Integrity of the Intestinal Barrier: The Involvement of Epithelial Cells and Microbiota—A Mutual Relationship

The gastrointestinal tract, which is constantly exposed to a multitude of stimuli, is considered responsible for maintaining the homeostasis of the host. It is inhabited by billions of microorganisms, the gut microbiota, which form a mutualistic relationship with the host. Although the microbiota is generally recognized as beneficial, at the same time, together with pathogens, they are a permanent threat to the host. Various populations of epithelial cells provide the first line of chemical and physical defense against external factors acting as the interface between luminal microorganisms and immunocompetent cells in lamina propria. In this review, we focus on some essential, innate mechanisms protecting mucosal integrity, thus responsible for maintaining intestine homeostasis. The characteristics of decisive cell populations involved in maintaining the barrier arrangement, based on mucus secretion, formation of intercellular junctions as well as production of antimicrobial peptides, responsible for shaping the gut microbiota, are presented. We emphasize the importance of cross-talk between gut microbiota and epithelial cells as a factor vital for the maintenance of the homeostasis of the GI tract. Finally, we discuss how the imbalance of these regulations leads to the compromised barrier integrity and dysbiosis considered to contribute to inflammatory disorders and metabolic diseases.

Inhibiting TLR7 Expression in the Retinal Pigment Epithelium Suppresses Experimental Autoimmune Uveitis

Experimental autoimmune uveitis (EAU), a model of human uveitis, is an organ-specific, T cell-mediated autoimmune disease. Autoreactive T cells can penetrate the blood-retinal barrier, which is a physical defense composed of tight junction-linked retinal pigment epithelial (RPE) cells. RPE cells serve as antigen-presenting cells (APCs) in the eye since they express MHC class I and II and Toll-like receptors (TLRs). Although previous studies have shown that supplementation with TLR agonists exacerbates uveitis, little is known about how TLR signaling in the RPE contributes to the development of uveitis. In this study, we isolated the RPE from EAU mice, which were induced by active immunization (aEAU) or adoptive transfer of antigen-specific T cells (tEAU). The expression of TLRs on RPE was determined, and both aEAU and tEAU mice exhibited induced tlr7 expression. The TLR7 agonist R848 was shown to induce aggressive disease progression, along with significantly elevated levels of the uveopathogenic cytokine IL-17. Furthermore, not only IL-17 but also R848 appeared to enhance the inflammatory response and to impair the barrier function of the RPE, indicating that TLR7 signaling is involved in the pathogenesis of EAU by affecting the behaviors of the RPE and consequently allowing the infiltration of autoreactive T cells intraocularly. Finally, local application of shRNA against TLR7 delivered by recombinant AAV effectively inhibited disease severity and reduced IFN-γ and IL-17. Our findings highlight an immunomodulatory role of RPE TLR7 in EAU development and provide a potential therapeutic strategy for autoimmune uveitis.

Integration of genome-wide association studies and gene coexpression networks unveils promising soybean resistance genes against five common fungal pathogens

Soybean is one of the most important legume crops worldwide. However, soybean yield is dramatically affected by fungal diseases, leading to economic losses of billions of dollars yearly. Here, we integrated publicly available genome-wide association studies and transcriptomic data to prioritize candidate genes associated with resistance to Cadophora gregata, Fusarium graminearum, Fusarium virguliforme, Macrophomina phaseolina, and Phakopsora pachyrhizi. We identified 188, 56, 11, 8, and 3 high-confidence candidates for resistance to F. virguliforme, F. graminearum, C. gregata, M. phaseolina and P. pachyrhizi, respectively. The prioritized candidate genes are highly conserved in the pangenome of cultivated soybeans and are heavily biased towards fungal species-specific defense responses. The vast majority of the prioritized candidate resistance genes are related to plant immunity processes, such as recognition, signaling, oxidative stress, systemic acquired resistance, and physical defense. Based on the number of resistance alleles, we selected the five most resistant accessions against each fungal species in the soybean USDA germplasm. Interestingly, the most resistant accessions do not reach the maximum theoretical resistance potential. Hence, they can be further improved to increase resistance in breeding programs or through genetic engineering. Finally, the coexpression network generated here is available in a user-friendly web application (https://soyfungigcn.venanciogroup.uenf.br/) and an R/Shiny package (https://github.com/almeidasilvaf/SoyFungiGCN) that serve as a public resource to explore soybean-pathogenic fungi interactions at the transcriptional level.

145 Conservation Grazing by Goats in Eastern Oak-hickory Forest: Browsing Behavior and Forage Selectivity

Oak-hickory (Quercus spp. and Carya spp.) recruitment and regeneration are negatively impacted by non-natives species. Goats can provide an initial control of non-native vegetation; however, browsing behavior and preference should be studied before their introduction in the forest. Our objective was to analyze goats’ browsing behavior and preference in an eastern oak-hickory forest. We hypothesized that browsing behavior and preference are dependent on browse biomass composition and grazing duration. This study was conducted in Coshocton, Ohio, and contained two units with nine experimental plots each. Vegetation was surveyed during the summer of 2019 for biomass composition. Cluster analyses were performed using species identity and biomass. Vegetation nutritional analysis including fiber, protein, and sugar was completed. Two stocking rate densities were implemented: high (1,019 goat×day×ha-1) and low (509×goat×day×ha-1). Goats browsed for four or two days for a total of 36 days. Continuous observation was implemented to determine browsing behavior. Forage selectivity was measured using Jacob’s Selectivity Index (JSI). A linear mixed model was calculated using clusters, species, and/or grazing duration (fixed effect), experimental plots (random effect), and JSI (dependent variable). Spicebush (Lindera benzoin), multiflora rose (Rosa multiflora Thunb.), privet (Ligustrum spp.), and oriental bittersweet (Celastrus orbiculatus Thunb.) composed more than 70% of goats’ diet. Species identity, including nutritional components and physical defense mechanisms, was the most important factor in browsing selectivity. Goats preferred spicebush and privet and avoided multiflora irrespective clusters (P 0.001). In the first two days of browsing, goats favored spicebush and privet, avoided multiflora, and had a neutral preference for bittersweet (P-value 0.001). In the last two days, privet was selected and other species had a neutral selection (P 0.001). Goats will target spicebush and privet and avoid multiflora; therefore, an extended browsing period is recommended. Future studies should evaluate the effect of chemical defense mechanisms on browsing.

Lignin concentrations in phloem and outer bark are not associated with resistance to mountain pine beetle among high elevation pines

A key component in understanding plant-insect interactions is the nature of host defenses. Research on defense traits among Pinus species has focused on specialized metabolites and axial resin ducts, but the role of lignin in defense within diverse systems is unclear. We investigated lignin levels in the outer bark and phloem of P. longaeva, P. balfouriana, and P. flexilis; tree species growing at high elevations in the western United States known to differ in susceptibility to mountain pine beetle (Dendroctonus ponderosae; MPB). Pinus longaeva and P. balfouriana are attacked by MPB less frequently than P. flexilis, and MPB brood production in P. longaeva is limited. Because greater lignification of feeding tissues has been shown to provide defense against bark beetles in related genera, such as Picea, we hypothesized that P. longaeva and P. balfouriana would have greater lignin concentrations than P. flexilis. Contrary to expectations, we found that the more MPB-susceptible P. flexilis had greater phloem lignin levels than the less susceptible P. longaeva and P. balfouriana. No differences in outer bark lignin levels among the species were found. We conclude that lignification in Pinus phloem and outer bark is likely not adaptive as a physical defense against MPB.

Integration of genome-wide association studies and gene coexpression networks unveils promising soybean resistance genes against five common fungal pathogens

ABSTRACTSoybean is one of the most important legume crops worldwide. However, soybean yield is dramatically affected by fungal diseases, leading to economic losses of billions of dollars yearly. Here, we integrated publicly available genome-wide association studies and transcriptomic data to prioritize candidate genes associated with resistance to Cadophora gregata, Fusarium graminearum, Fusarium virguliforme, Macrophomina phaseolina, and Phakopsora pachyrhizi. We identified 188, 56, 11, 8, and 3 high-confidence candidates for resistance to F. virguliforme, F. graminearum, C. gregata, M. phaseolina and P. pachyrhizi, respectively. The prioritized candidate genes are highly conserved in the pangenome of cultivated soybeans and are heavily biased towards fungal species-specific defense response. The vast majority of the prioritized candidate resistance genes are related to plant immunity processes, such as recognition, signaling, oxidative stress, systemic acquired resistance, and physical defense. Based on the number of resistance alleles, we selected the five most resistant accessions against each fungal species in the soybean USDA germplasm. Interestingly, the most resistant accessions do not reach the maximum theoretical resistance potential. Hence, they can be further improved to increase resistance in breeding programs or through genetic engineering. Finally, the coexpression network generated here is available in a user-friendly web application (https://soyfungigcn.venanciogroup.uenf.br/) and an R/Shiny package (https://github.com/almeidasilvaf/SoyFungiGCN) that serve as a public resource to explore soybean-pathogenic fungi interactions at the transcriptional level.

Surviving or thriving: trade-offs between growth, defense, and reproduction in a native versus in an invasive Rubus

Little is known about limits to reproduction in plants, especially as to how their other life history functions (growth and defense) may constrain reproductive investment. Understanding these constraints can help researchers refine best practices for cultivating species like Rubus (Family: Rosaceae) that produce nutritious fruits as well as controlling invasive species. Here, we sought to elucidate potential trade-offs between growth, defense, and reproduction in native Rubus allegheniensis (common blackberry) and invasive R. phoenicolasius (wineberry) while accounting for the effects of varying insect herbivory and resource availability levels. We observed traits related to physical defense (e.g., prickle intensity), growth (e.g., cane length), and floral reproduction (e.g., ripe fruits) as well as carbon availability (e.g., canopy cover). We then used multiple regressions to characterize relationships between these variables for both species. We found potential evidence for two induced defenses in the invasive wineberry. Also, five models returned significant results indicative of trade-offs between reproduction and growth, reproduction and defense, and defense and growth in one or both species. Our results highlight the importance of understanding the defensive strategies utilized by these species because inducible defenses may result in trade-offs that could reduce yields and also increase the invasive potential of Rubus species.

Squash Varieties Domesticated for Different Purposes Differ in Chemical and Physical Defense Against Leaf and Root Herbivores

Plant domestication often reduces plant defenses by selection on chemical and physical defense traits. Thus, it is expected that herbivorous insects perform better on crop varieties than on their wild relatives. However, recent studies show that this pattern is not ubiquitous. We examined how varieties of squash (Cucurbita spp.) domesticated for different purposes (for consumption or as ornamentals), differ in plant defense traits and in their interactions with a leaf and a root herbivore. Two types of defenses were measured: cucurbitacins, which are toxic phytochemicals, and trichomes that are physical barriers for most herbivores. We addressed the following questions: (i) what is the variation in cucurbitacin content and leaf trichome density among varieties? (ii) does purpose of domestication explain differences in defense traits among varieties? and (iii) are herbivore feeding preferences and performance altered by the defense traits of squash varieties? We found great variation in cucurbitacin content among varieties, but not according to their purpose of domestication. Cucurbitacins were found mostly in cotyledons and roots and in very small quantities in the leaves. In contrast, trichome density was greater on the varieties selected for consumption than on the ornamental varieties. The performance of a leaf herbivore (Spodoptera latifascia) and a root herbivore (Diabrotica balteata), was not different among squash varieties. Moreover, in a choice experiment, larvae of the root herbivore preferred to feed on squash varieties with high cucurbitacin content. Whereas, in the field, native leaf herbivores preferred to feed on varieties selected for consumption. Our results contribute to a better understanding on how varietal selection may differentially affect plant defenses. This knowledge could help in the development of crop varieties with enhanced insect resistance.

The Effectiveness of Physical and Chemical Defense Responses of Wild Emmer Wheat Against Aphids Depends on Leaf Position and Genotype

The bird cherry-oat aphid (Rhopalosiphum padi) is one of the most destructive insect pests in wheat production. To reduce aphid damage, wheat plants have evolved various chemical and physical defense mechanisms. Although these mechanisms have been frequently reported, much less is known about their effectiveness. The tetraploid wild emmer wheat (WEW; Triticum turgidum ssp. dicoccoides), one of the progenitors of domesticated wheat, possesses untapped resources from its numerous desirable traits, including insect resistance. The goal of this research was to determine the effectiveness of trichomes (physical defense) and benzoxazinoids (BXDs; chemical defense) in aphid resistance by exploiting the natural diversity of WEW. We integrated a large dataset composed of trichome density and BXD abundance across wheat genotypes, different leaf positions, conditions (constitutive and aphid-induced), and tissues (whole leaf and phloem sap). First, we evaluated aphid reproduction on 203 wheat accessions and found large variation in this trait. Then, we chose eight WEW genotypes and one domesticated durum wheat cultivar for detailed quantification of the defense mechanisms across three leaves. We discovered that these defense mechanisms are influenced by both leaf position and genotype, where aphid reproduction was the highest on leaf-1 (the oldest), and trichome density was the lowest. We compared the changes in trichome density and BXD levels upon aphid infestation and found only minor changes relative to untreated plants. This suggests that the defense mechanisms in the whole leaf are primarily anticipatory and unlikely to contribute to aphid-induced defense. Next, we quantified BXD levels in the phloem sap and detected a significant induction of two compounds upon aphid infestation. Moreover, evaluating aphid feeding patterns showed that aphids prefer to feed on the oldest leaf. These findings revealed the dynamic response at the whole leaf and phloem levels that altered aphid feeding and reproduction. Overall, they suggested that trichomes and the BXD 2,4-dihydroxy-7- methoxy-1,4-benzoxazin-3-one (DIMBOA) levels are the main factors determining aphid resistance, while trichomes are more effective than BXDs. Accessions from the WEW germplasm, rich with trichomes and BXDs, can be used as new genetic sources to improve the resistance of elite wheat cultivars.