Phytopathology最新文献

4'-Phosphopantetheinyl transferases (PPTases) play important roles in the posttranslational modifications of bacterial carrier proteins, which are involved in various metabolic pathways. Here, we found that RsacpS and RspcpS encoded a functional AcpS-type and Sfp-type PPTase, respectively, in Ralstonia solanacearum GMI1000, and both are capable of modifying R. solanacearum AcpP1, AcpP2, AcpP3, and AcpP5 proteins. RspcpS is located on the megaplasmid, which does not affect strain growth and fatty acid synthesis but significantly contributes to the virulence of R. solanacearum and preferentially participates in secondary metabolism. We found that deletion of RspcpS did not affect the abilities of cellulose degradation, biofilm formation, and resistance to NaCl, sodium dodecyl sulfate, and H₂O₂ and attenuated R. solanacearum pathogenicity only in the assay of soil-drenching infection but not stem injection of tomato. It is hypothesized that RsPcpS plays a role in cell viability in complex environments and in the process during which the strain recognizes and approaches plants. These results suggest that both RsAcpS and RsPcpS may be potential targets for controlling diseases caused by R. solanacearum.

Bacterial blight (BB) caused by Xanthomonas oryzae pv. oryzae is one of the epidemic diseases in rice. Rapid changes in the pathogenicity of the X. oryzae pv. oryzae pathogen demand the identification and characterization of novel BB resistance genes. Here, we report the transfer and mapping of a new BB resistance gene from Oryza rufipogon acc. CR100098A. Inheritance studies on the BC₂F₂ population, BC₂F₃ progenies, and backcross-derived recombinant inbred lines derived from a cross between Pusa44/O. rufipogon acc. CR100098A//2^*PR114 showed that a single recessive gene confers resistance in O. rufipogon acc. CR100098A. Bulked segregant analysis using 203 simple sequence repeat (SSR) markers localized the BB resistance gene on chromosome 11 bracketed between two SSR markers, RM27235 and RM2136. Using PR114 and O. rufipogon acc. CR100098A genotyping by sequencing data, 86 KASP markers within the bracketed region were designed and tested for bulked segregant analysis. Only five KASP markers showed polymorphism between parents, and three were associated with the target gene. Seventy-seven new SSR markers were designed from the same interval. A total of 33 polymorphic markers were analyzed on the whole population and mapped the BB gene in an interval of 2.8 cM flanked by SSR markers PAU11_65 and PAU11_44 within a physical distance of 376.3 kb. The BB resistance gene mapped in this study is putatively new and designated as xa49(t). Fourteen putative candidate genes were identified within the xa49(t) region having a role in biotic stress resistance. The linked markers to the xa49(t) gene were validated in other rice cultivars for its successful deployment in BB resistance breeding.

Dollar spot is a destructive foliar disease of amenity turfgrass caused by Clarireedia spp. fungi, mainly C. jacksonii, on the Northern United States region's cool-season grass. Oxalic acid (OA) is an important pathogenicity factor in related fungal plant pathogens such as Sclerotinia sclerotiorum; however, the role of OA in the pathogenic development of C. jacksonii remains unclear due to its recalcitrance to genetic manipulation. To overcome these challenges, a CRISPR/Cas9-mediated homologous recombination approach was developed. Using this novel approach, the oxaloacetate acetylhydrolase (oah) gene that is required for the biosynthesis of OA was deleted from a C. jacksonii wild-type (WT) strain. Two independent knockout mutants, ΔCjoah-1 and ΔCjoah-2, were generated and inoculated on potted creeping bentgrass along with a WT isolate and a genome sequenced isolate LWC-10. After 12 days, bentgrass inoculated with the mutants ΔCjoah-1 and ΔCjoah-2 exhibited 59.41% lower dollar spot severity compared with the WT and LWC-10 isolates. OA production and environmental acidification were significantly reduced in both mutants when compared with the WT and LWC-10. Surprisingly, stromal formation was also severely undermined in the mutants in vitro, suggesting a critical developmental role of OA independent of plant infection. These results demonstrate that OA plays a significant role in C. jacksonii virulence and provide novel directions for future management of dollar spot. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.

Commercialized genetically modified (GM) papaya cultivars have protected papaya from the devastating disease caused by papaya ringspot virus (PRSV). However, papaya leaf distortion mosaic virus (PLDMV), which causes similar infection symptoms but is serologically distinct from PRSV, was found to be a competitive threat to the papaya industry. Our study surveyed the occurrence of PRSV and PLDMV, as well as the transgenic markers of the 35S promoter from cauliflower mosaic virus and the neomycin phosphotransferase II gene in feral papaya plants, which were found frequently growing outside of cultivated papaya fields on Hainan Island. In total, 123 feral papayas, comprising 62 (50.4%) GM plants and 61 (49.6%) non-GM ones, were sampled. Among them, 23 (18.7%) were positive for PRSV, 49 (39.8%) were positive for PLDMV (including five plants co-infected by PRSV and PLDMV), and 56 (45.5%) were free of either virus. In traditional papaya-growing regions, we detected fewer PRSV-infected plants (2 in 33, 6%) than in other regions (21 in 90, 23%). However, overall, whether plants were transgenic or not made no difference to PRSV incidence (P = 0.230), with 9 PRSV-infected plants among 62 GM papayas and 14 among 61 non-GM papayas. Phylogenetic and genetic differentiation analysis showed a clear correlation between PRSV and PLDMV populations and their geographic origins. Negative selection was estimated for the selected gene regions of both viruses. Notably, PLDMV has deviated from neutral evolution and experienced population expansion, exhibiting increased genetic diversity, and is becoming the predominant threat to papaya in Hainan.

The image-based detection and classification of plant diseases has become increasingly important to the development of precision agriculture. We consider the case of tomato, a high-value crop supporting the livelihoods of many farmers around the world. Many biotic and abiotic plant health issues impede the efficient production of this crop, and laboratory-based diagnostics are inaccessible in many remote regions. Early detection of these plant health issues is essential for efficient and accurate response, prompting exploration of alternatives for field detection. Considering the availability of low-cost smartphones, artificial intelligence-based classification facilitated by mobile phone imagery can be a practical option. This study introduces a smartphone-attachable 30× microscopic lens, used to produce the novel tomato microimaging data set of 8,500 images representing 34 tomato plant conditions on the upper and lower sides of leaves as well as on the surface of tomato fruits. We introduce TOMMicroNet, a 14-layer convolutional neural network (CNN) trained to classify biotic and abiotic plant health issues, and we compare it against six existing pretrained CNN models. We compared two separate pipelines of grouping data for training TOMMicroNet, either presenting all data at once or separating the data into subsets based on the three parts of the plant. Comparing configurations based on cross-validation and F1 scores, we determined that TOMMicroNet attained the highest performance when trained on the complete data set, with 95% classification accuracy on both training and external data sets. Given TOMMicroNet's capabilities when presented with unfamiliar data, this approach has potential for the identification of plant health issues.

Understanding the ecology of pathogens is important for disease management. Recently a devastating canker disease was found on red alder (Alnus rubra) planted as landscape trees. Bacteria were isolated from two groups of symptomatic trees located approximately 1 kilometer apart and one strain from each group was used to complete Koch's postulates. Results showed that these bacteria can not only cause disease on red alder but also on two other alder species. Unexpectedly, analyses of genome sequences of bacterial strains identified them as Lonsdalea quercina, a pathogenic species previously known to cause dieback of oak species, but not alder. Additionally, a core genome phylogeny clustered bacterial strains isolated from red alder within a subclade of L. quercina strains isolated from symptomatic oak trees. Consistent with the close phylogenetic relationship, there was no obvious evidence for divergence in genome composition of strains isolated from red alder and oak. Altogether, findings indicate that L. quercina is a potential threat to Alnus species.

A major resistance gene of the melon accession PI414723 to zucchini yellow mosaic virus (ZYMV) is located at the Zym-1 locus on chromosome 2, but the underlying defense mechanisms are poorly understood. The physiological responses and expression of selected genes at Zym-1 were assessed in PI414723 and in the susceptible genotype Védrantais. Viral titers and the expression of genes related to systemic acquired resistance (SAR) were evaluated in inoculated (Inoc) and non-inoculated (Non-Inoc) portions of the cotyledons at 3, 7 and 10 days after inoculation (dai) and in apical leaves at 10 dai. ZYMV was detected in both portions of the cotyledons but not in the apical leaves of PI414723 plants. Also, ZYMV was recovered in a susceptible zucchini only from Inoc portions at 3 dai. By contrast, in Védrantais ZYMV was detected and recovered from all tissues at high concentrations. Starchy local lesions and accumulation of transcripts of the SAR marker genes PR1 and PR4 were also detected in the resistant genotype. Plus, transcripts of one candidate resistance gene analog previously located at Zym-1 and of two melon homologs of restricted tobacco etch virus movement 2 (RTM2) genes located close to Zym-1, accumulated only in PI414723. It is proposed that resistance results from the combined action of the R gene, involved in restricting ZYMV replication after a supposed recognition event and of the RTM genes which impact viral systemic movement to distal apical tissues.

Fusarium oxysporum f. sp. lactucae (FOLac) is a soil- and seedborne fungal pathogen that causes Fusarium wilt of lettuce, an important disease threatening global lettuce production. Based on pathogenicity on differential lettuce cultivars, four races (1-4) have been identified, with race 1 the only race detected in the United States, and the closely related, emerging race 4 known only in Europe. The development of race-specific diagnostic tools is hindered by insufficient genomic data to distinguish between the two races and FOLac from other F. oxysporum formae speciales and nonpathogenic isolates. Here, we describe a systematic approach for developing diagnostic markers for FOLac race 1 that utilized a comprehensive sequence database of F. oxysporum to identify 15 unique genomic sequences. Marker specificity was validated through an exhaustive screening process against genomic data from 797 F. oxysporum isolates representing 64 formae speciales and various plants and non-plant substrates. One of the unique sequences was used to develop a TaqMan quantitative polymerase chain reaction assay and a recombinase polymerase amplification assay, both exhibiting 100% sensitivity and specificity when tested against purified DNA from 171 F. oxysporum isolates and 69 lettuce samples. The relationship between qPCR C_t values and colony forming units (CFU)/g values was also determined. This study not only introduces a new marker for FOLac race 1 diagnostics and soil quantitation, but also underscores the value of an extensive genomic database and screening software pipeline for developing molecular diagnostics for F. oxysporum formae speciales and other fungal taxa.

Due to a lack of understanding of the disease epidemiology and comprehensive control measures, tea leaf spot caused by Didymella segeticola has a significant negative impact on tea yield and quality in the tea plantations of Southwest China. Phenazine-1-carboxamide (PCN) is a phenazine compound derived from Pseudomonas species, which exhibits antimicrobial activity against various pathogens. However, its inhibitory mechanism is not yet clear. The current study evaluated the inhibitory activity of PCN against various phytopathogenic fungi and found that PCN has inhibitory activity against multiple pathogens, with a half-maximal effective concentration (EC₅₀) value for D. segeticola of 16.11 μg/mL in vitro and a maximum in-vivo curative activity of 72.28% toward tea leaf spot. Morphological changes in the hyphae after exposure to PCN were observed through microstructure and ultrastructure analysis, and indicated that PCN causes abnormalities in the hyphae, such as cytoplasmic coagulation, shortened hyphal inter-septum distances, and unclear boundaries of organelles. Transcriptomic analysis revealed that PCN upregulated the expression of genes related with energy metabolism. PCN significantly reduced the ATP concentration in the hyphae and decreased mitochondrial membrane potential. Molecular docking analysis indicated that PCN binds to one of the candidate target proteins, pyruvate dehydrogenase, with lower free energy of -10.7 kcal/mol. This study indicated that PCN can interfere with energy metabolism, reducing ATP generation, ultimately affecting hyphal growth. Overall, PCN shows potential for future application in the control of tea leaf spot due to its excellent antifungal activity and unique mode of action.

Tea leaf spot caused by Didymella segeticola is a disease that has recently been discovered in the tea plantations of Southwest China, and which has a significant negative impact on the yield and quality of tea leaves. Wuyiencin is a nucleotide antimicrobial that is effective against a range of fungal diseases. However, its mode of action is still unclear. The current study found that wuyiencin inhibited the mycelial growth of D. segeticola in vitro. Meanwhile, in vivo experiments confirmed that wuyiencin had a significant curative effect on tea leaf spot. Microscopic observation represented it damaged the organelles and nucleus in fungal cells. Reverse transcription quantitative PCR assays revealed that mycelium treated with wuyiencin at the half-maximal effective concentration (EC₅₀) dosage for 1 hour exhibited 3.23 times lower expression of Threonine dehydratase (Td) gene, which is responsible for producing pyruvate. The wild type (WT) strain had a 1.77-fold higher pyruvate concentration than that in the td mutant (P < 0.05). The td mutant was more sensitive than the WT to wuyiencin treatment, with the EC₅₀ value in the td mutant being 30.01 μg/ml, compared with 82.34 μg/ml in the WT. Molecular docking demonstrated that wuyiencin bound to Td, with a binding energy of -10.47 kcal/mol. Compared with the WT strain, wuyiencin significantly reduced ATP concentration of the td mutant strain at dosages of 80.0 and 160.0 µg/ml. In total, wuyiencin reduced Td activity, inhibited pyruvate production, and decreased ATP content in the phytopathogenic fungus, ultimately disturbing the growth of the mycelium.