Interaction between Plants and Fungi and Oomycetes

White rot, caused by Coniella vitis, is a devastating disease in grapevine (Vitis vinifera) that seriously affects yield and quality. Breeding resistant grapevine varieties is a highly economical, environmentally friendly, and effective strategy to protect against the disease; however, this strategy requires a comprehensive understanding of the genes and pathways related to resistance. In this study, we sequenced the transcriptome of V. vinifera L. cv. GF, a highly resistant variety, at six time points after C. vitis inoculation. A transcriptome analysis showed that the salicylic acid (SA) signaling pathway was activated in response to C. vitis. Transient silencing of the VvTGA8 gene in the cv. GF greatly increased susceptibility to C. vitis. Subcellular localization studies showed that the VvTGA8 gene is localized in the nucleus. Heterologous expression of VvTGA8 in Solanum lycopersicum improved resistance to C. vitis and increased levels of the SA signaling pathway marker genes SlPR1 and SlPR2 significantly. To explore the mechanism by which VvTGA8 mediates disease resistance, we silenced SlICS1, a key gene in the SA synthesis pathway, through virus-induced gene silencing to inhibit SA synthesis in a VvTGA8 overexpression line, resulting in significantly weakened resistance to C. vitis and decreased expression levels of SlPR1 and SlPR2. We conclude that VvTGA8 is involved in SA signaling pathway, which activates the expression of pathogenesis-related genes in the nucleus, thereby mediating resistance to C. vitis in grapevine. This study provides an excellent target gene for disease-resistant breeding and gene editing in grapevine. Full article

Fusarium crown rot (FCR), caused by the fungal pathogen Fusarium pseudograminearum (Fp), is a major constraint to cereal production worldwide. The pathogen restricts the movement of solutes within the plant due to mycelial colonisation of vascular tissue. Yield loss and quality downgrades are exacerbated by this disease under water stress conditions. Plant root systems are adaptive and can alter their architecture to optimise production in response to changes in environment and plant health. This plasticity of root systems typically favours resource acquisition of primarily water and nutrients. This study examined the impact of FCR on the root system architecture of multiple commercial bread and durum wheat varieties. Root system growth was recorded in-crop in large transparent rhizoboxes allowing visualization of root architecture over time. Furthermore, electrical resistivity tomography was used to quantify spatial root activity vertically down the soil profile. Results demonstrated a significant reduction in the total root length and network area with the inoculation of FCR. Electrical resistivity measurements indicated that the spatial pattern of water use for each cultivar was influenced differently from infection with FCR over the growing season. Specifically temporal water use can be correlated with FCR tolerance of the varieties marking this investigation the first to link root architecture and water use as tolerance mechanisms to FCR infection. This research has implications for more targeted selection of FCR tolerance characteristics in breeding programs along with improved specific varietal management in-crop. Full article

The effects of the bioherbicidal activity of the fungal phytopathogen, Albifimbria verrucaria (AV), formerly Myrothecium verrucaria, on glyphosate-resistant and –susceptible Conyza canadensis (horseweed) were examined in greenhouse and field studies. Spray applications of mycelial formulations of AV infected both glyphosate-resistant and -susceptible C. canadensis plants at various growth stages. Young plants in the rosette leaf stage of growth were controlled more efficaciously than were older plants that had bolted or that were in the inflorescence stage; nevertheless, severe injury and mortality also occurred in mature plants. The results indicate that this bioherbicidal fungus can infect and control C. canadensis, thereby demonstrating the potential of this fungus as a bioherbicidal agent against this troublesome weed, which has become resistant to various herbicides. Full article

Oidiodendron maius G.L. Barron is a recognized fungal species capable of forming ericoid mycorrhiza with various positive effects on host plants; therefore, newly found and previously uncharacterized O. maius strains may be valuable for heather plants’ controlled mycorrhization. Characteristics of the O. maius F3860 strain were studied, i.e., mycelium growth on various nutrient media and the ability to secrete auxins and enzymes. O. maius F3860 grew rapidly on malt extract agar and potato dextrose agar. It was also able to grow on nutrient media suitable for heather plant cultivation. The presence of the flavonoids rutin and quercetin increased the mycelium growth rate compared to the control, starting from the 8th to the 13th days of cultivation. The ability to secrete auxins was confirmed with bioassay and thin-layer chromatography, and their content, as well as phytase activity, was estimated spectrophotometrically. Both in nutrient media with tryptophan and without it, O. maius F3860 secreted about 6 μg IAA/mL growth medium. O. maius F3860 possessed extracellular phytase, protease, and phenol oxidase activities. The investigation indicates O. maius F3860’s promise for heather seedling inoculation as an approach to increase their fitness. Full article

The development of new anti-ureolytic compounds is of great interest due to the newly discovered role of urease inhibitors in crop protection. Purine degradation and the generation of ammonium by urease are required for the full virulence of biotrophic and hemibiotrophic fungal plant pathogens. Accordingly, chemicals displaying urease inhibitor activity may be used as a novel class of fungicides. Several urease inhibitors belonging to different chemical classes are known, and some compounds have been developed as urea fertilizer additives. We tested whether the natural urease inhibitors p-benzoquinone (p-HQ) and hydroquinone (HQ), as well as the synthetic inhibitors isopropoxy carbonyl phosphoric acid amide (iCPAA), benzyloxy carbonyl phosphoric acid amide (bCPAA), and dipropyl-hexamino-1,3 diphosphazenium chloride (DDC), prevent or delay plant infection caused by pathogens differing in lifestyles and host plants. p-BQ, HQ, and DCC not only protected maize from infection by the hemibiotroph C. graminicola, but also inhibited the infection process of biotrophs such as the wheat powdery mildew fungus Blumeria graminis f. sp. tritici and the broad bean rust fungus Uromyces viciae-fabae. Interestingly, the natural quinone-based compounds even reduced the symptom severity of the necrotrophic fungi, i.e., the grey mold pathogen B. cinerea and the Southern Leaf Spot fungus C. heterostrophus, to some extent. The urease inhibitors p-BQ, HQ, and DCC interfered with appressorial penetration and confirmed the appropriateness of urease inhibitors as novel fungicidal agents. Full article

Bellucia imperialis is one of the most abundant pioneer tree species in anthropized areas of the Central Amazon, and has ecological importance for the environmental resilience of phosphorus (P)-depleted areas. Thus, we investigated whether B. imperialis depends on symbiosis with arbuscular mycorrhizal fungi (AMF) to grow and establish under the edaphic stresses of low nutrient content and low surface moisture retention capacity of the substrate. We tried three AMF inoculation treatments: (1) CON—no mycorrhizae; (2) MIX—with AMF from pure collection cultures, and (3) NAT—with native AMF, combined with five doses of P via a nutrient solution. All CON treatment seedlings died without AMF, showing the high mycorrhizal dependence of B. imperialis. Increasing P doses significantly decreased the leaf area and shoot and root biomass growth for both the NAT and MIX treatments. Increasing P doses did not affect spore number or mycorrhizal colonization, but decreased the diversity of AMF communities. Some species of the AMF community showed plasticity, enabling them to withstand shortages of and excess P. B. imperialis was shown to be sensitive to excess P, promiscuous, dependent on AMF, and tolerant of scarce nutritional resources, highlighting the need to inoculate seedlings to reforest impacted areas. Full article

The phytopathogen Phytophthora parasitica, from the Oomycetes class, known to be the tobacco black shank agent, can induce devastating diseases in various crop, plant and forest ecosystems. The genus Phytophthora has been studied at the cellular level, suggesting that different developmental steps are induced by the expression of some specific genes. However, these studies have only been carried out on certain species, such as Phytophthora infestans and Phytophthora cactorum. As for Phytophthora parasitica, which can be considered as one of the top ten oomycete pathogens due to the economic impact and effect it has on food security, even less functional analyses and transcriptomics data are available. To date, little is known about the protein expression of Phytophthora parasitica, information that is essential for achieving a better understanding of this species. In this study, we aimed to gain insight into the proteomics of the mycelium of the Phytophthora parasitica strain INRA 310 by addressing the following questions: (i) how many predicted proteins can be detected on the mycelium of P. parasitica INRA 310, and (ii) what proteins can be detected? The proteomics experiments were performed on the mycelium of the strain Phytophthora parasitica INRA310, using the nanoliquid chromatography-MS/MS technique. A total of 219 proteins were identified, including ten unknown proteins and 209 proteins involved in lipid, carbohydrate, nucleotide, energy production and other metabolic pathways. This proteomics study is, to our knowledge, the first to be performed on the mycelium of Phytophthora parasitica INRA 310. It gives a brief first insight into its in vitro-expressed proteins. This work may be the first step before further, more comprehensive studies are undertaken with the aim of better understanding the biology of this species and its pathogenicity. Full article

ABC transporters play important roles in plant growth and resistance to abiotic and biotic stresses. Here, we showed that the TaABCG36 gene positively regulates leaf rust resistance in the wheat line Thatcher + Lr19 (TcLr19) when challenged with an avirulent pathotype of Puccinia triticina (Pt). The TaABCG36 gene was cloned from genomic DNA and cDNA from wheat line TcLr19. The clone was 6730 bp in gDNA and 4365 bp in cDNA for this gene. It encoded an ABC transporter with 1454 amino acids in length. BLASTp analysis indicated a considerable identity ABC transporter G family member 36 with Aegilops tauschii subsp. strangulata, Triticum dicoccoides, and T. aestivum; thus, we named the gene TaABCG36. TaABCG36 was proved to be a plasma transmembrane protein by bioinformatic analysis and subcellular localization of the TaABCG36–GFP fusion protein. The expression of TaABCG36 in wheat leaves reached a peak at 72 h post-inoculation by Pt avirulence pathotype, and the expression was also induced by phytohormone treatments of salicylic acid (SA), abscisic acid (ABA), and methyl jasmonate (MeJA). Three fragments (V1–V3) of the TaABCG36 gene were introduced to the BSMV-VIGS vector and, thus, silenced the expression of TaABCG36 in the wheat line TcLr19. All the three BSMV:VIGS-infected plants showed reaction type “3” to Pt pathotype THTS, which was fully avirulent on TcLr19 (infection type “0”). Histopathological observation showed that silencing of TaABCG36 facilitated the formation of haustorial mother cells (HMC) and mycelial growth, implying that TaABCG36 plays a positive role in the response of TcLr19 against THTS. These results provide molecular insight into the interaction between Pt and its wheat host and identify a potential target for engineering resistance in wheat to damaging pathogen of Pt. Full article

Due to at least 3 years of cultivation, Panax ginseng (ginseng) is susceptible to being attacked by pathogens which severely affect its quality and yield. Compared with other diseases of ginseng, Phytophthora blight caused by Phytophthora cactorum (P. cactorum) can spread rapidly and destroy almost the entire plant of ginseng, such as leaves, stems, and roots. However, little research was focused on this area, and how P. cactorum affected the metabolic profile of ginseng is still obscure. In the current study, we conducted a comprehensive analysis of metabolomics and transcriptomics to compare the differences in health and P. cactorum-affected ginseng leaves and stems. Metabolome analysis revealed that 110 and 113 significant differential metabolites were observably disturbed separately in ginseng leaves and stems. Transcriptome analysis demonstrated that 6424 and 9508 genes had remarkable variation in ginseng leaves and stems. Using conjoint analysis, we also revealed the changes in pathways “Alanine, aspartate and glutamate metabolism”, “Glycine, serine and threonine metabolism”, and “Biosynthesis of unsaturated fatty acids” and “Plant hormone signal transduction” in ginseng response to the P. cactorum. The current work provides an overview of the alteration of metabolic profile and gene expression profiles in ginseng leaves and stems in response to P. cactorum affection, which may help to further screen out the mechanism of plant-pathogen interaction at the molecular level. Full article

Blight caused by Phytophthora pathogens has a devastating impact on crop production. Phytophthora species secrete an array of effectors, such as Phytophthora cactorum-Fragaria (PcF)/small cysteine-rich (SCR) phytotoxic proteins, to facilitate their infections. Understanding host responses to such proteins is essential to developing next-generation crop resistance. Our previous work identified a small, 8.1 kDa protein, SCR96, as an important virulence factor in Phytophthora cactorum. Host responses to SCR96 remain obscure. Here, we analyzed the effect of SCR96 on the resistance of tomato treated with this recombinant protein purified from yeast cells. A temporal transcriptome analysis of tomato leaves infiltrated with 500 nM SCR96 for 0, 3, 6, and 12 h was performed using RNA-Seq. In total, 36,779 genes, including 2704 novel ones, were detected, of which 32,640 (88.7%) were annotated. As a whole, 5929 non-redundant genes were found to be significantly co-upregulated in SCR96-treated leaves (3, 6, 12 h) compared to the control (0 h). The combination of annotation, enrichment, and clustering analyses showed significant changes in expression beginning at 3 h after treatment in genes associated with defense and metabolism pathways, as well as temporal transcriptional accumulation patterns. Noticeably, the expression levels of resistance-related genes encoding receptor-like kinases/proteins, resistance proteins, mitogen-activated protein kinases (MAPKs), transcription factors, pathogenesis-related proteins, and transport proteins were significantly affected by SCR96. Quantitative reverse transcription PCR (qRT-PCR) validated the transcript changes in the 12 selected genes. Our analysis provides novel information that can help delineate the molecular mechanism and components of plant responses to effectors, which will be useful for the development of resistant crops. Full article

The interactions between plants and microorganisms, which are widely present in the microbial-dominated rhizosphere, have been studied. This association is highly beneficial to the organisms involved, as plants benefit soil microorganisms by providing them with metabolites, while microorganisms promote plant growth and development by promoting nutrient uptake and/or protecting the plant from biotic and abiotic stresses. Piriformospora indica, an endophytic fungus of Sebacinales, colonizes the roots of a wide range of host plants and establishes various benefits for the plants. In this work, an interaction between barley and the P. indica was established to elucidate microRNA (miRNA)-based regulatory changes in miRNA profiles and gene expression that occurred during the symbiosis. Growth promotion and vigorous root development were confirmed in barley colonized by P. indica. The genome-wide expression profile analysis of miRNAs in barley root showed that 7,798,928, 6,418,039 and 7,136,192 clean reads were obtained from the libraries of mock, 3 dai and 7 dai roots, respectively. Sequencing of the barley genome yielded in 81 novel miRNA and 450 differently expressed genes (DEGs). Additionally, 11, 24, 6 differentially expressed microRNAs (DEMs) in barley were found in the three comparison groups, including 3 dai vs. mock, 7 dai vs. mock and 7 dai vs. 3 dai, respectively. The predicted target genes of these miRNAs are mainly involved in transcription, cell division, auxin signal perception and transduction, photosynthesis and hormone stimulus. Transcriptome analysis of P. indica identified 667 and 594 differentially expressed genes (DEG) at 3 dai and 7 dai. Annotation and GO (Gene Ontology) analysis indicated that the DEGs with the greatest changes were concentrated in oxidoreductase activity, ion transmembrane transporter activity. It implies that reprogramming of fundamental miRNA and gene expression occurs both in barley and P. indica. Analysis of global changes in miRNA profiles of barley colonized with P. indica revealed that several putative endogenous barley miRNAs expressed upon colonization belonging to known micro RNA families involved in growth and developmental regulation. Full article