Effects of Moisture Content and Temperature on Storage of Metarhizium flavoviride Conidia

The effects of moisture content and temperature on the medium-term (3-4 months) storage of conidia of Metarhizium flavoviride were investigated. Conidia harvested after 24 days of culturing on rice showed greater tolerance to long storage than conidia from 12-day cultures. The moisture content of the conidia was of greatest importance; at harvest from the culture, conidial moisture contents could be 40%, while the optimal moisture content for storage was found to be 4-5%. Dried conidia stored in oil benefited from the addition of dried silica gel, as did conidia stored as powder. A range of mineral oils proved satisfactory for storage, and when dried silica gel was added to suspensions, germination levels were 79.8% after 105 days at 28-32 C. Dried conidia stored in oil maintained germination levels of up to 96 and 85% after 80 days at 10-14 C and 28-32 C respectively. Dried conidia stored as powder retained germination levels of 95% at 10-14 C, but only up to 27% at 28-32 C. In another experiment, dried conidia maintained greater than 90% germination over 128 days, with or without silica gel at 10 - 14 C or -15 - -18 C.     

Effect of drying on conidial viability of Penicillium frequentans, a biological control agent against peach brown rot disease caused by Moniliniaspp

The effects of drying methods (freeze-, spray-, and fluid bed-drying) on viability of Penicillium frequentans conidia were compared. Viability, estimated by germination of fluid bed- and freeze-dried conidia, was similar to that of fresh conidia. Skimmed milk alone, or in combination with other protectants, was added to conidia before freeze-drying. After the freeze-drying process, all protectants used, except glycerol improved conidial viability. Freeze-dried P. frequentans conidia did not maintain viability after 30 days of storage at room temperature, while conidia dried by fluid bed-drying showed 28% viability following 180 days after drying. This work also demonstrated a relationship between conidial viability after 1 year of storage at room temperature, moisture content after fluid bed-drying and initial weight of sample. Conidial moisture contents must be reduced to 5-15% for optimal storage at room temperature. P. frequentans conidia dried by fluid bed-drying were as effective as fresh conidia in controlling brown rot of peaches.     

The effects of some storage conditions on viability of Lecanicillium lecanii conidia to whitefly (Homoptera: Trialeurodes vaporariorum)

A study on the survival of Lecanicillium lecanii conidia in storage at room temperature was carried out. Firstly, drying methods of conidia powder were compared. Vacuum-freeze drying (VFD) was more suitable for drying conidia as compared to vacuum drying (VD) at room temperature. Vacuum-freeze drying for 24-h resulted in a water content of 5.4%, and a viability, determined as germination of conidia in 2% glucose solution after16 h, was 90.3% and the infection in greenhouse whitefly, Trialeurodes vaporariorum was about 94.7% at a dose of 1×10⁸ conidia/mL. Secondly, the factors influencing viability of conidia stored at room temperature were evaluated in the laboratory. Temperature was the most critical factor influencing conidial storage stability, among the tested factors affecting survival of conidia stored at room temperature for 6 months. Both conidial germination and infection of hosts decreased with storage temperature increasing from 15 to 35°C, and at 35°C the survival of stored conidia for 6 months was near zero. The moisture content of the conidial powder was another major factor influencing viability of stored conidia at room temperature. Conidial powder dried to about 5% moisture content showed higher viability than non-dried conidial powder. For the carriers, clay and charcoal were more suitable for storage of L. lecanii conidia at room temperature. At a room temperature of 25°C, L. lecanii conidia which were dried to 5% water content and mixed with clay or charcoal could retain about 50% survival after 6 months' storage.     

不同含水量和温度下贮存中球孢白僵菌分生孢子活力与内贮营养的衰变

将球孢白僵菌(Beauveriabassiana)BBSG8702的未干燥孢子粉(含水量58.9±1.6%)和真空冷冻干燥孢子粉(含水量7.4±0.9%)置于4℃和20℃下贮存1个月,每隔5d取样测定活孢率和孢子内贮总糖和蛋白含量,发现含水量和贮存温度交互影响孢子的活力以及内贮总糖和蛋白质的代谢水平,各组合中的活孢率一般与内贮总糖和蛋白质代谢水平均存在显著或极显著相关性.在1个月的贮存期间,4℃下冻干粉总糖含量下降13.4%,蛋白质含量下降39.2%,清水中的萌发率下降32.0%,营养液中的萌发率仅下降6.7%,而未干燥孢子粉的相同指标分别下降42.4%、66.3%、96.4%和99%;在20℃下,冻干粉的上述指标分别下降了14.1%、38.2%、55.8%和 10.4%,而未干燥孢子粉则分别下降了 43.2%、65.4%、99.4%和98.4%. 显然,含水量影响活孢率和内贮营养衰变的幅度,而温度影响衰变的速度,但内贮营养的耗尽并不立即引起孢子失活,在供给外源营养之后孢子仍能萌发.将含水量降至4.0±0.9%的冻干粉贮存1年,4℃下活孢率由初始的99.0%下降至90.2%,而20℃下贮存的前165d活孢率下降较为缓慢,但此后急剧下降,至第240d时几乎全部失活.模拟分析表明,低含水量冻干粉在4℃和20℃下贮存的半衰期(即活孢率减少一半所需的时间)分别为1006d和197d.这些结果说明,白僵菌纯孢粉的含水量     

Wheat Seed Colonized with Atoxigenic Aspergillus flavus: Characterization and Production of a Biopesticide for Aflatoxin Control

Biocontrol of aflatoxin contamination using atoxigenic Aspergillus flavus to competitively exclude aflatoxin-producing strains has previously been reported, and is currently in the third year of commercial-scale tests (treating 50-200 ha per annum). Wheat seed colonized with atoxigenic A. flavus has been used in the commercial trials. Requirements for production of this colonized wheat seed are described and the spore yield of wheat is compared to other substrates. The study suggests that the most cost-effective inoculum production would require colonization of wheat (106 conidia kg -1 of wheat seed) at 25% (w/w) moisture for 18 h at 31 C. To prevent fungal growth and associated wheat aggregation in storage, seed had to be dried below 15% (w/w) moisture, although a moisture content of 35% (w/w) did not reduce viability in sealed containers stored at 18-25 C over an 8-month period. The dry biopesticide had multi-year stability without refrigeration and withstood temperatures of 70 C for 20 min. Sporulation of the product occurred within 3 days at 31 C and 100% relative humidity with yields averaging 4.9 X 109 conidia g -1 by day 7.     

The effect of environmental parameters on growth,cholinesterase activity and glutathione S-transferase activity in the earthworm (Apporectodea caliginosa)

The effect of age and environmental parameters on growth, cholinesterase (ChE), and glutathione S-transferase (GST) activities were assessed in juvenile Apporectodea caliginosa earthworms. Earthworms were maintained in three types of soil- loam, sandy, and clay- at a range of moisture contents (15-30%), and temperatures (5-20 C). Earthworm age (1-3 months) had no significant affect on ChE activity. Growth rates were influenced by all environmental parameters tested and these effects were interrelated. Optimum conditions for growth appeared to be in loam or sandy soil with 25-30% moisture at 10-15 C. The GST activity was influenced by soil temperature and activity was significantly higher at 15 C than at 5 C and 10 C. Soil type also influenced GST activity and this influence was dependent on moisture content. In sandy soil GST activity was significantly lower at 30% moisture than at lower moisture contents, in loam soil GST activity was significantly higher at 15% moisture than at higher moisture contents, while in clay soil GST activity was not affected by moisture content. These results indicate that in field experiments when evaluating GST activity soil temperature and soil type need to be consistent between control and 'contaminated sites'. ChE activity was only affected by temperature, so this should be considered when comparing control and treatedareas.     

The effect of environmental parameters on growth, cholinesterase activity and glutathione S-transferase activity in the earthworm (Apporectodea caliginosa)

The effect of age and environmental parameters on growth, cholinesterase (ChE), and glutathione S-transferase (GST) activities were assessed in juvenile Apporectodea caliginosa earthworms. Earthworms were maintained in three types of soil- loam, sandy, and clay- at a range of moisture contents (15-30%), and temperatures (5-20 C). Earthworm age (1-3 months) had no significant affect on ChE activity. Growth rates were influenced by all environmental parameters tested and these effects were interrelated. Optimum conditions for growth appeared to be in loam or sandy soil with 25-30% moisture at 10-15 C. The GST activity was influenced by soil temperature and activity was significantly higher at 15 C than at 5 C and 10 C. Soil type also influenced GST activity and this influence was dependent on moisture content. In sandy soil GST activity was significantly lower at 30% moisture than at lower moisture contents, in loam soil GST activity was significantly higher at 15% moisture than at higher moisture contents, while in clay soil GST activity was not affected by moisture content. These results indicate that in field experiments when evaluating GST activity soil temperature and soil type need to be consistent between control and 'contaminated sites'. ChE activity was only affected by temperature, so this should be considered when comparing control and treatedareas.     

Produktion von Ochratoxin A und B durch<Emphasis Type="Italic">Penicillium verrucosum</Emphasis> auf Gerste in Abhängigkeit der Parameter Wasseraktivität,Wassergehalt und Temperatur

The ochratoxin A and B (OTA, OTB) production by a toxigenic isolate ofPenicillium verrucosum grown on brewing barley up to six weeks was studied at a storage temperature of 25 °C and different moisture and water activity conditions. Sorption isothermes for barley were prepared at temperatures of 10°C, 15°C and 25°C. OTA was produced after 2 weeks of storage at moisture contents of ≥19%, which is equivalent to water activities (a_w) of 0.83 (adsorptive) and 0.82 (desorptive) at 25 °C. Increased OTA concentrations (5.8-fold and 16.1-fold) were noticed when the moisture contents were adjusted to 20% (a_{w [ads]} 25 °C=0.86) and 21% (a_{w [ads]} [ 25 °C=0.88), respectively. An increase was also shown during storage of 4 and 6 weeks (1.2-fold and 2.4-fold, respectively). Production of OTB was shown to occur at moisture contents ≥18% (a_{w [ads]} 25 °C=0.81). The findings document that OTA and OTB are not produced byP. verrucosum grown on barley stored below 18% moisture content.     

Formation of Fusarium metabolites in barley grain

Summary The influence of moisture content and temperature during storage of grain on the formation of Fusarium metabolites was studied. Naturally and artificially contaminated barley grain samples were stored at 15%, 25%, and 30% moisture contents, and at temperatures of + 5°C + 25°C, and + 30°C. Time of storage varied between one week and six months and the occurrence of Fusarium species and metabolites was analysed. The only Fusarium metabolite detected was zearalenone. The extent of Fusarium contamination decreased during storage whilst the concentration of zearalenone increased. To avoid the danger of mycotoxicoses, grains must be dryed immediately after harvest and then stored at a low temperature.     

Effects of Rehydration on the Conidial Viability of Metarhizium flavoviride Mycopesticide Formulations

The rehydration of dried conidia of Metarhizium flavoviride was investigated in an attempt to increase speed of kill of locusts and grasshoppers by formulations of this fungus. Conidia were dried to 4-5% moisture content with no apparent adverse effects on viability, but rapid rehydration (by putting dried conidia directly in free water) reduced viability. Rehydration in an atmosphere of high humidity allowed dry conidia to absorb sufficient moisture to avoid imbibition damage. Rehydrating and pre-germinating conidia prior to spraying (in an oil-based formulation) on to the desert locust, Schistocerca gregaria, did not decrease the time to death, suggesting that moisture uptake by dry conidia on the desert locust cuticle is easily achieved.     

Effects of stabilizers on shelf-life of Epicoccum nigrum formulations and their relationship with biocontrol of postharvest brown rot by Monilinia of peaches

AIM: To find a formulation of Epicoccum nigrum conidia that maintains a high viability over time and which proves efficient to biocontrol peach rot caused by Monilinia spp. METHODS AND RESULTS: We tested the effect of stabilizers and desiccants on the shelf-life of Epicoccum nigrum conidia. Conidial samples were dried for 40 min at 40 degrees C in a fluidized bed-dryer to obtain moisture contents <15%. The toxicity of additives was tested by assaying production of conidia in fermentations and germinability of the produced conidia: 50% PEG300, 10%-5% KCl (stabilizers) and 95.24% Cl(2)Ca (desiccant) significantly (P = 0.05) reduced conidial germination. To enhance shelf-life of dried conidia, nontoxic stabilizers were added at the following different stages of the production-drying process: (i) to substrate contained in bags before production, (ii) to conidial centrifuge pellets obtained after production, before filtering and drying, (iii) to conidial centrifuge pellets obtained after production, before adding talc and drying, and (iv) to conidial centrifuge pellets obtained after production, before adding silica powder and drying. Conidial germinability was tested at 0, 180 and 365 days after storage at room temperature. Shelf-life of formulations retaining the highest viability were conidia produced with 1% KCl or 50% PEG 8000, conidia dried with 2.5% methylcellulose, and conidia dried with 1% KCl + silica powder. All these formulations improved the shelf-life of E. nigrum conidia and significantly reduced brown rot on peaches. CONCLUSIONS: Our results show that additives improve the shelf-life of E. nigrum and assist controlling brown rot on peaches. SIGNIFICANCE AND IMPACT OF THE STUDY: New improved formulations of a biocontrol agent have been obtained which will improve the control of Monilinia on peach.     

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.     

顽拗性竹柏种子的贮藏特性

文章对竹柏(Podocarpus nagi)种子的脱水耐性和贮藏特性进行了研究,结果表明:竹柏种子成熟时初始含水量约为(35±0.7)%,种子对脱水敏感,其最低安全含水量约为(16.86±0.73)%,具有顽拗性种子的典型特征;湿藏和半干藏都可以作为短期保存竹柏种子的方法,且以4℃保存优于15℃保存,但不管种子含水量如何,零下低温保存对竹柏种子都是致命的;半干藏法保存实验中,未进行脱水处理的种子(对照)在4℃贮藏6个月,种子萌发率没有发生明显下降,但贮藏期延长到9个月时,临界含水量的种子萌发力保存最高;不管贮藏介质的含水量高低,也无论贮藏在4℃还是15℃,湿藏种子在9个月的贮藏期内萌发率均没有明显的降低,但当贮藏到12个月时,15℃湿藏种子的萌发率显著高于4℃贮藏的种子,但15℃湿藏的种子在贮藏到3个月时即发现种子在贮藏期间萌发,且随着贮藏介质含水量的升高和贮藏期的延长,萌发的种子增多;竹柏的离体胚经过2 h硅胶快速脱水至含水量7%后再冷冻即可获得90%以上的融后存活率,且超低温保存1年的离体胚解冻后,与只保存1周的存活率没有明显差异,表明超低温长期保存竹柏种子是可行的。本研究可以为进一步探究顽拗性种子的短期贮藏和长期保存提供理论基础和基础资料。     

Influence of two formulation types and moisture levels on the storage stability and insecticidal activity of Beauveria bassiana

The formulation of mycopesticides may require a physical separation of conidia from the substrate and subsequent drying. In the present study, Beauveria bassiana conidia produced by solid-state fermentation were harvested either through a dry or washing protocol. Washed conidia were used to design a water-dispersible granule (WG) formulation, whereas sieved conidia were mixed with an emulsifiable oil to achieve an oil-based formulation (OD). Potential harmful effects caused by the formulation type on the storage stability and insecticidal activity against Hypothenemus hampei were assessed. As expected, the time for initial conidial germination to drop 50% (GT₅₀) in all treatments was deeply influenced by storage temperatures, which varied from over 180 days at 4 °C to less than 90 days at 35°C. In all four tested temperatures, GT₅₀s for unformulated dry conidia were significantly higher than for those formulated as WG, and the latter was similar to conidia formulated as OD in the two highest temperatures. Residual water content in the OD formulation (1,600 vs. 340?ppm) had a negative influence on conidial survival under storage, whereas WG granules immediately dried after the washing protocol showed conidial germination similar to granules exposed to a slower dehydration regime. Mortality of H. hampei adults exposed to different concentrations of B. bassiana formulated as WG was slightly lower (10–15%) than either the OD or the unformulated conidia. In brief, we have demonstrated that formulation type and their moisture level can affect the storage stability and insecticidal activity of B. bassiana conidia toward the coffee berry borer. Of particular importance, we have shown that drying oils prior to formulation could improve the storage of mycopesticides, an approach that may find industrial applications.     

Viability of Lettuce Seeds: II. SURVIVAL AND OXYGEN UPTAKE IN OSMOTICALLY CONTROLLED STORAGE

Lettuce seeds were stored under aerobic conditions at varioustemperatures from 20 ?C to 35 ?C, under anaerobic conditionsat 35 ?C, and at various moisture contents ranging from 17%to 44%, controlled osmotically by polyethyleneglycol. Survivalof these seeds was compared with others maintained over a similarrange of temperatures and moisture contents but in which themoisture contents were controlled by hermetic storage. Irrespectiveof the method of controlling moisture content, under anaerobicconditions the trend (shown previously in drier seeds) of decreasein longevity with increase in hydration continued up to about27% moisture content, but above this value there was no furtherchange in longevity. In contrast, under aerobic conditions longevityincreased exponentially up to 44% moisture content, this increasestarting at about 15% moisture content at 20 ?C and at about24% at 35 ?C. Under all circumstances longevity was less athigher temperatures. On a water substrate, thermodormant seeds showed a rapid increasein rate of oxygen uptake, followed by a rapid decline, and thena slower decline to a steady state. Similar phases were shownin seeds of various moisture contents controlled osmoticallydown to 17%, although the corresponding rates were lower withdecrease in moisture content and temperature, and the phaseswere extended at lower temperatures. The implications of these results are discussed in relationto the conditions needed to sustain sub-cellular repair andturnover as postulated by Villiers. Key words: Lettuce, Lactuca sativa L, Seed viability, Seed storage, Respiration     

Effect of soil temperature and moisture on survival of eggs and first-instar larvae of Delia radicum

Edaphic factors such as soil temperature and moisture influence soil-dwelling insects, whose most vulnerable stages typically are eggs and young larvae. In this study, the survival of eggs and first-instar larvae of the cabbage maggot, Delia radicum L., was measured under laboratory conditions after exposure to a range of soil temperatures and moistures. When eggs were exposed to constant temperature (20-29°C) and humidity (5-200% [wt:wt]), temperature had no significant effect on survival, whereas humidity <25% [wt:wt] caused egg mortality. The gradual exposure of eggs to high temperatures resulted in low mortality below 33°C, but <5% of eggs survived at 40°C. When first-instar larvae were exposed to constant temperature (17-29°C) and humidity (5-100% [wt:wt]), both factors as well as their interaction had a significant effect on larval survival, which was nil at 5% (wt:wt) for all temperatures but increased from 21.9 to 42.8% at 17°C and from 34.1 to 55.0% at 29°C, for soil moisture contents of 15% and 100% (wt:wt), respectively. Eggs of D. radicum are resistant to low soil moisture and high temperature conditions. Larval survival tends to increase with an increase in soil temperature and moisture. It is suggested that soil temperature be integrated into insect development simulation models instead of air temperature, to build more effective models for cabbage maggot management.     

Effects of long-term storage at different temperatures on conidia of Botrytis cinerea Pers.: Fr.

Survival of Botrytis cinerea conidia was studied after storage without pretreatments at different temperatures (-80 degrees C, -20 degrees C, 4 degrees C and 21 degrees C). Germination tests performed during 3 years showed that viability at 21 degrees C was completely lost after 1 month. Conidia stored for 30 months at -80 degrees C, -20 degrees C and 4 degrees C were able to germinate, respectively, at 79%, 8% and 0.2%. Changes in adenylate level, energy charge and respiration (O(2) consumption) made on each set of conidia were correlated to the germination rate. The 30-month-old stored conidia showed differences in pathogenicity tests on apples. While the pathogenic aggressiveness of conidia stored at -80 degrees C was almost the same as for fresh conidia, it decreased with increasing temperature of storage. An ultrastructural study made on conidia stored for 30 months at -80 degrees C has shown the emergence of a new wall layer in a retraction zone of the cytoplasm by comparison to fresh conidia. However, the integrity of the cytoplasmic content was maintained. The effects of low temperature storage, maintenance of cell integrity and pathogenicity of conidia of B. cinerea are discussed.     

Fluidized-bed drying and storage stability of Cryptococcus flavescens OH 182.9, a biocontrol agent of Fusarium head blight

A method to produce dried granules of Cryptococcus flavescens (formerly Cryptococcus nodaensis) OH 182.9 was developed and the granules evaluated for storage stability. Small spherical granules were produced and dried using a fluidized-bed dryer. A drying and survival curve was produced for the process of fluidized-bed drying at 30°C. The granules were dried to different moisture contents (4, 7, 9 and 12%) and evaluated for storage stability at 4°C for up to a year. These different moisture contents granules had the following respective water activities (0.22, 0.38, 0.47 and 0.57 a _w). The results show the storage stability varied significantly across this moisture content range. The 9% moisture content sample had the best short-term stability (up to 4 months), while 4% moisture content had the best long-term survival (1 year). A desorption isotherm of C. flavescens was determined and modeled. The results of the storage stability and drying studies are interpreted in context of the desorption isotherm.     

Desiccation studies in relation to the storage of Araucaria seed

Relationships between seed moisture content (fresh weight basis) and germination were examined for nine Araucaria species by desiccation under mild environmental conditions. The lowest safe moisture content, below which germination percentage begins to decline, was estimated in each case. Seeds can be grouped into three moisture content categories: the first group (including A. araucana, A. angustifolia, A. hunsteinii and A. bidwillii) cannot be safely dried to below 25–40%; the second group (including A. columnaris, A. rulei, A. nemorosa and A. scopulorum) cannot be dried to below about 12% without damage; the third category contains A. cunninghamii, which can be dried to 2% without damage. Seeds in the first group should be stored at 0–5 °C with moisture contents above the lowest-safe value. Provided freezing damage does not exceed 10%, seeds in the second group should be kept at - 18°C or lower with about 7% moisture content for long-term storage and at 0–5 °C with about 12% moisture content in the short term. Seed of A. cunninghamii is best retained at near 5% moisture content and in -18°C or lower. The lowest-safe moisture content was found to be associated with seed size and weight, higher moisture content values coinciding with greater size and weight of seed. Food reserve materials also differed among the groups; seeds of the first group were mainly starchy, whilst those in the other categories possessed a high lipid content.     

Viability loss of neem (Azadirachta indica) seeds associated with membrane phase behaviour.

Storage of neem (Azadirachta indica) seeds is difficult because of their sensitivity to chilling stress at moisture contents (MC) > or =10% or imbibitional stress below 10% MC. The hypothesis was tested that an elevated gel-to-liquid crystalline phase transition temperature (Tm) of membranes is responsible for this storage behaviour. To this end a spin probe technique, Fourier transform infrared microspectroscopy, and electron microscopy were used. The in situ Tm of hydrated membranes was between 10 degrees C and 15 degrees C, coinciding with the critical minimum temperature for germination. During storage, viability of fresh embryos was lost within two weeks at 5 degrees C, but remained high at 25 degrees C. The loss of viability coincided with an increased leakage of K+ from the embryos upon imbibition and with an increased proportion of cells with injured plasma membranes. Freeze-fracture replicas of plasma membranes from chilled, hydrated axes showed lateral phase separation and signs of the inverted hexagonal phase. Dehydrated embryos were sensitive to soaking in water, particularly at low temperatures, but fresh embryos were not. After soaking dry embryos at 5 degrees C (4 h) plus 1 d of further incubation at 25 degrees C, the axis cells were structurally disorganized and did not become turgid. In contrast, cells had a healthy appearance and were turgid after soaking at 35 degrees C. Imbibitional stress was associated with the loss of plasma membrane integrity in a limited number of cells, which expanded during further incubation of the embryos at 25 degrees C. It is suggested that the injuries brought about by storage or imbibition at sub-optimal temperatures in tropical seeds whose membranes have a high intrinsic Tm (10-15 degrees C), are caused by gel phase formation.