A putative effector UvHrip1 inhibits BAX-triggered cell death in Nicotiana benthamiana, and infection of Ustilaginoidea virens suppresses defense-related genes expression

Rice false smut (RFS), caused by Ustilaginoidea virens, is one of the most detrimental rice fungal diseases and pose a severe threat to rice production and quality. Effectors in U. virens often act as a set of essential virulence factors that play crucial roles in the interaction between host and the pathogen. Thus, the functions of each effector in U. virens need to be further explored. Here, we performed multiple alignment analysis and demonstrated a small secreted hypersensitive response-inducing protein (hrip), named UvHrip1, was highly conserved in fungi. The predicted SP of UvHrip1 was functional, which guided SUC secreted from yeast and was recognized by plant cells. The localization of UvHrip1 was mainly in the nucleus and cytoplasm monitored through the GFP fusion protein in Nicotiana benthamiana cells. uvhrip1 was drastically up-regulated in the susceptible cultivar LYP9 of rice during the pathogen infection, while did not in the resistant cultivar IR28. We also proved that UvHrip1 suppressed the mammalian BAX-induced necrosis-like defense symptoms in N. benthamiana. Furthermore, patterns of expression of defense-related genes, OsPR1#012 and OsPR10b, were regulated over U. virens infection in rice. Collectively, our data demonstrated that infection of U. virens suppresses defense-related genes expression and UvHrip1 was most likely a core effector in regulating plant immunity.

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The Conserved Effector UvHrip1 Interacts with OsHGW and Infection of Ustilaginoidea virens Regulates Defense- and Heading Date-Related Signaling Pathway

International Journal of Molecular Sciences ◽

10.3390/ijms21093376 ◽

2020 ◽

Vol 21 (9) ◽

pp. 3376

Author(s):

Songhong Wei ◽

Yingling Wang ◽

Jianming Zhou ◽

Shibo Xiang ◽

Wenxian Sun ◽

...

Keyword(s):

Fusion Protein ◽

Virulence Factors ◽

Molecular Mechanisms ◽

Expression Patterns ◽

Heading Date ◽

Ustilaginoidea Virens ◽

Bifc Assay ◽

Gfp Fusion Protein ◽

False Smut ◽

Insight Into

Ustilaginoidea virens, which causes rice false smut (RFS), is one of the most detrimental rice fungal diseases and poses a severe threat to rice production and quality. Effectors in U. virens often act as a group of essential virulence factors that play crucial roles in the interaction between host and the pathogen. Thus, the functions of individual effectors in U. virens need to be further explored. Here, we demonstrated a small secreted hypersensitive response-inducing protein (hrip), named UvHrip1, which was highly conserved in U. virens isolates. UvHrip1 was also proven to suppress necrosis-like defense symptoms in N. benthamiana induced by the oomycete elicitor INF1. The localization of UvHrip1 was mainly in the nuclei and cytoplasm via monitoring the UvHrip1-GFP fusion protein in rice cells. Furthermore, Y2H and BiFC assay demonstrated that UvHrip1 interacted with OsHGW, which is a critical regulator in heading date and grain weight signaling pathways in rice. Expression patterns of defense- and heading date-related genes, OsPR1#051 and OsMYB21, were down-regulated over U. virens infection in rice. Collectively, our data provide a theory for gaining an insight into the molecular mechanisms underlying the UvHrip1 virulence function.

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Structure of CARB-4 and AER-1 CarbenicillinHydrolyzing β-Lactamases

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.42.8.1966 ◽

1998 ◽

Vol 42 (8) ◽

pp. 1966-1972 ◽

Cited By ~ 15

Author(s):

François Sanschagrin ◽

Noureddine Bejaoui ◽

Roger C. Levesque

Keyword(s):

Amino Acids ◽

Pseudomonas Aeruginosa ◽

Dna Sequences ◽

Active Site ◽

Multiple Alignment ◽

Dna Homology ◽

Class A ◽

Canonical Amino Acid ◽

Multiple Alignment Analysis ◽

Alignment Analysis

ABSTRACT We determined the nucleotide sequences ofblaCARB-4 encoding CARB-4 and deduced a polypeptide of 288 amino acids. The gene was characterized as a variant of group 2c carbenicillin-hydrolyzing β-lactamases such as PSE-4, PSE-1, and CARB-3. The level of DNA homology between thebla genes for these β-lactamases varied from 98.7 to 99.9%, while that between these genes andblaCARB-4 encoding CARB-4 was 86.3%. TheblaCARB-4 gene was acquired from some other source because it has a G+C content of 39.1%, compared to a G+C content of 67% for typical Pseudomonas aeruginosa genes. DNA sequencing revealed that blaAER-1 shared 60.8% DNA identity with blaPSE-3 encoding PSE-3. The deduced AER-1 β-lactamase peptide was compared to class A, B, C, and D enzymes and had 57.6% identity with PSE-3, including an STHK tetrad at the active site. For CARB-4 and AER-1, conserved canonical amino acid boxes typical of class A β-lactamases were identified in a multiple alignment. Analysis of the DNA sequences flankingblaCARB-4 and blaAER-1 confirmed the importance of gene cassettes acquired via integrons inbla gene distribution.

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Comparative analysis of ITS1 nucleotide sequence reveals distinct genetic difference between Brugia malayi from Northeast Borneo and Thailand

Parasitology ◽

10.1017/s0031182012001242 ◽

2012 ◽

Vol 140 (1) ◽

pp. 39-45 ◽

Cited By ~ 2

Author(s):

MUN-YIK FONG ◽

RAHMAH NOORDIN ◽

YEE-LING LAU ◽

FEI-WEN CHEONG ◽

MUHAMMAD HAFIZNUR YUNUS ◽

...

Keyword(s):

Phylogenetic Trees ◽

Genetic Difference ◽

Brugia Malayi ◽

Wide Distribution ◽

Multiple Alignment ◽

Molecular Techniques ◽

Human Blood Samples ◽

Multiple Alignment Analysis ◽

Alignment Analysis ◽

Parasitic Worms

SUMMARYBrugia malayi is one of the parasitic worms which causes lymphatic filariasis in humans. Its geographical distribution includes a large part of Asia. Despite its wide distribution, very little is known about the genetic variation and molecular epidemiology of this species. In this study, the internal transcribed spacer 1 (ITS1) nucleotide sequences of B. malayi from microfilaria-positive human blood samples in Northeast Borneo Island were determined, and compared with published ITS1 sequences of B. malayi isolated from cats and humans in Thailand. Multiple alignment analysis revealed that B. malayi ITS1 sequences from Northeast Borneo were more similar to each other than to those from Thailand. Phylogenetic trees inferred using Neighbour-Joining and Maximum Parsimony methods showed similar topology, with 2 distinct B. malayi clusters. The first cluster consisted of Northeast Borneo B. malayi isolates, whereas the second consisted of the Thailand isolates. The findings of this study suggest that B. malayi in Borneo Island has diverged significantly from those of mainland Asia, and this has implications for the diagnosis of B. malayi infection across the region using ITS1-based molecular techniques.

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Multiple alignment analysis on phylogenetic tree of the spread of SARS epidemic using distance method

Journal of Physics Conference Series ◽

10.1088/1742-6596/890/1/012080 ◽

2017 ◽

Vol 890 ◽

pp. 012080 ◽

Cited By ~ 2

Author(s):

S Amiroch ◽

M S Pradana ◽

M I Irawan ◽

I Mukhlash

Keyword(s):

Phylogenetic Tree ◽

Multiple Alignment ◽

Distance Method ◽

Multiple Alignment Analysis ◽

Alignment Analysis

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Combination of multiple alignment analysis and surface mapping paves a way for a detailed pathway reconstruction--The case of VHL (von Hippel-Lindau) protein and angiogenesis regulatory pathway

Protein Science ◽

10.1110/ps.03454904 ◽

2004 ◽

Vol 13 (3) ◽

pp. 786-796 ◽

Cited By ~ 9

Author(s):

S. Sikora

Keyword(s):

Multiple Alignment ◽

Regulatory Pathway ◽

Surface Mapping ◽

Von Hippel Lindau ◽

Pathway Reconstruction ◽

Multiple Alignment Analysis ◽

Alignment Analysis

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A novel transcription factor UvCGBP1 regulates development and virulence of rice false smut fungus Ustilaginoidea virens

Virulence ◽

10.1080/21505594.2021.1936768 ◽

2021 ◽

Vol 12 (1) ◽

pp. 1563-1579

Author(s):

Xiaoyang Chen ◽

Pingping Li ◽

Hao Liu ◽

Xiaolin Chen ◽

Junbin Huang ◽

...

Keyword(s):

Transcription Factor ◽

Smut Fungus ◽

Ustilaginoidea Virens ◽

Rice False Smut ◽

False Smut

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Quantitative Proteomics Analysis Reveals the Function of the Putative Ester Cyclase UvEC1 in the Pathogenicity of the Rice False Smut Fungus Ustilaginoidea virens

International Journal of Molecular Sciences ◽

10.3390/ijms22084069 ◽

2021 ◽

Vol 22 (8) ◽

pp. 4069

Author(s):

Xiaoyang Chen ◽

Zhangxin Pei ◽

Pingping Li ◽

Xiabing Li ◽

Yuhang Duan ◽

...

Keyword(s):

Quantitative Proteomics ◽

Fungal Pathogens ◽

Protein Localization ◽

Proteomics Data ◽

Ustilaginoidea Virens ◽

Rice False Smut ◽

Oxidative Stresses ◽

Tandem Mass Tag ◽

Cyclase Domain ◽

False Smut

Rice false smut is a fungal disease distributed worldwide and caused by Ustilaginoidea virens. In this study, we identified a putative ester cyclase (named as UvEC1) as being significantly upregulated during U. virens infection. UvEC1 contained a SnoaL-like polyketide cyclase domain, but the functions of ketone cyclases such as SnoaL in plant fungal pathogens remain unclear. Deletion of UvEC1 caused defects in vegetative growth and conidiation. UvEC1 was also required for response to hyperosmotic and oxidative stresses and for maintenance of cell wall integrity. Importantly, ΔUvEC1 mutants exhibited reduced virulence. We performed a tandem mass tag (TMT)-based quantitative proteomic analysis to identify differentially accumulating proteins (DAPs) between the ΔUvEC1-1 mutant and the wild-type isolate HWD-2. Proteomics data revealed that UvEC1 has a variety of effects on metabolism, protein localization, catalytic activity, binding, toxin biosynthesis and the spliceosome. Taken together, our findings suggest that UvEC1 is critical for the development and virulence of U. virens.

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A vector system for fast-forward studies of the HOPZ-ACTIVATED RESISTANCE1 (ZAR1) resistosome in the model plant Nicotiana benthamiana

Plant Physiology ◽

10.1093/plphys/kiab471 ◽

2021 ◽

Author(s):

Adeline Harant ◽

Hsuan Pai ◽

Toshiyuki Sakai ◽

Sophien Kamoun ◽

Hiroaki Adachi

Keyword(s):

Nicotiana Benthamiana ◽

Cell Biology ◽

Plant Immunity ◽

Transient Gene Expression ◽

Coiled Coil ◽

Experimental System ◽

Vector System ◽

In Planta ◽

Functional Studies

Abstract Nicotiana benthamiana has emerged as a complementary experimental system to Arabidopsis thaliana. It enables fast-forward in vivo analyses primarily through transient gene expression and is particularly popular in the study of plant immunity. Recently, our understanding of nucleotide-binding leucine-rich repeat (NLR) plant immune receptors has greatly advanced following the discovery of the Arabidopsis HOPZ-ACTIVATED RESISTANCE1 (ZAR1) resistosome. Here, we describe a vector system of 72 plasmids that enables functional studies of the ZAR1 resistosome in N. benthamiana. We showed that ZAR1 stands out among the coiled coil class of NLRs (CC-NLRs) for being highly conserved across distantly related dicot plant species and confirmed NbZAR1 as the N. benthamiana ortholog of Arabidopsis ZAR1. Effector-activated and autoactive NbZAR1 trigger the cell death response in N. benthamiana and this activity is dependent on a functional N-terminal α1 helix. C-terminally tagged NbZAR1 remains functional in N. benthamiana, thus enabling cell biology and biochemical studies in this plant system. We conclude that the NbZAR1 open source pZA plasmid collection forms an additional experimental system to Arabidopsis for in planta resistosome studies.

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