Inoculation Treatments Research Articles

Arbuscular mycorrhizal fungi alter carbon metabolism and invertase genes expressions of Populus simonii × P. nigra under drought stress.

Arbuscular mycorrhizal fungi (AMF) play a crucial role in regulating the allocation of carbon between source and sink tissues in plants and in regulating their stress responses by changing the sucrose biosynthesis, transportation, and catabolism in plants. Invertase, a key enzyme for plant development, participates in the response of plants to drought stress by regulating sucrose metabolism. However, the detailed mechanisms by which INV genes respond to drought stress in mycorrhizal plants remain unclear. This study examined the sugar content, enzyme activity, and expression profiles of INV genes of Populus simonii × P. nigra (PsnINVs) under two inoculation treatments (inoculation or non-inoculation) and two water conditions (well-watered or drought stress). Results showed that under drought stress, AMF up-regulated the expressions of PsnA/NINV1, PsnA/NINV2, PsnA/NINV3, and PsnA/NINV5 in leaves, which may be related to the enhancement of photosynthetic capacity. Additionally, AMF up-regulated the expressions of PsnA/NINV6, PsnA/NINV10, and PsnA/NINV12 in leaves, which may be related to enhancing osmotic regulation ability and drought tolerance.

Mycorrhizal inoculation alters rhizosphere fungal community and root morphology to improve drought tolerance of resource-acquisitive but not resource-conservative Quercus species

The mutualistic association between mycorrhizal fungi and plants is well known to improve plant drought resistance. However, the influence of external inoculants on the interactions between soil microbial communities and plant functional traits and how these interactions improve plant drought tolerance under drought stress remain unclear. We conducted a pot inoculation experiment to assess the effect of two inoculated fungal strains (Clitopolius hobsonii NL-19 and C. sp. HSL-YX-7-A) on morphological traits and rhizosphere fungal communities of eight Quercus species seedlings. Plant and soil fungal traits were measured 2 months after inoculation and then underwent a short-term (33 days) drought rewetting treatment. The biomass of all tree species reduced significantly under drought stress followed by rewetting (drought treatment) compared with that in the control treatment (well-watered); however, inoculated mycorrhizal fungi (drought + mycorrhizal inoculation treatment) increased the biomass of five Quercus species (growth-promoting species (GPS) exhibiting a significant increase in biomass under mycorrhizal inoculation treatment than under the uninoculated treatment during drought stress, characterized by a resource-use acquisitive strategy) and had no effect on the other three Quercus species (non-growth-promoting species (NGPS) exhibiting no significant difference in biomass between mycorrhizal inoculation and uninoculated treatments during drought stress, characterized by a resource-use conservative strategy). The leaf traits of the two species groups (GPS and NGPS) varied little among the three treatments. The values of most root traits (e.g., specific root length and root length) of GPS increased significantly under drought and drought + mycorrhizal inoculation treatments compared with those in the control, whereas relatively minor variations were observed in NGPS among the three treatments. The fungal community composition and functional groups (primary trophic modes) in the drought treatment were altered for NGPS but not for GPS; additionally, they changed in the drought + mycorrhizal inoculation treatment for GPS but not for NGPS when compared with those in the drought treatment. Furthermore, the drought tolerance of GPS was improved directly by fungal functional groups (i.e., Pathotroph-Saprotroph- Symbiotrophs proliferation) and indirectly by root traits (e.g., increased specific root length), whereas NGPS may adapt to drought stress through other pathways (e.g., physiological and biochemical regulation). Overall, drought and mycorrhizal inoculation affected the root traits and fungal community structure of Quercus seedlings, which rely on plant resource-use strategies. Our results provide new insights into the relationship between plant resource-use strategies and soil microbes for improving plant performance under drought stress.

The potential of soil endemic microorganisms in ameliorating the physicochemical properties of soil subjected to a freeze-thaw cycle

Establishing soil biological crusts will result in the long-term restoration of ecosystems. Nonetheless, little research has been conducted to demonstrate the influence of soil endemic microorganisms on suppressing the adverse effects of freezing-thawing on soil properties. This current study evaluated the formation of biological crusts, the enhancement of physical and chemical characteristics of soil, and surface soil stability by inoculating native bacteria and cyanobacteria into the soil during a freezing-thawing cycle. The soil was collected from the Badranlou Region in North Khorasan Province, Northern Iran, and native bacteria and cyanobacteria were isolated, identified, chosen, and cultured. The native treatments of bacteria and cyanobacteria were then inoculated in individual bacteria, cyanobacteria, and combined inoculation of cyanobacteria and bacteria onto an experimental soil in six replications. After 60 days, they were subjected to freezing-thawing conditions to have maximum effect on the soil environment, and finally, the soil surface properties were statistically compared. The results of the significant effect (p<0.001) of inoculation treatments on the physical and chemical properties of the study soil revealed that the carbon, nitrogen, organic matter, phosphorus, potassium, and surface soil stability in bacterial treatment compared to the control treatment increased by 68, 39, 68, 17, 25, 99 %, respectively. While under cyanobacterial treatment, they rose by 83, 61, 83, 18, 73, and 172 %, respectively. The combination inoculation treatment of bacteria and cyanobacteria enhanced the study variables by 73, 66, 73, 25, 58, and 189 %, respectively. Compared to control plots, the soil bulk density in bacterial, cyanobacterial, and compound inoculation treatments was substantially reduced (p<0.001) by 9, 15, and 12 %, respectively. The soil stability, carbon, and organic matter were among the most essential properties of the soil, and they best showed the difference between the various treatments applied. It confirmed the region's potential restoration by inoculating native soil microorganisms.

Enhancing drought resistance in Pinus tabuliformis seedlings through root symbiotic fungi inoculation.

Drought constitutes a major abiotic stress factor adversely affecting plant growth and productivity. Plant-microbe symbiotic associations have evolved regulatory mechanisms to adapt to environmental stress conditions. However, the interactive effects of different fungi on host growth and stress tolerance under drought conditions remain unclear. This study explored the effects of varying polyethylene glycol (PEG-6000) concentrations (0%, 15%, 25%, and 35%) on the growth and physiological responses of two ectomycorrhizal fungi (Suillus granulatus (Sg) and Pisolithus tinctorius (Pt)) and two dark septate endophytes (Pleotrichocladium opacum (Po) and Pseudopyrenochaeta sp. (Ps)) isolated from the root system of Pinus tabuliformis. Specifically, the study aimed to evaluate six inoculation treatments, including no inoculation (CK), single inoculations with Sg, Pt, Po, Ps, and a mixed inoculation (Sg: Pt : Po: Ps = 1:1:1:1), on the growth and physiological characteristics of P. tabuliformis seedlings under different water regimes: well-watered at 70% ± 5%, light drought at 50% ± 5%, and severe drought at 30% ± 5% of the maximum field water holding capacity. All four fungi exhibited the capacity to cope with drought stress by enhancing antioxidant activities and regulating osmotic balance. Upon successful root colonization, they increased plant height, shoot biomass, root biomass, total biomass, and mycorrhizal growth response in P. tabuliformis seedlings. Under drought stress conditions, fungal inoculation improved seedling drought resistance by increasing superoxide dismutase and catalase activities, free proline and soluble protein contents, and promoting nitrogen and phosphorus uptake. Notably, mixed inoculation treatments significantly enhanced antioxidant capacity, osmotic adjustment, and nutrient acquisition abilities, leading to superior growth promotion effects under drought stress compared to single inoculation treatments. All four fungi tolerated PEG-induced drought stress, with increased antioxidant enzyme activities and osmotic adjustment substances and they promoted the growth and enhanced drought resistance of P. tabuliformis seedlings.

Differential Strategies of Ectomycorrhizal Development between Suillus luteus and Pinus massoniana in Response to Nutrient Changes.

Ectomycorrhizal fungi employ different strategies for mycelial growth and host colonization under varying nutrient conditions. However, key genes associated with mycorrhizal interaction should be influenced solely by the inoculation treatment and not by nutrient variations. To utilize subtle nutrient differences and rapidly screen for key genes related to the interaction between Suillus luteus and Pinus massoniana, we performed an inoculation experiment using culture bottles containing high- and low-nutrient media. Interestingly, S. luteus LS88 promoted the growth of P. massoniana seedlings without mature ectomycorrhiza, and the impact of LS88 inoculation on P. massoniana roots was greater than that of nutrient changes. In this study, the resequenced genome of the LS88 strain was utilized for transcriptome analysis of the strain. The analysis indicated that a unique gene encoding glutathione S-transferase (GST) in LS88 is likely involved in colonizing P. massoniana roots. In this study, the GST gene expression was independent of nutrient levels. It was probably induced by P. massoniana and could be used as a marker for S. luteus colonization degree.

Impact of gastrointestinal inoculation and benznidazole treatment on infection by Trypanosoma cruzi (Y strain, DTU TcII) in Swiss mice

In Brazil, where Chagas disease is endemic, the most frequent form of transmission of the parasite is the oral route, associated with greater severity and worse response to benznidazole (BZ), the drug used in its treatment. This study aimed to evaluate the impact of gastrointestinal infection (GI) and BZ treatment on the parasitological and histopathological parameters in mice inoculated with a strain of T. cruzi II. Swiss mice were inoculated by GI and intraperitoneal (IP) routes with 2x106 culture-derived metacyclic trypomastigotes of the Y strain (TcII) of T. cruzi and were treated with BZ in the acute phase of the infection. Fresh blood examination, qPCR, histopathological and biochemical evaluations (enzymatic dosages and oxidative stress-OS) were performed. BZ treatment of uninfected animals caused changes in the liver, increased the activity of aspartate aminotransferase and alanine aminotransferase enzymes and OS, showing that the drug alone affects this organ. Inflammation and necrosis in the cardiac tissue were less intense and deaths occurred later in animals inoculated via the GI route than the animals inoculated via the IP route. BZ reduced the intensity of tissue lesions and avoided lethality in animals inoculated via the GI route, and decreased parasitemia and OS in those inoculated via both routes. Although BZ alone caused liver damage, it was less intense than that caused by both routes of inoculation. Infection with the Y strain of T. cruzi II via the GI route proved to be less virulent and pathogenic and responded better to treatment than the infection acquired via the IP route.

Effects of earthworm and arbuscular mycorrhizal fungal inoculation on carbon component accumulation and allocation in rocky desertification soils.

Exploring the responses of carbon component accumulation and allocation to arbuscular mycocorrhizal fungi (AM) and earthworm inoculation can provide reference for improving carbon sequestration potential and bioremediation efficiency in rocky desertification soils. In this study, we chose Fraxinus malacophylla as the host plant to inoculate with Funneliformis mosseae (FM), earthworm (E), and E+FM, using no earthworm and mycorrhizae addition as CK to examine the spatiotemporal variations in soil carbon components (i.e., total organic carbon, microbial biomass carbon, easily oxidized organic carbon, and recalcitrant organic carbon) and their allocation (i.e., microbial biomass carbon/total organic carbon, easily oxidized organic carbon/total organic carbon, and recalcitrant organic carbon/total organic carbon). The results showed that 1) The respective and interactive inoculation of E and AM significantly promoted the accumulation of each carbon component. In contrast with the control, the average carbon component levels under three inoculation treatments were ranked as E+FM>E>FM. The three inoculation treatments significantly promoted soil microbial carbon/total organic carbon (30.5%-68.5%) and easily oxidized carbon/total organic carbon (31.2%-39.2%), but decreased recalcitrant organic carbon/total organic carbon (2.9%-16.2%). 2) The spatiotemporal variation in accumulation and allocation of soil carbon components varied between the inoculation treatments. The maximum value of each carbon component occurred in June. The increase in each carbon component was significantly higher in E+FM (33.0%-122.1%) than that in E (31.2%-95.4%) and FM (9.2%-41.3%). The maximum value of microbial biomass carbon/total organic carbon and easily oxidized organic carbon/total organic carbon was observed in June, while that of recalcitrant organic carbon/total organic carbon was recorded in December. In contrast with CK, the amplitude of variation in the proportion of carbon components in total organic carbon under the three inoculation treatments was ranked as E+FM>E>FM. The accumulation and allocation of all carbon components decreased (9.7%-146.2%) along the soil profile. The level of carbon components in the E treatment decreased the smallest. The microbial biomass carbon/total organic carbon and easily oxidized carbon/total organic carbon decreased the least and the recalcitrant organic carbon/total organic carbon decreased the greatest under the E+FM treatment. 3) Changes in soil physicochemical properties under the three inoculation treatments significantly affected the accumulation and allocation of organic carbon components. Soil pH was negatively correlated with carbon component accumulation and allocation, whereas other soil variables were positively correlated with them. 4) The results of principal component analysis showed that soil water content, total nitrogen, and total phosphorus were the main factors driving carbon component accumulation, while soil water content, total phosphorus, and pH were the main factors controlling carbon component allocation. Therefore, we concluded that the earthworms, AM fungi and their interaction affected the accumulation and allocation of carbon components in Yunnan rocky desertification soils, which would primarily depend on the changes of soil water content, acid-base property, as well as nitrogen and phosphorus conditions.

Additive manufacturing of high-strength low-alloy AISI 4340 steel with an optimal strength-ductility-toughness trade-off

Additive manufacturing (AM) has revolutionised steel part fabrication, yet not all steels are amenable to its unique solidification features, characterised by cyclic and rapid heating/cooling, and directional solidification. These conditions often result in challenges such as columnar grain formation, microstructural heterogeneity, and consequently, inferior mechanical performance, brittleness and severe anisotropy in particular. Recent studies have adopted inoculation or post-fabrication treatments to address this issue, but often entailing extra cost and processing time. This study aims to verify that some steels such as AISI 4340 steel are inherently compatible to AM, producing components that are innately robust and ready for use in the as-built state. The medium carbon content and low alloying element concentration enable AM processing to produce a uniform and refined bainite microstructure with minimal elemental segregation, avoiding the formation of unstable retained austenite. The high AM-processability of this steel is demonstrated by achieving high densification (>99.9 %) across a broad processing window, which allows precise microstructural control via proper tunning the processing parameter modifications, inducing a transition from upper bainite to lower bainite dominance, to tailor mechanical properties for specific applications. The as AM-fabricated AISI 4340 steel exhibits a good combination of strength, ductility, and toughness, manifested by a yield strength range from 1240 to 1370 MPa, an ultimate tensile strength from 1360 to 1740 MPa, an elongation from 7 % to 14 %, and an impact toughness range of 11–44 J. The mechanical properties of the AM-fabricated 4340 steel are comparable to those of the wrought counterpart and superior to the majority of other AM-fabricated steels. This research reveals the high potential of AM to process high-strength low-alloy steels.

Phyto-treatment of tannery industry effluents under combined application of citric acid and chromium-reducing bacterial strain through Lemna minor L.: A lab scale study

Contamination of agricultural soils with heavy metals (HMs) poses a significant environmental threat, especially because industrial discharges often irrigate agricultural lands. A prominent source of HM(s) pollution occurs from tannery effluents containing high concentrations of chromium (Cr) in both Cr3+ and Cr6+ forms along with other toxic materials. Cr is known for its carcinogenic and mutagenic properties in biological systems. Microbe-assisted phytoremediation has emerged as a promising and environmentally friendly approach for detoxifying Cr-contaminated environments. This study aimed to evaluate the performance of citric acid (CA) and a Cr-reducing bacterial strain (Staphylococcus aureus) on the phytoextraction potential of Lemna minor within a Constructed Wetland System treated with tannery wastewater. Various combinations of tannery wastewater (0, 50, and 100 %), CA (0, 5 and 10 mM), and microbial inoculants were applied to the test plants. The mitigative effects of Staphylococcus aureus strain K1 were examined in combination with different concentrations of CA (0, 5, 10 mM). Data on growth and yield attributes highlighted the beneficial effects of bacterial inoculation and CA in ameliorating Cr toxicity in L. minor, as evidenced by increased foliar chlorophyll and carotenoid contents, enhanced antioxidant enzyme activities (SOD, POD, APX, CAT), and improved nutrient uptake. Specifically, CA application resulted in an enhancement of Cr ranging from 12% to 15% and 23%–31% in concentration, and 134%–141% and 322%–337% in Cr accumulation, respectively. When combined with the S. aureus inoculation treatment, CA application (5 and 10 mM) further increased the concentration and accumulation of Cr in L. minor. The enhancement in Cr ranged from 12% to 23% and 27%–41% in concentration, 68%–75%, and 179%–185% in accumulation, respectively. These results demonstrated that L. minor is an effective choice for environmentally friendly Cr remediation due to its continued ability to grow in polluted wastewater. This study suggested that microbial-assisted phytoextraction combined with chelating agents such as CA could be a practical and effective approach for remediating tannery effluents.

Ecological phage therapy: Can bacteriophages help rapidly restore the soil microbiome?

Soil microbiota underpin ecosystem functionality yet are rarely targeted during ecosystem restoration. Soil microbiota recovery following native plant revegetation can take years to decades, while the effectiveness of soil inoculation treatments on microbiomes remains poorly explored. Therefore, innovative restoration treatments that target soil microbiota represent an opportunity to accelerate restoration outcomes. Here, we introduce the concept of ecological phage therapy-the application of phage for the targeted reduction of the most abundant and dominant bacterial taxa present in degraded ecosystems. We propose that naturally occurring bacteriophages-viruses that infect bacteria-could help rapidly shift soil microbiota towards target communities. Bacteriophages sculpt the microbiome by lysis of specific bacteria, and if followed by the addition of reference soil microbiota, such treatments could facilitate rapid reshaping of soil microbiota. Here, we experimentally tested this concept in a pilot study. We collected five replicate pre-treatment degraded soil samples, then three replicate soil samples 48 hours after phage, bacteria, and control treatments. Bacterial 16S rDNA sequencing showed that phage-treated soils had reduced bacterial diversity; however, when we combined ecological phage therapy with reference soil inoculation, we did not see a shift in soil bacterial community composition from degraded soil towards a reference-like community. Our pilot study provides early evidence that ecological phage therapy could help accelerate the reshaping of soil microbiota with the ultimate aim of reducing timeframes for ecosystem recovery. We recommend the next steps for ecological phage therapy be (a) developing appropriate risk assessment and management frameworks, and (b) focussing research effort on its practical application to maximise its accessibility to restoration practitioners.

Assessing inoculation's effectiveness in motivating resistance to conspiracy propaganda in Finnish and United States samples.

This study tested the motivational power of inoculation to foster resistance to conspiracy propaganda (9/11 Truth Movement), comparing inoculation effects across United States and Finnish study participants. We used a 2 inoculation (treatment vs. control) × 2 national culture (American vs. Finnish) independent groups design (N = 319), while examining the effects of motivational threat and thinking modes-analytic vs. intuitive-on the inoculation process. To test the effectiveness of the inoculation strategy, we used an excerpt from a conspiracy film Loose Change as a counterattitudinal attack message. Our results indicated that inoculation was effective at motivating resistance regardless of national culture. Inoculation effects emerged mostly as a direct effect on resistance and two indirect effects wherein motivational threat mediated the relationship between inoculation and resistance as well as inoculation and analytic mode of message processing. Although we found that an increase in analytic mode of processing facilitated resistance and intuitive processing increased conspiracy-theory endorsement, the indirect effects between inoculation and resistance via message processing modes were not significant. Finally, the data revealed national culture differences in analytic mode and cultural-context differences mostly pertaining to the relationships between thinking styles, media literacy, and modes of thinking. These results offer important theoretical implications for inoculation scholarship and suggest viable practical solutions for efforts to mitigate misinformation and conspiratorial beliefs.

Microbial inoculum effects on the rumen epithelial transcriptome and rumen epimural metatranscriptome in calves

Manipulation of the rumen microbial ecosystem in early life may affect ruminal fermentation and enhance the productive performance of dairy cows. The objective of this experiment was to evaluate the effects of dosing three different types of microbial inoculum on the rumen epithelium tissue (RE) transcriptome and the rumen epimural metatranscriptome (REM) in dairy calves. For this objective, 15 Holstein bull calves were enrolled in the study at birth and assigned to three different intraruminal inoculum treatments dosed orally once weekly from three to six weeks of age. The inoculum treatments were prepared from rumen contents collected from rumen fistulated lactating cows and were either autoclaved (control; ARF), processed by differential centrifugation to create the bacterial-enriched inoculum (BE), or through gravimetric separation to create the protozoal-enriched inoculum (PE). Calves were fed 2.5 L/d pasteurized waste milk 3x/d from 0 to 7 weeks of age and texturized starter until euthanasia at 9 weeks of age, when the RE tissues were collected for transcriptome and microbial metatranscriptome analyses, from four randomly selected calves from each treatment. The different types of inoculum altered the RE transcriptome and REM. Compared to ARF, 9 genes were upregulated in the RE of BE and 92 in PE, whereas between BE and PE there were 13 genes upregulated in BE and 114 in PE. Gene ontology analysis identified enriched GO terms in biological process category between PE and ARF, with no enrichment between BE and ARF. The RE functional signature showed different KEGG pathways related to BE and ARF, and no specific KEGG pathway for PE. We observed a lower alpha diversity index for RE microbiome in ARF (observed genera and Chao1 (p < 0.05)). Five microbial genera showed a significant correlation with the changes in host gene expression: Roseburia (25 genes), Entamoeba (two genes); Anaerosinus, Lachnospira, and Succiniclasticum were each related to one gene. sPLS-DA analysis showed that RE microbial communities differ among the treatments, although the taxonomic and functional microbial profiles show different distributions. Co-expression Differential Network Analysis indicated that both BE and PE had an impact on the abundance of KEGG modules related to acyl-CoA synthesis, type VI secretion, and methanogenesis, while PE had a significant impact on KEGGs related to ectoine biosynthesis and D-xylose transport. Our study indicated that artificial dosing with different microbial inocula in early life alters not only the RE transcriptome, but also affects the REM and its functions.

Disruption of pollination by herbivores is rescued by nectar yeasts

Abstract Attracting pollinators to achieve successful reproduction is a key challenge for wild plants that may be disturbed by complex multispecies interactions in nature. Pairwise plant–pollinator interactions have traditionally been studied for decades, while ignoring other ecological players may obscure a comprehensive understanding on how plants recruit partners or combat enemies in the pollination process. Hence, integrated studies considering the inherent complexity of ecological interactions are needed, which may open up new perspectives for deciphering intricate systems and predicting ecological consequences. We examined the presence of nectar yeasts using a combination of high‐throughput sequencing, cultivation and microscopy and quantified floral herbivory by evaluating the incidence of flowers with visible holes in 13 natural populations of Iris bulleyana in the Hengduan Mountains of southwest China during 2017–2022. We combined yeast inoculation and herbivore manipulation treatments to illustrate the isolated and combined impacts of two contrasting nectarivorous organisms, the ascomycetous yeast Metschnikowia reukaufii and adult sawflies, on pollinator visitation and plant reproductive success in two populations. In the lab, we first employed gas chromatography–mass spectrometry to profile the volatile metabolites of yeast‐inoculated nectar relative to control, followed by a behavioural bioassay to test the preference of honeybees for these microbial volatiles. Yeasts commonly inhabited floral nectar and insect herbivores frequently bit holes in the perianth tube to consume nectar and nectaries. Nectar yeasts indirectly facilitated plant reproduction through increased pollinator visits, probably because of microbial metabolism as honeybees preferred nectar volatiles produced by yeasts in behavioural bioassays. Insect herbivores increased total floral visits but reduced legitimate visits by inducing legitimate‐to‐robbing behavioural changes of honeybees, thus leading to lower seed production. The detrimental effect of herbivory was mitigated by the presence of yeasts, which diminished the relative proportion of robbing visits and thereby ‘rescued’ flowers from reproductive failure. Synthesis: Overall, we found contrasting effects of non‐pollinator species, including both micro‐ and macro‐organisms, on plant–pollinator interactions in a biodiversity hotspot, where pollination deficit may be a ubiquitous phenomenon. Our findings suggest that both microbial and herbivory effects are likely to be important in explaining the exact causes of pollen limitation in species‐rich areas, highlighting the biological context dependence of species interactions in natural ecosystems.

Genome-wide analysis of R2R3-MYB transcription factors in poplar and functional validation of PagMYB147 in defense against Melampsora magnusiana

Main conclusionTranscription of PagMYB147 was induced in poplar infected by Melampsora magnusiana, and a decline in its expression levels increases the host's susceptibility, whereas its overexpression promotes resistance to rust disease.Poplars are valuable tree species with diverse industrial and silvicultural applications. The R2R3-MYB subfamily of transcription factors plays a crucial role in response to biotic stresses. However, the functional studies on poplar R2R3-MYB genes in resistance to leaf rust disease are still insufficient. We identified 191 putative R2R3-MYB genes in the Populus trichocarpa genome. A phylogenetic analysis grouped poplar R2R3-MYBs and Arabidopsis R2R3-MYBs into 33 subgroups. We detected 12 tandem duplication events and 148 segmental duplication events, with the latter likely being the main contributor to the expansion of poplar R2R3-MYB genes. The promoter regions of these genes contained numerous cis-acting regulatory elements associated with response to stress and phytohormones. Analyses of RNA-Seq data identified a multiple R2R3-MYB genes response to Melampsora magnusiana (Mmag). Among them, PagMYB147 was significantly up-regulated under Mmag inoculation, salicylic acid (SA) and methyl jasmonate (MeJA) treatment, and its encoded product was primarily localized to the cell nucleus. Silencing of PagMYB147 exacerbated the severity of Mmag infection, likely because of decreased reactive oxygen species (ROS) production and phenylalanine ammonia-lyase (PAL) enzyme activity, and up-regulation of genes related to ROS scavenging and down-regulation of genes related to PAL, SA and JA signaling pathway. In contrast, plants overexpressing PagMYB147 showed the opposite ROS accumulation, PAL enzyme activity, SA and JA-related gene expressions, and improved Mmag resistance. Our findings suggest that PagMYB147 acts as a positive regulatory factor, affecting resistance in poplar to Mmag by its involvement in the regulation of ROS homeostasis, SA and JA signaling pathway.

Cinnamic acid derivatives played a major role in the resistance of peach leaves to Xanthomonas arboricola pv. pruni infection

AbstractPeach bacterial shot hole is a common global disease of peach trees caused by the pathogen Xanthomonas arboricola pv. pruni (Xap). To preliminarily explore the resistance mechanism of peach bacterial shot hole, leaves of the resistant variety ‘Yanbao’ and the susceptible variety ‘Frederic’ were used as test materials. The contents of total phenols, total flavonoids and lignin were analysed after 1 day of inoculation treatment, and the activities of defence enzymes and cell wall degradation enzymes in the leaves were determined. Furthermore, the resistant substances were screened by combining broad‐target metabolomics technology and in vitro antibacterial activity. The results showed that the contents of total phenols, total flavonoids and lignin in peach leaves were related to resistance. The resistant varieties resisted infection by inhibiting the activities of polygalacturonase (PG) and pectin methyl trans‐eliminase (PMTE) produced by Xap in peach leaves and increasing the activities of 4‐coumarate:coenzyme A ligase (4CL), cinnamic acid‐4‐hydroxylase (C4H), phenylalanine ammonia lyase (PAL) and peroxidase (POD). A total of six resistant compounds were screened from 85 differential metabolites. The minimum inhibitory concentrations of methyl ferulate (MF), p‐coumaric acid (p‐CA) and methyl caffeate (MC) on Xap were 0.0625, 00625 and 0.125 mg/mL, respectively, while that of the positive control drug oxytetracycline (OTC) was 0.125 mg/mL. The antibacterial activities of MF and p‐CA were better than that of OTC, and MF was equivalent to OTC. In addition, homoeriodictyol, hesperetin and fraxin have certain antibacterial effects. Before inoculation treatment, the contents of MF, p‐CA and MC in ‘Yanbao’ were 1.13, 1.24 and 1.96 times higher than those in ‘Frederic’, respectively, while they were 1.29, 1.80 and 1.94 times after inoculation treatment, respectively. During the process of pathogen infection, MF and p‐CA in the leaves of resistant peach varieties played a role in inducing resistance, while MC was a component of resistance. The above three cinnamic acid derivatives play an important roles in the resistance of peach bacterial shot hole and are potential control drugs.

Impact of sequential and concurrent inoculations of Meloidogyne incognita and Fusarium oxysporum f.sp. vasinfectum on the growth performance of diverse okra cultivars

The present study was conducted to assess the effects of inoculation methods viz. concurrent and sequential inoculations of the root-knot nematode, Meloidogyne incognita, and the wilt-causing fungus, Fusarium oxysporum f.sp. vasinfectum, on various growth parameters of nine okra cultivars. The study aimed to identify cultivars that exhibited robust growth parameters in response to these two pathogens. Significant disparities were found in shoot lengths among cultivars and inoculation methods (F=1033.86, P0.001), with Arka Anamika, Irka-1, and Irka-2 experiencing the most considerable reductions. In contrast, Pusa Swami and Green Star had the least reduction across all inoculation treatments. Sequential inoculation led to shoot length reductions ranging from 76.00% to 90.25%, averaging 87.07%. Concurrent inoculation resulted in reductions between 82.50% and 95.98%, averaging 90.29%. For nematode inoculation followed by fungus after 15 days, the reductions ranged from 81.74% to 95.34%, with an average of 89.21%. Regarding root length, concurrent inoculation generally had a less detrimental effect on root lengths compared to sequential inoculation. The reduction in root length varied from 0.01% to 24.72%, with concurrent inoculation showing less reduction than sequential methods. Similarly, concurrent inoculation typically resulted in lesser reductions in shoot weights compared to sequential inoculations, with reductions ranging widely among cultivars. Unlike shoot weight, concurrent inoculation did not consistently increase root weight compared to sequential methods. In conclusion, the study suggests that concurrent inoculation of M. incognita and F. oxysporum f.sp. vasinfectum is less harmful to the growth parameters of okra cultivars than sequential inoculation, with some variations among specific cultivars.

Sequential Fermentation in Red Wine cv. Babić Production: The Influence of Torulaspora delbrueckii and Lachancea thermotolerans Yeasts on the Aromatic and Sensory Profile.

This research aimed to analyze the impact of two different non-Saccharomyces yeast species on the aromatic profile of red wines made from the cv. Babić (Vitis vinifera L.) red grape variety. The grapes were obtained from two positions in the Middle and South of Dalmatia. This study compared a control treatment with the Saccharomyces cerevisiae (Sc) strain as a type of sequential inoculation treatment with Lachancea thermotolerans (Lt x Sc) and Torulaspora delbrueckii (Td x Sc). The focus was on the basic wine parameters and volatile aromatic compound concentrations determined using the SPME-Arrow-GC/MS method. The results revealed significant differences in cis-linalool oxide, geraniol, neric acid, and nerol, which contribute to the sensory profile with floral and rose-like aromas; some ethyl esters, such as ethyl furoate, ethyl hexanoate, ethyl lactate, ethyl 2-hydroxy-3-methylbutanoate, ethyl 3-hydroxy butanoate, diethyl glutarate, and diethyl succinate, contribute to the aromatic profile with fruity, buttery, overripe, or aging aromas. A sensory evaluation of wines confirmed that Td x Sc treatments exhibited particularly positive aromatic properties together with a more intense fullness, harmony, aftertaste, and overall impression.

Sweet Cherry Plants Prioritize Their Response to Cope with Summer Drought, Overshadowing the Defense Response to Pseudomonas syringae pv. syringae.

Disease severity and drought due to climate change present significant challenges to orchard productivity. This study examines the effects of spring inoculation with Pseudomonas syringae pv. syringae (Pss) on sweet cherry plants, cvs. Bing and Santina with varying defense responses, assessing plant growth, physiological variables (water potential, gas exchange, and plant hydraulic conductance), and the levels of abscisic acid (ABA) and salicylic acid (SA) under two summer irrigation levels. Pss inoculation elicited a more pronounced response in 'Santina' compared to 'Bing' at 14 days post-inoculation (dpi), and those plants inoculated with Pss exhibited a slower leaf growth and reduced transpiration compared to control plants during 60 dpi. During differential irrigations, leaf area was reduced 14% and 44% in Pss inoculated plants of 'Bing' and 'Santina' respectively, under well-watered (WW) conditions, without changes in plant water status or gas exchange. Conversely, water-deficit (WD) conditions led to gas exchange limitations and a 43% decrease in plant biomass compared to that under WW conditions, with no differences between inoculation treatments. ABA levels were lower under WW than under WD at 90 dpi, while SA levels were significantly higher in Pss-inoculated plants under WW conditions. These findings underscore the influence on plant growth during summer in sweet cherry cultivars that showed a differential response to Pss inoculations and how the relationship between ABA and SA changes in plant drought level responses.

Species interactions and bacterial inoculation enhance plant growth and shape rhizosphere bacterial community structure in faba bean – wheat intercropping under water and P limitations

Grain legumes / cereals intercropping systems with microbial inoculants, hold promise for improving crop productivity under stressful conditions. However, the tripartite interaction involving intercropping system, associated microbiota and stress combining water deficit and low phosphorus (P) availability remains understudied. This study evaluated the impact of three bacterial consortia (C4, C6, and Cref containing Rhizobium and PSB strains) including single Rhizobium (Rhizobium laguerreae) on the agro-physiological performance of wheat (Triticum durum) and faba bean (Vicia faba) grown as intercrops or sole-crops under P and water deficient conditions. Inoculation, especially with C6, significantly improved shoot and root biomasses of both wheat (up to 66 and 81 %) and faba bean (up to 54 and 266 %) intercrops compared to single Rhizobium inoculation and control treatments. Intercropping generally outperformed sole-cropping in above-ground physiology, root morphological traits, shoot and root P content, with a notable effect in response to C6 exhibiting low microbial biomass P. Changes in bacterial community structure were primarily driven by cropping pattern and water regime rather than bacterial inoculation. Intercropping maintained bacterial diversity but shifted community structure, favoring Proteobacteria. Overall, inoculating intercropped wheat and faba bean with Rhizobium-containing consortia induced beneficial below-ground interspecies interactions under water and P-limiting conditions.

Uncertainty and inoculation: Instilling resistance to anti-vaccination conspiracy propaganda

ABSTRACT Employing inoculation theory as a theoretical framework, we conducted an independent-groups experiment (N = 150), comparing the effects of an inoculation treatment to a control condition and examining uncertainty as a mediator of inoculation effects. To test inoculation effectiveness, we used an excerpt from an anti-vaccination conspiracy film Vaxxed as a counterattitudinal attack message. The results indicated that inoculation roused motivational defenses to protect initial attitudes and, subsequently, was effective at conferring resistance to conspiracy propaganda. Furthermore, relative to the control condition, inoculation treatment decreased attitudinal uncertainty. Importantly, and as predicted, within the inoculation condition, attitudinal uncertainty emerged as a mediator of the resistance process: For inoculated participants, an increase in motivational threat to defend one’s attitudes fostered by the inoculation treatment reduced uncertainty, and attitudinal certainty, subsequently, facilitated resistance against the message advocated in the conspiracy film, damaging to public health. These results have theoretical implications for research on inoculation and offer practical import for attempts at mitigating conspiratorial ideation.