scholarly journals Characterization of two catalase-peroxidase-encoding genes in Fusarium verticillioides reveals differential responses to in vitro versus in planta oxidative challenges

2017 ◽  
Vol 19 (5) ◽  
pp. 1127-1139 ◽  
Author(s):  
Shan Gao ◽  
Scott E. Gold ◽  
Anthony E. Glenn
Keyword(s):  
Fusarium Verticillioides ◽  
In Planta ◽  
Catalase Peroxidase ◽  
Differential Responses ◽  
Encoding Genes

scholarly journals Characterization of Two BAHD Acetyltransferases Highly Expressed in the Flowers of Jasminum sambac (L.) Aiton

Plants ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 13
Author(s):  
Yuting Wang ◽  
Hongliang Zhang ◽  
Chao Wan ◽  
Xian He ◽  
Jinfeng Huang ◽  
...  
Keyword(s):  
Steady State ◽  
Ectopic Expression ◽  
Catalytic Reactions ◽  
Total Emission ◽  
Emission Pattern ◽  
Jasminum Sambac ◽  
The Tropics ◽  
Encoding Genes

Volatile benzenoid compounds are found in diverse aromatic bouquets emitted by most moth-pollinated flowers. The night-blooming Jasminum sambac is widely cultivated worldwide in the tropics and subtropics for ornamental and industrial purposes owing to its fragrant flowers. Benzylacetate is a characteristic constituent in jasmine scent which makes up to approximately 20–30% of the total emission in the headspace or extract, but the biosynthesis enzymes and the encoding genes have not yet been described. Here, we identify two cytosolic BAHD acyltransferases specifically expressed in the petals with a positive correlation closely to the emission pattern of the volatile benzenoids. Both JsBEAT1 and JsBEAT2 could use benzylalcohol and acetate-CoA as substrates to make benzylacetate in vitro. The recombinant GST-JsBEAT1 has an estimated apparent Km of 447.3 μM for benzylalcohol and 546.0 μM for acetate-CoA, whereas in the instance of the His-JsBEAT2, the Km values are marginally lower, being 278.7 and 317.3 μM, respectively. However, the catalytic reactions by the GST-JsBEAT1 are more efficient than that by the His-JsBEAT2, based on the steady-state kcat parameters. Furthermore, ectopic expression of JsBEAT1 and JsBEAT2 in the transgenic P. hybrida plants, driven by a flower-specific promotor, significantly enhances the biosynthesis of benzylbenzoate and benzylacetate, as well as the total VOCs.

scholarly journals Characterization of Lysinibacillus fusiformis strain S4C11: In vitro, in planta, and in silico analyses reveal a plant-beneficial microbe

2021 ◽  
Vol 244 ◽  
pp. 126665
Author(s):  
Alessandro Passera ◽  
Marzia Rossato ◽  
John S. Oliver ◽  
Giovanna Battelli ◽  
Gul-I-Rayna Shahzad ◽  
...  
Keyword(s):  
In Silico ◽  
In Planta ◽  
Lysinibacillus Fusiformis ◽  
In Silico Analyses

scholarly journals Ralstonia solanacearum Needs Flp Pili for Virulence on Potato

2012 ◽  
Vol 25 (4) ◽  
pp. 546-556 ◽  
Author(s):  
Charles K. Wairuri ◽  
Jacquie E. van der Waals ◽  
Antoinette van Schalkwyk ◽  
Jacques Theron
Keyword(s):  
Ralstonia Solanacearum ◽  
Type Iv Pili ◽  
Periodontal Pathogen ◽  
Wild Type ◽  
In Planta ◽  
Phytopathogenic Bacterium ◽  
Type Iv ◽  
Plant Pathogenesis

Type IV pili are virulence factors in various bacteria. Several subclasses of type IV pili have been described according to the characteristics of the structural prepilin subunit. Although type IVa pili have been implicated in the virulence of Ralstonia solanacearum, type IVb pili have not previously been described in this plant pathogen. Here, we report the characterization of two distinct tad loci in the R. solanacearum genome. The tad genes encode functions necessary for biogenesis of the Flp subfamily of type IVb pili initially described for the periodontal pathogen Aggregatibacter actinomycetemcomitans. To determine the role of the tad loci in R. solanacearum virulence, we mutated the tadA2 gene located in the megaplasmid that encodes a predicted NTPase previously reported to function as the energizer for Flp pilus biogenesis. Characterization of the tadA2 mutant revealed that it was not growth impaired in vitro or in planta, produced wild-type levels of exopolysaccharide galactosamine, and exhibited swimming and twitching motility comparable with the wild-type strain. However, the tadA2 mutant was impaired in its ability to cause wilting of potato plants. This is the first report where type IVb pili in a phytopathogenic bacterium contribute significantly to plant pathogenesis.

scholarly journals In Vitro Antifungal Susceptibility and Molecular Characterization of Clinical Isolates of Fusarium verticillioides (F. moniliforme) and Fusarium thapsinum

2008 ◽  
Vol 52 (6) ◽  
pp. 2228-2231 ◽  
Author(s):  
Mónica Azor ◽  
Josepa Gené ◽  
Josep Cano ◽  
Deanna A. Sutton ◽  
Annette W. Fothergill ◽  
...  
Keyword(s):  
Active Drug ◽  
Fusarium Verticillioides ◽  
Antifungal Susceptibility ◽  
Antifungal Drugs ◽  
Clinical Isolates ◽  
Microdilution Method ◽  
Fusarium Thapsinum ◽  
In Vitro Antifungal Susceptibility

ABSTRACT A microdilution method was used to test 11 antifungal drugs against clinical isolates of Fusarium thapsinum and three different phylogenetic clades of Fusarium verticillioides that were characterized by sequencing a region of the β-tubulin gene. Terbinafine was the most-active drug against both species, followed by posaconazole against F. verticillioides.

scholarly journals Chromosome-Based Genetic Complementation System for Xylella fastidiosa

2009 ◽  
Vol 75 (6) ◽  
pp. 1679-1687 ◽  
Author(s):  
Ayumi Matsumoto ◽  
Glenn M. Young ◽  
Michele M. Igo
Keyword(s):  
Dna Sequences ◽  
Xylella Fastidiosa ◽  
Cell Physiology ◽  
Multiple Cloning Site ◽  
Wild Type ◽  
In Planta ◽  
Catalase Peroxidase ◽  
The Stability

ABSTRACT Xylella fastidiosa is a xylem-limited, gram-negative bacterium that causes Pierce's disease of grapevine. Here, we describe the construction of four vectors that facilitate the insertion of genes into a neutral site (NS1) in the X. fastidiosa chromosome. These vectors carry a colE1-like (pMB1) replicon and DNA sequences from NS1 flanking a multiple-cloning site and a resistance marker for one of the following antibiotics: chloramphenicol, erythromycin, gentamicin, or kanamycin. In X. fastidiosa, vectors with colE1-like (pMB1) replicons have been found to result primarily in the recovery of double recombinants rather than single recombinants. Thus, the ease of obtaining double recombinants and the stability of the resulting insertions at NS1 in the absence of selective pressure are the major advantages of this system. Based on in vitro and in planta characterizations, strains carrying insertions within NS1 are indistinguishable from wild-type X. fastidiosa in terms of growth rate, biofilm formation, and pathogenicity. To illustrate the usefulness of this system for complementation analysis, we constructed a strain carrying a mutation in the X. fastidiosa cpeB gene, which is predicted to encode a catalase/peroxidase, and showed that the sensitivity of this mutant to hydrogen peroxide could be overcome by the introduction of a wild-type copy of cpeB at NS1. Thus, this chromosome-based complementation system provides a valuable genetic tool for investigating the role of specific genes in X. fastidiosa cell physiology and virulence.

scholarly journals Functional Characterization of RING-Type E3 Ubiquitin Ligases In Vitro and In Planta

10.3791/60533 ◽  
2019 ◽  
Author(s):  
Joanna Kud ◽  
Wenjie Wang ◽  
Yulin Yuan ◽  
Allan Caplan ◽  
Joseph C. Kuhl ◽  
...  
Keyword(s):  
Functional Characterization ◽  
Ubiquitin Ligases ◽  
E3 Ubiquitin Ligases ◽  
In Planta ◽  
Ring Type

Biochemical and histological characterization of succulent plant Tacitus bellus response to Fusarium verticillioides infection in vitro

2020 ◽  
Vol 244 ◽  
pp. 153086
Author(s):  
Tijana Cvetić Antić ◽  
Dušica Janošević ◽  
Vuk M. Maksimović ◽  
Miroslav Živić ◽  
Snežana Budimir ◽  
...  
Keyword(s):  
Fusarium Verticillioides ◽  
Succulent Plant

scholarly journals Functional Characterization of BrF3'H, Which Determines the Typical Flavonoid Profile of Purple Chinese Cabbage

2021 ◽  
Vol 12 ◽  
Author(s):  
Sangkyu Park ◽  
Hyo Lee ◽  
Myung Ki Min ◽  
Jihee Ha ◽  
Jaeeun Song ◽  
...  
Keyword(s):  
Chinese Cabbage ◽  
Expression Patterns ◽  
Functional Characterization ◽  
Phenylpropanoid Pathway ◽  
Amino Acid Sequences ◽  
In Planta ◽  
Normal Green ◽  
Wide Range

Flavonols and anthocyanins are the two major classes of flavonoids in Brassica rapa. To elucidate the flavonoid biosynthetic pathway in Chinese cabbage (B. rapa L. subsp. pekinensis), we analyzed flavonoid contents in two varieties of Chinese cabbage with normal green (5546) and purple (8267) leaves. The 8267 variety accumulates significantly higher levels of quercetin, isorhamnetin, and cyanidin than the 5546 variety, indicating that 3′-dihydroxylated flavonoids are more prevalent in the purple than in the green variety. Gene expression analysis showed that the expression patterns of most phenylpropanoid pathway genes did not correspond to the flavonoid accumulation patterns in 5546 and 8267 varieties, except for BrPAL1.2 while most early and late flavonoid biosynthetic genes are highly expressed in 8267 variety. In particular, the flavanone 3′-hydroxylase BrF3′H (Bra009312) is expressed almost exclusively in 8267. We isolated the coding sequences of BrF3′H from the two varieties and found that both sequences encode identical amino acid sequences and are highly conserved with F3'H genes from other species. An in vitro enzymatic assay demonstrated that the recombinant BrF3′H protein catalyzes the 3′-hydroxylation of a wide range of 4′-hydroxylated flavonoid substrates. Kinetic analysis showed that kaempferol is the most preferred substrate and dihydrokaempferol (DHK) is the poorest substrate for recombinant BrF3′H among those tested. Transient expression of BrF3′H in Nicotiana benthamiana followed by infiltration of naringenin and DHK as substrates resulted in eriodictyol and quercetin production in the infiltrated leaves, demonstrating the functionality of BrF3′H in planta. As the first functional characterization of BrF3′H, our study provides insight into the molecular mechanism underlying purple coloration in Chinese cabbage.

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