Development of a Model to Predict the Effect of Temperature and Moisture on Fungal Spore Longevity

A model was developed to quantify the effect of temperatureand moisture content on the longevity of conidia of the entomopathogenicfungusMetarhizium flavoviride . This model incorporated a negativesemi-logarithmic relation between longevity and temperatureand a negative logarithmic relation between longevity and moisturecontent. Replacing the latter with a negative semi-logarithmicrelation between longevity and the equilibrium relative humidityof the conidia was also effective. The latter model was appliedsuccessfully to observations on the survival of conidia of afurther four entomopathogenic fungi (Metarhizium anisopliae,Beauveria bassiana ,Beauveria brongniartii, andPaecilomycesfarinosus ) and ascospores, conidia or uredospores of four phytopathogenicfungi (Alternaria porri ,Helminthosporium oryzae ,Uromyces appendiculatus, andSclerotinia sclerotiorum ) across a wide range of differenttemperatures and relative humidities. The sensitivity of sporelongevity to both temperature and equilibrium relative humidity,and the upper and lower relative humidity limits to the applicationof the model, varied considerably between entomopathogenic andphytopathogenic fungi, among species within each group, andamong different strains within certain species. Metarhizium flavoviride W. Gams & J. Roszypal; entomopathogenic fungi; phytopathogenic fungi; spore survival; storage environment; conidia; uredospore; ascospore     

Low Moisture Content Limits to Relations Between Seed Longevity and Moisture

Discontinuities at low moisture contents in the otherwise logarithmicrelations between seed longevity and seed moisture content (%,f. wt basis) in hermetic storage at 65 °C were detectedat 2–0% in groundnut (Arachis hypogaea L.), 3·5%in onion (Allium cepa L.), 4·5% in sugar beet (Beta vulgarisL.), 4·6% in barley (Hordeum vulgare L.), 5·3%in chickpea (Cicer arietinum L.) and wheat (Triticum aestinumL.), and 5·6% in cowpea [Vigna unguiculata (L.) Walp.].In contrast, the equilibrium relative humidity of seeds at thesevalues was similar, varying between 9·9% (onion and sugarbeet) and 11·5% (wheat). The mean value was 10·5%r.h. (s.e. 0.2). There was no significant (P > 0·05)effect of further reduction in seed moisture content below thesecritical values on longevity, except in wheat (P < 0·005),in which there was a further increase in longevity. In soyabean [Glycine max (L) Merrill], the logarithmic relation continueddown to the lowest moisture content investigated, 3·3%(11·4% equilibrium relative humidity). Above the criticalvalue, seed longevity in groundnut showed the least sensitivityto increase in percentage moisture content, while barley showedthe greatest, the values of the viability constant C_w (slopeof the negative logarithmic relation between longevity and moisture)being 4·089 (s.e. 0·278) and 5·966 (s.e.0·325), respectively. These differences in the valueof C_w among the eight crops were significant P < 0·005),whereas the relative sensitivity of seed longevity to changein equilibrium relative humidity above the critical moisturecontent did not differ significantly among the eight (P >0·10) and was equivalent to a doubling of longevity foreach 8·7% reduction in equilibrium relative humidity.Accordingly it is concluded that the relative effect of waterpotential on seed longevity can be considered to be the samefor these and probably also for many other orthodox species. Barley, chickpea, cowpea, groundnut, onion, soya bean, sugar beet, wheat, seed storage, seed longevity, seed moisture content, viability equation, water relations     

A Model of the Effect of Temperature and Moisture on Pollen Longevity in Air-dry Storage Environments

Data on the survival of pollen ofTypha latifoliaL. stored forup to 261 d over seven different saturated salt solutions (providing0.5 to 66% relative humidity) and six different constant temperatures(from -5 to +45 °C) were analysed to quantify the effectof air-dry storage environment on pollen longevity. Pollen survivalcurves conformed much more closely to negative cumulative normaldistributions than to negative exponential relations. Estimatesofp₅₀(storage period required to reduce pollen viability to50%), provided by negative cumulative normal distributions,were available from 37 different storage environments in whichpollen viability was reduced below 50%. Once observations at0.5% and 5.5% relative humidity were excluded from analysis,there was a negative logarithmic relation between these estimatesof longevity and pollen moisture content (%, wet basis) anda curvilinear semi-logarithmic relation between longevity andtemperature. When the negative logarithmic relation betweenlongevity and moisture content was replaced by a negative semi-logarithmicrelation between longevity and the relative humidity of thestorage environment the resultant model was less satisfactory,principally because pollen longevity over saturated solutionsof calcium nitrate (43–62% relative humidity) and sodiumnitrite (60–66% relative humidity) were consistently greaterand smaller, respectively, than fitted values. Notwithstandingthese errors, comparison between the fitted relations and observationsat the two lowest relative humidities provided estimates ofthe lower-relative-humidity limits to these relations. Theseprovisional estimates varied with storage temperature beinglowest at 25 °C (<5.5% relative humidity). However, therewas no linear trend to that variation (P>0.25): the meanestimate was 11.9 (s.e.=1.4)%. The considerable similaritiesamong models of pollen longevity in air-dry storage, and theirestimated lower limits, and those developed previously for orthodoxseeds and spores are discussed.Copyright 1999 Annals of BotanyCompany. Typha latifoliaL., pollen, storage, survival, longevity, relative humidity, moisture content, temperature.     

Moisture Content and the Longevity of Seeds of Phaseolus vulgaris

The lower limit to the negative logarithmic relation betweenseed longevity and moisture was determined in bean (Phaseolusvulgaris L.). Sub-lots of seed were hermetically stored at 65°C and 11 moisture contents (3.26–13.6% f. wt) forup to 80 d, tested for germination and the seedlings evaluated.In accordance with the seed viability equation, there was anegative logarithmic relation between moisture content (%, f.wt) and longevity. Two different criteria for estimating theslope constant of this relation gave similar values of 4.76(s.e. 0.26) and 4.82 (s.e. 0.24). The calculated lower moisturecontent limits to the relation were 5.7 and 5.6%, respectively,values at equilibrium with 10.6–10.8% relative humidity(rh). Further drying to 3.26% had little additional effect onlongevity, but initial germination was slightly reduced. Theresults are discussed in relation to water potential and comparedwith other crops. Arguments against transforming germinationvalues to disregard the seeds initially failing to germinateare emphasized. Common bean, Phaseolus vulgaris L. cv. Provider, seed storage, seed longevity, seed moisture, viability equation, water relations     

Studies on Prolonged Storage of Beauveria bassiana Conidia: Effects of Temperature and Relative Humidity on Conidial Viability and Virulence against Chickpea Borer,Helicoverpa armigera

Unformulated conidia of Beauveria bassiana were stored at five different temperatures (0°, 10°, 20°, 30° and 40°C) at six different relative humidities (RH) (0, 33, 53, 75, 85 and 98%). Conidial viabilities and virulence against third instar larvae of Helicoverpa armigera were determined over a 24‐month period. Conidia survived longest at lower temperatures (0–20°C) and lower RH levels (0–53% RH). At higher temperatures (30–40°C) conidia did not survive. When the temperature was decreased from 30°C to 0°C, at nearly all RH levels the longevity of conidia increased. Conidia remained virulent for third instar larvae of H. armigera under favourable storage conditions for 24 months.     

Saturated salt solutions for humidity control and the survival of dry powder and oil formulations of Beauveria bassiana conidia

Oil-based formulated conidia sprayed on steel plates and conidia powder (control) of Beauveria bassiana isolate IMI 386243 were stored at temperatures from 10 to 40 degrees C in desiccators over saturated salt solutions providing relative humidities from 32 to 88%, or in hermetic storage at 40 degrees C, and moisture contents in equilibrium with 33 or 77% relative humidity. The negative semi-logarithmic relation (P<0.005) between conidia longevity (at 40 degrees C) and equilibrium relative humidity did not differ (P>0.25) between formulated conidia and conidia powder. Despite this, certain saturated salts provided consistently greater longevity (NaCl) and others consistently shorter longevity (KCl) for formulated conidia compared to conidia powder. These results, analysis of previous data, and comparison with hermetic storage, indicate that storage of conidia over saturated salt solutions provides inconsistent responses to environment and so may be problematic for bio-pesticide research. In hermetic storage, oil formulation was not deleterious to longevity and in the more moist environment enhanced survival periods.     

The Low-moisture-content Limit to the Negative Logarithmic Relation Between Seed Longevity and Moisture Content in Three Subspecies of Rice

The lower limit to the negative logarithmic relation betweenseed longevity and moisture content was determined in threesubspecies of rice (Oryza satwa L.) by storing seeds of fivecultivars at 65 °C with 11 different moisture contents (1.5–15.3%f. wt) for various periods up to 150 d and then testing forgermination. The estimates of the low-moisture-content limit(m_c) were 4.3% for subsp. indica, 4.4% for subsp. japonica,and 4.5% for subsp. javanica. These moisture contents were inequilibrium with 10.5—12.0% r.h. No significant effectof moisture content between 1.5% and m_c on longevity was detected(P > 0.05), while between m_c and 15.3% there were negativelogarithmic relations between longevity and moisture content.There were no significant differences in the relations betweenlongevity and moisture either above or below m_c between thetwo japonica cultivars or between the two javanica cultivars(P > 0.10). There was also no significant difference in theslope of the negative logarithmic relation between longevityand moisture above m_c among the three subspecies (P > 0.25).However, there were significant differences in the standarddeviation of the frequency distribution of seed deaths in timeat any one moisture content, both above and below m_c; this isa measure of seed longevity which is independent of pre-storageenvironment, and the differences observed show that there aregenetically determined differences in longevity among the threesubspecies (P < 0.005), indica being the longest and japonicathe shortest lived. The results provide no evidence for intra-specificvariation in m_c and support the view that the maximum seed storagemoisture content which provides the maximum longevity is thatwhich is in equilibrium with about 10–11% r.h. It is concludedthat while the seed viability constant C_w of the seed viabilityequation is species specific and therefore applies to most,if not all, cultivars of rice, variation in the value of K_Eis the source of the differences in potential longevity of thethree subspecies. Rice, Oryza sativa L, seed storage, seed longevity, seed moisture, viability equation     

Fluctuating Temperature and the Longevity of Conidia of Metarhizium flavoviride in Storage

Conidia of Metarhizium flavoviride were hermetically stored at 13.7% moisture content with four constant (20, 30, 40 and 50 C) and six fluctuating temperature regimes (20:30, 20:40, 20:50, 30:40, 30:50 and 40 C:50 C, all 24 h:24 h) for up to 119 days. Survival of conidia stored at both constant and fluctuating temperatures conformed to cumulative negative normal distributions and all 10 survival curves could be constrained to a common origin. The effect of constant storage temperature on conidia longevity was quantified satisfactorily by a negative curvilinear semi-logarithmic relation developed previously, in which the estimates of the constants CH and CQ were 0.0176 (SE 0.0013) and 0.000703 (SE 0.000019), respectively. The fitted relation at constant temperatures showed that Q10 for loss in conidia viability increased the warmer the temperature regime. The effect of the cooler temperature of each fluctuating temperature regime on conidia longevity was small, since the effective temperature of each regime for loss in conidia viability was always much warmer than the mean. Conidia were also stored in two further regimes: at 30 C for 21 or 35 days before transfer to 50 C. The standard deviations of the conidia survival curves at 50 C were unaffected by the duration of previous storage at 30 C. Thus change in temperature per se had no effect on conidia survival: conidia survival curve slopes were solely dependent upon the contemporary storage environment. Approaches are developed in order to predict loss of conidia viability in fluctuating temperature storage environments, and the predictions compared against independent observations.     

Studies on the prolonged storage of Metarhizium anisopliae conidia: effect of growth substrate on conidial survival and virulence against mosquitoes

Metarhizium anisopliae was grown on six complex mycological media and on three types of rice at three moisture levels to determine the effect of growth substrate on conidial yield, viability, and virulence against mosquitoes immediately after spore maturation and after the storage of conidia at four different temperature-relative humidity (RH) combinations over a 1-year period. Conidial yields varied with the mycological media, but the viability and virulence of conidia against mosquitoes produced on all substrates were similar when spores were stored under the same conditions. The storage conditions were more critical to spore survival and virulence than the substrate upon which conidia were produced. The comparison of rice types for conidial production indicated that conidial yield, viability, and virulence to mosquitoes were more dependent upon the moisture level during growth and on the storage conditions that upon the rice used. The best storage conditions among those tested for the retention of both spore viability and virulence against mosquitoes were 19°C–97% RH and 4°C–0% RH.     

Effect of relative humidity,temperature and fungicide on germination of conidia of Cercospora canescens caused the Cercospora leaf spot disease in mungbean

The aim of the present investigation was to determine the impact of relative humidity (RH) and temperature on conidial germination, nuclear position and effect of important fungicides on growth and conidial germination of Cercospora canescens. Germination of conidia was observed at RH range 92–100% at 5–35°C. Significant interaction between temperature and RH indicated that higher humidity and high temperature promoted quick germination both in the presence and absence of free moisture. Although in absence of free moisture at 92–95% RH higher temperatures 25–35°C promoted quick evaporation of moisture and no conidial germination. Number of germtube was increased significantly at the optimum temperature 25–30°C and higher humidity (98–100%). But higher temperature 25–35°C with lower RH did not support the conidial germination. This finding is very important for disease forecasting using meteorological data. The spray of Carbendazim as contact fungicide may not be useful since it is not effective against the conidia of C. canescens. Triadimefon did not inhibit the conidia germination but completely inhibited mycelium development at 50 μg/ml. Propriconazole inhibited both conidia germination and mycelial development. Therefore, Propiconazole may be taken as protective as well as curative spray. In non-systemic fungicide, Copper oxychloride gave anticipated result by inhibiting both conidial germination and mycelium development. Therefore, copper oxychloride can be used as protectant fungicides for Cercospora leaf spot caused by C. canescens.     

Storage Behaviour of Salix alba and Salix matsudana Seeds

Effects of dehydration, storage temperature and humidificationon germination of Salix alba andS. matsudana seeds were studied.Newly released seeds showed 100% germination before and afterdehydration to 11–12% moisture content. Germination ofthe high vigour lot (100% initial normal germination) was notaffected by dehydration to 6.7% moisture content but germinationdecreased with further dehydration to 4.3%. The lower vigourlot (75% initial normal germination) was more susceptible todehydration and germination decreased following dehydrationto 6.7% moisture content. Dry seeds of both species survivedimmersion in liquid nitrogen without loss of viability. Thegermination of seeds stored with 9% moisture content decreasedto 35–40% in 5 months at -20°C or in 2 months at 5°C.However, at 25°C seeds entirely lost viability within 2weeks. Seeds showed improved performance when stored at -70°C> - 20°C > 5°C > 25°C and tolerated dehydrationto a moisture content in equilibrium with 15% relative humidity.Results suggest that they are orthodox in storage behaviouralthough they are short-lived. Humidification treatment of lowvigour seed lots resulted in a remarkable increase in germinationpercentage. Copyright 2000 Annals of Botany Company Salix alba, Salix matsudana, willow, seed storage behaviour, dehydration, humidification, cryopreservation     

A Low-Moisture-Content Limit to Logarithmic Relations Between Seed Moisture Content and Longevity

Seeds of quinoa (Chenopodium quinoa Willd.), sunflower (Helianthusannuus L.) and linseed (Linum usitatissimum L.) showed negativelogarithmic relations between longevity and moisture contentsbetween 4.4 and 15.4, 3.2 and 13.0, and 3.2 and 15.5%, respectively,in hermetic storage at 65 °C. However, between 1.8 and 3.1,1.1 and 1.9, and 1.1 and 2.1%, respectively, longevity did notvary. The critical moisture content, below which further reductionin moisture content no longer increased longevity in hermeticstorage at 65 °C, for each species was 4.1, 2.04 and 2.7%,respectively. Quinoa, Chenopodium quinoa Willd., sunflower, Helianthus annuus L., linseed, Linum usitatissimum L., seed storage, improved viability equation, seed longevity, seed moisture content     

The Effect of Temperature and Moisture Content on the Longevity of Seed of Ulmus carpinifolia and Terminalia brassii

Seeds of the Smooth-leafed Elm (Uimus carpinifolia) and of thetropical forest tree Terb (Terminalia brassii) were stored hermeticallyand sampled at intervals for periods of up to two years. Bothspecies possess ‘orthodox’ seed (increasing longevityis observed as either moisture content or temperature are reduced)within the temperature ranges from — 13 to 52°C (Elm)and from —4 to 42°C (Terb) and within the moisturecontent ranges from 3 to 19 per cent (Elm) and from 5 to 14per cent (Terb) on a fresh weight basis. Elm seed stored at—75°C showed the expected relationship between longevityand moisture content, but did not differ significantly in longevityfrom seed kept at — 13°C when moisture contents wereheld constant. Probit analysis of the relationship between germinationpercentage and time was performed for each storage environment,yielding a slope from which the standard deviation of the distributionof seed deaths over time () was calculated. Standard deviationvalues were used in turn to determine the values of constantsin a viability equation which had previously been applied toseed of barley, chickpea, cowpea and soybean. The equation,which gave a good fit to the results obtained, can be used topredict viability for seed in storage over a wide range of environmentalconditions. Some limitations to the applicability of the viability equationwere defined. At 22 per cent and higher moisture contents Elmseed survived longer than predicted. Furthermore, all Elm andTerb seed was killed quickly on placing in —75°C at22 and 20 per cent moisture content respectively, but high viabilitywas retained for several days at 19 and 17 per cent respectively.Practical implications of the results are discussed. Uimus carpinifolia Gleditsch, Smooth-leafed Elm, Terminalia brassii Exell, Terb, seed longevity, seed storage, moisture content, temperature     

Seed Viability Constants for Groundnut

This research determined constants for a viability equationto predict the longevity of groundnut seeds and to improve themanagement of seedlot storage throughout the trading period.Seeds of the Brazilian cultivar ‘Tatu’ (Valenciabunch type) were tested. Nine moisture content levels (rangingfrom 2.4 to 12.8%) and three storage temperatures (40, 50 and65 °C) were used. Sub-samples for each moisture content-storagetemperature combination were sealed in laminated aluminium-foilpackets and stored in incubators until complete survival curveswere obtained. A reliable equation was obtained to predict groundnutseed longevity through the constantsK_E=6.177,C_W=3.426,C_H=0.0304andC_Q=0.000453.Copyright 1998 Annals of Botany Company Arachis hypogaeaL., seed longevity, seed storage, viability equation.     

Effect of high temperatures and artificial sunlight on the viability of conidia of Metarhizium anisopliae

Studies on the heat resistance of the conidia of Metarhizium anisopliae showed a clear correlation to the actual moisture conditions. The medium lethal temperature for 30 min of exposure in a suspension was 42°C, but 50.5°C at 100% RH, 57.5°C at 76% RH, and 68.8°C at 33% RH. The experiments on the effect of artificial sunlight indicated an extrapolated half-life of the conidia under field conditions of 1 hr, 40 min for 24 hr incubation at 25°C in the dark following exposure and about 2 hr, 45 min for 48 hr incubation.     

Survival and Vigour of Lettuce (Lactuca sativa L.) and Sunflower (Helianthus annuus L.) Seeds Stored at Low and Very-low Moisture Contents

Seeds of lettuce (Lactuca sativa L.) and sunflower (Helianthusannuus L.) were stored hermetically at 35 °C with 11 differentmoisture contents between 1·3 and 6·9%, and between1·3 and 7·1% of fresh mass, respectively. Germinationand vigour (mean germination time, root length, seedling dryweight) were determined after storage for 0, 8, or 16 weeks(sunflower) or 0, 8, 16, or 48 weeks (lettuce) in these environmentsfollowed by various humidification treatments (to avoid imbibitioninjury). The range of seed storage moisture contents over whichdeterioration was minimized depended upon the criterion of deteriorationused, and varied somewhat between species. Comparison of theseranges for seeds stored for the longest durations showed thatfor some criteria seed performance was poorer (P < 0·05)at both the lowest and highest moisture contents investigatedthan at certain of the intermediate storage moisture contents(e.g, most rapid germination occurred in sunflower followingstorage at 2·2-4·7% moisture content), whereasfor other criteria all the drier storage moisture contents weresuperior to the more moist (e,g. greatest seedling growth occurredin sunflower following storage at 1·3-5·1% moisturecontent). But none of these results suggested that lettuce andsunflower seeds stored hermetically at 2·5-3·0%or 2·2-2·5% moisture content, respectively, wereless vigorous than at any other moisture content tested. Inboth species, these storage moisture contents are in equilibriumwith about 8-10% relative humidity (r.h.) at 20 °C, whichis similar to and indeed marginally less than the 10-13% r.h.recommended following earlier studies on the longevity of seedsin hermetic storage at much warmer temperatures. Thus, theseresults show no evidence that the optimum seed moisture contentfor storage increases with decrease in temperature, at leastover the range 35-65 °C, as has been suggested elsewhere.We conclude that the international recommendation for the long-termseed storage for genetic conservation at 5 ± 1% moisturecontent should not be revised upwardly, and that in situationswhere refrigeration cannot be provided storage at even lowermoisture contents is worthy of further investigation for thoseseeds in which desiccation at 20 °C to equilibrium at 10%r.h. results in moisture contents well below 5%.Copyright 1995,1999 Academic Press Helianthus annuus L., sunflower, Lactuca sativa L., lettuce, desiccation, seed storage, seed vigour     

Effects of Culture Age, Washing and Storage Conditions on Desiccation Tolerance of Colletotrichum truncatum Conidia

Colletotrichum truncatum conidia produced from a one week-old culture in a liquid semi-defined medium with a C:N ratio of 5:1 were more tolerant of desiccation than those harvested from two or three week-old cultures. Conidia washed with 20% (w/v) sucrose germinated better than unwashed conidia or those washed in 10% (w/v) sucrose, 10 and 20% (w/v) glucose or fructose, 0.1% (w/v) soluble starch, 0.9% (w/v) NaCl or deionized water. Washing with sucrose (20% w/v) also resulted in significantly longer germ tubes than those produced by unwashed conidia or conidia washed with deionized water or NaCl (0.9% w/v). Conidia washed twice in sucrose showed greater desiccation tolerance during storage at 15% relative humidity (RH) and 15°C than at 30% RH and 15 or 25°C or at 15% RH and 25, 5 or -10°C.     

Factors affecting sporulation and germination of Pandora nouryi (Entomophthorales: Entomophthoraecae), a pathogen of Myzus persicae (Homoptera: Aphididae)

Pandora nouryi discharged large numbers of primary conidia between 8 and 25°C from cadavers on the surface of water-agar. At 8°C conidial discharge lasted for 120 h, but most conidia were produced within 48 h when temperature was >15°C. Saturated humidity alone was not enough to allow for sporulation to occur freely and where RH?<?95%, no conidia were discharged. Light did not affect the pattern of conidial production nor the total number of conidia. Germination percentages of conidia on the surface of water-agar were 40 and 66% at 8 and 30°C, respectively, and were significantly lower than that at 15–25°C where germination was >95%. Conidia on leaves germinated well when RH?>?74%, while no germination occurred when RH?<?100% on cover slips. All eight insecticides tested entirely inhibited conidial germination at recommended doses (R), in particular, both the organophosphorus pesticides Lorsben (chlorpyrifos) and the organochlorine pesticides Thiodan (endosulfan) completely inhibited conidial germination even at 0.2R dose.     

Biotic and abiotic factors affecting stability of Beauveria bassiana conidia in soil

Beauveria bassiana conidia were stored in sterile and nonsterile soil under various temperature, relative humidity, soil water content, and pH regimes. Survival of the conidia was primarily dependent on temperature and soil water content. Conidia half-lives ranged from 14 days at 25°C and 75% water saturation to 276 days at 10°C and 25% water saturation. Conidia held at ?15°C exhibited little or no loss in viability regardless of water content, relative humidity, or pH. Conidia were not recoverable after 10 days from soils held at 55°C. Conidia survival in nonsterile soil that was amended with carbon sources, nitrogen sources, or combinations of carbon and nitrogen was greatly decreased and loss was often complete in less than 22 days whereas sterile soil treated in the same manner showed dramatic increases in number, demonstrating that B. bassiana is capable of growth in sterile soil. The obvious fungistatic effect in amended nonsterile soils was possibly related to Penicillium urticae which was routinely isolated from the soils and is shown to produce a water-soluble inhibitor of B. bassiana. The fungistatic effect was shown to be an active inhibition rather than due to competition.     

The Effect of Moisture Content on the Low Temperature Responses of Araucaria hunsteinii Seed and Embryos

The effect of moisture content on the low temperature responsesof 'recalcitrant' Araucaria hunsteinii seed and excised embryoswas investigated in relation to germination and storage. At6°C seed longevity was decreased as mean moisture contentwas reduced from 45 to 30% (fresh weight basis); the time predictedfor the loss of one probit of germination,