scholarly journals The Sclerotinia sclerotiorum pac1 Gene Is Required for Sclerotial Development and Virulence

2003 ◽  
Vol 16 (9) ◽  
pp. 785-795 ◽  
Author(s):  
Jeffrey A. Rollins
Keyword(s):  
Oxalic Acid ◽  
Sclerotinia Sclerotiorum ◽  
Acid Production ◽  
Plant Pathogens ◽  
Gene Replacement ◽  
Loss Of Function ◽  
Physiological Processes ◽  
Gene Transcripts ◽  
Sclerotial Development

The synergistic activities of oxalic acid and endopolygalacturonases are thought to be essential for full virulence of Sclerotinia sclerotiorum and other oxalate-producing plant pathogens. Both oxalic acid production and endopolygalacturonase activity are regulated by ambient pH. Since many gene products with pH-sensitive activities are regulated by the PacC transcription factor in Aspergillus nidulans, we functionally characterized a pacC gene homolog, pac1, from S. sclerotiorum. Mutants with loss-of-function alleles of the pac1 locus were created by targeted gene replacement. In vitro mycelial growth of these pac1 mutants was normal at acidic pH, but growth was inhibited as culture medium pH was increased. Development and maturation of sclerotia in culture was also aberrant in these pac1 replacement mutants. Although oxalic acid production remained alkaline pH-responsive, the kinetics and magnitude of oxalate accumulation were dramatically altered. Additionally, maximal accumulation of endopolygalacturonase gene transcripts (pg1) was shifted to higher ambient pH. Virulence in loss-of-function pac1 mutants was dramatically reduced in infection assays with tomato and Arabidopsis. Based on these results, pac1 appears to be necessary for the appropriate regulation of physiological processes important for pathogenesis and development of S. sclerotiorum.

scholarly journals Pathogenicity and Sclerotial Development of Sclerotinia sclerotiorum: Involvement of Oxalic Acid and Chitin Synthesis

1995 ◽  
Author(s):  
Martin B. Dickman ◽  
Oded Yarden
Keyword(s):  
Oxalic Acid ◽  
Sclerotinia Sclerotiorum ◽  
Plant Pathogens ◽  
Detailed Examination ◽  
Genetically Engineered ◽  
Molecular Techniques ◽  
Chitin Synthesis ◽  
Dry Bean ◽  
Fungal Plant Pathogens ◽  
Sclerotial Development

Sclerotinia sclerotiorum (Lib.) de Bary is among the world's most successful and omnivorous fungal plant pathogens. Included in the nearly 400 species of plants reported as hosts to this fungus are canola, alfalfa, soybean, sunflower, dry bean and potato. The general inability to develop resistant germplasm with these economically important crops to this pathogen has focused attention on the need for a more detailed examination of the pathogenic determinants involved in disease development. A mechanistic understanding of the successful strategy(ies) used by S. sclerotiorum in colonizing host plants and their linkage to fungal development may provide targets and/or novel approaches with which to design resistant crop plants. This proposal involved experiments which were successful in generating genetically-engineered plants harboring resistance to S. sclerotiorum, the establishment and improvement of molecular tools for the study of this pathogen and the analysis of the linkage between pathogenicity, sclerotial morphogenesis and two biosynthetic pathways: oxalic acid production and chitin synthesis. The highly collaborative project has improved our understanding of S. sclerotiorum pathogenicity, established reliable molecular techniques to facilitate experimental manipilation and generated transgenic plants which are resistant to this econimically important fungus.

scholarly journals Oxalic Acid Is an Elicitor of Plant Programmed Cell Death during Sclerotinia sclerotiorum Disease Development

2008 ◽  
Vol 21 (5) ◽  
pp. 605-612 ◽  
Author(s):  
Kyoung Su Kim ◽  
Ji-Young Min ◽  
Martin B. Dickman
Keyword(s):  
Cell Death ◽  
Programmed Cell Death ◽  
Oxalic Acid ◽  
Sclerotinia Sclerotiorum ◽  
Plant Pathogens ◽  
Broad Host Range ◽  
Disease Development ◽  
Oxygen Species ◽  
Pathogenicity Determinant ◽  
Sclerotial Development

Accumulating evidence supports the idea that necrotrophic plant pathogens interact with their hosts by controlling cell death. Sclerotinia sclerotiorum is a necrotrophic ascomycete fungus with a broad host range (>400 species). Previously, we established that oxalic acid (OA) is an important pathogenicity determinant of this fungus. In this report, we describe a mechanism by which oxalate contributes to the pathogenic success of this fungus; namely, that OA induces a programmed cell death (PCD) response in plant tissue that is required for disease development. This response exhibits features associated with mammalian apoptosis, including DNA laddering and TUNEL reactive cells. Fungal mutants deficient in OA production are nonpathogenic, and apoptotic-like characteristics are not observed following plant inoculation. The induction of PCD by OA is independent of the pH-reducing abilities of this organic acid, which is required for sclerotial development. Moreover, oxalate also induces increased reactive oxygen species (ROS) levels in the plant, which correlate to PCD. When ROS induction is inhibited, apoptotic-like cell death induced by OA does not occur. Taken together, we show that Sclerotinia spp.-secreted OA is an elicitor of PCD in plants and is responsible for induction of apoptotic-like features in the plant during disease development. This PCD is essential for fungal pathogenicity and involves ROS. Thus, OA appears to function by triggering in the plant pathways responsible for PCD. Further, OA secretion by Sclerotinia spp. is not directly toxic but, more subtly, may function as a signaling molecule.

scholarly journals Transcriptional Profiling of Diffusible Lipopeptides and Fungal Virulence Genes During Bacillus amyloliquefaciens EZ1509-Mediated Suppression of Sclerotinia sclerotiorum

2020 ◽  
Vol 110 (2) ◽  
pp. 317-326 ◽  
Author(s):  
Ayaz Farzand ◽  
Anam Moosa ◽  
Muhammad Zubair ◽  
Abdur Rashid Khan ◽  
Muhammad Ayaz ◽  
...  
Keyword(s):  
Antifungal Activity ◽  
Oxalic Acid ◽  
Sclerotinia Sclerotiorum ◽  
Bacillus Amyloliquefaciens ◽  
Virulence Genes ◽  
Mass Spectrometry Analysis ◽  
Dual Culture ◽  
Biosynthetic Genes ◽  
In Vitro Interaction

Sclerotinia sclerotiorum is a devastating necrotrophic pathogen that infects multiple crops, and its control is an unremitting challenge. In this work, we attempted to gain insights into the pivotal role of lipopeptides (LPs) in the antifungal activity of Bacillus amyloliquefaciens EZ1509. In a comparative study involving five Bacillus strains, B. amyloliquefaciens EZ1509 harboring four LPs biosynthetic genes (viz. surfactin, iturin, fengycin, and bacilysin) exhibited promising antifungal activity against S. sclerotiorum in a dual-culture assay. Our data demonstrated a remarkable upsurge in LPs biosynthetic gene expression through quantitative reverse transcription PCR during in vitro interaction assay with S. sclerotiorum. Maximum upregulation in LPs biosynthetic genes was observed on the second and third days of in vitro interaction, with iturin and fengycin being the highly expressed genes. Subsequently, Matrix-assisted laser desorption/ionization-time of flight-mass spectrometry analysis confirmed the presence of LPs in the inhibition zone. Scanning electron microscope analysis showed disintegration, shrinkage, plasmolysis, and breakdown of fungal hyphae. During in planta evaluation, S. sclerotiorum previously challenged with EZ1509 showed significant suppression in pathogenicity on detached leaves of tobacco and rapeseed. The oxalic acid synthesis was also significantly reduced in S. sclerotiorum previously confronted with antagonistic bacterium. The expression of major virulence genes of S. sclerotiorum, including endopolygalacturonase-3, oxalic acid hydrolase, and endopolygalacturonase-6, was significantly downregulated during in vitro confrontation with EZ1509.

scholarly journals Pathogenicity Determinants of Sclerotinia sclerotiorum and Their Association to Its Aggressiveness on Brassica juncea

2021 ◽  
Vol 37 (4) ◽  
pp. 365-374
Author(s):  
Rupeet Gill ◽  
Prabhjodh S. Sandhu ◽  
Sanjula Sharma ◽  
Pankaj Sharma
Keyword(s):  
Oxalic Acid ◽  
Disease Progression ◽  
Brassica Juncea ◽  
Sclerotinia Sclerotiorum ◽  
Acid Production ◽  
White Rot ◽  
Geographical Regions ◽  
High Level ◽  
First Time

White rot or stem rot caused by Sclerotinia sclerotiorum is one of the most destructive fungal diseases that have become a serious threat to the successful cultivation of oilseed Brassicas. The study was designed with an aim to investigate the association between the pathogenic aggressiveness and pathogenicity determinants of this pathogen specifically in Brassica for the first time. For this, a total of 58 isolates of S. sclerotiorum from different geographical regions were collected and purified. These isolates were inoculated on a Brassica juncea cv. RL-1359 and they exhibited high level of variation in their disease progression. The isolates were grouped and then 24 isolates were selected for the biochemical analysis of pathogenicity determinants. The isolates varied significantly with respect to their total organic acids, oxalic acid production and pectin methyl esterase and polygalacturonase activity. The oxalic acid production corresponded to the disease progression of the isolates; the isolates with higher oxalic acid production were the more aggressive ones and vice-versa. This is, in our knowledge, the first study to establish a correlation between oxalic acid production and pathogenic aggressiveness of S. sclerotiorum on B. juncea. However, the pectinases’ enzyme activity did not follow the trend as of disease progression. These suggest an indispensable role of oxalic acid in pathogenicity of the fungus and the potential to be used as biochemical marker for preliminary assessment of pathogenic aggressiveness of various isolates before incorporating them in a breeding program.

Sclerotinia sclerotiorum: An Evaluation of Virulence Theories

2018 ◽  
Vol 56 (1) ◽  
pp. 311-338 ◽  
Author(s):  
Liangsheng Xu ◽  
Guoqing Li ◽  
Daohong Jiang ◽  
Weidong Chen
Keyword(s):  
Oxalic Acid ◽  
Sclerotinia Sclerotiorum ◽  
Acid Production ◽  
Control Strategies ◽  
Critical Evaluation ◽  
Necessary Condition ◽  
Acidic Ph ◽  
Ph Sensing ◽  
Induced Mutants ◽  
Sclerotial Formation

Oxalic acid production in Sclerotinia sclerotiorum has long been associated with virulence. Research involving UV-induced, genetically undefined mutants that concomitantly lost oxalate accumulation, sclerotial formation, and pathogenicity supported the conclusion that oxalate is an essential pathogenicity determinant of S. sclerotiorum. However, recent investigations showed that genetically defined mutants that lost oxalic acid production but accumulated fumaric acid could cause disease on many plants and substantiated the conclusion that acidic pH, not oxalic acid per se, is the necessary condition for disease development. Critical evaluation of available evidence showed that the UV-induced mutants harbored previously unrecognized confounding genetic defects in saprophytic growth and pH responsiveness, warranting reevaluation of the conclusions about virulence based on the UV-induced mutants. Furthermore, analyses of the evidence suggested a hypothesis for the existence of an unrecognized regulator responsive to acidic pH. Identifying the unknown pH regulator would offer a new avenue for investigating pH sensing/regulation in S. sclerotiorum and novel targets for intervention in disease control strategies.

scholarly journals Transfection of Sclerotinia sclerotiorum withIn VitroTranscripts of a Naturally Occurring Interspecific Recombinant of Sclerotinia sclerotiorum Hypovirus 2 Significantly Reduces Virulence of the Fungus

2015 ◽  
Vol 89 (9) ◽  
pp. 5060-5071 ◽  
Author(s):  
Shin-Yi Lee Marzano ◽  
Houston A. Hobbs ◽  
Berlin D. Nelson ◽  
Glen L. Hartman ◽  
Darin M. Eastburn ◽  
...  
Keyword(s):  
Sclerotinia Sclerotiorum ◽  
Plant Pathogens ◽  
High Throughput Sequencing ◽  
Infectious Clone ◽  
White Mold ◽  
Content Type ◽  
Fungal Plant Pathogens ◽  
Rapid Amplification ◽  
Valsa Ceratosperma

ABSTRACTA recombinant strain ofSclerotinia sclerotiorumhypovirus 2 (SsHV2) was identified from a North AmericanSclerotinia sclerotiorumisolate (328) from lettuce (Lactuca sativaL.) by high-throughput sequencing of total RNA. The 5′- and 3′-terminal regions of the genome were determined by rapid amplification of cDNA ends. The assembled nucleotide sequence was up to 92% identical to two recently reported SsHV2 strains but contained a deletion near its 5′ terminus of more than 1.2 kb relative to the other SsHV2 strains and an insertion of 524 nucleotides (nt) that was distantly related toValsa ceratospermahypovirus 1. This suggests that the new isolate is a heterologous recombinant of SsHV2 with a yet-uncharacterized hypovirus. We named the new strainSclerotinia sclerotiorumhypovirus 2 Lactuca (SsHV2L) and deposited the sequence in GenBank with accession numberKF898354.Sclerotinia sclerotiorumisolate 328 was coinfected with a strain ofSclerotinia sclerotiorumendornavirus 1 and was debilitated compared to cultures of the same isolate that had been cured of virus infection by cycloheximide treatment and hyphal tipping. To determine whether SsHV2L alone could induce hypovirulence inS. sclerotiorum, a full-length cDNA of the 14,538-nt viral genome was cloned. Transcripts corresponding to the viral RNA were synthesizedin vitroand transfected into a virus-free isolate ofS. sclerotiorum, DK3. Isolate DK3 transfected with SsHV2L was hypovirulent on soybean and lettuce and exhibited delayed maturation of sclerotia relative to virus-free DK3, completing Koch's postulates for the association of hypovirulence with SsHV2L.IMPORTANCEA cosmopolitan fungus,Sclerotinia sclerotioruminfects more than 400 plant species and causes a plant disease known as white mold that produces significant yield losses in major crops annually. Mycoviruses have been used successfully to reduce losses caused by fungal plant pathogens, but definitive relationships between hypovirus infections and hypovirulence inS. sclerotiorumwere lacking. By establishing a cause-and-effect relationship betweenSclerotinia sclerotiorumhypovirus Lactuca (SsHV2L) infection and the reduction in host virulence, we showed direct evidence that hypoviruses have the potential to reduce the severity of white mold disease. In addition to intraspecific recombination, this study showed that recent interspecific recombination is an important factor shaping viral genomes. The construction of an infectious clone of SsHV2L allows future exploration of the interactions between SsHV2L andS. sclerotiorum, a widespread fungal pathogen of plants.

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