Aspens (Populus tremula and its hybrids), economically and ecologically important fast-growing trees, are often damaged by Phellinus tremulae, a rot-causing fungus. Plant-associated bacteria can be used to increase plant growth and resistance; however, no systematic studies relating the activity of symbiotic bacteria to aspen resistance against Phellinus tremulae have been conducted so far. The present pioneer study investigated the responses of two Populus tremula and two P. tremula × P. tremuloides genotypes to in vitro inoculations with, first, either Pseudomonas sp. or Paenibacillus sp. bacteria (isolated originally from hybrid aspen tissue cultures and being most closely related to Pseudomonas oryzihabitans and Paenibacillus tundrae, respectively) and, in the subsequent stage, with Phellinus tremulae. Both morphological parameters of in vitro-grown plants and biochemical content of their leaves, including photosynthesis pigments and secondary metabolites, were analyzed. It was found that both Populus tremula × P. tremuloides genotypes, whose development in vitro was significantly damaged by Phellinus tremulae, were characterized by certain responses to the studied bacteria: decreased shoot development by both Paenibacillus sp. and Pseudomonas sp. and increased phenol content by Pseudomonas sp. In turn, these responses were lacking in both Populus tremula genotypes that showed in vitro resistance to the fungus. Moreover, these genotypes showed positive long-term growth responses to bacterial inoculation, even synergistic with the subsequent fungal inoculation. Hence, the studied bacteria were demonstrated as a potential tool for the improved in vitro propagation of fungus-resistant aspen genotypes.

European ash (Fraxinus excelsior) is highly affected by the pathogenic fungus Hymenoscyphus fraxineus in all of Europe. Increases in plant’s secondary metabolite (SM) production is often linked tol enhanced resistance to stress, both biotic and abiotic. Moreover, plant-associated bacteria have been shown to enhance SM production in inoculated plants. Thus, our hypothesis is that bacteria may boost ash SM production, hence priming the tree’s metabolism and facilitating higher levels of resilience to H. fraxineus. We tested three different ash genotypes and used Paenibacillus sp. and Pseudomonas sp. for inoculation in vitro. Total phenol (TPC), total flavonoid (TFC) and carotenoid contents were measured, as well as the chlorophyll a/b ratio and morphometric growth parameters, in a two-stage trial, whereby seedlings were inoculated with the bacteria during the first stage and with H. fraxineus during the second stage. While the tested bacteria did not positively affect the morphometric growth parameters of ash seedlings, they had a statistically significant effect on TPC, TFC, the chlorophyll a/b ratio and carotenoid content in both stages, thus confirming our hypothesis. Specifically, in ash genotype 64, both bacteria elicited an increase in carotenoid content, TPC and TFC during both stages. Additionally, Pseudomonas sp. inoculated seedlings demonstrated an increase in phenolics after infection with the fungus in both genotypes 64 and 87. Our results indicate that next to genetic selection of the most resilient planting material for ash reforestation, plant-associated bacteria could also be used to boost ash SM production.

Phenotypic plasticity, in response to adverse conditions, determines plant productivity and survival. The aim of this study was to test if two highly productive Populus genotypes, characterised by different in vitro etiolation patterns, differ also in their responses to hormones gibberellin (GA) and abscisic acid (ABA), and to a GA biosynthesis inhibitor paclobutrazol (PBZ). The experiments on shoot cultures of ‘Hybrida 275′ (abbr. H275; Populus maximowiczii × P. trichocarpa) and IBL 91/78 (Populus tremula × P. alba) were conducted by either modulating the physical in vitro environment or by adding specific chemicals to the nutrient medium. Our results revealed two main sets of differences between the studied genotypes in environmental and hormonal regulation of growth responses. First, the genotype H275 responded to darkness with PBZ-inhibitable shoot elongation; in contrast, the elongation of IBL 91/78 shoots was not affected either by darkness or PBZ treatment. Secondly, the explants of H275 were unable to recover their growth if it was inhibited with ABA; in contrast, those of IBL 91/78 recovered so well after the temporal inhibition by ABA that, when rooted subsequently, they developed longer shoots and roots than without a previous ABA treatment. Our results indicate that GA catabolism and repressive signalling provide an important pathway to control growth and physiological adaptation in response to immediate or impending adverse conditions. These observations can help breeders define robust criteria for identifying genotypes with high resistance and productivity and highlight where genotypes exhibit susceptibility to stress.

Bacterial biostimulants are an eco-friendly alternative to chemical fertilizers. However, before their introduction into open ecosystems, broad-scope studies need to be carried out. Paenibacillus sp. was shown to positively affect poplar root growth. It was hypothesized that alongside these improvements, the Paenibacillus sp. inoculant may affect its host’s secondary metabolism. Populus tremula × P. alba microshoots were inoculated in vitro. Microshoots were tested for chlorophyll, carotenoid, total flavonoid (TFC), total phenol content (TPC) and free radical scavenging capacity during primary growth after 4, 6 and 8 weeks. The results showed that the inoculation decreased shoot phenolics and free radical scavenging capacity after 6 and 8 weeks. Chlorophyll b amounts increased after 6 and 8 weeks. Carotenoid content decreased after 6 weeks, while chlorophyll a and carotenoid levels increased after 8 weeks. Correlation and principal component analyses showed that the inoculant changed the way in which the photosynthesis pigment content relates to TPC, TFC and radical scavenging activity. Overall, these data suggest that the inoculant does statistically significantly affect Populus tree secondary metabolism in the later stages of the initial growth period. This effect may potentially be compensatory in nature.

Phenotypic plasticity in response to adverse conditions determines plant productivity and survival. The aim of this study was to test if two highly productive Populus genotypes, characterized by different in vitro etiolation patterns, differ also in their responses to hormones gibberellin (GA) and abscisic acid (ABA), and to a GA biosynthesis inhibitor paclobutrazol (PBZ). The experiments on shoot cultures of ‘Hybrida 275’ (abbr. H275; Populus maximowiczii × P. trichocarpa) and IBL 91/78 (Populus tremula × P. alba) were conducted either by modulating the physical in vitro environment or by adding specific chemicals to the nutrient medium. Our results show that there are significant differences between the studied genotypes in environmental and hormonal regulation of growth responses. The genotype H275, which responded to darkness with PBZ-inhibitable shoot elongation, was unable to recover its growth after treatment with ABA. In contrast, the genotype IBL 91/78, whose shoot elongation was not affected either by darkness or PBZ treatment, recovered so well after the ABA treatment that, when rooted subsequently, it developed longer shoots and roots than without ABA treatment. Our results indicate that GA catabolism and repressive signaling provide an important pathway to control growth and physiological adaptation in response to immediate or impending adverse conditions. These observations can help breeders define robust criteria for identifying genotypes with high resistance and productivity and highlight where genotypes exhibit susceptibility to stress.