The Mevalonate Pathway Is Important for Growth, Spore Production, and the Virulence of Phytophthora sojae

The mevalonate (MVA) pathway in eukaryotic organisms produces isoprenoids, sterols, ubiquinone, and dolichols. These molecules are vital for diverse cellular functions, ranging from signaling to membrane integrity, and from post-translational modification to energy homeostasis. However, information on the MVA pathway in Phytophthora species is limited. In this study, we identified the MVA pathway genes and reconstructed the complete pathway in Phytophthora sojae in silico. We characterized the function of the MVA pathway of P. sojae by treatment with enzyme inhibitor lovastatin, deletion of the geranylgeranyl diphosphate synthase gene (PsBTS1), and transcriptome profiling analysis. The MVA pathway is ubiquitously conserved in Phytophthora species. Under lovastatin treatment, mycelial growth, spore production, and virulence of P. sojae were inhibited but the zoospore encystment rate increased. Heterozygous mutants of PsBTS1 showed slow growth, abnormal colony characteristics, and mycelial morphology. Mutants showed decreased numbers of sporangia and oospores as well as reduced virulence. RNA sequencing analysis identified the essential genes in sporangia formation were influenced by the enzyme inhibitor lovastatin. Our findings elucidate the role of the MVA pathway in P. sojae and provide new insights into the molecular mechanisms underlying the development, reproduction, and virulence of P. sojae and possibly other oomycetes. Our results also provide potential chemical targets for management of plant Phytophthora diseases.

Temperature-dependent modelling and spatial prediction reveal suitable geographical areas for deployment of two Metarhizium anisopliae isolates for Tuta absoluta management

Tuta absoluta is one of the most devastating pests of Solanaceae crops in Africa. We previously demonstrated the efficacy of Metarhizium anisopliae isolates ICIPE 18, ICIPE 20 and ICIPE 665 against adult T. absoluta. However, adequate strain selection and accurate spatial prediction are fundamental to optimize their efficacy and formulations before field deployment. This study therefore assessed the thermotolerance, conidial yield and virulence (between 15 and 35 °C) of these potent isolates. Over 90% of conidia germinated at 20, 25 and 30 °C while no germination occurred at 15 °C. Growth of the three isolates occurred at all temperatures, but was slower at 15, 33 and 35 °C as compared to 20, 25 and 30 °C. Optimum temperatures for mycelial growth and spore production were 30 and 25 °C, respectively. Furthermore, ICIPE 18 produced higher amount of spores than ICIPE 20 and ICIPE 665. The highest mortality occurred at 30 °C for all the three isolates, while the LT50 values of ICIPE 18 and ICIPE 20 were significantly lower at 25 and 30 °C compared to those of ICIPE 665. Subsequently, several nonlinear equations were fitted to the mortality data to model the virulence of ICIPE 18 and ICIPE 20 against adult T. absoluta using the Entomopathogenic Fungi Application (EPFA) software. Spatial prediction revealed suitable locations for ICIPE 18 and ICIPE 20 deployment against T. absoluta in Kenya, Tanzania and Uganda. Our findings suggest that ICIPE 18 and ICIPE 20 could be considered as effective candidate biopesticides for an improved T. absoluta management based on temperature and location-specific approach.

Massive Ulva Green Tides Caused by Inhibition of Biomass Allocation to Sporulation

The green seaweed Ulva spp. constitute major primary producers in marine coastal ecosystems. Some Ulva populations have declined in response to ocean warming, whereas others cause massive blooms as a floating form of large thalli mostly composed of uniform somatic cells even under high temperature conditions—a phenomenon called “green tide”. Such differences in population responses can be attributed to the fate of cells between alternative courses, somatic cell division (vegetative growth), and sporic cell division (spore production). In the present review, I attempt to link natural population dynamics to the findings of physiological in vitro research. Consequently, it is elucidated that the inhibition of biomass allocation to sporulation is an important key property for Ulva to cause a huge green tide.

Sand Particle Size and Phosphorus Amount Affect Rhizophagus irregularis Spore Production Using In Vitro Propagated Spore as a Starter Inoculum in Rhizosphere of Maize (Zea mays) Plantlets

Microbial inoculants, particularly arbuscular mycorrhizal (AM) fungi, have great potential for sustainable crop management. In this study, monoxenic culture of indigenous R. irregularis was developed and used as a tool to determine the minimum phosphorus (P) level for maximum spore production under the in vitro conditions. This type of starter AM fungal inoculum was then applied to an in vivo substrate-based mass-cultivation system. Spore production, colonization rate, and plant growth were examined in maize (Zea mays L.) plant inoculated with the monoxenic culture of R. irregularis in sand graded by particle size with varying P levels in nutrient treatments. In the in vitro culture, the growth medium supplemented with 20 µM P generated the maximum number of spores (400 spores/mL media) of R. irregularis. In the in vivo system, the highest sporulation (≈500 spores g−1 sand) occurred when we added a half-strength Hoagland solution (20 µM P) in the sand with particle size between 500 µm and 710 µm and omitted P after seven weeks. However, the highest colonization occurred when we added a half-strength Hoagland solution in the sand with particle sizes between 710 µm and 1000 µm and omitted P after seven weeks. This study suggests that substrate particle size and P reduction and regulation might have a strong influence on the maximization of sporulation and colonization of R. irregularis in sand substrate-based culture.

Context, competition, and symbiont-induced bet-hedging between Dictyostelium discoideum and Paraburkholderia

Symbiotic interactions change with environmental context. We investigated context-dependence and bet-hedging in the symbiosis between social amoeba hosts and Paraburkholderia bacteria, where the context is the abundance of host food bacteria. Paraburkholderia have been shown to harm hosts dispersed to food-rich environments, but aid hosts dispersed to food-poor environments by allowing hosts to carry food bacteria. Through measuring symbiont density and host spore production, we show that this food context matters in three other ways. First, it matters for symbionts, who suffer a greater cost from competition with food bacteria in the food-rich context. Second, it matters for host-symbiont conflict, changing how symbiont density negatively impacts host spore production. Third, data-based simulations show in some cases this context-dependence can lead to a symbiont-induced bet-hedging advantage for hosts. These results show how food context can have many consequences for the Dictyostelium-Paraburkholderia symbiosis and suggest that symbionts can induce bet-hedging in hosts.

Influence of experimental storage conditions of spores of Floating fern (Salvinia natans (L.) All., Salviniaceae Martinov) on the reproduction of the species

Salvinia natans is a heterospore annual aquatic fern from the Salviniaceae family. Increasing the spores’ viability by storage simulations is the basis for creating fern spore banks. In this aspect, S. natans appears to be an unconventional subject. Between October and August 2019–2020 we conducted an experiment on the storage of S. natans spore production for 4 months in 12 combinations of abiotic factors (lighting water content – temperature) and subsequent sporesgermination during the spring – summer under the same conditions of natural temperature regime, natural light and thepresence of water. The spore production viability was assessed by the time of megasporangia germination and the numberof new individuals in each storage option. We established that the of the S. natans reproduction occurs successfully witha natural combination of abiotic environmental factors – a gradually changing temperature with winter freezing, naturallight, and the presence of sporangia in water. Storage has a beneficial effect during stratification (+ 5 °C, water) and in thecombination of factors “natural temperature – dark – dry”. Early germination and the appearance of the maximum number of new individuals in these three wintering options indicate that S. natans adapts to the seasonal climate with low winter temperatures and warm summers. Dormancy of S. natans spores belongs to the forced type. Dry storage delayed germination of megasporangia for a period of two weeks to a month. The dormancy of S. natans spores belongs to the forcedtype, since the content of sporangia in water in the light and at room temperature showed the possibility of the appearanceof new individuals in October-November.

Primary Characterization of a Life-Cycle Mutant akasusabi of the Red Alga Neopyropia yezoensis

Gametophyte-to-sporophyte transition in the haploid-diploid life cycle depends on fertilization of male and female gametes. We describe here a mutant of the marine red seaweed Neopyropia yezoensis, designated akasusabi (aks), where the gametophyte-to-sporophyte transition occurs independently of fertilization. Although conchocelis filaments were produced from carpospores, severe defects in the maturation of carposporangia via mitosis to generate conchospores were observed. In the aks mutant, however, somatic cells of gametophytic thalli were able to produce conchocelis filaments without fertilization. Thus, apogamy occurs in aks. In addition, aks was highly sensitive to wounding that promotes both asexual and apogamous reproductive responses by producing spores, which develop either into blades or conchocelis filaments, indicating that aks responds to wounding by enhanced reproduction. These findings indicated that the aks mutation enables the transformation of vegetative cells to carpospores to produce sporophytes by apogamy and wound-inducible life cycle trade-off, stimulating a reset of the timing of reproduction during the life cycle. Therefore, AKS is involved in regulations of the gametophyte-to-sporophyte transition and asexual spore production in N. yezoensis.

Sporulation environment drives phenotypic variation in the pathogen Aspergillus fumigatus

Aspergillus fumigatus causes more than 300,000 life-threatening infections annually and is widespread across varied environments with a single colony producing thousands of conidia, genetically-identical dormant spores. Conidia are easily wind-dispersed to new environments where they can germinate and, if inhaled by susceptible hosts, cause disease. Using high-throughput single-cell analysis via flow cytometry we analyzed conidia produced and germinated in nine environmentally- and medically-relevant conditions (complete medium, minimal medium, high temperature, excess copper, excess iron, limited iron, excess salt, excess reactive oxygen species, and limited zinc). We found that germination phenotypes vary among genetically-identical individuals, that the environment of spore production determines the size of spores and the degree of germination heterogeneity, and that the environment of spore production impacts virulence in a Galleria mellonella host.

Biological control of Western flower thrips, Frankliniella occidentalis using a self-sustaining granular fungal treatment

Western flower thrips (WFT), Frankliniella occidentalis, is one of the most destructive pests of vegetables, fruits and ornamental crops worldwide, causing extensive damage by direct feeding of the crop and transmitting economically important viruses. Despite the successes of biocontrol agents to control WFT, more efficient and cost-effective ways must be found to encourage grower adoption of integrated pest management. A sustainable fungal treatment was developed to preserve fungal inoculum in potting soil and reduce thrips populations. Combining cooked, oven-dried millet with BotaniGard® (a commercial form of Beauveria bassiana strain GHA) to potting soil increased spore production and persistence of the fungus in the soil. In treated pots with millet, spore concentrations were 3–4 times greater after 30 days compared with spore yields at 10 days. The number of WFT adults was significantly lower in the marigold pots treated with GHA mix + millet than untreated controls, 12% and 10% in treated pots and 70% and 68% in untreated pots in sterile and non-sterile soil, respectively. Incorporation of millet in the potting mix enhanced the effect of the fungal treatments by providing a nutritive substrate on which the fungus could become established. This method is relatively inexpensive and easy for growers to use in greenhouses because granular formulations of B. bassiana are not commercially available.