scholarly journals Host Specificity and Differential Pathogenicity of Pectobacterium Strains from Dicot and Monocot Hosts

2020 ◽  
Vol 8 (10) ◽  
pp. 1479 ◽  
Author(s):  
Nirmal Khadka ◽  
Janak Raj Joshi ◽  
Noam Reznik ◽  
Nofar Chriker ◽  
Adi Nudel ◽  
...  

Recent phylogenetic studies have transferred certain isolates from monocot plants previously included in the heterogeneous group of Pectobacteriumcarotovorum (Pc) to a species level termed Pectobacterium aroidearum. The specificity of Pectobacterium associated infections had received less attention, and may be of high scientific and economic importance. Here, we have characterized differential responses of Pectobacterium isolates from potato (WPP14) and calla lily (PC16) on two typical hosts: Brassica oleracea var. capitata (cabbage) a dicot host; and Zantedeschia aethiopica (calla lily) a monocot host. The results revealed clear host specific responses following infection with the two bacterial strains. This was demonstrated by differential production of volatile organic compounds (VOCs) and the expression of plant defense-related genes (pal, PR-1, lox2, ast). A related pattern was observed in bacterial responses to each of the host’s extract, with differential expression of virulence-related determinants and genes associated with quorum-sensing and plant cell wall-degrading enzymes. The differences were associated with each strain’s competence on its respective host.

2016 ◽  
Vol 13 (2) ◽  
pp. 81-88
Author(s):  
Nazmun Nahar ◽  
Md. Manjurul Haque ◽  
Md. Abu Ashraf Khan ◽  
Md. Ashraful Haque

Soft-rotting bacteria caused diseases by secreting the plant cell-wall-degrading enzymes mainly pectate lyase (Pel). In this study, a total of 20 bacterial strains were isolated from soft rotted fruits and vegetables. Among them, 11 isolates produced soft rot symptom on potato tubers. All these isolates were partially characterized. When these isolates were tested for their Pel production in different media, the isolate P1 produced significantly higher amount of Pel than other isolates in M63 glycerol minimal medium. Interestingly, the isolate P1 grown on M63 glycerol minimal medium supplementing with 0.4% of polygalacturonic acid (PGA) and M63 glycerol minimal medium plus 0.4% of PGA and 1% of various plant extracts, Pel production was induced in M63 glycerol minimal medium containing 0.4% of PGA and hyper-induced in M63 glycerol minimal medium plus 0.4% of PGA and 1% of various plant extracts. However, Pel synthesis was not hyper-induced in high concentrations of potato extract. Thus, not only plant extracts but also concentrations of plant extract may be important for hyper-induction of Pel.The Agriculturists 2015; 13(2) 81-88


2017 ◽  
Vol 30 (11) ◽  
pp. 886-895 ◽  
Author(s):  
Maria Chiara Paccanaro ◽  
Luca Sella ◽  
Carla Castiglioni ◽  
Francesca Giacomello ◽  
Ana Lilia Martínez-Rocha ◽  
...  

Endo-polygalacturonases (PGs) and xylanases have been shown to play an important role during pathogenesis of some fungal pathogens of dicot plants, while their role in monocot pathogens is less defined. Pg1 and xyr1 genes of the wheat pathogen Fusarium graminearum encode the main PG and the major regulator of xylanase production, respectively. Single- and double-disrupted mutants for these genes were obtained to assess their contribution to fungal infection. Compared with wild-type strain, the ∆pg mutant showed a nearly abolished PG activity, slight reduced virulence on soybean seedlings, but no significant difference in disease symptoms on wheat spikes; the ∆xyr mutant was strongly reduced in xylanase activity and moderately reduced in cellulase activity but was as virulent as wild type on both soybean and wheat plants. Consequently, the ΔpgΔxyr double mutant was impaired in xylanase, PG, and cellulase activities but, differently from single mutants, was significantly reduced in virulence on both plants. These findings demonstrate that the concurrent presence of PG, xylanase, and cellulase activities is necessary for full virulence. The observation that the uronides released from wheat cell wall after a F. graminearum PG treatment were largely increased by the fungal xylanases suggests that these enzymes act synergistically in deconstructing the plant cell wall.


2020 ◽  
Author(s):  
Pallavi Kumari ◽  
Tali Sayas ◽  
Patricia Bucki ◽  
Sigal Brown Miyara ◽  
Maya Kleiman

AbstractStudying the interactions between microorganisms and plant roots is crucial for understanding a variety of phenomena concerning crop yield and health. The role of root surface properties in these interactions, is rarely addressed. To this end, we previously built a synthetic system, from the inert polymer polydimethyl siloxane (PDMS), mimicking the root surface microstructure, using a replication technique. This replica enables the study of isolated effects of surface structure on microorganism-plant interactions. Since the root surface is composed mostly of cellulose, using cellulose-like materials as our replica, instead of PDMS, is the next logical step. This will enable following the hydrolysis of such surfaces as a result of microorganisms secreting Plant Cell Wall Degrading Enzymes (PCWDE), and in particular, cellulase. Visualization of such hydrolysis in a synthetic system can assist in studying the localization and activity of microorganisms and how they correlate with surface microtopography, separately from chemical plant signals.In this work, we modified the known carboxymethyl cellulase (CMC) hydrolysis visualization method to enable real-time tracking of cellulase activity of microorganisms on the surface. Surface was formed from pure CMC, rather than CMC incorporated in agar as is often done, and by that, eliminating diffusion issues. Acridine orange dye, which is compatible, at low concentrations, with microorganisms, as opposed to other routinely used dyes, was incorporated into the film. The dye disassociated from the film when hydrolysis occurred, forming a halo surrounding the point of hydrolysis. This enabled real-time visualization since the common need for post hydrolysis dyeing was negated. Using Root Knot Nematode (RKN) as a model organism that penetrates the plant root, we showed it was possible to follow microorganism cellulase secretion on the surface in the form of CMC film hydrolysis. Furthermore, the addition of natural additives, in the form of root extract was also shown to be an option and resulted in an increased RKN response. We tested our newly developed method by changing temperature and pH conditions and by characterization of the hydrolyzed surface using both Scanning Electron Microscopy (SEM) and Atomic Force Microscopy (AFM).This method will be implemented in the future on a root surface microstructure replica. We believe the combination of this new method with our previously developed root surface microstructure replication technique can open a new avenue of research in the field of plant root-microorganism interactions.


Plant Disease ◽  
2014 ◽  
Vol 98 (6) ◽  
pp. 854-854 ◽  
Author(s):  
B.-J. Li ◽  
H.-Y. Ben ◽  
Y.-X. Shi ◽  
X.-W. Xie ◽  
A.-L. Chai

Zantedeschia aethiopica (L.) Spreng. (calla lily), belonging to family Araceae, is a popular ornamental plant in China. In the summer of 2010, leaves of calla lily with typical symptoms of necrotic lesions were observed in a commercial glasshouse in Beijing, China (116°20′ E, 39°44′ N). The initial symptoms were circular to subcircular, 1 to 3 mm, and dark brown lesions on the leaf lamina. Under high humidity, lesions expanded rapidly to 5 to 10 mm with distinct concentric zones and produced black sporodochia, especially on the backs of leaves. Later, the infected leaves were developing a combination of leaf lesions, yellowing, and falling off; as a result, the aesthetic value of the plant was significantly impacted. Leaf samples were used in pathogen isolation. Symptomatic leaf tissues were cut into small pieces and surface sterilized with 70% ethanol for 30 s and then in 0.1% mercuric chloride solution for 1 to 3 min. After being washed in sterile distilled water three times, the pieces were plated on potato dextrose agar (PDA) and incubated at 25°C in darkness for 7 days (5). Initial colonies of isolates were white, floccose mycelium and developed dark green to black concentric rings that were sporodochia bearing viscid spore masses after incubating 5 days. Conidiophores branched repeatedly. Conidiogenous cells were hyaline, clavate, and 10.0 to 16.0 × 1.4 to 2.0 μm. Conidia were hyaline, cylindrical, both rounded ends, and 6.0 to 8.2 × 1.9 to 2.4 μm. Morphological characteristics of the fungus were consistent with the description of Myrothecium roridum Tode ex Fr. (3,4). To confirm the pathogenicity, three healthy plants of calla lily were inoculated with a conidial suspension (1 × 106 conidia per ml) brushed from a 7-day-old culture of the fungus. Control plants were sprayed with sterile water. The inoculated plants were individual with clear plastic bags and placed in a glass cabinet at 25°C. After 7 days, all inoculated leaves developed symptoms similar to the original samples, but control plants remained disease free. Re-isolation and identification confirmed Koch's postulates. For molecular identification, genomic DNA of a representative isolate (MTL07081001) was extracted by modified CTAB method (1), and the rDNA-ITS region was amplified by using primers ITS1 (5-TCCGTAGGTGAACCTGCGG-3) and ITS4 (5-TCCTCCGCTTATTGATATGC-3). The 465-bp amplicon (GenBank Accession No. KF761293) was 100% identity to the sequence of M. roridum (JF724158.1) from GenBank. M. roridum has an extensive host range, covering 294 host plants (2). To our knowledge, this is the first record of leaf spot caused by M. roridum on calla lily in China. References: (1) F. M. Ausubel et al. Current Protocols in Molecular Biology. John Wiley & Sons Inc, New York, 1994. (2) D. F. Farr and A. Y. Rossman, Fungal Databases. Syst. Mycol. Microbiol. Lab., ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , October 2013. (3) M. T. Mmbaga et al. Plant Dis. 94:1266, 2010. (4) Y. X. Zhang et al. Plant Dis. 95:1030, 2011. (5) L. Zhu et al. J. Phytopathol. 161:59, 2013.


2018 ◽  
Vol 108 (10) ◽  
pp. 1206-1211 ◽  
Author(s):  
Takeo Shimizu ◽  
Satoko Kanematsu ◽  
Hajime Yaegashi

Understanding the molecular mechanisms of pathogenesis is useful in developing effective control methods for fungal diseases. The white root rot fungus Rosellinia necatrix is a soilborne pathogen that causes serious economic losses in various crops, including fruit trees, worldwide. Here, using next-generation sequencing techniques, we first produced a 44-Mb draft genome sequence of R. necatrix strain W97, an isolate from Japan, in which 12,444 protein-coding genes were predicted. To survey differentially expressed genes (DEGs) associated with the pathogenesis of the fungus, the hypovirulent W97 strain infected with Rosellinia necatrix megabirnavirus 1 (RnMBV1) was used for a comprehensive transcriptome analysis. In total, 545 and 615 genes are up- and down-regulated, respectively, in R. necatrix infected with RnMBV1. Gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses of the DEGs suggested that primary and secondary metabolism would be greatly disturbed in R. necatrix infected with RnMBV1. The genes encoding transcriptional regulators, plant cell wall-degrading enzymes, and toxin production, such as cytochalasin E, were also found in the DEGs. The genetic resources provided in this study will accelerate the discovery of genes associated with pathogenesis and other biological characteristics of R. necatrix, thus contributing to disease control.


Pathogens ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 976
Author(s):  
Lakshmipriya Perincherry ◽  
Chaima Ajmi ◽  
Souheib Oueslati ◽  
Agnieszka Waśkiewicz ◽  
Łukasz Stępień

Being pathogenic fungi, Fusarium produce various extracellular cell wall-degrading enzymes (CWDEs) that degrade the polysaccharides in the plant cell wall. They also produce mycotoxins that contaminate grains, thereby posing a serious threat to animals and human beings. Exposure to mycotoxins occurs through ingestion of contaminated grains, inhalation and through skin absorption, thereby causing mycotoxicoses. The toxins weaken the host plant, allowing the pathogen to invade successfully, with the efficiency varying from strain to strain and depending on the plant infected. Fusariumoxysporum predominantly produces moniliformin and cyclodepsipeptides, whereas F. proliferatum produces fumonisins. The aim of the study was to understand the role of various substrates and pea plant extracts in inducing the production of CWDEs and mycotoxins. Additionally, to monitor the differences in their levels when susceptible and resistant pea plant extracts were supplemented. The cultures of F. proliferatum and F. oxysporum strains were supplemented with various potential inducers of CWDEs. During the initial days after the addition of substrates, the fungus cocultivated with pea extracts and other carbon substrates showed increased activities of β-glucosidase, xylanase, exo-1,4-glucanase and lipase. The highest inhibition of mycelium growth (57%) was found in the cultures of F. proliferatum strain PEA1 upon the addition of cv. Sokolik extract. The lowest fumonisin content was exhibited by the cultures with the pea extracts and oat bran added, and this can be related to the secondary metabolites and antioxidants present in these substrates.


2020 ◽  
Vol 117 (11) ◽  
pp. 6003-6013 ◽  
Author(s):  
Vincent W. Wu ◽  
Nils Thieme ◽  
Lori B. Huberman ◽  
Axel Dietschmann ◽  
David J. Kowbel ◽  
...  

Filamentous fungi, such asNeurospora crassa, are very efficient in deconstructing plant biomass by the secretion of an arsenal of plant cell wall-degrading enzymes, by remodeling metabolism to accommodate production of secreted enzymes, and by enabling transport and intracellular utilization of plant biomass components. Although a number of enzymes and transcriptional regulators involved in plant biomass utilization have been identified, how filamentous fungi sense and integrate nutritional information encoded in the plant cell wall into a regulatory hierarchy for optimal utilization of complex carbon sources is not understood. Here, we performed transcriptional profiling ofN. crassaon 40 different carbon sources, including plant biomass, to provide data on how fungi sense simple to complex carbohydrates. From these data, we identified regulatory factors inN. crassaand characterized one (PDR-2) associated with pectin utilization and one with pectin/hemicellulose utilization (ARA-1). Using in vitro DNA affinity purification sequencing (DAP-seq), we identified direct targets of transcription factors involved in regulating genes encoding plant cell wall-degrading enzymes. In particular, our data clarified the role of the transcription factor VIB-1 in the regulation of genes encoding plant cell wall-degrading enzymes and nutrient scavenging and revealed a major role of the carbon catabolite repressor CRE-1 in regulating the expression of major facilitator transporter genes. These data contribute to a more complete understanding of cross talk between transcription factors and their target genes, which are involved in regulating nutrient sensing and plant biomass utilization on a global level.


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