scholarly journals UPLC-QTOF-MS-Based Untargeted Study of Metabolites Secreted by Sclerotinia Sclerotiorum Pathogen Under Axenic Condition

Author(s):  
Navin Chandra Gupta ◽  
Shaweta Arora ◽  
Aditi Kundu ◽  
Pankaj Sharma ◽  
Mahesh Rao ◽  
...  

Abstract The stem rot disease has emerged globally as a major disease threat to the productivity and seed quality of oilseed Brassica. The generalist causal pathogen Sclerotinia sclerotiorum (Lib.) de Bary shows large variability in their aggressiveness and pathogenicity. Revealing the metabolic profile and signaling components of the pathogen in host-pathogen interaction are fundamental in understanding host resistance to the disease. In this study, the metabolites released by the pathogenic strains of S. sclerotiorum under the axenic culture have been identified using the untargeted high-resolution UPLC-QTOF-ESI-MS/MS. The analysis of the ethyl acetate extracts of the S. sclerotiorum culture revealed ten major secondary metabolites namely, sclerin, sclerotinin-B, sclerone, melanin, bostrycoidin, botcinin-D, botcinin-A, gliovirin, scleramide, and botcinic acid. The later six metabolites are being reported for the first time in the culture extract of the S. sclerotiorum pathogen. The phylogenetic analysis based on the overlapping and unique informative peaks in the chromatograms, the six S. sclerotiorum strains were grouped into three major clads. The clustering based on metabolic profiles does not substantiate the diversity based on morphology or virulence differences on the host. The findings of the study signified the metabolites secreted under the axenic conditions are varies based on their growth and developmental stages and may not necessarily be the determining factors for their differential aggressiveness and virulence over the host.

Marine Drugs ◽  
2021 ◽  
Vol 19 (2) ◽  
pp. 109
Author(s):  
Hani A. Alhadrami ◽  
Bathini Thissera ◽  
Marwa H. A. Hassan ◽  
Fathy A. Behery ◽  
Che Julius Ngwa ◽  
...  

Coculture is a productive technique to trigger microbes’ biosynthetic capacity by mimicking the natural habitats’ features principally by competition for food and space and interspecies cross-talks. Mixed cultivation of two Red Sea-derived actinobacteria, Actinokineospora spheciospongiae strain EG49 and Rhodococcus sp. UR59, resulted in the induction of several non-traced metabolites in their axenic cultures, which were detected using LC–HRMS metabolomics analysis. Antimalarial guided isolation of the cocultured fermentation led to the isolation of the angucyclines actinosporins E (1), H (2), G (3), tetragulol (5) and the anthraquinone capillasterquinone B (6), which were not reported under axenic conditions. Interestingly, actinosporins were previously induced when the axenic culture of the Actinokineospora spheciospongiae strain EG49 was treated with signalling molecule N-acetyl-d-glucosamine (GluNAc); this finding confirmed the effectiveness of coculture in the discovery of microbial metabolites yet to be discovered in the axenic fermentation with the potential that could be comparable to adding chemical signalling molecules in the fermentation flask. The isolated angucycline and anthraquinone compounds exhibited in vitro antimalarial activity and good biding affinity against lysyl-tRNA synthetase (PfKRS1), highlighting their potential developability as new antimalarial structural motif.


Agronomy ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 268
Author(s):  
Pedro Miranda-Fuentes ◽  
Ana B. García-Carneros ◽  
Leire Molinero-Ruiz

The management of downy mildew (Plasmopara halstedii) in sunflower, is heavily dependent on genetic resistance, whilst entomopathogenic fungi (EF) can reduce other sunflower diseases. In this work, we characterized P. halstedii from Spain and other countries collected in the past few years. Twenty-three races were identified (the most frequent in Spain being 310, 304, 705 and 715), with an increasing proportion of highly virulent races. Five isolates from countries other than Spain overcame the resistance in RHA-340. In addition, we assessed the efficacy of five EF against downy mildew and their effects on sunflower growth in axenic conditions. None of the entomopathogens reduced disease severity, nor did they have any effect on plant growth when applied together with P. halstedii. In contrast, three EF reduced some of the plant growth variables in the absence of the pathogen. Microbiological and molecular diagnostics suggest that the axenic system and the short experimental time used in this study did not favor the successful establishment of EF in the plants or their potential biocontrol effect. Our results show a shift in P. halstedii racial patterns and suggest that soil as a growth substrate and long infection times are needed for EF effectiveness against downy mildew.


2019 ◽  
Author(s):  
Fatemeh Maghuly ◽  
Tamas Deak ◽  
Klemens Vierlinger ◽  
Stephan Pabinger ◽  
Hakim Tafer ◽  
...  

Abstract Background: Jatropha curcas, a tropical shrub, is a promising biofuel crop, which produces seeds with a high content of oil and protein. To better understand the development of its seeds to improve Jatropha`s agronomic performance, a two-step approach was performed: 1) generation of the entire transcriptome of six different maturation stages of J. curcas seeds using 454-Roche sequencing of a cDNA library, 2) comparison of transcriptional expression levels in six different developmental stages of seeds using a custom Agilent 8x60K oligonucleotide microarray. Results: A total of 793,875 high-quality reads were assembled into 19,841 unique full-length contigs, of which 13,705 could be annotated with Gene Ontology (GO) terms. Microarray data analysis identified 9,111 probes (out of 57,842 probes), which were differentially expressed between the six developmental stages. The expression results were validated for 70 randomly selected putative genes. Result from cluster analyses showed that transcripts related to sucrose, fatty acid, flavonoid, phenylpropanoid, lignin, hormone biosynthesis were over-represented in the early stage, while lipid storage, seed dormancy and maturation in the late stage. Generally, the expression of the most over-represented transcripts decrease in the last stage of seed maturation. Further, expression analyses of different maturation stages of J. curcas seed showed that most changes in transcript abundance occurred between the two last stages, suggesting that the timing of metabolic pathways during seed maturation in J. curcas is in late stages. The co-expression result showed a high degree of connectivity between genes that play essential role in fatty acid biosynthesis and nutrient mobilization. Furthermore, seed development and hormone pathways are significantly well connected. Conclusion: The obtained results revealed DESs regulating important pathways related to seed maturation, which could contribute to understanding the complex regulatory network during seed development. This study provides detailed information on transcription changes during J. curcas seed development and provides a starting point for a genomic survey of seed quality traits. The current results highlighted specific genes and processes relevant to the molecular mechanisms involved in Jatropha seed development, and it is anticipated that this data can be delivered to other Euphorbiaceae species of economic value.


2016 ◽  
Vol 96 (2) ◽  
pp. 243-251 ◽  
Author(s):  
Dorota Remlein-Starosta ◽  
Joanna Krzymińska ◽  
Jolanta Kowalska ◽  
Jan Bocianowski

Virginia mallow is a plant from the central and eastern states of North America. A large amount of high quality biomass obtained from V. mallow led to interest in this plant for bioenergy purposes. Unfortunately, high incidence of white rot disease caused by Sclerotinia sclerotiorum destroyed plantations of this plant. In previous attempts, various control strategies were not successful and all treatments failed (including the use of Coniothurium minitans). The aim of the study was to find and evaluate yeast-like fungi for biocontrol of S. sclerotiorum, evaluate the optimal thermal condition for biocontrol efficacy, and describe possible modes of action of potential biocontrol organisms. In dual bioassay test with 19 strains of yeast-like fungi with S. sclerotiorum, antagonistic potential were obtained. At different temperatures variations in antagonistic activity of yeast-like fungi were observed. At all tested temperatures (i.e., 4, 12, and 23°C), positive results were obtained. Extracellular enzymes were produced by the majority of antagonistic yeast, such as: amylases (the most frequent), chitinases, proteases, pectinases and xylanases. In this research, reduction in white rot symptoms on V. mallow stems was significant (P < 0.05). The highest inhibition of disease was observed after treatment with strains (114/64) Candida albidus and (117/10) Pichia anomala.


Plant Disease ◽  
2008 ◽  
Vol 92 (5) ◽  
pp. 763-766 ◽  
Author(s):  
G. D. Bates ◽  
C. S. Rothrock ◽  
J. C. Rupe

Archer, a maturity group I soybean cultivar with demonstrated flood tolerance and resistance to Pythium ultimum, was compared with Hutcheson, a widely planted maturity group V cultivar in Arkansas, for resistance to P. ultimum, P. irregulare, P. aphanidermatum, P. vexans, and group HS. Emergence and establishment assays demonstrated that Archer had greater emergence and fewer disease symptoms after 10 days for all pathogenic species of Pythium than did Hutcheson. Archer also demonstrated higher root weights and fewer disease symptoms compared with Hutcheson in assays conducted for 6 weeks. Similar results were found using two different seed lots of the cultivars, suggesting that the results were not due to seed quality differences. This study indicates that resistance in Archer to Pythium damping-off and root rot compared with Hutcheson is robust, with efficacy over a number of Pythium spp. and a range of plant developmental stages.


Plant Disease ◽  
2005 ◽  
Vol 89 (5) ◽  
pp. 530-530 ◽  
Author(s):  
S. Gaetán ◽  
M. Madia

Canola (Brassica napus) was introduced as an alternative crop for wheat in Argentina. During 2003, typical symptoms of stem rot disease were observed on canola plants in two commercial fields located at Bragado, in northern Buenos Aires Province in Argentina. Average disease incidence across four canola cultivars was 21% (range = 13 to 29%). Symptoms included chlorosis and wilting of foliage and necrosis of basal stems. The disease appeared singly or in patches consisting of 4- to 5-month-old plants. The first visible symptom noticed was chlorosis and wilting of the foliage beginning from the basal leaves. Infection of the main stem at ground level typically was followed by a grayish white discoloration that progressed above the soil line to the apex. In advanced stages of the disease, stems and branches became bleached and eventually died. Black and irregularly shaped sclerotia (average size 5.5 × 2.8 mm) inside necrotic stem tissue were the typical signs of the pathogen. From September to October 2003, four samples consisting of six affected plants per sample were arbitrarily collected from two commercial fields located at Bragado. Sclerotia were taken from diseased stems, dipped in 70% ethanol, surface sterilized with 1% sodium hypochlorite for 1 min, and rinsed in sterile water. Each sclerotium was blotted dry on sterile Whatman's filter paper and placed on potato dextrose agar. Plates were incubated in the dark at 25°C for 2 to 3 days, followed by incubation under 12-h NUV light/12-h dark for 6 to 8 days. Six resulting colonies were identified as Sclerotinia sclerotiorum (Lib.) de Bary on the basis of taxonomic characteristics of the plant pathogenic species of Sclerotinia (3). Koch's postulates for three fungal isolates from infected plants were carried out on 6-week-old canola plants (cvs. Eclipse, Impulse, Master, and Mistral) by placing a colonized agar disk into wounds made in the basal stem region with a sterile scalpel. Pathogenicity tests, which included five inoculated and three control plants potted in a sterilized soil mix (soil/sand, 3:1), were conducted in a greenhouse at 23 to 26°C and 75% relative humidity with no supplemental light. Characteristic symptoms identical to the original observations developed within 12 days after inoculation on 100% of the inoculated plants for three isolates. Symptoms included wilted foliage, collapsed plants, and plant death. White mycelium and sclerotia developed on infected tissues, and the pathogen was successfully reisolated from symptomatic plants in all instances. Control plants, which were treated similarly except that the agar disk did not contain fungal growth, remained healthy. The experiment was repeated, and the results were identical to the first inoculations. Canola stem rot disease incited by S. sclerotiorum was first reported in Argentina during 1995 at experimental field plots in Buenos Aires. S. sclerotiorum, which has been reported to cause disease in canola in Canada (2) and the United States (1,4), currently represents a serious problem to the main canola cultivars grown in Argentina. To our knowledge, this is the first report of the occurrence of S. sclerotiorum causing a high incidence of stem rot in commercial crops of canola in Argentina. References: (1) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St. Paul, MN, 1989. (2) L. B. Jamaux et al. Plant Pathol. 44:22, 1995. (3) L. M. Kohn. Phytopathology 69:881, 1979. (4) D. V. Phillips et al. Phytopathology 92:785, 2002.


2020 ◽  
Vol 8 (4) ◽  
pp. 881-885
Author(s):  
Vinod Kumar Bairwa ◽  
Shailesh Godika ◽  
Jitendra Sharma ◽  
Rohit Kumar Nayak ◽  
Nitisha Gahlot ◽  
...  

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