scholarly journals Quantitative detection of the rice false smut pathogen Ustilaginoidea virens by real-time PCR

2013 ◽  
Vol 12 (4) ◽  
pp. 6433-6441 ◽  
Author(s):  
H. Li ◽  
D.H. Ni ◽  
Y.B. Duan ◽  
Y. Chen ◽  
J. Li ◽  
...  
2010 ◽  
Vol 128 (2) ◽  
pp. 221-232 ◽  
Author(s):  
Taketo Ashizawa ◽  
Mami Takahashi ◽  
Jouji Moriwaki ◽  
Kazuyuki Hirayae

2020 ◽  
Author(s):  
Yufu Wang ◽  
Jinsong Qu ◽  
Yueran Liu ◽  
Geng Sun ◽  
Zuoqian Wang ◽  
...  

Abstract Background The necrosis- and ethylene-inducing peptide 1 (Nep1)-like proteins (NLPs) are a broadly distributed superfamily of protiens in plant-associated microorganisms, including bacteria, fungi, and oomycetes. NLPs are considered to be important virulence factors, and some have been well studied. However, the role of NLPs in Ustilaginoidea virens , the agent of rice false smut fungus, remains to be uncovered.Results A protein containing NLP-specific NPP1 domain was identified in U. virens and named UvNLP. Phylogenetic analysis revealed that UvNLP was a type 2 NLP, and real-time PCR revealed that UvNLP was highly expressed at 3 days post-inoculation. A yeast secretion assay demonstrated that UvNLP contained a functional signal peptide (SP). Furthermore, transient expression of UvNLP in Nicotiana benthamiana induced weak cell death, and replacement of the SP enhanced cell death. Real-time PCR indicated that UvNLP induced the expression of plant defence related genes.Conclusions We identified a secreted protein UvNLP in U. virens that was infection related and could induce plant cell death and defence responses. This study provided evidence that NLPs act as proteinaceous MAMPs, giving insight into the elucidation of the pathogenicity mechanism of U. virens. The UvNLP might be used as a potential plant defence inducer for disease control in practice.


Virulence ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 1563-1579
Author(s):  
Xiaoyang Chen ◽  
Pingping Li ◽  
Hao Liu ◽  
Xiaolin Chen ◽  
Junbin Huang ◽  
...  

2021 ◽  
Vol 22 (8) ◽  
pp. 4069
Author(s):  
Xiaoyang Chen ◽  
Zhangxin Pei ◽  
Pingping Li ◽  
Xiabing Li ◽  
Yuhang Duan ◽  
...  

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.


2005 ◽  
Vol 71 (7) ◽  
pp. 3911-3916 ◽  
Author(s):  
Mark G. Wise ◽  
Gregory R. Siragusa

ABSTRACT Strains of Clostridium perfringens are a frequent cause of food-borne disease and gas gangrene and are also associated with necrotic enteritis in chickens. To detect and quantify the levels of C. perfringens in the chicken gastrointestinal tract, a quantitative real-time PCR assay utilizing a fluorogenic, hydrolysis-type probe was developed and utilized to assay material retrieved from the broiler chicken cecum and ileum. Primers and probe were selected following an alignment of 16S rDNA sequences from members of cluster I of the genus Clostridium, and proved to be specific for C. perfringens. The assay could detect approximately 50 fg of C. perfringens genomic DNA and approximately 20 cells in pure culture. Measurements of the analytical sensitivity determined with spiked intestinal contents indicated that the consistent limit of detection with ileal samples was approximately 102 CFU/g of ileal material, but only about 104 CFU/g of cecal samples. The decreased sensitivity with the cecal samples was due to the presence of an unidentified chemical PCR inhibitor(s) in the cecal DNA purifications. The assay was utilized to rapidly detect and quantify C. perfringens levels in the gut tract of broiler chickens reared without supplementary growth-promoting antibiotics that manifested symptoms of necrotic enteritis. The results illustrated that quantitative real-time PCR correlates well with quantification via standard plate counts in samples taken from the ileal region of the gastrointestinal tract.


2005 ◽  
Vol 68 (6) ◽  
pp. 1217-1221 ◽  
Author(s):  
PAVEL KRCMAR ◽  
EVA RENCOVA

A sensitive and rapid method for the quantitative detection of bovine-, ovine-, swine-, and chicken-specific mitochondrial DNA sequences based on real-time PCR has been developed. The specificity of the primers and probes for real-time PCR has been tested using DNA samples of other vertebrate species that may also be present in rendered products. The quantitative detection was performed with dual-labeled probes (TaqMan) using absolute quantification with external standards of single species meat-and-bone meals. This method facilitates the detection of 0.01% of the target species–derived material in concentrate feed mixtures and fish meals.


2017 ◽  
Vol 150 (3) ◽  
pp. 669-677 ◽  
Author(s):  
Mingli Yong ◽  
Qide Deng ◽  
Linlin Fan ◽  
Jiankun Miao ◽  
Chaohui Lai ◽  
...  

2003 ◽  
Vol 38 (3) ◽  
pp. 265-271 ◽  
Author(s):  
Chengbo Yang ◽  
Yuan Jiang ◽  
Kehe Huang ◽  
Changqing Zhu ◽  
Yulong Yin

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