We recently demonstrated that nitric oxide (NO) accumulation and PHYTOGB1 transcriptional regulation are early components of the regulatory pathway that is activated in tomato roots during the onset of the mycorrhizal symbiosis between Rhizophagus irregularis and tomato roots. We further showed that the mycorrhizal interaction was associated with a specific NO-related signature, different from that triggered by the pathogen Fusarium oxysporum. Here, we extend our investigation by exploring the NO- and PHYTOGB1-related root responses elicited by another root mutualistic endosymbiotic fungus: Trichoderma harzianum T-78. By using T-78 in vitro-grown cultures, we found that T-78 triggered an early and transient burst of NO in tomato roots during the first hours after the interaction. T-78 also elicited the early upregulation of PHYTOGB1, which was maintained during the analyzed timespan. By using glass-house bioassays, we found that in a well-established tomato-T-78 symbiosis, NO root levels were maintained at basal level while PHYTOGB1 expression remained upregulated. Our results demonstrate that the T-78 symbiosis is associated with a rapid and transient burst of NO in the host roots and the transcriptional activation of PHYTOGB1 from early stages of the interaction until the establishment of the symbiosis, most likely to control NO levels and favor the mutualistic symbiosis.

Priming of plant defences provides increased plant protection against herbivores and reduces the allocation costs of defence. Defence priming in woody plants remains obscure, in particular, due to plant development traits such as endogenous rhythmic growth. By using bioassays with oak microcuttings (Quercus robur) and combining transcriptomic and metabolomic analyses, we investigated how leaf herbivory by Lymantria dispar and root inoculation with the ectomycorrhizal fungus Piloderma croceum prime oak defences. We further investigated how defence priming is modulated by the rhythmic growth of the oaks. A first herbivory challenge in oak leaves primed newly grown leaves for enhanced induction of jasmonic acid (JA)‐related direct defences, or enhanced emission of volatiles, depending on the specific growth stage at which the plants were first challenged. Root inoculation with Piloderma abolished the enhanced induction of JA‐related defences and volatile emission. Synthesis: Our results indicate that a first herbivore attack primes direct and indirect defences of newly formed oak leaves and that the specific display of defence priming is regulated by rhythmic growth, and modulated by the interaction Piloderma. Our results show that the priming memory in oaks can be transmitted to the next growth cycle, to the leaves of the new shoot unit.

Summary: Microbe‐induced resistance (MIR) encompasses a broad range of plant responses that mediate both direct and indirect defenses against herbivores. However, previous studies have addressed MIR in relation to either direct or indirect defenses in isolation, overlooking potential conflicts or synergies that may arise between MIR‐elicited defense strategies. We hypothesize that in multitrophic contexts, MIR‐driven elicitation of direct and indirect defenses interacts, thereby shaping overall MIR phenotypes. To test this, we used two MIR‐eliciting fungi: an arbuscular mycorrhizal and Trichoderma species; and the tri‐trophic system including tomato plants, the herbivore Manduca sexta , and the parasitoid Cotesia congregata . We conducted a series of MIR bioassays, including performance and behavioral assays, and explored the main mechanisms underlying MIR phenotypes. We found that MIR involves a dynamic elicitation of direct and indirect defense‐related traits that plastically adjust to herbivory time. Upon short‐term herbivory, MIR primed jasmonate‐regulated defenses, thereby enhancing the mortality of early‐instar larvae. After a longer period of herbivory, MIR, especially triggered by Trichoderma , boosted green leaf volatile emissions, enhancing parasitoid attraction. Parasitoid performance was improved by MIR. Our study revealed that MIR enhances resistance by priming both direct and indirect defenses at different herbivory stages. This fosters complementary and synergistic responses, enhancing MIR phenotypes.

One century after the first report of Dutch elm disease (DED), there is still no practical solution for this problem threatening European and American elms (Ulmus spp.). The long breeding cycles needed to select resistant genotypes and the lack of efficient treatments keep disease incidence at high levels. In this work, the expression of defense-related genes to the causal agent of DED, Ophiostoma novo-ulmi Brasier, was analyzed in in vitro clonal plantlets from two DED-resistant and two DED-susceptible Ulmus minor Mill. trees. In addition, the effect of the inoculation of an endophytic pink-pigmented yeast (Cystobasidium sp.) on the plant’s defense system was tested both individually and in combination with O. novo-ulmi. The multifactorial nature of the resistance to DED was confirmed, as no common molecular response was found in the two resistant genotypes. However, the in vitro experimental system allowed discrimination of the susceptible from the resistant genotypes, showing higher levels of oxidative damage and phenolic compounds in the susceptible genotypes after pathogen inoculation. Inoculation of the endophyte before O. novo-ulmi attenuated the plant molecular response induced by the pathogen and moderated oxidative stress levels. Niche competition, endophyte–pathogen antagonism and molecular crosstalk between the host and the endophyte are discussed as possible mechanisms of stress reduction. In sum, our results confirm the complex and heterogeneous nature of DED resistance mechanisms and highlight the possibility of using certain endophytic yeasts as biological tools to improve tree resilience against biotic stress.