scholarly journals Endopolygalacturonase Genes from Colletotrichum lindemuthianum: Cloning of CLPG2 and Comparison of Its Expression to That of CLPG1 During Saprophytic and Parasitic Growth of the Fungus

1997 ◽  
Vol 10 (6) ◽  
pp. 769-775 ◽  
Author(s):  
Sylvie Centis ◽  
Isabelle Guillas ◽  
Nathalie Séjalon ◽  
Marie-Thérèse Esquerré-Tugayé ◽  
Bernard Dumas
Keyword(s):  
Synthetic Medium ◽  
Amino Acid Level ◽  
Single Copy ◽  
Colletotrichum Lindemuthianum ◽  
In Planta ◽  
Gene Encoding ◽  
Host Cell Wall ◽  
Extensive Degradation ◽  
Necrotrophic Stage

Following the previous isolation of CLPG1, a gene encoding an endopolygalacturonase (endoPG) secreted into the culture filtrate of Colletotrichum lindemuthianum, we have isolated and sequenced an additional endoPG gene, CLPG2. This gene is present as a single copy in the genome of the fungus. At the amino acid level, CLPG2 shows 61% identity to CLPG1 and between 37 to 59% identity to other fungal endoPGs. RNA blot analyses of endoPG gene expression were followed with specific probes during in vitro culture of the fungus. When conidia were used to inoculate a synthetic medium containing pectin as sole carbon source, only CLPG1 was found to be expressed after 3 days of culture. However, transferring the mycelium grown on glucose for 4 days to a pectin-containing medium allowed the detection of CLPG1 and CLPG2 transcripts as early as 12 h after transfer on this substrate. Expression of CLPG2 was transient while that of CLPG1 was more prolonged. Immunocytological localization of endoPG in C. lindemuthianum-infected bean tissues with antibodies against CLPG1 confirmed that the protein is produced in planta and is associated with extensive degradation of the host cell wall. Detection of endoPG transcripts by reverse transcription-polymerase chain reaction revealed that CLPG1, but not CLPG2, is expressed at the beginning of the necrotrophic stage of infection. These results show that the two endoPG genes are differentially expressed and that CLPG1 encodes the major secreted endoPG both during saprophytic growth and during plant infection.

scholarly journals The Multifunctional β-Oxidation Enzyme Is Required for Full Symptom Development by the Biotrophic Maize Pathogen Ustilago maydis

2006 ◽  
Vol 5 (12) ◽  
pp. 2047-2061 ◽  
Author(s):  
Jana Klose ◽  
James W. Kronstad
Keyword(s):  
Fatty Acids ◽  
Ustilago Maydis ◽  
Long Chain Fatty Acids ◽  
In Planta ◽  
Gene Encoding ◽  
Metabolic Role ◽  
Fungal Phytopathogen ◽  
Long Chain ◽  
Chain Fatty Acids

ABSTRACT The transition from yeast-like to filamentous growth in the biotrophic fungal phytopathogen Ustilago maydis is a crucial event for pathogenesis. Previously, we showed that fatty acids induce filamentation in U. maydis and that the resulting hyphal cells resemble the infectious filaments observed in planta. To explore the potential metabolic role of lipids in the morphological transition and in pathogenic development in host tissue, we deleted the mfe2 gene encoding the multifunctional enzyme that catalyzes the second and third reactions in β-oxidation of fatty acids in peroxisomes. The growth of the strains defective in mfe2 was attenuated on long-chain fatty acids and abolished on very-long-chain fatty acids. The mfe2 gene was not generally required for the production of filaments during mating in vitro, but loss of the gene blocked extensive proliferation of fungal filaments in planta. Consistent with this observation, mfe2 mutants exhibited significantly reduced virulence in that only 27% of infected seedlings produced tumors compared to 88% tumor production upon infection by wild-type strains. Similarly, a defect in virulence was observed in developing ears upon infection of mature maize plants. Specifically, the absence of the mfe2 gene delayed the development of teliospores within mature tumor tissue. Overall, these results indicate that the ability to utilize host lipids contributes to the pathogenic development of U. maydis.

scholarly journals Disruption of the ech42 (Endochitinase-Encoding) Gene Affects Biocontrol Activity in Trichoderma harzianum P1

1999 ◽  
Vol 12 (5) ◽  
pp. 419-429 ◽  
Author(s):  
S. L. Woo ◽  
B. Donzelli ◽  
F. Scala ◽  
R. Mach ◽  
G. E. Harman ◽  
...  
Keyword(s):  
Antifungal Activity ◽  
Trichoderma Harzianum ◽  
Pythium Ultimum ◽  
Single Copy ◽  
Wild Type ◽  
Gene Encoding ◽  
Culture Filtrates ◽  
In Vitro Antifungal Activity ◽  
Better Than

The biocontrol strain P1 of Trichoderma harzianum was genetically modified by targeted disruption of the single-copy ech42 gene encoding for the secreted 42-kDa endochitinase (CHIT42). Stable mutants in which ech42 was interrupted, and unable to produce CHIT42, were obtained and characterized. These mutants lacked the ech42 transcript, the protein, and endochitinase activity in culture filtrates, and they were unable to clear a medium containing colloidal chitin. Other chitinolytic and glucanolytic enzymes expressed during mycoparasitism were not affected by the disruption of ech42. The disrupted mutant D11 grew and sporulated similarly to the wild type. In vitro antifungal activity of the ech42 disruptant culture filtrates against Botrytis cinerea and Rhizoctonia solani was reduced about 40%, compared with wild type; antifungal activity was fully restored by adding an equivalent amount of CHIT42 as secreted by P1. The mutant exhibited the same biocontrol effect against Pythium ultimum as strain P1, but the antagonism against B. cinerea on bean leaves by the mutant was significantly reduced (33% less biocontrol), compared with strain P1. Conversely, the endochitinase-deficient mutant performed better than the wild type (16% improvement of survival) in biocontrol experiments in soil infested with the soilborne fungus R. solani. These results indicate that the antagonistic interaction between the T. harzianum strain and various fungal hosts is based on different mechanisms.

Use of Green Fluorescent Protein To Detect Expression of an Endopolygalacturonase Gene of Colletotrichum lindemuthianum during Bean Infection

1999 ◽  
Vol 65 (4) ◽  
pp. 1769-1771 ◽  
Author(s):  
Bernard Dumas ◽  
Sylvie Centis ◽  
Nathalie Sarrazin ◽  
Marie-Thérèse Esquerré-Tugayé
Keyword(s):  
Green Fluorescent Protein ◽  
Fluorescence Microscopy ◽  
Fluorescent Protein ◽  
Colletotrichum Lindemuthianum ◽  
Fungal Genome ◽  
Appressorium Formation ◽  
Gene Encoding ◽  
Green Fluorescent ◽  
Microscopy Examination

ABSTRACT The 5′ noncoding region of clpg2, an endopolygalacturonase gene of the bean pathogenColletotrichum lindemuthianum, was fused to the coding sequence of a gene encoding a green fluorescent protein (GFP), and the construct was introduced into the fungal genome. Detection of GFP accumulation by fluorescence microscopy examination revealed thatclpg2 was expressed at the early stages of germination of the conidia and during appressorium formation both in vitro and on the host plant.

scholarly journals Antagonism of the Two-Needle Pine Stem Rust Fungi Cronartium flaccidum and Peridermium pini by Cladosporium tenuissimum In Vitro and In Planta

2001 ◽  
Vol 91 (5) ◽  
pp. 457-468 ◽  
Author(s):  
Salvatore Moricca ◽  
Alessandro Ragazzi ◽  
Keith Richard Mitchelson ◽  
Gemma Assante
Keyword(s):  
Cell Wall ◽  
Spore Wall ◽  
Conidial Suspension ◽  
In Planta ◽  
Pine Seedlings ◽  
Host Cell Wall ◽  
Peridermium Pini ◽  
Hyphal Tip ◽  
Gain Access

Selected isolates of Cladosporium tenuissimum were tested for their ability to inhibit in vitro aeciospore germination of the two-needle pine stem rusts Cronartium flaccidum and Peridermium pini and to suppress disease development in planta. The antagonistic fungus displayed a number of disease-suppressive mechanisms. Aeciospore germination on water agar slides was reduced at 12, 18, and 24 h when a conidial suspension (1.5 × 107 conidia per ml) of the Cladosporium tenuissimum isolates was added. When the aeciospores were incubated in same-strength conidial suspensions for 1, 11, 21, and 31 days, viability was reduced at 20 and 4°C. Light and scanning electron microscopy showed that rust spores were directly parasitized by Cladosporium tenuissimum and that the antagonist had evolved several strategies to breach the spore wall and gain access to the underlying tissues. Penetration occurred with or without appressoria. The hyperparasite exerted a mechanical force to destroy the spore structures (spinules, cell wall) by direct contact, penetrated the aeciospores and subsequently proliferated within them. However, an enzymatic action could also be involved. This was shown by the dissolution of the host cell wall that comes in contact with the mycelium of the mycoparasite, by the lack of indentation in the host wall at the contact site, and by the minimal swelling at the infecting hyphal tip. Culture filtrates of the hyperparasite inhibited germination of rust propagules. A compound purified from the filtrates was characterized by chemical and spectroscopic analysis as cladosporol, a known β-1,3-glucan biosynthesis inhibitor. Conidia of Cladosporium tenuissimum reduced rust development on new infected pine seedlings over 2 years under greenhouse conditions. Because the fungus is an aggressive mycoparasite, produces fungicidal metabolites, and can survive and multiply in forest ecosystems without rusts, it seems a promising agent for the biological control of pine stem rusts in Europe.

scholarly journals Genomic differences between the new Fusarium oxysporum f. sp. apii (Foa) race 4 on celery, the less virulent Foa races 2 and 3, and the avirulent on celery f. sp. coriandrii

BMC Genomics ◽  
2020 ◽  
Vol 21 (1) ◽  
Author(s):  
Peter Henry ◽  
Sukhwinder Kaur ◽  
Quyen Anh Tran Pham ◽  
Radwan Barakat ◽  
Samuel Brinker ◽  
...  
Keyword(s):  
Species Complex ◽  
Pcr Primers ◽  
Single Copy ◽  
Preliminary Evidence ◽  
In Planta ◽  
Conserved Genes ◽  
Compatibility Group ◽  
Race 2 ◽  
Race 4

Abstract Background Members of the F. oxysporium species complex (FOSC) in the f. sp. apii (Foa) are pathogenic on celery and those in f. sp. coriandrii (Foci) are pathogenic on coriander (=cilantro). Foci was first reported in California in 2005; a new and highly aggressive race 4 of Foa was observed in 2013 in California. Preliminary evidence indicated that Foa can also cause disease on coriander, albeit are less virulent than Foci. Comparative genomics was used to investigate the evolutionary relationships between Foa race 4, Foa race 3, and the Foci, which are all in FOSC Clade 2, and Foa race 2, which is in FOSC Clade 3. Results A phylogenetic analysis of 2718 single-copy conserved genes and mitochondrial DNA sequence indicated that Foa races 3 and 4 and the Foci are monophyletic within FOSC Clade 2; these strains also are in a single somatic compatibility group. However, in the accessory genomes, the Foci versus Foa races 3 and 4 differ in multiple contigs. Based on significantly increased expression of Foa race 4 genes in planta vs. in vitro, we identified 23 putative effectors and 13 possible pathogenicity factors. PCR primers for diagnosis of either Foa race 2 or 4 and the Foci were identified. Finally, mixtures of conidia that were pre-stained with different fluorochromes indicated that Foa race 4 formed conidial anastomosis tubes (CATs) with Foci. Foa race 4 and Foa race 2, which are in different somatic compatibility groups, did not form CATs with each other. Conclusions There was no evidence that Foa race 2 was involved in the recent evolution of Foa race 4; Foa race 2 and 4 are CAT-incompatible. Although Foa races 3 and 4 and the Foci are closely related, there is no evidence that either Foci contributed to the evolution of Foa race 4, or that Foa race 4 was the recent recipient of a multi-gene chromosomal segment from another strain. However, horizontal chromosome transfer could account for the major difference in the accessory genomes of Foa race 4 and the Foci and for their differences in host range.

scholarly journals Detection of a Single-Copy Plasmid, pXFSL21, in Xylella fastidiosa Strain Stag’s Leap with Two Toxin-Antitoxin Systems Using Next-Generation Sequencing

2019 ◽  
Vol 109 (2) ◽  
pp. 240-247 ◽  
Author(s):  
Christopher Van Horn ◽  
Fengnian Wu ◽  
Zheng Zheng ◽  
Zehan Dai ◽  
Jianchi Chen
Keyword(s):  
Next Generation Sequencing ◽  
Xylella Fastidiosa ◽  
Bacterial Genome ◽  
Illumina Miseq ◽  
Single Copy ◽  
Next Generation ◽  
In Planta ◽  
Term Survival ◽  
Generation Sequencing

Plasmids are important genetic elements contributing to bacterial evolution and environmental adaptation. Xylella fastidiosa is a nutritionally fastidious Gram-negative bacterium causing economically devastating diseases such as Pierce’s disease (PD) of grapevine. In this study, the plasmid status of a highly virulent PD strain, Stag’s Leap, originally isolated from Napa Valley, CA, was studied using sequencing and bioinformatics tools. DNA samples extracted from a pure culture in periwinkle wilt medium (in vitro DNA) and a PD-symptomatic grapevine artificially inoculated in the greenhouse (in planta DNA) were subject to next-generation sequencing (NGS) analyses (Illumina MiSeq or HiSeq). Sequence analyses and polymerase chain reaction experiments revealed the presence of a circular plasmid, pXFSL21, of 21,665 bp. This plasmid existed as a single copy per bacterial genome under both in vitro and in planta conditions. Two toxin-antitoxin (T-A) systems (ydcD-ydcE and higB-higA) were detected in pXFSL21, a possible mechanism for the long-term survival of this single-copy plasmid in the bacterial population. BLAST searches against the GenBank database (version 222) detected homologs of the two T-A systems in chromosomes or plasmids of some X. fastidiosa strains. However, double T-A systems were found only in pXFSL21. pXFSL21 was not found in other known PD strains and, therefore, could serve as a molecular marker for strain Stag’s Leap monitoring and tracking. The NGS-based technique outlined in this article provides an effective tool for identifying single- or low-copy-number plasmids in fastidious prokaryotes.

Transgenic expression of a gene encoding a synthetic antimicrobial peptide results in inhibition of fungal growth in vitro and in planta

Plant Science ◽  
2000 ◽  
Vol 154 (2) ◽  
pp. 171-181 ◽  
Author(s):  
Jeffrey W. Cary ◽  
Kanniah Rajasekaran ◽  
Jesse M. Jaynes ◽  
Thomas E. Cleveland
Keyword(s):  
Antimicrobial Peptide ◽  
Fungal Growth ◽  
In Planta ◽  
Gene Encoding ◽  
Transgenic Expression

scholarly journals Two differentially expressed interleukin-11 receptor genes in the mouse genome

1996 ◽  
Vol 320 (2) ◽  
pp. 359-363 ◽  
Author(s):  
Petra BILINSKI ◽  
Mark A. HALL ◽  
Herbert NEUHAUS ◽  
Cornelia GISSEL ◽  
John K. HEATH ◽  
...  
Keyword(s):  
Mouse Genome ◽  
Amino Acid Level ◽  
Cell Types ◽  
Ligand Specificity ◽  
Gene Encoding ◽  
Cell Proliferation And Differentiation ◽  
Proliferation And Differentiation ◽  
Interleukin 11

Interleukin-11 (IL-11) is a multifunctional cytokine involved in the regulation of cell proliferation and differentiation in a variety of cell types and tissues in vitro and in vivo. The effects of IL-11 were shown to be mediated by the IL-11 receptor (hereafter referred to as IL-11Rα), which is a ligand-binding subunit and provides ligand specificity in a functional multimeric signal-transduction complex with gp130. Here we show that the mouse genome contains a second gene encoding an IL-11-binding protein, referred to as IL-11Rβ. The structure of the IL-11Rβ gene is highly similar to that of IL-11Rα, and IL-11Rβ exhibits 99% sequence identity with IL-11Rα at the amino acid level. IL-11Rβ is co-expressed with IL-11Rα, albeit at lower levels, in embryos and in various adult tissues. IL-11Rβ transcripts are abundant in testis, and, in contrast with IL-11Rα, absent from skeletal muscle. IL-11Rβ expressed in vitro binds IL-11 with high affinity, suggesting that the mouse genome contains a second functional IL-11R.

scholarly journals Cloning and Disruption of pgx4 Encoding an In Planta Expressed Exopolygalacturonase from Fusarium oxysporum

2000 ◽  
Vol 13 (4) ◽  
pp. 359-365 ◽  
Author(s):  
F. I. García-Maceira ◽  
Antonio Di Pietro ◽  
M. Isabel G. Roncero
Keyword(s):  
Amino Acid ◽  
Fusarium Oxysporum ◽  
Vascular Tissue ◽  
Single Copy ◽  
Gene Replacement ◽  
Amino Acid Sequences ◽  
In Planta ◽  
Calculated Molecular Mass ◽  
Tomato Plants

Fusarium oxysporum f. sp. lycopersici, the causal agent of tomato vascular wilt, produces an array of pectinolytic enzymes, including at least two exo-α1,4-polygalac-turonases (exoPGs). A gene encoding an exoPG, pgx4, was isolated with degenerate polymerase chain reaction primers derived from amino acid sequences conserved in two fungal exoPGs. pgx4 encodes a 454 amino acid polypeptide with nine potential N-glycosylation sites and a putative 21 amino acid N-terminal signal peptide. The deduced mature protein has a calculated molecular mass of 47.9 kDa, a pI of 8.0, and 51 and 49% identity with the exoPGs of Cochliobolus carbonum and Aspergillus tubingensis, respectively. The gene is present in a single copy in different formae speciales of F. oxysporum. Expression of pgx4 was detected during in vitro growth on pectin, polygalacturonic acid, and tomato vascular tissue and in roots and stems of tomato plants infected by F. oxysporum f. sp. lycopersici. Two mutants of F. oxy-sporum f. sp. lycopersici with a copy of pgx4 inactivated by gene replacement were as virulent on tomato plants as the wild-type strain.

scholarly journals A novel cyclic peptide (Naturido) modulates glia–neuron interactions in vitro and reverses ageing-related deficits in senescence-accelerated mice

PLoS ONE ◽  
2021 ◽  
Vol 16 (1) ◽  
pp. e0245235
Author(s):  
Shinichi Ishiguro ◽  
Tetsuro Shinada ◽  
Zhou Wu ◽  
Mayumi Karimazawa ◽  
Michimasa Uchidate ◽  
...  
Keyword(s):  
Hippocampal Neurons ◽  
Cyclic Peptide ◽  
Single Copy ◽  
Learning Ability ◽  
Gene Encoding ◽  
Medicinal Fungus ◽  
Neuron Interactions ◽  
Senescence Accelerated Mice

The use of agents that target both glia and neurons may represent a new strategy for the treatment of ageing disorders. Here, we confirmed the presence of the novel cyclic peptide Naturido that originates from a medicinal fungus (Isaria japonica) grown on domestic silkworm (Bombyx mori). We found that Naturido significantly enhanced astrocyte proliferation and activated the single copy gene encoding the neuropeptide VGF and the neuron-derived NGF gene. The addition of the peptide to the culture medium of primary hippocampal neurons increased dendrite length, dendrite number and axon length. Furthermore, the addition of the peptide to primary microglial cultures shifted CGA-activated microglia towards anti-inflammatory and neuroprotective phenotypes. These findings of in vitro glia–neuron interactions led us to evaluate the effects of oral administration of the peptide on brain function and hair ageing in senescence-accelerated mice (SAMP8). In vivo analyses revealed that spatial learning ability and hair quality were improved in Naturido-treated mice compared with untreated mice, to the same level observed in the normal ageing control (SAMR1). These data suggest that Naturido may be a promising glia–neuron modulator for the treatment of not only senescence, but also Alzheimer’s disease and other neurodegenerative diseases.

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