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1.
J Agric Food Chem ; 72(38): 20763-20774, 2024 Sep 25.
Artículo en Inglés | MEDLINE | ID: mdl-39271247

RESUMEN

Trichoderma longibrachiatum is a filamentous fungus used as a biological control agent against different plant diseases. The multifunctional secondary metabolites synthesized by Trichoderma, called peptaibols, have emerged as key elicitors in plant innate immunity. This study obtained a high-quality genome sequence for the T. longibrachiatum strain 40418 and identified two peptaibol biosynthetic gene clusters using knockout techniques. The two gene cluster products were confirmed as trilongin AIV a (11-residue) and trilongin BI (20-residue) using liquid chromatography coupled with tandem mass spectrometry. Further investigations revealed that these peptaibols induce plant resistance to Pseudomonas syringae pv tomato (Pst) DC3000 infection while triggering plant immunity and cell death. Notably, the two peptaibols exhibit synergistic effects in plant-microbe signaling interactions, with trilongin BI having a predominant role. Moreover, the induction of tomato resistance against Meloidogyne incognita showed similarly promising results.


Asunto(s)
Resistencia a la Enfermedad , Peptaiboles , Enfermedades de las Plantas , Pseudomonas syringae , Solanum lycopersicum , Trichoderma , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/parasitología , Enfermedades de las Plantas/prevención & control , Solanum lycopersicum/microbiología , Solanum lycopersicum/inmunología , Trichoderma/química , Trichoderma/metabolismo , Trichoderma/genética , Peptaiboles/farmacología , Peptaiboles/química , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Tylenchoidea/efectos de los fármacos , Inmunidad de la Planta , Animales
2.
Plants (Basel) ; 13(16)2024 Aug 15.
Artículo en Inglés | MEDLINE | ID: mdl-39204709

RESUMEN

In plants, WRKY transcription factors play a crucial role in plant growth, development, and response to abiotic and biotic stress. Cowpea (Vigna unguiculata) is an important legume crop. However, cowpea Fusarium wilt (CFW), caused by Fusarium oxysporum f. sp. tracheiphilum (Fot), poses a serious threat to its production. In this study, we systematically identified members of the cowpea WRKY (VuWRKY) gene family and analyzed their expression patterns under CFW stress. A total of 91 WRKY transcription factors were identified in the cowpea genome. Phylogenetic and synteny analyses indicated that the expansion of VuWRKY genes in cowpea is primarily due to recent duplication events. Transcriptome analysis of cowpea inoculated with Fo revealed 31 differentially expressed VuWRKY genes, underscoring their role in the response to CFW infection. Four differentially expressed WRKY genes were selected for validation. Subcellular localization and Western blot assays showed their nuclear localization and normal expression in N. benthamiana. Additionally, yeast one-hybrid assays demonstrated that VuWRKY2 can bind to the promoter region of the Catalase (CAT) gene, indicating its potential role in transcriptional regulation. This study establishes a foundation for further exploration of the role and regulatory mechanisms of VuWRKY genes in response to CFW stress.

3.
Sci Total Environ ; 951: 175443, 2024 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-39134273

RESUMEN

To reveal the outstanding high-emission problems that occur when heavy-duty diesel vehicles (HDDV) pass uphill and downhill, this study proposes a method to depict the nitrogen oxides (NOx) and carbon dioxide (CO2) high-emission driving behaviors caused by slopes from the perspective of engine principles. By calculating emission and grade data of HDDV based on on-board diagnostic (OBD) data and digital elevation model (DEM) data, the 262 short trips including uphill, flat-road and downhill are firstly obtained through the rule-based short trip segmentation method, and the significant correlation between the road grade and emissions of the short trips is verified by Kendall's Tau and K-means clustering. Secondly, by comparing the distribution changes of three speed categories (acceleration state, constant speed state and deceleration state), the differences in HDDV operating states under different grade levels are discussed. Finally, the machine learning models (Random Forest, XGBoost and Elastic Net), are used to develop the NOx and CO2 emission estimation model, identifying high-emission driving behaviors, particularly during uphill driving, which showed the highest proportion of high-emission. Explained by the feature importance and SHapley Additive exPlanations (SHAP) model that large accelerator pedal opening, frequent aggressive acceleration, and high engine load have positive effects both on NOx and CO2 emissions. The difference is in the air-fuel ratio that the engine in the rich or slightly lean burning state will increase CO2 emissions and the lean burning state will increase NOx emissions. In addition, due to the uncertainty of the actual uphill, drivers often undergo a rapid "deceleration-uniform-acceleration" process, which significantly contributes to high NOx and CO2 emissions from the engine perspective. The findings provide insights for designing driving strategies in slope scenarios and offer a novel perspective on depicting driving behaviors.

4.
Microbiome ; 12(1): 160, 2024 Aug 30.
Artículo en Inglés | MEDLINE | ID: mdl-39215347

RESUMEN

BACKGROUND: Cabbage Fusarium wilt (CFW) is a devastating disease caused by the soil-borne fungus Fusarium oxysporum f. sp. conglutinans (Foc). One of the optimal measures for managing CFW is the employment of tolerant/resistant cabbage varieties. However, the interplay between plant genotypes and the pathogen Foc in shaping the rhizosphere microbial community, and the consequent influence of these microbial assemblages on biological resistance, remains inadequately understood. RESULTS: Based on amplicon metabarcoding data, we observed distinct differences in the fungal alpha diversity index (Shannon index) and beta diversity index (unweighted Bray-Curtis dissimilarity) within the rhizosphere of the YR (resistant to Foc) and ZG (susceptible to Foc) cabbage varieties, irrespective of Foc inoculation. Notably, the Shannon diversity shifts in the resistant YR variety were more pronounced following Foc inoculation. Disease-resistant plant variety demonstrate a higher propensity for harboring beneficial microorganisms, such as Pseudomonas, and exhibit superior capabilities in evading harmful microorganisms, in contrast to their disease-susceptible counterparts. Furthermore, the network analysis was performed on rhizosphere-associated microorganisms, including both bacteria and fungi. The networks of association recovered from YR exhibited greater complexity, robustness, and density, regardless of Foc inoculation. Following Foc infection in the YR rhizosphere, there was a notable increase in the dominant bacterium NA13, which is also a hub taxon in the microbial network. Reintroducing NA13 into the soil significantly improved disease resistance in the susceptible ZG variety, by directly inhibiting Foc and triggering defense mechanisms in the roots. CONCLUSIONS: The rhizosphere microbial communities of these two cabbage varieties are markedly distinct, with the introduction of the pathogen eliciting significant alterations in their microbial networks which is correlated with susceptibility or resistance to soil-borne pathogens. Furthermore, we identified a rhizobacteria species that significantly boosts disease resistance in susceptible cabbages. Our results indicated that the induction of resistance genes leading to varied responses in microbial communities to pathogens may partly explain the differing susceptibilities of the cabbage varieties tested to CFW. Video Abstract.


Asunto(s)
Brassica , Resistencia a la Enfermedad , Fusarium , Microbiota , Enfermedades de las Plantas , Rizosfera , Microbiología del Suelo , Brassica/microbiología , Resistencia a la Enfermedad/genética , Enfermedades de las Plantas/microbiología , Fusarium/genética , Microbiota/genética , Bacterias/clasificación , Bacterias/genética , Raíces de Plantas/microbiología , Hongos/genética , Hongos/clasificación
5.
Commun Biol ; 7(1): 757, 2024 Jun 22.
Artículo en Inglés | MEDLINE | ID: mdl-38909167

RESUMEN

N-methyltransferase (NMT)-catalyzed methylation at the termini of nonribosomal peptides (NRPs) has rarely been reported. Here, we discover a fungal NMT LcsG for the iterative terminal N-methylation of a family of NRPs, leucinostatins. Gene deletion results suggest that LcsG is essential for leucinostatins methylation. Results from in vitro assays and HRESI-MS-MS analysis reveal the methylation sites as NH2, NHCH3 and N(CH3)2 in the C-terminus of various leucinostatins. LcsG catalysis yields new lipopeptides, some of which demonstrate effective antibiotic properties against the human pathogen Cryptococcus neoformans and the plant pathogen Phytophthora infestans. Multiple sequence alignments and site-directed mutagenesis of LcsG indicate the presence of a highly conserved SAM-binding pocket, along with two possible active site residues (D368 and D395). Molecular dynamics simulations show that the targeted N can dock between these two residues. Thus, this study suggests a method for increasing the variety of natural bioactivity of NPRs and a possible catalytic mechanism underlying the N-methylation of NRPs.


Asunto(s)
Cryptococcus neoformans , Hypocreales , Metiltransferasas , Metiltransferasas/metabolismo , Metiltransferasas/genética , Metiltransferasas/química , Metilación , Hypocreales/enzimología , Hypocreales/genética , Cryptococcus neoformans/enzimología , Cryptococcus neoformans/genética , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Proteínas Fúngicas/química , Simulación de Dinámica Molecular , Phytophthora infestans/enzimología , Phytophthora infestans/genética , Secuencia de Aminoácidos , Mutagénesis Sitio-Dirigida , Dominio Catalítico , Péptidos Catiónicos Antimicrobianos
6.
Int J Mol Sci ; 25(8)2024 Apr 12.
Artículo en Inglés | MEDLINE | ID: mdl-38673861

RESUMEN

Plant-parasitic nematodes (PPNs) are among the most serious phytopathogens and cause widespread and serious damage in major crops. In this study, using a genome mining method, we identified nonribosomal peptide synthetase (NRPS)-like enzymes in genomes of plant-parasitic nematodes, which are conserved with two consecutive reducing domains at the N-terminus (A-T-R1-R2) and homologous to fungal NRPS-like ATRR. We experimentally investigated the roles of the NRPS-like enzyme (MiATRR) in nematode (Meloidogyne incognita) parasitism. Heterologous expression of Miatrr in Saccharomyces cerevisiae can overcome the growth inhibition caused by high concentrations of glycine betaine. RT-qPCR detection shows that Miatrr is significantly upregulated at the early parasitic life stage (J2s in plants) of M. incognita. Host-derived Miatrr RNA interference (RNAi) in Arabidopsis thaliana can significantly decrease the number of galls and egg masses of M. incognita, as well as retard development and reduce the body size of the nematode. Although exogenous glycine betaine and choline have no obvious impact on the survival of free-living M. incognita J2s (pre-parasitic J2s), they impact the performance of the nematode in planta, especially in Miatrr-RNAi plants. Following application of exogenous glycine betaine and choline in the rhizosphere soil of A. thaliana, the numbers of galls and egg masses were obviously reduced by glycine betaine but increased by choline. Based on the knowledge about the function of fungal NRPS-like ATRR and the roles of glycine betaine in host plants and nematodes, we suggest that MiATRR is involved in nematode-plant interaction by acting as a glycine betaine reductase, converting glycine betaine to choline. This may be a universal strategy in plant-parasitic nematodes utilizing NRPS-like ATRR to promote their parasitism on host plants.


Asunto(s)
Arabidopsis , Betaína , Péptido Sintasas , Tylenchoidea , Betaína/metabolismo , Animales , Tylenchoidea/metabolismo , Tylenchoidea/genética , Arabidopsis/parasitología , Arabidopsis/metabolismo , Arabidopsis/genética , Péptido Sintasas/metabolismo , Péptido Sintasas/genética , Interacciones Huésped-Parásitos , Enfermedades de las Plantas/parasitología , Proteínas del Helminto/metabolismo , Proteínas del Helminto/genética , Nematodos/metabolismo , Nematodos/genética
7.
Front Microbiol ; 15: 1385255, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38638906

RESUMEN

Chemical study of the nematicidal biocontrol fungus Pochonia chlamydosporia PC-170 led to discovery of six resorcylic acid lactones (RALs), including three nematicidal glycosylated RALs, monocillin VI glycoside (1), colletogloeolactone A (2) and monocillin II glycoside (3), and three antibacterial non-glycosylated RALs, monocillin VI (4), monocillin IV (5) and monocillin II (6). The planar structure of the new compound monocillin VI glycoside (1) was elucidated using HRESIMS and NMR data, and its monosaccharide configuration was further determined through sugar hydrolysis experiment and GC-MS analysis method. Furthermore, their two biosynthetic-related PKS genes, pchE and pchI, were identified through the gene knockout experiment. The glycosylated RALs 1-3 exhibited nematicidal activity against Meloidogyne incognita, with LC50 values of 94, 152 and 64 µg/mL, respectively, and thus had great potential in the development of new nematicidal natural products to control M. incognita in the future.

8.
Front Microbiol ; 15: 1342584, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38362502

RESUMEN

Trichoderma harzianum exhibits a strong biological control effect on many important plant pathogens, such as Fusarium oxysporum, Botrytis cinerea, and Meloidogyne. However, its biocontrol effectiveness is weakened or reduced under salt stress. The aim of this study was to investigate the molecular response of T. harzianum to salt stress at the whole-genome level. Here, we present a 44.47 Mb near-complete genome assembly of the T. harzianum qt40003 strain for the first time, which was assembled de novo with 7.59 Gb Nanopore sequencing long reads (~170-fold) and 5.2 Gb Illumina short reads (~116-fold). The assembled qt40003 genome contains 12 contigs, with a contig N50 of 4.81 Mb, in which four of the 12 contigs were entirely reconstructed in a single chromosome from telomere to telomere. The qt40003 genome contains 4.27 Mb of repeat sequences and 12,238 protein-coding genes with a BUSCO completeness of 97.5%, indicating the high accuracy and completeness of our gene annotations. Genome-wide transcriptomic analysis was used to investigate gene expression changes related to salt stress in qt40003 at 0, 2% (T2), and 4% (T4) sodium chloride concentrations. A total of 2,937 and 3,527 differentially expressed genes (DEGs) were obtained under T2 and T4 conditions, respectively. GO enrichment analysis showed that the T2-treatment DEGs were highly enriched in detoxification (p < 0.001), while the T4 DEGs were mainly enriched in cell components, mostly in cellular detoxification, cell surface, and cell wall. KEGG metabolic pathway analysis showed that 91 and 173 DEGs were significantly enriched in the T2 and T4 treatments, respectively (p < 0.01), mainly in the glutathione metabolism pathway. We further experimentally analyzed the differentially expressed glutathione transferase genes in the glutathione metabolic pathway, most of which were downregulated (13/15). In addition, we screened 13 genes related to active oxygen clearance, including six upregulated and seven downregulated genes, alongside five fungal hydrophobic proteins, of which two genes were highly expressed. Our study provides high-quality genome information for the use of T. harzianum for biological control and offers significant insights into the molecular responses of T. harzianum under salt-stress conditions.

9.
J Fungi (Basel) ; 10(1)2024 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-38248977

RESUMEN

The endoparasitic fungus Hirsutella rhossiliensis is an important biocontrol agent of cyst nematodes in nature. To determine the potential parasitism of the fungus on a non-natural host, the pinewood nematode (Bursaphelenchus xylophilus) living in pine trees and the endophytic ability of the fungus on plants, in this paper, we first constructed and utilized a green fluorescent protein (GFP)-tagged H. rhossiliensis HR02 transformant to observe the fungal infection process on B. xylophilus and its colonization on Arabidopsis roots. Then, we compared the fungal parasitism on three species of nematodes with different lifestyles, and we found that the fungal parasitism is correlated with nematode species and stages. The parasitic effect of H. rhossiliensis on adults of B. xylophilus is similar to that on second-stage juveniles (J2) of the root-knot nematode Meloidogyne incognita after 24 h of inoculation, although the virulence of the fungus to second-stage juveniles of M. incognita is stronger than that to those of B. xylophilus and Caenorhabditis elegans. Moreover, the endophytism of H. rhossiliensis was confirmed. By applying an appropriate concentration of H. rhossiliensis conidial suspension (5 × 106 spores/mL) in rhizosphere soil, it was found that the endophytic fungus can promote A. thaliana growth and reproduction, as well as improve host resistance against M. incognita. Our results provide a deeper understanding of the fungus H. rhossiliensis as a promising biocontrol agent against plant-parasitic nematodes.

10.
Plant Commun ; 5(2): 100723, 2024 Feb 12.
Artículo en Inglés | MEDLINE | ID: mdl-37742073

RESUMEN

Root-knot nematodes (RKNs) cause huge agricultural losses every year. They secrete a repertoire of effectors to facilitate parasitism through the induction of plant-derived giant feeding cells, which serve as their sole source of nutrients. However, the mode of action of these effectors and their targeted host proteins remain largely unknown. In this study, we investigated the role of the effector Mi2G02 in Meloidogyne incognita parasitism. Host-derived Mi2G02 RNA interference in Arabidopsis thaliana affected giant cell development, whereas ectopic expression of Mi2G02 promoted root growth and increased plant susceptibility to M. incognita. We used various combinations of approaches to study the specific interactions between Mi2G02 and A. thaliana GT-3a, a trihelix transcription factor. GT-3a knockout in A. thaliana affected feeding-site development, resulting in production of fewer egg masses, whereas GT-3a overexpression in A. thaliana increased susceptibility to M. incognita and also root growth. Moreover, we demonstrated that Mi2G02 plays a role in maintaining GT-3a protein stabilization by inhibiting the 26S proteasome-dependent pathway, leading to suppression of TOZ and RAD23C expression and thus promoting nematode parasitism. This work enhances our understanding of how a pathogen effector manipulates the role and regulation of a transcription factor by interfering with a proteolysis pathway to reprogram gene expression for development of nematode feeding cells.


Asunto(s)
Arabidopsis , Nematodos , Animales , Factores de Transcripción/genética , Arabidopsis/genética , Plantas Modificadas Genéticamente , Interferencia de ARN , Nematodos/genética
11.
Front Plant Sci ; 14: 1163271, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37324672

RESUMEN

Introduction: Root-knot nematodes (RKN) disease is a devastating disease in Cucumis crops production. Existing studies have shown that resistant and susceptible crops are enriched with different rhizosphere microorganisms, and microorganisms enriched in resistant crops can antagonize pathogenic bacteria. However, the characteristics of rhizosphere microbial communities of Cucumis crops after RKN infestation remain largely unknown. Methods: In this study, we compared the changes in rhizosphere bacterial communities between highly RKN-resistant Cucumis metuliferus (cm3) and highly RKN-susceptible Cucumis sativus (cuc) after RKN infection through a pot experiment. Results: The results showed that the strongest response of rhizosphere bacterial communities of Cucumis crops to RKN infestation occurred during early growth, as evidenced by changes in species diversity and community composition. However, the more stable structure of the rhizosphere bacterial community in cm3 was reflected in less changes in species diversity and community composition after RKN infestation, forming a more complex and positively co-occurrence network than cuc. Moreover, we observed that both cm3 and cuc recruited bacteria after RKN infestation, but the bacteria enriched in cm3 were more abundant including beneficial bacteria Acidobacteria, Nocardioidaceae and Sphingomonadales. In addition, the cuc was enriched with beneficial bacteria Actinobacteria, Bacilli and Cyanobacteria. We also found that more antagonistic bacteria than cuc were screened in cm3 after RKN infestation and most of them were Pseudomonas (Proteobacteria, Pseudomonadaceae), and Proteobacteria were also enriched in cm3 after RKN infestation. We hypothesized that the cooperation between Pseudomonas and the beneficial bacteria in cm3 could inhibit the infestation of RKN. Discussion: Thus, our results provide valuable insights into the role of rhizosphere bacterial communities on RKN diseases of Cucumis crops, and further studies are needed to clarify the bacterial communities that suppress RKN in Cucumis crops rhizosphere.

12.
J Fungi (Basel) ; 9(5)2023 May 19.
Artículo en Inglés | MEDLINE | ID: mdl-37233306

RESUMEN

Trichoderma hamatum is a filamentous fungus that serves as a biological control agent for multiple phytopathogens and as an important resource promising for fungicides. However, the lack of adequate knockout technologies has hindered gene function and biocontrol mechanism research of this species. This study obtained a genome assembly of T. hamatum T21, with a 41.4 Mb genome sequence comprising 8170 genes. Based on genomic information, we established a CRISPR/Cas9 system with dual sgRNAs targets and dual screening markers. CRISPR/Cas9 plasmid and donor DNA recombinant plasmid were constructed for disruption of the Thpyr4 and Thpks1 genes. The result indicates the consistency between phenotypic characterization and molecular identification of the knockout strains. The knockout efficiencies of Thpyr4 and Thpks1 were 100% and 89.1%, respectively. Moreover, sequencing revealed fragment deletions between dual sgRNA target sites or GFP gene insertions presented in knockout strains. The situations were caused by different DNA repair mechanisms, nonhomologous end joining (NHEJ), and homologous recombination (HR). Overall, we have successfully constructed an efficient and convenient CRISPR/Cas9 system in T. hamatum for the first time, which has important scientific significance and application value for studies on functional genomics of Trichoderma and other filamentous fungi.

13.
Plant Biotechnol J ; 21(7): 1496-1509, 2023 07.
Artículo en Inglés | MEDLINE | ID: mdl-37074757

RESUMEN

Root-knot nematodes (RKNs) are infamous plant pathogens in tomato production, causing considerable losses in agriculture worldwide. Mi-1 is the only commercially available RKN-resistance gene; however, the resistance is inactivated when the soil temperature is over 28 °C. Mi-9 in wild tomato (Solanum arcanum LA2157) has stable resistance to RKNs under high temperature but has not been cloned and applied. In this study, a chromosome-scale genome assembly of S. arcanum LA2157 was constructed through Nanopore and Hi-C sequencing. Based on molecular markers of Mi-9 and comparative genomic analysis, the localization region and candidate Mi-9 genes cluster consisting of seven nucleotide-binding sites and leucine-rich repeat (NBS-LRR) genes were located. Transcriptional expression profiles confirmed that five of the seven candidate genes were expressed in root tissue. Moreover, virus-induced gene silencing of the Sarc_034200 gene resulted in increased susceptibility of S. arcanum LA2157 to Meloidogyne incognita, and genetic transformation of the Sarc_034200 gene in susceptible Solanum pimpinellifolium conferred significant resistance to M. incognita at 25 °C and 30 °C and showed hypersensitive responses at nematode infection sites. This suggested that Sarc_034200 is the Mi-9 gene. In summary, we cloned, confirmed and applied the heat-stable RKN-resistance gene Mi-9, which is of great significance to tomato breeding for nematode resistance.


Asunto(s)
Solanum lycopersicum , Solanum , Tylenchoidea , Animales , Solanum/genética , Calor , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Fitomejoramiento , Solanum lycopersicum/genética , Cromosomas/metabolismo , Raíces de Plantas/genética , Enfermedades de las Plantas/genética
14.
Front Plant Sci ; 14: 1143171, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37021316

RESUMEN

Root-knot nematode (RKN) is a major factor that limits the growth and productivity of important Cucumis crops, such as cucumber and melon, which lack RKN-resistance genes in their genome. Cucumis metuliferus is a wild Cucumis species that displays a high degree of RKN-resistance. WRKY transcription factors were involved in plant response to biotic stresses. However, little is known on the function of WRKY genes in response to RKN infection in Cucumis crops. In this study, Cucumis metuliferus 60 WRKY genes (CmWRKY) were identified in the C. metuliferus genome, and their conserved domains were classified into three main groups based on multiple sequence alignment and phylogenetic analysis. Synteny analysis indicated that the WRKY genes were highly conserved in Cucumis crops. Transcriptome data from of C. metuliferus roots inoculated with RKN revealed that 16 CmWRKY genes showed differential expression, of which 13 genes were upregulated and three genes were downregulated, indicating that these CmWRKY genes are important to C. metuliferus response to RKN infection. Two differentially expression CmWRKY genes (CmWRKY10 and CmWRKY28) were selected for further functional analysis. Both CmWRKY genes were localized in nucleus, indicating they may play roles in transcriptional regulation. This study provides a foundation for further research on the function of CmWRKY genes in RKN stress resistance and elucidation of the regulatory mechanism.

15.
J Appl Microbiol ; 134(2)2023 Feb 16.
Artículo en Inglés | MEDLINE | ID: mdl-36611228

RESUMEN

AIMS: Root-knot nematodes (RKNs) are plant pathogens that cause huge economic losses worldwide. The biological management of RKNs may be a sustainable alternative to chemical control methods. Here, the biocontrol potential of Methylorubrum rhodesianum M520 against the RKN Meloidogyne incognita was investigated to theoretically support its application as a biocontrol agent in field production. METHODS AND RESULTS: In-vitro assays showed 91.9% mortality of M. incognita second-stage juveniles in the presence of strain M520 and that the hatching rate of M. incognita eggs was 21.7% lower than that of eggs treated with sterile water. In pot experiments, the M520 treatment caused 70.8% reduction in root-knots and increased plant shoot length and stem and root fresh weights, compared to control plant values. In split-root experiments, cucumber roots treated with M520 showed 25.6% decrease in root gall number, compared to that in control roots. CONCLUSION: M520 has multiple mechanisms against RKNs and might be used as a biocontrol agent against M. incognita in cucumber, laying a foundation for further studying M520 biocontrol against RKNs.


Asunto(s)
Cucumis sativus , Methylobacteriaceae , Tylenchida , Tylenchoidea , Animales , Raíces de Plantas
16.
J Fungi (Basel) ; 9(1)2023 Jan 08.
Artículo en Inglés | MEDLINE | ID: mdl-36675913

RESUMEN

An effective selection marker is necessary for genetic engineering and functional genomics research in the post-genomic era. Isaria javanica is an important entomopathogenic fungus with a broad host range and prospective biocontrol potentials. Given that no antibiotic marker is available currently in this fungus, developing an effective selection marker is necessary. In this study, by applying overlap PCR and split-marker deletion strategy, combining PEG-mediated protoplasm transformation method, the uridine auxotrophy gene (ura3) in the I. javanica genome was knocked out. Then, using this transformation system, the pH response transcription factor gene (IjpacC) was disrupted successfully. Loss of IjpacC gene results in an obvious decrease in conidial production, but little impact on mycelial growth. The virulence of the ΔIjpacC mutant on caterpillars is similar to that of the wild-type strain. RT-qPCR detection shows that expression level of an acidic-expressed S53 gene (IF1G_06234) in ΔIjpacC mutant is more significantly upregulated than in the wild-type strain during the fungal infection on caterpillars. Our results indicate that a markerless transformation system based upon complementation of uridine auxotrophy is successfully developed in I. javanica, which is useful for exploring gene function and for genetic engineering to enhance biological control potential of the fungus.

17.
Int J Mol Sci ; 24(2)2023 Jan 04.
Artículo en Inglés | MEDLINE | ID: mdl-36674475

RESUMEN

Fusarium oxysporum f. sp. phaseoli, the causal agent of cowpea fusarium wilt, is a serious threat to cowpea production in China. In this study, a sample of cowpea fusarium wilt was identified as Fusarium oxysporum f. sp. phaseoli using the methods of morphological characters and molecular detection. We further reported the first genome assembly for Fusarium oxysporum f. sp. phaseoli, with 53.7 Mb genome sequence comprising 14,694 genes. Comparative genomic analysis among five Fusarium oxysporum genomes showed that four accessory chromosomes in the five Fusarium oxysporum display similar characteristics, with low sequence similarity (55.35%, vs. overall average of 81.76%), low gene density (2.18 genes/10 kb vs. 3.02 genes/Mb) and highly transposable element density (TEs) (15.01/100 kb vs. 4.89/100 kb), indicating that variable accessory chromosomes are the main source of Fusarium oxysporum evolution. We identified a total of 100 Fusarium oxysporum f. sp. phaseoli-specific effectors in the genome and found 13 specific effector genes located in large insertion or deletion regions, suggesting that insertion or deletion events can cause the emergence of species-specific effectors in Fusarium oxysporum. Our genome assembly of Fusarium oxysporum f. sp. phaseoli provides a valuable resource for the study of cowpea fusarium wilt, and the comparative genomic study of Fusarium oxysporum could contribute to the knowledge of genome and effector-associated pathogenicity evolution in Fusarium oxysporum study.


Asunto(s)
Fusarium , Fusarium/genética , Enfermedades de las Plantas , Genoma Fúngico
18.
Artículo en Inglés | MEDLINE | ID: mdl-36554518

RESUMEN

With increasing water resources stress under climate change, it is of great importance to deeply understand the spatio-temporal variation of crop water requirements and their response to climate change for achieving better water resources management and grain production. However, the quantitative evaluation of climate change impacts on crop water requirements and the identification of determining factors should be further explored to reveal the influencing mechanism and actual effects thoroughly. In this study, the water requirements of winter wheat and summer maize from 1981 to 2019 in the lower reaches of the Yellow River Basin were estimated based on the Penman-Monteith model and crop coefficient method using daily meteorological data. Combined with trends test, sensitivity and contribution analysis, the impacts of different meteorological factors on crop water requirement variation were explored, and the dominant factors were then identified. The results indicated that the temperature increased significantly (a significance level of 0.05 was considered), whereas the sunshine duration, relative humidity and wind speed decreased significantly from 1981 to 2019 in the study area. The total water requirements of winter wheat and summer maize presented a significant decreasing trend (-1.36 mm/a) from 1981 to 2019 with a multi-year average value of 936.7 mm. The crop water requirements of winter wheat was higher than that of summer maize, with multi-year average values of 546.6 mm and 390.1 mm, respectively. In terms of spatial distribution patterns, the crop water requirement in the north was generally higher than that in the south. The water requirements of winter wheat and summer maize were most sensitive to wind speed, and were less sensitive to the minimum temperature and relative humidity. Wind speed was the leading factor of crop water requirement variation with the highest contribution rate of 116.26% among the considered meteorological factors. The results of this study will provide important support for strengthening the capacity to cope with climate change and realizing sustainable utilization of agricultural water resources in the lower reaches of the Yellow River Basin.


Asunto(s)
Triticum , Zea mays , Triticum/fisiología , Zea mays/fisiología , Cambio Climático , Ríos , Productos Agrícolas , Agua , China
19.
Sci Data ; 9(1): 748, 2022 12 03.
Artículo en Inglés | MEDLINE | ID: mdl-36463249

RESUMEN

The potato tuberworm, Phthorimaea operculella Zeller, is an oligophagous pest feeding on crops mainly belonging to the family Solanaceae. It is one of the most destructive pests of potato worldwide and attacks foliage and tubers in the field and in storage. However, the lack of a high-quality reference genome has hindered the association of phenotypic traits with their genetic basis. Here, we report on the genome assembly of P. operculella at the chromosomal level. Using Illumina, Nanopore and Hi-C sequencing, a 648.2 Mb genome was generated from 665 contigs, with an N50 length of 3.2 Mb, and 92.0% (596/648.2 Mb) of the assembly was anchored to 29 chromosomes. In total, 16619 genes were annotated, and 92.4% of BUSCO genes were fully represented. The chromosome-level genome of P. operculella will provide a significant resource for understanding the genetic basis for the biological study of this insect, and for promoting the integrative management of this pest in future.


Asunto(s)
Cromosomas , Mariposas Nocturnas , Solanum tuberosum , Secuenciación de Nucleótidos de Alto Rendimiento , Tubérculos de la Planta/parasitología , Solanum tuberosum/parasitología , Mariposas Nocturnas/genética , Animales
20.
J Fungi (Basel) ; 8(10)2022 Oct 16.
Artículo en Inglés | MEDLINE | ID: mdl-36294654

RESUMEN

Fusarium oxysporum f. sp. conglutinans (FOC), the causal agent of cabbage fusarium wilt, is a serious threat to cabbage production in northern China, and most Chinese FOC isolates were identified as FOC race 1 (FOC1). To better understand the genetic diversity of FOC1 in northern China, we collected FOC isolates from five provinces in northern China and identified them as FOC1 through pathogenicity and race test. To evaluate the genome-level diversity of FOC1, we performed a genome assembly for a FOC1 isolate (FoYQ-1) collected from Yanqing, Beijing, where cabbage fusarium wilt was first reported in China. Using resequencing data of FOC1 isolates, we conducted a genome-wide SNP (single nucleotide polymorphism) analysis to investigate the genetic diversity and population structure of FOC1 isolates in northern China. Our study indicated that Chinese FOC1 can be grouped into four populations and revealed that the genetic diversity of FOC1 were closely associated with geographical locations. Our study further suggests that genetic differentiation occurred when FOC1 spread to the northwest provinces from Beijing Province in China. The FOC1 genetic diversity based on whole-genome SNPs could deepen our understanding of FOC1 variation and provide clues for the control of cabbage fusarium wilt in China.

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