Cigar tobacco, an economically important cash crop, holds a substantial role within the fiscal framework of the national economy. This crop, however, is characterized by a marked vulnerability to pathogenic bacteria, culminating in consequential financial loss throughout its cultivation phase. Plant growth-promoting rhizobacteria (PGPR), a salient class of advantageous bacterial flora, are recognized for their ability to enhance plant growth, inhibit deleterious pathogens, and synthesize compounds that either have a direct impact on plant morphogenesis or activate otherwise ineffectual soil components. Through these mechanisms, PGPR augments the soil’s nutritional profile, making it more receptive to plant uptake, thus stimulating vegetative growth. The Bacillus subtilis microbial fertilizer, the prime exemplar of PGPR, demonstrates not only a pathogen-suppressive effect but also an induction of the plant’s innate disease resistance mechanism. This bolsters the plant’s resilience to disease fosters a probiotic milieu within the soil, and catalyzes the formation of agglomerate structures, all of which contribute to enhanced soil fertility and moisture retention, increased soil friability, and the facilitation of root expansion. In this study, a controlled pot experiment was conducted to elucidate the mechanism through which inter-root probiotics rehabilitate the soil’s ecosystem and foster crop growth in cigar tobacco seedlings afflicted with root black rot bacteria. Four treatments were instituted, including CK: a blank control (no microbial application); A: probiotic only (Bacillus subtilis microbial fertilizer); B: both pathogenic and probiotic (the Bacillus subtilis microbial fertilizer together with root black rot pathogen); C: pathogenic only (the root black rot pathogen). Our empirical findings delineate that the presence of pathogenic bacteria deteriorates the soil environment, thereby constraining the transmutation of soil nutrients and their subsequent assimilation by plants. This severely impedes the vegetative development of cigar plants. By contrast, the application of a PGPR microbial fertilizer modified the soil microbial community structure, exhibiting an antagonistic interaction with the indigenous pathogenic bacterial species. Relative to the CK treatment, the application of the Bacillus subtilis microbial fertilizer was found to invigorate the catalytic conversion of soil enzymes, incrementing the peroxidase, acid phosphatase, urease, and sucrase activity by 12.98%, 19.55%, 13.57%, and 17.91%, respectively. Meanwhile, it was observed to ameliorate the soil’s physicochemical attributes, enhancing the available content of nitrogen, phosphorus, and potassium by 4.52%, 6.52%, and 15.14%, respectively, along with the augmentation of soil organic matter content by 17.33%. The fortification of soil physicochemical properties and the enrichment of soil fertility, as a result of the PGPR microbial fertilizer application, translated into a robust 57.23% enhancement of root vigor and a 60.47% extension of the root length of cigar tobacco seedlings. These soil amendments subsequently fueled an uptick in the growth parameters of cigar plants, including increases in plant height, stem girth, leaf count, maximal leaf dimensions, and both the fresh and dry weight of cigar tobacco. Full article

Antioxidant molecules play a crucial role in maintaining redox homeostasis, eliminating oxidative damage, and regulating pathogenesis in phytopathogenic fungi. These antioxidants function through a complex regulatory network involving enzymatic scavengers such as the superoxide dismutases (SODs), catalases, thioredoxins, and glutathiones, as well as non-enzymatic molecules. Specifically, the thioredoxin and glutathione systems (AaTrr1, AaTsa1, AaGpx3, and AaGlr1) mediated by NADPH oxidase represent vital thiol antioxidant systems utilized to regulate the detoxification of reactive oxygen species (ROS) in Alternaria alternata. This present study investigated the regulatory roles of AaBemA, a component of the fungal NADPH oxidase (Nox) complex, in the tangerine pathotype of A. alternata. Loss-of-function genetic analysis demonstrated that AaBemA is essential for the accumulation of cellular hydrogen peroxide (H₂O₂). Mutant strains with defective AaBemA displayed higher sensitivity to H₂O₂ and the ROS-generating oxidant tert-butyl-hydroperoxide. These phenotypes closely resembled those previously observed in AaNoxB, AaTrr1, AaTsa1, AaGlr1, and AaYap1 mutants, suggesting a potential interconnection among them. Notably, the defective phenotype of ΔAaBemA could be restored through genetic complementation with wildtype AaBemA gene. Subcellular localization of a functional AaBemA fused with green fluorescent protein (GFP) gene under confocal microscope revealed a widespread distribution of green fluorescence in the cytoplasm and cell membrane, indicating high expression of AaBemA during fungal growth in A. alternata. Intriguingly, inactivation of AaBemA did not affect the morphological phenotype and pathogenicity, indicating that the AaBemA is dispensable for the aforementioned phenotypes. To explore the molecular mechanisms underlying the regulation of ROS stress response, we sequenced the whole transcriptomes of A. alternata wildtype and ROS-sensitive mutants (ΔAaBemA, ΔAaNoxB, ΔAaGlr1, ΔAaTrr1, ΔAaTsa1, ΔAaYap1) which displayed considerable sensitivity to oxidants. Comparative transcriptome analysis revealed significant influence on the gene-expression pattern of numerous genes related to glutathione metabolism, cellular oxidant detoxification, cellular response to abiotic stimulus, and cellular response to osmotic stress. The gene-expression data and pathways related to various essential metabolic processes and ROS tolerance enabled us to propose a NADPH oxidase-mediated regulatory network involving NADPH oxidase (AaNoxB, AaBemA), AaYap1, glutaredoxin (AaGPx3, AaGlr1), and thioredoxin systems (AaTrr1, AaTsa1) in responding to ROS stress in A. alternata. Full article

Fusarium wilt of banana caused by the soil-borne fungi Fusarium oxysporum f. sp. cubense, Tropical Race 4 (Foc TR4) (Syn. Fusarium odoratissimum), is a major threat to the global banana industry. Aiming to identify predisposing soil factors for Fusarium wilt of banana (FWB) TR4, the 23 physical and chemical soil properties were studied in three commercial banana farms in La Guajira, Colombia. Disturbed and undisturbed soil samples were collected from areas affected by the disease (affected plots) and disease-free areas (healthy plots). Five repetitions per farm were considered, with a total sample of n = 30. The data were analyzed using one-way analysis of variance (ANOVA). Subsequently, the debiased sparse partial correlation (DSPC) algorithm was applied. Organic matter (OM), pH, calcium (Ca), magnesium (Mg), zinc (Zn), and cation-exchange capacity on the exchange complex (ECEC), showed significant differences between the affected and healthy plots. In addition, the bulk density and saturated hydraulic conductivity (HC) were associated suggesting that physical attributes, such as soil compaction and poor drainage, create favorable conditions for FWB. According to the DSPC algorithm, the HC variable presented a grade of 5 and an intermediation of 14.67, which indicates that it has significant associations with BD, sand, porosity, ECEC, and OM and plays a critical role in the connection of other variables in the network and the differentiation of healthy and affected plots. These findings establish a baseline of information under field conditions in Colombia, which can be used to design soil management strategies to mitigate the detrimental effects of Foc TR4 by creating less favorable conditions for the pathogen. Full article

Polyphenol oxidase (PPO) is the key enzyme in the melanogenesis pathway of enzymatic browning that converts phenolic substrates to quinones and eventually polymerizes to form melanin. In this study, the genome-wide characterization of the Abppo gene family was performed, and six Abppo genes were identified. These genes were divided into three groups based on sequence alignment and phylogenetic analysis, with members of the same group possessing similar motif structures. Expression analysis showed that the Abppo genes demonstrate diverse expression patterns at different growth stages and postharvest storage. In addition, the expression of Abppo genes could be significantly induced by abscisic acid, salicylic acid, methyl jasmonate, and gibberellic acid 3, indicating their potential roles in response to abiotic stresses. These results provide insights into the potential functions of the Abppo gene family, offering a theoretical reference in the future for mushroom breeders. Full article

The present study provides a comprehensive overview of the Bacillus velezensis strain Htq6, and its potential applications in plant disease control. Htq6 is an endophytic bacterium derived from walnut, which was found to possess a strong inhibitory effect on a wide range of plant pathogenic microorganisms and was identified as a good plant disease control agent. The entire genome of the Bacillus velezensis Htq6 was sequenced, and a comparative genomic analysis was conducted with various Bacillus species in order to better understand the mechanism of the strain’s biological control. At the same time, a new classification result was presented. Additionally, transcriptome analysis was performed to explore the response mechanism of tomato gray mold fungus after treatment with the fermentation liquid of Bacillus velezensis Htq6. The study analyzed the distribution of various secondary metabolite gene clusters in the Bacillus model strains and employed RNA-Seq technology to obtain transcriptome expression profiles. Furthermore, the cell wall, cell membrane, and antioxidant-related genes of Botrytis cinerea were analyzed, providing insight into the antibacterial mechanism of biocontrol bacteria and the stress response mechanism of Botrytis cinerea. The results of the research are promising, and could potentially lead to the development of an effective biocontrol agent for the prevention and control of various plant diseases. Full article

Soilless cultivation has increased in China. Like vegetables or ornamental plants, tomatoes can be cultivated in soilless culture systems. Fusarium wilt (FW) is an economic tomato disease. The nature and amount of volatile organic compounds in Trichoderma asperellum PT-15 were investigated, and 6-pentyl-α-pyrone (6-PP) was detected. Furthermore, the effect of 6-PP on Fusarium oxysporum HF-26 was evaluated. Results revealed that 25 mg/L 6-PP was the optimal concentration inhibiting F. oxysporum HF-26 and that the content of fusaric acid decreased considerably compared to that of the control. FUB1, FUB4, and FUB10 toxin synthesis and transport genes were downregulated. Additionally, VelA, velB, and LaeA genes were downregulated, reducing F. oxysporum mycelial growth and hyphae formation. 6-PP was added to the soilless culture solution in a greenhouse experiment. The results showed that an antifungal–nutrient solution containing 25 mg/L 6-PP significantly suppressed FW with 70.71% efficacy and a 27.23% disease index (DI), which were higher efficacy and lower DI than that of the control. Furthermore, treatment with an antifungal–nutrient solution containing 6-PP increased the levels of defence enzymes 24 h post-inoculation (hpi) compared with those at other time periods. The relative expression levels of the PR1, NPR1, PR2, and PR5 genes were considerably upregulated at 24 hpi. Full article

Host-pathogen interactions are the result of the continuously evolving dynamics of the genomic interphases between pathogens and the host plants. Alternaria brown spot (ABS) caused by the pathogen Alternaria alternata is a serious threat to tangerine production. Although recent studies have made significant advances in the characterization of A. alternata virulence factors, a gap exists in the regulation of virulent genes throughout the course of A. alternata infection on host plants. To gain a better understanding of the dynamic defense transcriptome in Alternaria alternata during Infection, we performed a comparative transcriptome approach. After inoculation on citrus, we found that 2142, 1964, 2359 genes were up-regulated, and 1948, 1434, 1996 genes were down-regulated at 12 hours-post-inoculation (hpi), 24 hpi and 48 hpi, respectively. Among these genes, 1333 genes were up-regulated at three time points, and 1054 genes were down-regulated, indicating that most of the differentially expressed genes at the early stage of infection tended to remain differentially expressed at the later stage of infection. In addition to the genes that are known to be part of the infection network in plant-pathogen interactions, many novel genes related to plant-pathogen interaction were identified. Interestingly, our results indicate that A. alternata is able to rapidly alter its gene expression pattern during infection process, which is vital for the successful colonization of the pathogen. Moreover, this rapid alteration of gene expression is likely to be an adaptive mechanism, enabling the pathogen to quickly respond to any changes in the environment and adapt to the host’s defense system. This ability to modify gene expression quickly in the face of environmental changes could play a critical role in the successful establishment of infection. RT-qPCR analysis confirmed that the expression pattern of nine randomly selected genes from the peroxisome pathway were consistent with the RNA-seq data. Our study provided a comprehensive study of the expression of genes during A. alternata infection of citrus, which may facilitate the understanding of host-plant interactions in A. alternata. Full article

To analyze the genetic structure and genetic diversity of Colletotrichum gloeosporioides as the dominant Colletotrichum species on Citrus, the Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genetic diversity, including 63 strains isolated and selected from 8 different sites and 5 different citrus species, was studied. A total of 19 GAPDH haplotypes were identified by genetic analysis, and the main haplotype (haplotype 5) was distributed in 28 isolates, mainly from Citrus unchiu Hort. ex Tanaka (WG) and Citrus reticulata Blanco cv. Succosa (BDZ) in Huangyan (HY), Linhai (LH), and Jiande (JD) of Zhejiang province, and Mashui tangerine (MSJ) in Mengshan of Guangxi province (GX). Using the genetic differentiation index, Fst revealed significant genetic differentiation in C. gloeosporioides populations between Jiangxi province (JXGZ) and GX, HY, LH, JD, and Chun’an (CA) of Zhejiang province, and also revealed slightly less genetic differentiation for C. gloeosporioides populations between HY, LH, JD, GX, Shaanxi province (SX), and Quzhou (QZ) of Zhejiang province. In addition, Fst revealed great genetic differentiation between the C. gloeosporioides populations obtained from MSJ and Citrus paradise Macf (PTY), and also revealed weak genetic differentiation between the C. gloeosporioides populations obtained from Citrus sinensis Osbeck (QC), WG, and BDZ. The AMOVA test showed that the levels of genetic differentiation for C. gloeosporioides were 19% and 81% among and within geographic populations, respectively. It also showed that C. gloeosporioides had levels of genetic differentiation among and within host populations of 12% and 88%, respectively. The Mantel test showed that the genetic distance was not linearly correlated with geographical distance and the haplotype phylogenetic analysis showed that C. gloeosporioides from different regions and hosts were scattered in the phylogenetic tree, implying that the genetic differentiation was independent of host variety and geographical origin. We speculated that genetic differentiation may be mainly due to gene mutation, gene recombination, or gene migration within native populations and has nothing to do with natural selection triggered by geography or host variety. Full article

Alternaria brown spot disease is caused by the Alternaria alternata tangerine pathotype, which relies on ACT-toxin for infection. At present, all identified ACT-toxin biosynthesis-related genes are multi-copy genes. In this study, we summarized the advances in important host-specific toxins (HSTs), and listed key genes required for the pathogenicity of the A. alternata tangerine pathotype. Toxin virulence test results revealed that different citrus species displayed distinctly different tolerances to ACT-toxin. The extraction method of ACT-toxin crude extract was described in schematic form to make the method easier to understand. In addition, target gene disruption of two copies of ACTT5 (∆∆ACTT5) displayed significantly reduced virulence, indicating that ACTT5 is essential for the pathogenicity of the A. alternata tangerine pathotype. Full article

Popcorn cultivation has been growing in and has positively affected the Brazilian economy. However, these crops are grown with genotypes susceptible to diseases, generating high losses. Thus, studies aimed at obtaining resistant genotypes are particularly interesting, as popcorn is susceptible to several pathogens. The most efficient, environmentally correct, and economical method of disease control is using resistant cultivars. The present study aimed to evaluate the performance of inbred lines of popcorn and their respective testcross hybrids in terms of diseases caused by fungi. For this purpose, 15 S₇ inbred lines were crossed with five testers, four with a narrow genetic basis (inbred lines L270, L651, P1, and L70) and one with a broad genetic basis (open pollination variety PARA 172). The arrangement of treatments in incomplete blocks (lattice 10 × 10) with three replications was used. The testcross was efficient in the discrimination per se of the progenies and the testers. The inbred lines L685, L691, L696, and L684 and the PARA 172 tester showed potential resistance to P. polysora, B. maydis, and E. turcicum. The GT biplot method proved reliable in identifying efficient, responsive, and resistant inbred lines and revealing the hybrid 56 as the ideal genotype. Full article

Potatoes are among the four most important staple crops worldwide. Verticillium wilt in potatoes caused by Verticillium dahliae is a devastating disease that is difficult to control. To identify potential avenues for disease control, the pathogenicity of 72 V. dahliae isolates was tested here. We also tested the resistance to the most virulent isolate (Vd-36) induced by the attenuated isolate Gibellulopsis nigrescens Vn-1 in potatoes. Induction of Verticillium wilt resistance in potatoes was strongest when using attenuated isolate Vn-1 to inoculate potatoes with a spore suspension concentration of 1 × 10⁶ conidia mL⁻¹, followed by infection with isolate Vd-36 at 5 d intervals. After incubation of potatoes with the attenuated isolate Vn-1 followed by isolate Vd-36, reactive oxygen species (ROS) and hydrogen peroxide (H₂O₂) were produced and accumulated in potato leaves 12 h post-inoculation. The changes in respective defense enzymes, except phenylalanine ammonia-lyase, were consistent with the changes in ROS and H₂O₂ levels. Furthermore, the content of salicylic acid (SA) in inoculated plants was higher than that in the control, and biosynthesis-related genes StNPR1, StPR1b, StPR2, StPR5 were activated. However, there was no significant difference in the jasmonic acid and ethylene (JA/ET) content between the treatment and control groups. These results demonstrated that the attenuated isolate Vn-1 enhanced resistance to Verticillium wilt by inducing the SA signalling pathway and weakly activating the JA/ET signalling pathways in potatoes. Full article

6-pentyl-α-pyrone (6-PP) is a powerful Magnaporthiopsis maydis antifungal compound, recently discovered when the potent growth medium of Trichoderma asperellum was analyzed. Despite its high potential in plate assay, it was not inspected for plant treatment prevention. Late wilt disease, caused by the fungus M. maydis, threatens commercial maize production in high-risk areas. Thus, the search for control options against the pathogen is one of the top priorities in Israel, Egypt, and other countries. Disease-resistant maize genotypes can reduce the damages. Yet, aggressive variants of the fungus can overcome host resistance. The current study aimed at inspecting T. asperellum and its secreted metabolite, pure 6-PP, against the pathogen in plants over a full growth period. First, adding T. asperellum directly to seeds with sowing provides significant protection to sprouts (up to 42 days) in a growth room, with more than two-fold growth promotion and reduced pathogen root infection (detected by real-time PCR). The same procedure applied in a commercial field was less beneficial in rescuing the plants’ growth and yield. Still, it reduced the cobs’ symptoms by 11% and resulted in nine-fold lower levels of the pathogen’s DNA in the stem tissue. Second, the T. asperellum purified 6-PP compound (30 µg/seed) was used in seed coating and tested against the T. asperellum secretory metabolites’ crude (diluted to 50%). At the season’s end, these treatments improved plant biomass by 90–120% and cob weight by 60%. Moreover, the treatments significantly (p < 0.05) reduced the symptoms (up to 20%) and pathogen infection (94–98%). The current study’s results reveal the potential of 6-PP as a new fungicide against M. maydis. Such a treatment may protect maize plants from other soil diseases. Full article

Neofabraea spp. is one of the major causal agents of the postharvest decay of apple fruit. Bull’s eye rot can cause fruit rot, as well as tree canker. In Serbia, during February and March, 2017 and 2018, 56.3% of collected stored rotten apple fruits were found to be infected with Neofabraea spp. Using polymerase chain reaction, phylogenetic analysis, and morphological characterization, Neofabraeaalba was identified as the causal agent of Bull’s eye rot. Twenty-five selected isolates were analyzed by DNA sequencing of the internal transcribed spacer and 16 S mitochondrial ribosomal RNA. The phylogenetic analyses provided information on the distance between Neofabraea spp., while merged phylogenetic tree was required to give insight and clarify the relationships between isolates of Neofabraeaalba. N. alba demonstrated pathogenicity on apple fruit as well as on branches, with significantly different aggressiveness among the isolates. Isolates were able to produce larger cankers and acervuli on apple branches with abundant sporulation All identified isolates were separated into two morphotypes and described based on their macromorphological characteristics on culture media. It was noticed that different culture media, pH, and temperature could affect macromorphological characteristics and development. Representative isolates were able to grow at a temperature range of 0 to 25 °C, above which only one isolate was able to grow on culture media. All isolates showed growth at a pH range of 2–10, with significant differences detected between isolates at different pH values. The type of cultivation medium significantly affected the size and shape of conidia, while the presence of the microconidia was recorded within isolates. The aim of this study was to identify the causal agents of Bull’s eye rot on stored apples in Serbia in the period 2017–2018 and characterize the obtained isolates based on pathogenic, morphological, physiological, and molecular criteria. Full article

The coconut palm (Cocos nucifera L.) is a common crop in pantropical areas facing various challenges, one of them being the control of diseases and pests. Diseases such as bud rot caused by Phytophthora palmivora, lethal yellowing caused by phytoplasmas of the types 16SrIV-A, 16SrIV-D or 16SrIV-E, among others, and pests like the coconut palm weevil, Rhynchophorus vulneratus (Coleoptera: Curculionidae), and the horned beetle, Oryctes rhinocerus (Coleoptera: Scarabaeidae), are controlled by applying pesticides, pheromones and cultural control. These practices do not guarantee eradication since some causal agents have become resistant or are imbedded in infected tissues making them difficult to eradicate. This review condenses the current genomics, transcriptomics, proteomics and metabolomics studies which are being conducted with the aim of understanding the pathosystems associated with the coconut palm, highlighting the findings generated by omics studies that may become future targets for the control of diseases and pests in the coconut crop. Full article