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International Journal of Molecular... Jan 2024is the major pathogen responsible for the significant stem disease "blight" in × . The interacting proteins of the key pathogenic factor , BDUbc and BDSKL1, have...
is the major pathogen responsible for the significant stem disease "blight" in × . The interacting proteins of the key pathogenic factor , BDUbc and BDSKL1, have previously been obtained by two-hybrid, BiFC, GST pull-down yeast assays. However, the functions of these interacting proteins remain unknown. This study successfully obtained transgenic plants overexpressing , , and + via Agrobacterium-mediated gene overexpression. qRT-PCR analysis revealed significantly increased expression levels of and in the transgenic plants. After infection with the pathogenic spore suspension, the disease incidence and severity index significantly decreased across all three transgenic plants, accompanied by a marked increase in defense enzyme levels. Notably, the co-transformed plant, OE- + , demonstrated the lowest disease incidence and severity index among the transgenic variants. These results not only indicate that and are disease-resistant genes, but also that these two genes may exhibit a synergistic enhancement effect, which further improves the resistance to blight in × .
Topics: Bambusa; Agrobacterium; Biological Assay; Keratoconjunctivitis; Plants, Genetically Modified; Saccharomyces cerevisiae
PubMed: 38203739
DOI: 10.3390/ijms25010569 -
Biomolecules Mar 2023The study of interaction proteins of the pathogen effector protein is an important means to analyze the disease-resistance mechanism of shoot blight. To obtain the...
The study of interaction proteins of the pathogen effector protein is an important means to analyze the disease-resistance mechanism of shoot blight. To obtain the proteins interacting with the effector ApCE22 of , 27 proteins interacting with the effector ApCE22 were initially identified via a yeast two-hybrid assay, of which four interaction proteins were obtained after one-to-one validation. The B2 protein and the chaperone protein DnaJ chloroplast protein were then verified to interact with the ApCE22 effector protein by bimolecular fluorescence complementation and GST pull-down methods. Advanced structure prediction showed that the B2 protein contained the DCD functional domain related to plant development and cell death, and the DnaJ protein contained the DnaJ domain related to stress resistance. The results showed that both the B2 protein and DnaJ protein in were the target interaction proteins of the ApCE22 effector of and related to the stress resistance of the host . The successful identification of the pathogen effector interaction target protein in plays an important role in the mechanism of pathogen-host interaction, thus providing a theoretical basis for the control of shoot blight.
Topics: Bambusa; HSP40 Heat-Shock Proteins; Ascomycota; Host-Pathogen Interactions
PubMed: 37189340
DOI: 10.3390/biom13040590 -
Frontiers in Plant Science 2022is the main pathogen that causes blight. It secretes the cutinase transcription factor , which has been shown to play an important role in virulence. However,...
is the main pathogen that causes blight. It secretes the cutinase transcription factor , which has been shown to play an important role in virulence. However, knowledge about the interaction target genes of in remains limited. A cDNA library for the yeast two-hybrid system was constructed from shoots after 168 h treatment with . The library was identified as 1.20 × 10 cfu, with an average insert >1,000 bp in size and a 100% positive rate, providing a database for the subsequent molecular study of the interaction between and . The yeast two-hybrid (Y2H), bimolecular fluorescence complementation (BiFC), and glutathione-S-transferase (GST) pull-down assays were used to screen for and identify two interacting target proteins, and , providing a reliable theoretical basis to study the molecular mechanism underlying resistance in response to , which would, in turn, establish a platform to develop new strategies for the sustainable and effective control of the blight diseases of forest trees.
PubMed: 36186076
DOI: 10.3389/fpls.2022.991077 -
Biomolecules Sep 2022The shoot blight of × caused by made bamboo die in a large area, resulting in serious ecological and economic losses. Dual RNA-seq was used to sequence and analyze...
The shoot blight of × caused by made bamboo die in a large area, resulting in serious ecological and economic losses. Dual RNA-seq was used to sequence and analyze the transcriptome data of and × in the four periods after the pathogen infected the host and to screen the candidate effectors of the pathogen related to the infection. After the identification of the effectors by the tobacco transient expression system, the functions of these effectors were verified by gene knockout. Fifty-three differentially expressed candidate effectors were obtained by differential gene expression analysis and effector prediction. Among them, the effectors and can cause programmed cell death in tobacco. The disease index of × inoculated with mutant Δ and mutant Δ strains were 52.5% and 47.5%, respectively, which was significantly lower than that of the wild-type strains (80%), the complementary strain (77.5%), and the complementary strain (75%). The tolerance of the mutant Δ and mutant Δ strains to HO and NaCl stress was significantly lower than that of the wild-type strain and the complementary and complementary strains, but there was no difference in their tolerance to Congo red. Therefore, this study shows that the effectors and play an important role in virulence and response to HO and NaCl stress.
Topics: Ascomycota; Bambusa; Congo Red; Hydrogen Peroxide; Plant Diseases; Sodium Chloride; Nicotiana
PubMed: 36139102
DOI: 10.3390/biom12091264 -
International Journal of Molecular... Jun 2022shoot blight caused by is a fungal disease that has affected a large area in China in recent years. However, it is not clear which genes are responsible for the...
shoot blight caused by is a fungal disease that has affected a large area in China in recent years. However, it is not clear which genes are responsible for the disease resistance of × . Based on the analysis of transcriptome and proteome data, two genes, and , which may be involved in disease resistance, were screened. Two gene expression-interfering varieties, COF RNAi and CAD RNAi were successfully obtained using RNAi technology. Quantitative real-time fluorescence (qRT-PCR) results showed that gene, gene and seven related genes expression was down-regulated in the transformed varieties. After inoculating pathogen spore suspension, the incidence and disease index of cof-RNAi and cad-RNAi transformed plants increased significantly. At the same time, it was found that the content of total lignin and flavonoids in the two transformed varieties were significantly lower than that of the wild-type. The subcellular localization results showed that both CCoAOMT2 and CAD5 were localized in the nucleus and cytoplasm. The above results confirm that the and genes are involved in the resistance of to shoot blight through regulating the synthesis of lignin and flavonoids.
Topics: Bambusa; Disease Resistance; Flavonoids; Gene Expression Regulation, Plant; Lignin; Transcriptome
PubMed: 35743217
DOI: 10.3390/ijms23126760 -
Journal of Fungi (Basel, Switzerland) Nov 2021can cause branch wilting of , causing great economic losses and ecological damage. was sequenced in sterile deionized water (CK), rice tissue (T1) and (T2) fluid by...
can cause branch wilting of , causing great economic losses and ecological damage. was sequenced in sterile deionized water (CK), rice tissue (T1) and (T2) fluid by RNA-Seq, and the function of 1 and was verified by gene knockout. There were 424, 471 and 396 differentially expressed genes between the T2 and CK, T2 and T1, and CK and T1 groups, respectively. Thirty DEGs had verified the change in expression by fluorescent quantitative PCR. Twenty-nine DEGs were the same as the expression level in RNA-Seq. In addition, ΔApCtf1β 1 and ΔApCtf1β 2 showed weaker virulence by gene knockout, and the complementary strains Ctf1β 1 and Ctf1β 2 showed the same virulence as the wild-type strains. Relative growth inhibition of ΔApCtf1β 1 and ΔApCtf1β was significantly decreased by 21.4% and 19.2%, respectively, by adding HO compared to the estimates from the wild-type strain and decreased by 25% and 19.4%, respectively, by adding Congo red. The disease index of infected by two mutants was significantly lower than that of wild type. This suggested that genes are required for the stress response and virulence of .
PubMed: 34946984
DOI: 10.3390/jof7121001 -
PeerJ 2021is a fast-growing bamboo that is widely introduced in southern China and has great economic and ecological benefits. In recent years, a blight of × caused by has...
BACKGROUND
is a fast-growing bamboo that is widely introduced in southern China and has great economic and ecological benefits. In recent years, a blight of × caused by has led to much branch damage and even death of entire bamboo forests.
METHODS
To screen for resistance genes in × , transcriptome sequencing technology was used to compare the gene expression profiles of different varieties of × with variable resistance and the same varieties under different treatments. The Clusters of Orthologous Groups of Proteins (COG) database; the Gene Ontology (GO) database; and the Kyoto Encyclopedia of Genes and Genomes (KEGG) database were used to annotate and analyse the differentially expressed genes.
RESULTS
A total of 26,157 and 11,648 differentially expressed genes were obtained in the different varieties after inoculation with and the same varieties after inoculation or sterile water, respectively. There were 23 co-upregulated DGEs and 143 co-downregulated DEGs in #3 and #8, #6 and #8, #6 and #3. There were 50 co-upregulated DGEs and 24 co-downregulated DEGs in the same varieties after inoculation or sterile water. The results showed that many genes involved in cell wall composition synthesis, redox reactions and signal transduction were significantly different after pathogen infection. Twenty-one candidate genes for blight resistance, such as , , , and , were found. The qRT-PCR results were consistent with the sequencing results, verifying their authenticity. These results provide a foundation for the further exploration of resistance genes and their functions.
PubMed: 34721984
DOI: 10.7717/peerj.12301 -
Scientific Reports Dec 2019In this study, TMT (tandem mass tag)-labeled quantitative protein technology combined with LC-MS/MS (liquid chromatography-mass spectrometry/mass spectrometry) was used...
In this study, TMT (tandem mass tag)-labeled quantitative protein technology combined with LC-MS/MS (liquid chromatography-mass spectrometry/mass spectrometry) was used to isolate and identify the proteins of the hybrid bamboo (Bambusa pervariabilis × Dendrocalamopsis grandis) and the bamboo inoculated with the pathogenic fungi Arthrinium phaeospermum. A total of 3320 unique peptide fragments were identified after inoculation with either A. phaeospermum or sterile water, and 1791 proteins were quantified. A total of 102 differentially expressed proteins were obtained, of which 66 differential proteins were upregulated and 36 downregulated in the treatment group. Annotation and enrichment analysis of these peptides and proteins using the GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) databases with bioinformatics software showed that the differentially expressed protein functional annotation items were mainly concentrated on biological processes and cell components. The LC-PRM/MS (liquid chromatography-parallel reaction monitoring/mass spectrometry) quantitative analysis technique was used to quantitatively analyze 11 differential candidate proteins obtained by TMT combined with LC-MS/MS. The up-down trend of 10 differential proteins in the PRM results was consistent with that of the TMT quantitative analysis. The coincidence rate of the two results was 91%, which confirmed the reliability of the proteomic results. Therefore, the differentially expressed proteins and signaling pathways discovered here may be the further concern for the bamboo-pathogen interaction studies.
Topics: Bambusa; China; Chromatography, Liquid; Computational Biology; Crosses, Genetic; Gene Ontology; Hydrolysis; Peptides; Plant Diseases; Proteome; Proteomics; Reproducibility of Results; Tandem Mass Spectrometry; Up-Regulation; Water; Xylariales
PubMed: 31822726
DOI: 10.1038/s41598-019-55229-0 -
PeerJ 2019Water is an increasingly scarce resource while some crops, such as paddy rice, require large amounts of water to maintain grain production. A better understanding of...
BACKGROUND AND AIM
Water is an increasingly scarce resource while some crops, such as paddy rice, require large amounts of water to maintain grain production. A better understanding of rice drought adaptation and tolerance mechanisms could help to reduce this problem. There is evidence of a possible role of root-associated fungi in drought adaptation. Here, we analyzed the endospheric fungal microbiota composition in rice and its relation to plant genotype and drought.
METHODS
Fifteen rice genotypes ( ssp. indica) were grown in the field, under well-watered conditions or exposed to a drought period during flowering. The effect of genotype and treatment on the root fungal microbiota composition was analyzed by 18S ribosomal DNA high throughput sequencing. Grain yield was determined after plant maturation.
RESULTS
There was a host genotype effect on the fungal community composition. Drought altered the composition of the root-associated fungal community and increased fungal biodiversity. The majority of OTUs identified belonged to the Pezizomycotina subphylum and 37 of these significantly correlated with a higher plant yield under drought, one of them being assigned to .
CONCLUSION
This study shows that both plant genotype and drought affect the root-associated fungal community in rice and that some fungi correlate with improved drought tolerance. This work opens new opportunities for basic research on the understanding of how the host affects microbiota recruitment as well as the possible use of specific fungi to improve drought tolerance in rice.
PubMed: 31565550
DOI: 10.7717/peerj.7463 -
Metabolites Aug 2019McClure × (Q.H.Dai & X.l.Tao ex Keng f.) Ohrnb. blight is a widespread and dangerous forest fungus disease, and has been listed as a supplementary object of forest...
McClure × (Q.H.Dai & X.l.Tao ex Keng f.) Ohrnb. blight is a widespread and dangerous forest fungus disease, and has been listed as a supplementary object of forest phytosanitary measures. In order to study the control of × blight, this experiment was carried out. In this work, a toxin purified from the pathogen (Corda) Elli, which causes blight in × , with homologous heterogeneity, was used as an inducer to increase resistance to × . A functional analysis of the differentially expressed proteins after induction using a tandem mass tag labeling technique was combined with mass spectrometry and liquid chromatography mass spectrometry in order to effectively screen for the proteins related to the resistance of × to blight. After peptide labeling, a total of 3320 unique peptides and 1791 quantitative proteins were obtained by liquid chromatography mass spectrometry analysis. Annotation and enrichment analysis of these peptides and proteins using the Gene ontology and Kyoto Encyclopedia of Genes and Genomes databases with bioinformatics software show that the differentially expressed protein functional annotation items are mainly concentrated on biological processes and cell components. Several pathways that are prominent in the Kyoto Encyclopedia of Genes and Genomes annotation and enrichment include metabolic pathways, the citrate cycle, and phenylpropanoid biosynthesis. In the Protein-protein interaction networks four differentially expressed proteins-sucrose synthase, adenosine triphosphate-citrate synthase beta chain protein 1, peroxidase, and phenylalanine ammonia-lyase significantly interact with multiple proteins and significantly enrich metabolic pathways. To verify the results of tandem mass tag, the candidate proteins were further verified by parallel reaction monitoring, and the results were consistent with the tandem mass tag data analysis results. It is confirmed that the data obtained by tandem mass tag technology are reliable. Therefore, the differentially expressed proteins and signaling pathways discovered here is the primary concern for subsequent disease resistance studies.
PubMed: 31405188
DOI: 10.3390/metabo9080166