Exploring behavioural and physiological adaptations in mountain pine beetle in response to elevated ozone concentrations.

Elevated ozone (eO3) concentrations pose a threat to insect populations by potentially altering their behaviour and physiology. This study investigates the effects of eO3 concentrations on the mountain pine beetle which is a major tree-killing species of conifers in northwestern North America. We are particularly interested in understanding the effects of eO3 concentrations on beetle behaviour and physiology and possible transgenerational impacts on bark beetle broods. We conducted O3-enrichment experiments in a controlled laboratory setting using different O3 concentrations (100-200 ppb; projected for 2050-2100) and assessed various beetle responses, including CO2 respiration, mating behaviour, survival probability, locomotion, and attraction behaviour. Transgenerational impacts on the first and second generations were also analyzed by studying brood morphology, mating behaviour, survival, and pheromone production. We found that beetles exposed to eO3 concentrations had shorter oviposition galleries and reduced brood production. Beetle pheromones were also degraded by eO3 exposure. However, exposure to eO3 also prompted various adaptive responses in beetles. Despite reduced respiration, eO3 improved locomotor activity and the olfactory response of beetles. Surprisingly, beetle survival probability was also improved both in the parents and their broods. We also observed transgenerational plasticity in the broods of eO3-exposed parents, suggesting potential stress resistance mechanisms. This was evident by similar mating success, oviposition gallery length, and brood numbers produced in both control and eO3 concentration treatments. This study demonstrates the sensitivity of mountain pine beetles to increased O3 concentrations, contributing crucial insights into the ecological implications of eO3 concentrations on their populations. Overall, the outcome of this study contributes to informed climate change mitigation strategies and adaptive management practices for the development of resilient forests in response to emerging forest insect pests worldwide.

Unraveling the multifaceted effects of climatic factors on mountain pine beetle and its interaction with fungal symbionts.

Mountain pine beetles (MPBs) pose a substantial threat to North American pine forests, causing extensive tree mortality over large areas. Their tree-killing ability is closely linked to mass aggregation on host trees triggered via pheromones and dependence on their symbiotic fungi. However, the influence of a changing climate on the biology of MPBs and their co-evolved interactions with their fungal symbionts remains uncertain. To investigate this, male and female pairs of beetles were introduced into freshly cut logs from lodgepole pine trees and placed in controlled climate chambers with manipulated environmental conditions, including two levels of CO2 (ambient vs. 1000 ppm), O3 (ambient vs. 100 ppb) and humidity (33% vs. 65%). The beetle-infested logs were left in these chambers for 1 month and then returned to ambient conditions until brood emergence. Emerging broods were collected for further analysis. Additionally, three species of fungal symbionts (Grosmannia clavigera, Ophiostoma montium and Leptographium longiclavatum) were subjected to the same CO2 , O3 and humidity conditions for 5 days. Lower humidity promoted MPB reproduction and fungal growth. Elevated CO2 accelerated larval growth and emergence while improving brood pheromone production. Elevated O3 had a negative impact on MPB reproduction and brood fitness while improving its immune responses to an entomopathogenic fungus (Beauveria bassiana). It also inhibited fungal growth and reproduction, whereas elevated CO2 had varied (positive or negative) effects on fungal growth and ergosterol (proxy to fungal mass) production depending on the fungal species. Together, these findings suggest that climate change can potentially alter the interactions between MPBs and their fungal symbionts, highlighting the importance of understanding how climate change affects forest pests and their symbiotic relationships to develop effective management strategies in the future.

A Pine in Distress: How Infection by Different Pathogenic Fungi Affect Lodgepole Pine Chemical Defenses.

In North America, lodgepole pine is frequently subjected to attacks by various biotic agents that compromise its ability to defend against subsequent attacks by insect herbivores. We investigated whether infections of lodgepole pine by different pathogenic fungal species have varying effects on its defense chemistry. We selected two common pathogens, Atropellis canker, Atropellis piniphila, and western gall rust, Endocronartium harknessii, affecting mature lodgepole pine trees in western Canada. We also included three ophiostomatoid fungi Grosmannia clavigera, Ophiostoma montium, and Leptographium longiclavatum associated with the mountain pine beetle (Dendroctonus ponderosae), because they are commonly used to investigate induced defenses of host trees of bark beetles. We collected phloem samples from lodgepole pines infected with the rust or the canker and healthy lodgepole pines in the same stand. We also inoculated mature lodgepole pines with the three fungal symbionts and collected phloem samples 2 weeks later when the defense chemistry was at its highest level. Different fungal species differentially altered the terpene chemistry of lodgepole pine trees. E. harknessii and the fungal symbionts altered the terpene chemistry in a similar pattern while trees responded to the infection by the A. piniphila differently. Our study highlights the importance of considering specific biotic stress agents in tree susceptibility or resistance to the subsequent attacks by insect herbivores, such as mountain pine beetle.

Flavonoids and Devosia sp SL43 cell-free supernatant increase early plant growth under salt stress and optimal growth conditions.

Salt stress is a major threat to modern agriculture, significantly affecting plant growth and yield, and causing substantial economic losses. At this crucial time of increasing climate change conditions, soil salinity will continue to develop and become an even more serious challenge to crop agriculture. Hence, there is a pressing need for sustainable techniques in agricultural production that could meet the dual challenges of crop productivity and environmental instability. The use of biostimulants in agricultural production has greatly influenced plant health and global food production. In particular, the application of bioactive materials produced by beneficial microbes is becoming a common practice in agriculture and provides numerous benefits to plant growth and resistance to stressful conditions. In this research two biostimulants; a type of plant secondary metabolite (flavonoids) and a microbe-based material (CFS: Cell-Free Supernatant) containing active compounds secreted by a novel bacterial strain isolated from Amphecarpaea bracteata root nodules (Devosia sp - SL43), have been utilized to improve the growth and stress resistance of two major oil seed crops; canola and soybean, under optimal and salt stress conditions. Our findings suggested significant improvements in crop growth of canola and soybean following the application of both biostimulants. Under optimal growth conditions, soybean growth was significantly affected by foliar spray of flavonoids with increases in shoot fresh and dry weight, and leaf area, by 91, 99.5, and 73%, respectively. However, soybean growth was unaffected by flavonoids under salt stress. In contrast, CFS with a meaningful capacity to mitigate the negative effects of salinity stress improved soybean shoot fresh biomass, dry biomass, and leaf area by 128, 163 and 194%, respectively, under salt stress conditions. Canola was less responsive to both biostimulants, except for canola root variables which were substantially improved by flavonoid spray. Since this was the first assessment of these materials as foliar sprays, we strongly encourage further experimentation to confirm the findings reported here and to determine the full range of applicability of each of these potential technologies.

Seed Priming with Devosia sp. Cell-Free Supernatant (CFS) and Citrus Bioflavonoids Enhance Canola and Soybean Seed Germination.

Climate change, environmental pollution and associated abiotic stresses are beginning to meaningfully affect agricultural production worldwide. Salt stress is, however, one of the most important threats that significantly impairs plant growth and development. Plants in their early growth stages such as seed germination, seed emergence and early seedling growth are very sensitive to salt stress. Among the range of sustainable techniques adopted to improve seed germination and early plant growth is seed priming; however, with the use of ecofriendly substances, this is one of the most effective and economically viable techniques to improve seed tolerance against such environmental stresses. For instance, priming with appropriate non-synthetic compounds including microbial biostimulants are prominent ways to sustainably address these challenges. Therefore, in this research, by using the "priming technique", two biostimulants were tested for their potential as sustainable approaches to improve canola and soybean seed germination under salt stress and optimal growth conditions. Canola and soybean seeds were primed with flavonoids extracted from citrus fruits (flavopriming) and cell-free supernatant (CFS; produced by a novel strain of Devosia sp.-SL43), alone and in combination, and exposed to low-higher levels of salt stress and ideal growth conditions. Both biostimulants showed promising effects by significantly improving seed germination of soybean and canola under both ideal and stressful conditions. However, increases in seed germination were greater under salinity stress as flavonoids and CFS with stress amelioration effects showed substantial and statistically significant improvements in seed germination under varying levels of salt stress. In addition, combinations (mixtures) of both biostimulants were tested to determine if their effects might be more additive or multiplicative than the individual applications. However, results suggested incompatibility of both biostimulants as none of the combinations showed better results than that of the individual applications of either flavonoids or CFS. Conceivably, the use of flavonoids and this novel Devosia sp. CFS could be significant plant growth enhancers, perhaps much better than the few other biostimulants and bacterial-based compounds currently in use.

The Coevolution of Plants and Microbes Underpins Sustainable Agriculture.

Terrestrial plants evolution occurred in the presence of microbes, the phytomicrobiome. The rhizosphere microbial community is the most abundant and diverse subset of the phytomicrobiome and can include both beneficial and parasitic/pathogenic microbes. Prokaryotes of the phytomicrobiome have evolved relationships with plants that range from non-dependent interactions to dependent endosymbionts. The most extreme endosymbiotic examples are the chloroplasts and mitochondria, which have become organelles and integral parts of the plant, leading to some similarity in DNA sequence between plant tissues and cyanobacteria, the prokaryotic symbiont of ancestral plants. Microbes were associated with the precursors of land plants, green algae, and helped algae transition from aquatic to terrestrial environments. In the terrestrial setting the phytomicrobiome contributes to plant growth and development by (1) establishing symbiotic relationships between plant growth-promoting microbes, including rhizobacteria and mycorrhizal fungi, (2) conferring biotic stress resistance by producing antibiotic compounds, and (3) secreting microbe-to-plant signal compounds, such as phytohormones or their analogues, that regulate aspects of plant physiology, including stress resistance. As plants have evolved, they recruited microbes to assist in the adaptation to available growing environments. Microbes serve themselves by promoting plant growth, which in turn provides microbes with nutrition (root exudates, a source of reduced carbon) and a desirable habitat (the rhizosphere or within plant tissues). The outcome of this coevolution is the diverse and metabolically rich microbial community that now exists in the rhizosphere of terrestrial plants. The holobiont, the unit made up of the phytomicrobiome and the plant host, results from this wide range of coevolved relationships. We are just beginning to appreciate the many ways in which this complex and subtle coevolution acts in agricultural systems.

Plant Holobiont Theory: The Phytomicrobiome Plays a Central Role in Evolution and Success.

Under natural conditions, plants are always associated with a well-orchestrated community of microbes-the phytomicrobiome. The nature and degree of microbial effect on the plant host can be positive, neutral, or negative, and depends largely on the environment. The phytomicrobiome is integral for plant growth and function; microbes play a key role in plant nutrient acquisition, biotic and abiotic stress management, physiology regulation through microbe-to-plant signals, and growth regulation via the production of phytohormones. Relationships between the plant and phytomicrobiome members vary in intimacy, ranging from casual associations between roots and the rhizosphere microbial community, to endophytes that live between plant cells, to the endosymbiosis of microbes by the plant cell resulting in mitochondria and chloroplasts. If we consider these key organelles to also be members of the phytomicrobiome, how do we distinguish between the two? If we accept the mitochondria and chloroplasts as both members of the phytomicrobiome and the plant (entrained microbes), the influence of microbes on the evolution of plants becomes so profound that without microbes, the concept of the "plant" is not viable. This paper argues that the holobiont concept should take greater precedence in the plant sciences when referring to a host and its associated microbial community. The inclusivity of this concept accounts for the ambiguous nature of the entrained microbes and the wide range of functions played by the phytomicrobiome in plant holobiont homeostasis.