Effects of cold storage on life-history traits of Aphidius matricariae

Aphidius matricariae Haliday (Hymenoptera: Braconidae) is a polyphagous solitary endoparasitoid, attacking more than 40 species of aphids. This parasitoid is an important commercial product of many companies that produce biological control agents. Storage at low temperature increases the shelf life of many biocontrol agents, allowing companies to provide a steady and sufficient supply of insects for biocontrol programs. In the current study, the effects of cold storage of 1-day-old host mummies with A. matricariae for various time periods (5, 10, 15, 20, and 30 days) at 5 °C on the parasitoid’s key life-history traits were investigated. Parameters assessed after storage included adult emergence rate, offspring sex ratio, adult longevity, oviposition period, fecundity, and life-table parameters (R₀, r, λ, T, and DT). Our results showed that the mummies of A. matricariae could be stored at 5 °C for 5 days without loss of quality and for 10–15 days with minimal reduction in quality (e.g., some reduction in adult longevity and R₀). If parasitoids were stored for >15 days, quality was more strongly affected. In conclusion, A. matricariae pupae could be stored at 5 °C for up to 15 days without significant negative post-storage effects on fitness of the parasitoid. These results could be used to improve the planning of mass rearing and mass release of A. matricariae in augmentative biological control programs.     

Recurrent warming to improve cold storage of Trichogrammatids (Hymenoptera: Trichogrammatidae)

Cold storage can be used to slow development, facilitate accumulation of the organisms and accommodate fluctuating demand for augmentative biological control agents. Previous research suggested the possibility of improving cold storage of Trichogrammatids by recurrent warming, so we subjected Trichogramma ostriniae juveniles within Ephestia kuehniella host eggs to either 2°C constant or to 2°C with twice-weekly recurrent warming for 3 h to 20°C. Parasitoid subsamples were allowed to mature for 1–9 days before placement in cold storage for up to 8 weeks. Parasitism by parentals, progeny emergence and fecundity and longevity of progeny were measured weekly for 8 weeks. Relative to constant 2°C, recurrent warming generally improved emergence, fecundity and longevity, and all the response variables were affected by the interaction of temperature regimen, parasitoid maturity class, and cold storage duration. This implies the utility of recurrent warming to improve egg parasitoid performance and for extending the duration of cold storage.     

Side-effects of organic and synthetic pesticides on cold-stored diapausing prepupae of <Emphasis Type="Italic">Trichogramma cordubensis</Emphasis>

The side-effects of three insecticides (deltamethrin, lambda-cyhalothrin and Bacillus thuringiensis Berliner) and one fungicide (basic copper sulphate) were tested on cold-stored diapausing prepupae of Trichogramma cordubensis Vargas and Cabello (Hymenoptera: Trichogrammatidae). Pesticides were directly sprayed on parasitized host eggs (being the diapausing parasitoids in the prepupal stage) after cold storage (3°C) for three different periods (60, 120 and 180 days). Regardless of the period of cold storage, both pyrethroids reduced the emergence rates of T. cordubensis (both <25%) compared to the control (emergence varied from 83% to 89%). The most toxic pyrethroid was lambda-cyhalothrin; Trichogramma cordubensis adult emergence varied from 7% to 15%. Lambda-cyhalothrin also negatively affected the longevity and fecundity of parasitoids cold stored for 60 days. Bacillus thuringiensis Berliner subsp. kurstaki and basic copper sulphate had little or no adverse effect on emergence rates (generally >80%), longevity nor fecundity of T. cordubensis, indicating that these pesticides could be successfully integrated into pest management programs using wasps that were cold stored under diapause. Such integration would be valuable to pest management programs by reducing the costs of T. cordubensis mass rearing and by allowing producers to stockpile parasitoids for release in the growing season. However, since the emergence rate, longevity and fecundity of T. cordubensis generally decreased with increasing duration in cold storage, wasps to be used in integrated pest management programs should only be stored at 3°C for 60 days maximum. Handling Editor: Sam Cook.     

Differential response of fourTrichogramma species to low temperatures for short term storage

Differential response was noticed when 4 species ofTrichogramma egg parasitoids were stored at 2°, 5° and 10°C for 7 to 49 days. Pupal stage was found to be most appropriate for storage. Emergence ofTrichogramma achaeae Nagaraja and Nagarkatti andTrichogrammatoidea eldanae Viggiani was greater in comparison toTrichogramma chilonis Ishii andTrichogramma japonicum Ashmead at all 3 temperature regimes. Fecundity and longevity declined drastically after storage of 14 days at 2° and 5°C and 21 days at 10°C, when removed to room temperature. 10°C was found to be the best temperature for storage up to 49 days. Contribution No. 51004 of Biological Control Centre (NCIPM), Bangalore.     

Effect of long-term cold storage on the fitness of pre-wintering <Emphasis Type="Italic">Harmonia axyridis</Emphasis> (Pallas)

The multicolored Asian ladybird beetle, Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae), is considered an important generalist predator that can be used as a biological control agent against Hemiptera Sternorrhyncha, Thysanoptera, and the eggs and larvae of Lepidoptera, Coleoptera and Diptera. There are currently abundant natural resources of overwintering H. axyridis in Asia and North America. Given its potential as a biological control agent, methods can be developed to increase its effectiveness for pest control. The availability of an adequate cold storage method would enable the use of field-collected pre-wintering ladybirds for pest suppression in the following season. We studied the effect of cold storage (30, 60, 90, 120 and 150 days stored at −3, 0, 3 and 6°C) on survival, fecundity and predation in field-collected populations. The survival of both female and male ladybirds decreased significantly as storage duration increased at −3°C and 0°C. The ladybirds showed more than 80% survival when they were stored for 150 days at 3°C and 6°C. Long-term cold storage had different effects on the fecundity of H. axyridis at different temperatures. Prolonged cold storage at both 3°C and 6°C shortened pre-oviposition duration and had no adverse effect on reproductive capacity as compared to that of unstored individuals. The adults that experienced 90-day storage at 0°C had the shortest pre-oviposition duration and the largest reproductive capacity. The individuals that were stored for 150 days at 3°C consumed significantly more aphids than the unstored ones. Generally, 3–6°C is a suitable temperature for cold storage of the ladybird without any reduction in fitness. This study will help the exploitation and application of pre-wintering H. axyridis for the biological control of insect pests.     

The germination of grass seeds after storage at different temperatures in aluminium foil and manilla paper packets

The germination of seeds of three species of forage grasses, Lolium perenne, Festuca pratensis and Dactylis glomerata, was studied after storage for 3–5 years under five different storage conditions: in aluminium foil packets at —25°C, 0°C and laboratory temperature (c. 18°C), and in manilla paper packets at 0°C and laboratory temperature. With Lolium perenne and Festuca pratensis high germination values at 3 and 7 days were obtained from seed stored at — 25 °C and 0°C in foil packets (5% moisture), but at laboratory temperatures, seed from foil packets gave lower germination values than those from manilla paper packets. At all three temperatures Dactylis glomerata germination after 7 and 14 days was higher in seed stored in foil than in manilla packages. With all three species stored in manilla packets, germination was higher after laboratory than cold storage.     

球孢白僵菌对烟蚜茧蜂生命参数及控害效果的影响

在室内评价了球孢白僵菌对烟蚜茧蜂生命参数及控害效果的影响。分别在烟蚜茧蜂寄生桃蚜后不同时间进行高剂量（1900孢子/mm^{2）接菌,检测蚜虫感病率和寄生蜂形成的僵蚜率及僵蚜出蜂率。结果表明,球孢白僵菌对僵蚜率和僵蚜出蜂率的影响随接菌时间不同而变化。在烟蚜茧蜂寄生前1d、寄生当天和寄生后3d接菌,蚜虫感病率分别为59.6%、56.2%和34.8%;与对照相比,僵蚜率分别下降94%、59%和47%,僵蚜出蜂率分别减少83%、54%和49%。在寄生后5d或7d接菌,僵蚜率和僵蚜出蜂率不受明显影响,但蚜虫感病率降低到8.2%以下。对蚜尸内白僵菌菌体含量检测表明,随着烟蚜茧蜂寄生后接菌时间的推移,菌体数量迅速下降。寄生蜂寄生后5d或7d接菌,蚜尸内几乎检测不到菌体。直接喷雾接菌烟蚜茧蜂,成蜂寿命缩短4d左右,且81.8%的蜂尸受白僵菌感染。接菌后的寄生蜂对蚜虫寄生率几乎无影响,但寄生蜂在蚜虫体内的存活时间缩短了27.8%。}     

Effects of temperature on biology and life table parameters of the Asian citrus psyllid, Diaphorina citri Kuwayama (Homoptera: Psyllidae)

The development, survivorship, longevity, reproduction, and life table parameters of the Asian citrus psyllid, Diaphorina citri Kuwayama were evaluated at 10°C, 15°C, 20°C, 25°C, 28°C, 30°C and 33°C. The populations reared at 10°C and 33°C failed to develop. Between 15°C and 30°C, mean developmental period from egg to adult varied from 49.3 days at 15°C to 14.1 days at 28°C. The low‐temperature developmental thresholds for 1st through 5th instars were estimated at 11.7°C, 10.7°C, 10.1°C, 10.5°C and 10.9°C, respectively. A modified Logan model was used to describe the relationship between developmental rate and temperature. The survival of the 3rd through 5th nymphal instars at 15–28°C was essentially the same. The mean longevity of females increased with decreasing temperature within 15–30°C. The maximal longevity of individual females was recorded 117, 60, 56, 52 and 51 days at 15°C, 20°C, 25°C, 28°C and 30°C, respectively. The average number of eggs produced per female significantly increased with increasing temperature and reached a maximum of 748.3 eggs at 28°C (P<0.001). The population reared at 28°C had the highest intrinsic rate of increased (0.199) and net reproductive rate (292.2); and the shortest population doubling time (3.5 days) and mean generation time (28.6 days) compared with populations reared at 15–25°C. The optimum range of temperatures for D. citri population growth was 25–28°C.     

Long-term storage of sexual and asexual Trichogramma brassicae (Hymenoptera: Trichogrammatidae)

The effect of exposure duration to pre-storage (representing fall conditions) and storage (representing winter conditions) temperatures on the emergence rate in sexual and asexual Trichogramma brassicae Bezdenko (Hymenoptera: Trichogrammatidae) was studied under controlled laboratory conditions. Asexual T. brassicae was infected by Wolbachia, an endosymbiont bacterium. We considered 0, 30, 40, 50 and 60 days as exposure duration at 10°C and 0, 30, 60, 90 followed by 180 days as exposure duration at 4°C for both T. brassicae strains. Emergence rate was significantly affected by the presence of Wolbachia, the duration of exposure to 10°C, the duration of storage at 4°C and their interactions, but not by the interaction between strain × storage duration at 4°C. Emergence rate of adults was positively correlated with exposure duration at 10°C, but it gradually decreased as the cold storage duration (4°C) increased in both strains. In conclusion, we are able to store both asexual and sexual strains for longer periods of time at 4°C, but the emergence rate of sexual T. brassicae was greater than asexual in all the treatments. The presence of Wolbachia could have a negative effect on fitness of T. brassicae and reduces its emergence rate after long-term storage.     

Effects of cold storage on Thripobius javae (=T. semiluteus) (Hymenoptera: Eulophidae)

Thripobius javae (Girault) was introduced in 1995 from Israel into Italy to control the greenhouse thrips, Heliothrips haemorrhoidalis (Bouché). Following introduction, successive augmentative releases of this parasitoid gave unsatisfactory and contradictory results, mainly due to the difficulty in synchronising its availability in sufficient number at the time of release. Efficient storage of this biological control agent could improve its current production and use. The effects of different sets of storage techniques at a single temperature and with a combination of different temperatures and instars on several fitness traits (residual developmental time to adult emergence after the end of storage, pupal mortality, longevity with and without hosts and progeny of emerged adults) were evaluated in order to determine the best conditions for storing the parasitoid.

For the pupal stage, increasing storage up to 14 days, at 10°C, gave only a moderate reduction (33%) of a modified composite quality index of its fitness. In contrast, when adults were stored for more than 10 days, at 15°C, residual longevity and progeny were reduced significantly. A combination of two temperatures (10 and 15°C) for pupal storage and a combination of pupal (10°C) and adult (15°C) storage had detrimental effects on parasitoid fitness. Temperatures of storage lower than 15 and 10°C had detrimental effects on adults and pupae, respectively.     

Effect of water content and temperature on seed longevity of seven Brassicaceae species after 5 years of storage

Maximising seed longevity is crucial for genetic resource preservation and longevity of orthodox seeds is determined by environmental conditions (water content and temperature). The effect of water content (down to 0.01 g·H₂O·g^?1) on seed viability was studied at different temperatures for a 5‐year storage period in taxonomically related species. Seeds of seven Brassicaceae species (Brassica repanda, Eruca vesicaria, Malcolmia littorea, Moricandia arvensis, Rorippa nasturtium‐aquaticum, Sinapis alba, Sisymbrium runcinatum) were stored at 48 environments comprising a combination of eight water contents, from 0.21 to 0.01 g·H₂O·g^?1 DW and six temperatures (45, 35, 20, 5, ?25, ?170 °C). Survival curves were modelled and P50 calculated for those conditions where germination was reduced over the 5‐year assay period. Critical water content for storage of seeds of six species at 45 °C ranged from 0.02 to 0.03 g·H₂O·g^?1. The effect of extreme desiccation at 45 °C showed variability among species: three species showed damaging effects of drying below the critical water content, while for three species it was neither detrimental nor beneficial to seed longevity. Lipid content could be related to longevity, depending on the storage conditions. A variable seed longevity response to water content among taxonomically related species was found. The relative position of some of the species as long‐ or short‐lived at 45 °C varied depending on the humidity at which storage behaviour was evaluated. Therefore, predictions of survival under desiccated conditions based on results obtained at high humidity might be problematic for some species.     

Effect of cold storage of pupae parasitoid wasp,Habrobracon hebetor (Say) (Hymenoptera: Braconidae), on its efficiency

Cold storage is an appropriate way to extend the life of natural enemies such as insect parasitoids. Identification of risk factors for use in biological control and replacing it instead of chemical control is important, and therefore wasp species of genus Bracon have been used for controlling various pests in the past and present. The aim of this study was to determine the optimum temperature and time of storage of parasitoids pupae, Habrobracon hebetor (Say) in cold conditions. After in vitro rearing of the insect, the pupae of H. hebetor at the temperatures of 12, 9 and 4?°C for a period of six weeks in five replicates were stored. After these times, the effect of storage on the percentage of emerged adults and the longevity of male and female adults was evaluated on a weekly basis. The results of the survey conducted at 12?°C showed that the percentage of adults emerged from the first week has dropped by almost half. H. hebetor pupae could be stored for a week at 9?°C. At 4?°C, a significant reduction was caused in the efficacy of the stored pupae and is not recommended.     

Morphological and biological notes on Encarsia meritoria Gahan (Hymenoptera,Aphelinidae), a parasitoid of Trialeurodes vaporariorum (Westwood) (Homoptera,Aleyrodidae) new in Europe

Encarsia meritoria Gahan, a Neartic species recorded only in the USA, was found naturally occurring in Catalonia (north‐east Spain) in 1987. The morphology of immature stages, the rate of development in the range 12°C‐34°C, the longevity and fecundity at 24°C and some observations on its searching and host feeding behaviour are presented in this paper. Mean development times from egg laying to adult emergence ranged from 75 days at 12° C to 11 days at 28°C. The Lower Developmental Threshold computed from linear regression equations was 9°C. Females laid an average of 198 eggs (range 89–330) in an average of 34 days (longevity range 19–54 days). The intrinsic rate of increase at 24° C was 0.1717, slightly greater than the r_m of T. vaporariorum and smaller than the values reported for E. tricolor and E. formosa, which is not very promising for biological control. However, this may not be a definitive conclusion, because factors other than those considered in our experiments may play an important role infield conditions.     

Influence of larval rearing temperature on the quality of cold‐stored Oomyzus sokolowskii Kurdjumov (Hymenoptera: Eulophidae)

Oomyzus sokolowskii, an important parasitoid of Plutella xylostella, has great potential for use in biological control. Storage at suboptimal temperature is valuable for increasing the shelf‐life of insect parasitoids. In this study, O. sokolowskii larvae were reared at 30/25, 25/25 and 25/20°C light/dark (65 ± 5% RH, 16 : 8 h L : D) until pupation. The pupae were then cold‐stored at 4 ± 1°C (60 ± 5% RH, full darkness). The pupae were removed out from the storage at 10, 20, 30 and 40 days after storage (DAS) and maintained at 25 ± 2°C until adults emerged or pupae died. Quality of the emerging adults and their F₁ offspring were assessed. Incidence of parasitism by O. sokolowskii was higher at 30/25°C than at 25/20°C. Cold storage of O. sokolowskii pupae greatly affected the fitness of the parasitoid: adult emergence rates were lower in the 40 DAS treatment than in other treatments; when O. sokolowskii larvae developed at 25/25°C, female proportions of the emerged adults were lower in the 40 DAS treatment than in the 0 and 10 DAS treatments. Larval rearing temperature mildly affected the adult emergence rate, post‐storage developmental time and female proportion with a few exceptions. Number of parasitoids emerged per host pupa, and incidence of parasitism by the females were neither affected by larval rearing temperature nor cold storage duration. Trans‐generational effects on F₁ offspring were evident in adult emergence rate, egg‐adult developmental time and female proportion which were negatively affected by long duration of storage (40 days), but not by larval rearing temperature with a few exceptions. In conclusion, O. sokolowskii pupae could be stored at 4°C for up to 30 days without significant fitness loss.     

Viability of rape (Brassica napus L.) seeds following selection on the basis of newly-emerged radicles then subsequent drying and storage

Germinating rape seeds selected on the basis of newly-emerged radicles (1 ± 0.5 mm) were dried to an equilibrium moisture content (c. 11%) in air at 20°C and 80% relative humidity without loss of viability. Storage life of these low-moisture-content germinating (LMCG) seeds at 15°C was limited to 7 days before viability was significantly reduced. However, viability of LMCG seeds was maintained for 84 days in storage at -20°C. Longer periods in store reduced viability, but 96% of seeds still remained viable after 336 days at - 20°C. Increasing periods of storage at -20°C reduced the subsequent seed longevity at 15°C, indicating a reduction in vigour during storage. Storage under reduced pressure or in a nitrogen atmosphere had little significant effect on seed longevity. Reduction of moisture content below 11% using vacuum drying at a range of temperatures reduced seed vigour.     

Cold tolerance of coccons ofAllorhogas pyralophagus [Hym.: Braconidae]

Cold tolerance studies were conducted under laboratory conditions for coccons ofAllorhogas pyralophagus Marsh, a Mexican parasitoid of graminaceous borers. By storage, developmental time (from cocoon to adult emergence) could be extended by 2 to 6 times. However, cocoons stored for more than 14 days at 2°C failed to survive, while at 5 and 10°C, about 50% emergence was recorded for upto 21 days of storage. With respect to survival and adult longevity, 10°C seemed to be the most suitable storage temperature. Pre-emergence period was also significantly increased by storing cocoons for 21 to 35 days at this temperature. Sex-ratio of emerging adults was not significantly affected by storage. Fecundity was adversely affected in all the treatments except in the case of females emerging from cocoons stored at 5°C for 7 days. The progeny of parasitoids which emerged from cocoons stored at 5 and 10°C for 35 days consisted of only males. It is clear from the present study thatA. pyralophagus cocoons are more amenable to short-term storage. Contribution No. 285/87 of the Indian Institute of Horticultural Research, Bangalore-560 089.     

Prior frozen storage enhances the effect of edible coatings against Listeria monocytogenes on cold-smoked salmon during subsequent refrigerated storage

Aims: Listeria monocytogenes is a major safety concern for ready‐to‐eat foods. The overall objective of this study was to investigate whether prior frozen storage could enhance the efficacy of edible coatings against L. monocytogenes on cold‐smoked salmon during subsequent refrigerated storage. Methods and Results: A formulation consisting of sodium lactate (SL, 1·2–2·4%) and sodium diacetate (SD, 0·125–0·25%) or 2·5% Opti.Form (a commercial formulation of SL and SD) was incorporated into each of five edible coatings: alginate, κ‐carrageenan, pectin, gelatin and starch. The coatings were applied onto the surface of cold‐smoked salmon slices inoculated with L. monocytogenes at a level of 500 CFU cm^?2. In the first phase, the slices were first frozen at ?18°C for 6 days and stored at 22°C for 6 days. Alginate, gelatin and starch appeared to be the most effective carriers. In the second phase, cold‐smoked salmon slices were inoculated with L. monocytogenes, coated with alginate, gelatin or starch with or without the antimicrobials and stored frozen at ?18°C for 12 months. Every 2 months, samples were removed from the freezer and kept at 4°C for 30 days. Prior frozen storage at ?18°C substantially enhanced the antilisterial efficacy of the edible coatings with or without antimicrobials during the subsequent refrigerated storage. Conclusions: Plain coatings with ≥2 months frozen storage and antimicrobial edible coatings represent an effective intervention to inhibit the growth of L. monocytogenes on cold‐smoked salmon. Significance and Impact of the Study: This study demonstrates the effectiveness of the conjunct application of frozen storage and edible coatings to control the growth of L. monocytogenes to enhance the microbiological safety of cold‐smoked salmon.     

Life table parameters of an indigenous strain of Neoseiulus californicus McGregor (Acari: Phytoseiidae) when fed Tetranychus urticae Koch (Acari: Tetranychidae)

The life table of the indigenous Neoseiulus californicus was studied at different temperatures and 65 ± 5% relative humidity under conditions of 16 h light : 8 h dark (LD 16:8). The total developmental period from egg to adult varied from 3.0 to 14.0 days at 15 to 35°C. Survival to adulthood ranges from 86.21 to 93.94%, with the highest rate at 25°C. The lower threshold temperature from egg to adult stages of females and males was 10.84 and 10.72°C, respectively, and the thermal constant was 57.14 degree‐days (DD) for females and 56.18 DD for males. Total number of eggs laid by each female was the highest (70.38 eggs) at 25°C, whereas average daily fecundity was the highest (3.69 eggs/female/day) at 30°C. The net reproductive rate was the highest (48.49) at 25°C and lowest (26.18) at 30°C. Mean generation time decreased from 19.04 to 11.47 days with increasing temperature from 20 to 30°C. Both intrinsic rate of natural increase (0.284) and finite rate of increase (1.32) were maximum at 30°C. Adult longevity was the highest (42.75 days for females and 32.60 days for males) at 20°C and lowest (22.70 days for females and 15.30 days for males) at 30°C. Sex ratio was female biased and was the highest (78.08) at 25°C and lowest (70.24) at 30°C. Developmental data of five constant temperatures, temperature thresholds and thermal requirements may be used to predict the occurrence, number of generations and population dynamics of N. californicus as an important biocontrol agent of Tetranychus urticae.     

Age-specific fecundity and life table of Trissolcus semistriatus, an egg parasitoid of the sunn pest Eurygaster integriceps

The effects of temperature on age‐specific fecundity and life table parameters of the egg parasitoid Trissolcus semistriatus (Nees, 1834) (Hymenoptera: Scelionidae) were examined under four constant temperature conditions (17, 20, 26 and 32°C), using eggs of the sunn pest Eurygaster integriceps Puton, 1881 (Hemiptera: Scutelleridae), an important pest of wheat, as hosts. The intrinsic rate of increase increased linearly, while the mean generation time and the doubling time decreased with increases in temperature. The net reproductive rate, however, varied without clear correlation with temperature. Fecundity tended to be higher at higher temperatures. The total number of eggs per female was estimated as 52.0 and 116.4 eggs, respectively, at 17°C and 32°C, with the highest fecundity rate during the first day of oviposition. The oviposition rate fluctuated from 4.4 to 14.3 eggs per day. Oviposition and postoviposition periods and longevity decreased when temperature increased. Maximum longevity for females was 21.6 days at 20°C, and female parasitoids lived longer than males at all temperatures. The development period ranged from 7.1 days (32°C) to 35.6 days (17°C) for males and from 8.4 days (32°C) to 37.2 days (17°C) for females. The development of female T. semistriatus required 166.7 degree‐days (DD) above a theoretical threshold of 11.8°C and the development of males required 142.9 DD above 13.1°C. The numbers of generations per year for female and male T. semistriatus, given the temperature in Tekirdag, Turkey, were estimated to be 9.0 and 8.8, respectively. The potential of the egg parasitoid for the control of E. integriceps is discussed.     

Effects of pH,NH<Subscript>4</Subscript>-N concentration,temperature, and storage period on basidiospore germination in an ectomycorrhizal ammonia fungus <Emphasis Type="Italic">Hebeloma vinosophyllum</Emphasis>

Basidiospore germination in an ectomycorrhizal ammonia fungus Hebeloma vinosophyllum was stimulated by 10–500 mM NH₄Cl aqueous solution at pH 4.5–9.0, but not by pure water. The basidiospores germinated at 10°–35°C with an optimum at 25°–30°C. The highest germination percentage (83.0%) was observed in 100 mM NH₄Cl aqueous solution adjusted to pH 8.0 by KOH, when the basidiospores were incubated at a density of 10⁶ spores/ml at 30°C for 14 days. The percent germination value decreased with the increased duration of storage under both dry and wet conditions. Humidity and temperature affected the longevity of H. vinosophyllum basidiospores. The basidiospores maintained their germination ability longer under a dry condition than under a wet condition. The greatest longevity was accomplished by storage at 15°C under a dry condition.