Herrn Prof. Kloft gewährten Sachbeihilfe.     

Feeding of Burbot, Lota lota, at Different Temperatures

Daily food intake of adult burbot, Lota lota, fed on vendace, Coregonus albula, were estimated experimentally at four different water temperatures (2.4, 5.1, 10.8 and 23.4°C). Mean daily food intake (MDI; g d^–1) and relative daily food intake (RDI; g g^–1 d^–1) increased with temperature from 2.4 to 10.8°C and decreased at 23.4°C. Temperatures of maximum daily food intake values were 13.6°C for MDI and 14.4°C for RDI. No correlation between food intake values and burbot weight was observed. RDI values were used to estimate annual food consumption of burbot population. Annual food consumption estimates were 9.7kg ha^–1 and 24.3kg ha^–1 when burbot biomass was 2.0 or 5.0kg ha^–1, respectively.     

SelectingRhizobium phaseoli strains for use with beans (Phaseolus vulgaris L.) in Kenya: Tolerance of high temperature and antibiotic resistance

Forty one strains ofRhizobium phaseoli were screened for the ability to multiply at high temperatures on yeast extract-mannitol agar. Most strains were tolerant of 30°C, eight strains were tolerant of 45°C and two of 47°C although the rate of multiplication was reduced at 45–47°C. The high temperature-tolerant strains were isolated from Kenyan soils and were fast-growing. Seven of the eight strains tolerant of 45–47°C lost their infectiveness after incubation at high temperature but four strains tolerant of 40°C remained infective after incubation at that temperature.Thirty six strains were resistant to 200 g ml^–1 streptomycin sulphate and 29 strains to 200 g ml^–1 spectinomycin dihydrochloride. Eight strains were resistant to both antibiotics each at 200 g ml^–1. Two of the double-labelled antibiotic-resistant mutants lost their infectiveness onPhaseolus vulgaris. The response to acidity was unaltered and two of the mutants showed a decrease in temperature tolerance. The doublelabelled mutants were recoverable from two Kenyan soils.     

The life cycle of Laminaria abyssalis (Laminariales,Phaeophyta) in culture

Laminaria abyssalis occurs in deep water in tropical latitudes of the Brazilian coast (19° 23 S, 38° 28 W to 22° 54 S, 42° 13 09 W). Its life cycle has been completed in the laboratory in seven months using different conditions of light and temperature. The gametophytic stage required for growth the low photon flux density of 1.2 ± 0.3 µmol m^–2 s^–1 and 18 °C, while the juvenile and adult sporophytes needed 15 µmol m^–2 s^–1 and 18 °C. The sporophytes became fertile at 23 °C. Our results showed that light and temperature are the main factors regulating the growth and life history of this species under the culture conditions tested.     

Osteogenic cell contact with biomaterials influences phenotype expression

Relationship between (1) osteoblast adhesion and spreading, and (2) phenotype expression was investigated. Cellular adhesion and spreading were estimated after short time (24 h), whereas proliferation and other osteoblast functions – after 7 days. Primary human osteogenic cells were seeded on the samples of titanium (T), surgical steel (S) and tissue culture polystyrene (PS), and incubated at 37 °C. After 24 h a number of samples were stained with crystal violet and Hoechst; the average single cell area (spreading) and adhering cell number was measured on each sample. The remaining cultures were supplemented with dexamethasone (10 nM) and -glycerophosphate (5 mM), and incubation was continued for 7 days. The cells on each sample were counted and the following tests were performed: XTT mitochondrial activity assay, total protein content, alkaline phosphatase activity (ALP), Sirius Red test for collagen, osteocalcin and calcium concentration. After 24 h significantly greater cell spreading (p < 0.05) and number (p < 0.05) were on T than on S. After 7 days significantly higher on T than on S were: ALP activity (p < 0.000001), collagen (p < 0.0015) and calcium concentration (p < 0.03). XTT results were bigger on S than on T. In control – XTT results were higher than on the metals; collagen and ALP were lower than on T, and calcium level was significantly lower than on T and S (p < 0.025). After 7 days there were no differences in cell number between T and S. Cell number (24 h) correlated with ALP activity (7 days) on steel (coefficient of correlation, CC = 0.866) and titanium (CC = 0.742). The spreading correlated on steel and on titanium with calcium concentration (CC = 0.645 on S, CC = 0.696 on T) and collagen level (CC = –0.638 on S, CC = –0.69 on T). Conclusions: Better conditions for osteoblast phenotype expression on T after 7 days of culture coincided with greater adhesion and spreading of cells after 24 h on T, as compared with S. The initial contact of cells with underlying surface may influence osteoblast functions and possibly, bone regeneration and implant osteointegration in vivo. Early cell spreading may be an indicator of further expression of osteoblast phenotype and may be important for application of osteogenic cells in reconstructive surgery.     

Interpopulation and interyear variation in germination in Scotch thistle, Onopordum acanthium L., grown in a common garden: Genetics vs environment

Germination patterns were studied among three populations of Scotchthistle, Onopordum acanthium L. (Asteraceae). Cypselas(seeds) were collected twice during the growing season for four consecutiveyears (1996–1999) from plants grown in the same location, theEnvironmental Sciences Western Field Station (ESW) near London, Ontario.Cypselas from 1996 came from plants that had been transplanted as rosettes fromtheir original sites to ESW, while those for 1997–1999 came from plantsgrown from cypselas collected in 1996. Fresh cypselas were put to germinate at25°C, 14 hlight/10°C, 10 h dark for 55 days.Consistent and significant differences in final germination percentages amongpopulations were obtained over the four years. There were also differencesbetween collections within populations in 1996 and 1998. In addition, the finalgermination percentage (collectively) differed widely among years. There was agreater than 30 percent correlation between germination percentage and meandaily maximum temperature during cypsela maturation. Since there were similardifferences among populations every year, it appears that germination patternsof O. acanthium populations have a genetic basis as wellasan environmental one.     

Spontaneous excitatory postsynaptic currents in crayfish neuromuscular junctions in the absence and presence of serotonin and 3,4-diaminopyridine

Summary Spontaneous excitatory postsynaptic currents (sEPSCs) were recorded under voltage clamp in short fibres (l0.6mm) from opener muscles and the contractor epimeralis muscle of small crayfish. From the amplitude distributions of sEPSCs which could be approximated by a Gaussian function, a mean amplitudeã= –1.16 nA±0.28 (SE) was found for sEPSCs in 16 fibres of the claw opener voltage clamped toE=–60 mV (19–22 °C). In the opener of the first walking leg and in the contractor epimeralis muscleã=s-1.1 nA±0.21 (SE;n= 6, –100 mVE–60 mV, 5–10 °C) andã= –2.0 nA±0.2 (SE;n=4, E=–60 mV, 19–22 °C) were obtained. On average about 300–500 synaptic channels were estimated to open during a sEPSC. Giant sEPSCs (gsEPSCs) were also observed. The amplitudes of gsEPSCs were up to 14 times larger than the amplitude of an average normal sEPSC. Moreover, the lifetime of gsEPSCs was up to about 3 times longer than that of sEPSCs. Like sEPSCs, gsEPSCs could not be abolished by 0.1 mol/l tetrodotoxin. The rate at which sEPSCs and gsEPSCs occurred could be markedly enhanced by serotonin (1 mol/l) and 3,4-diaminopyridine (1 mol/l)Abbreviations sEPSCs spontaneous excitatory postsynaptic currents - gsEPSCs giant spontaneous excitatory postsynaptic currents - sIPSCs spontaneous inhibitory postsynaptic currents - gsIPSCs giant spontaneous inhibitory postsynaptic currents - 5-HT 5-hydroxytryptamine - 3,4-DAP 3,4-diaminopyridine - time constant of exponential decay of sEPSCs or gsEPSCs - t _B50 lifetime of sEPSCs or gsEPSCs given by the width of these currents at 50% of their amplitude; ã amplitude of sEPSCs or gsEPSCs - i current amplitude evoked by opening of single glutamate-activated channels - z number of channels open at the peak of an average sEPSC This investigation was supported by the Deutsche Forschungsgemeinschaft. Project Fi 305/1-3     

Integrating sub-lethal heating with Brassica amendments and summer irrigation for control of Macrophomina phaseolina

Effects of varying intensities of sub-lethal heating were ascertained in improving the efficiency of Brassica amendments and summer irrigation on survival of Macrophomina phaseolina, a dry root rot pathogen. Sub-lethal heating (45–55°C) of M. phaseolina infested dry soil reduced the viable propagules by only 12.8% in a period of 90 days. One summer irrigation without sub-lethal heating caused 33.9% reduction in M. phaseolinapropagules, which improved to 43.3% when it was combined with 60 days of sub-lethal heating. Addition of the Brassicaamendments to irrigated soil resulted in significant reduction (60.4–71.6%) in counts of M. phaseolinabut this reduction improved (89.4–96.1%) when sub-lethal heating was combined with amendments. Mustard oil-cake (0.18% w/w) was found to be the most effective with reduction but a 94% inoculum reduction by mustard pod straw (0.36% w/w) was also achieved at 0–30 cm soil depth under similar conditions. Moderate heat level could not exert detectable weakening effect on M. phaseolinapropagules. These results suggest a practical cultural control of soil-borne pathogens by combining sub-lethal heating, Brassica amendments with one summer irrigation.