scholarly journals Characterization of Sterol Demethylation Inhibitor-Resistant Isolates of Fusarium asiaticum and F. graminearum Collected from Wheat in China

2009 ◽  
Vol 99 (5) ◽  
pp. 487-497 ◽  
Author(s):  
Y. Yin ◽  
X. Liu ◽  
B. Li ◽  
Z. Ma
Keyword(s):  
Phytopathogenic Fungi ◽  
Cross Resistance ◽  
Pcr Analysis ◽  
Head Blight ◽  
Fusarium Asiaticum ◽  
Homologous Genes ◽  
Sterol Demethylation Inhibitors ◽  
Polymerase Chain ◽  
Dmi Resistance

Fusarium asiaticum and F. graminearum are the primary causal agents of Fusarium head blight (FHB) of wheat in China. In this study, sensitivities of 159 F. asiaticum and F. graminearum isolates to a benzimidazole fungicide carbendazim (MBC) and to sterol demethylation inhibitors (DMIs) tebuconazole and prochloraz were determined. Among the 159 isolates, 9 were resistant to MBC and designated as MBC-R isolates. Three showed resistance to tebuconazole and prochloraz and designated as DMI-R isolates. There was no cross-resistance between MBC and DMI. Genetic analysis by microsatellite-primed polymerase chain reaction (PCR) showed that MBC-R or DMI-R isolates had different genotypes, which indicated that they originated from different wild-type parents. Analysis of two 14α-demethylase (cyp51) homologous genes (cyp51A and cyp51B) showed that the F. asiaticum isolates could be distinguished from F. graminearum isolates based on the sequence of cyp51A. Analysis of deduced amino acid sequence of cyp51A and cyp51B suggested that no mutations were associated with DMI resistance. Real-time PCR analysis showed that the DMI resistance was not related to the expression of cyp51A and cyp51B in F. asiaticum and F. graminearum, but expressions of both genes were induced greatly by the tebuconazole. Results of this study indicated that cyp51A would be an informative marker for analysis of population structure of F. asiaticum and F. graminearum, and the existence of homologous cyp51 genes in F. asiaticum and F. graminearum could provide new insights into DMI resistance in phytopathogenic fungi.

Characterization of alpha-adrenoceptor gene expression in arterial and venous smooth muscle

1993 ◽  
Vol 265 (5) ◽  
pp. H1501-H1509 ◽  
Author(s):  
P. Ping ◽  
J. E. Faber
Keyword(s):  
Smooth Muscle ◽  
Messenger Rna ◽  
Vena Cava ◽  
Pcr Analysis ◽  
Relative Distribution ◽  
Alpha Adrenoceptor ◽  
Aortic Media ◽  
Unique Restriction ◽  
Polymerase Chain

Six genes coding for three unique alpha 1- (1A, 1B, 1C) and three unique alpha 2- (2A, 2B, 2C) adrenergic receptor (AR) subtypes have been cloned. Ligand binding and contractile studies have demonstrated that both alpha 1- and alpha 2-ARs can exist on vascular smooth muscle (VSM) cells, although less is known about the relative distribution and specific subtypes in different vascular segments. In the present study polymerase chain reaction (PCR) analysis was used to characterize the species of alpha-AR messenger RNA (mRNA) present in freshly isolated rat thoracic aortic media and vena cava and in cultured VSM cells (passage 2) derived from both sources. To prevent possible contamination of VSM mRNA, aortic media was separated from adventitia, and vessels were denuded of endothelial cells. Oligonucleotide primers specific for each of the six adrenergic genes were synthesized and used to probe for the presence of alpha-AR mRNA species after reverse transcription of total cellular RNA to cDNA. PCR-amplified AR transcripts were distinguished by the size of amplified DNA fragments and unique restriction endonuclease cleavage. Expression of alpha 1C- or alpha 2C-mRNA was not detected in vascular tissues or cultured VSM cells, although the alpha 2C-primers detected the expected alpha 2C expression in cerebral cortex. Only alpha 1A-mRNA was detected in aortic adventitia. VSM from aorta expressed alpha 1A-, alpha 1B-, and alpha 2A-mRNA, and this pattern was preserved in cultured aortic VSM. Vena cava also expressed both alpha 1A and alpha 1B; however only alpha 2B-mRNA was detected.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Characterization of Nivalenol-Producing Fusarium asiaticum That Causes Cereal Head Blight in Korea

2019 ◽  
Vol 35 (6) ◽  
pp. 543-552 ◽  
Author(s):  
Ja Yeong Jang ◽  
Seul Gi Baek ◽  
Jung-Hye Choi ◽  
Sosoo Kim ◽  
Jeomsoon Kim ◽  
...  
Keyword(s):  
Head Blight ◽  
Fusarium Asiaticum

scholarly journals Molecular Cloning and Characterization of the HOS1 Gene from ‘Muscat Hamburg’ Grapevine

2014 ◽  
Vol 139 (1) ◽  
pp. 54-62 ◽  
Author(s):  
Jitao Li ◽  
Nian Wang ◽  
Lina Wang ◽  
Haiping Xin ◽  
Shaohua Li
Keyword(s):  
Polymerase Chain Reaction ◽  
Arabidopsis Thaliana ◽  
Abscisic Acid ◽  
Pcr Analysis ◽  
Chain Reaction ◽  
The World ◽  
Drought Conditions ◽  
Polymerase Chain ◽  
Polyethylene Glycol 6000

Cold stress is an important factor that limits grape (Vitis sp.) production around the world. The high expression of osmotically responsive genes 1 (HOS1) protein acts as a repressor of cold-responsive genes in plants. To increase understanding of mechanism regulating cold tolerance in grape, we isolated and characterized a novel HOS1 gene, designated VvHOS1 from ‘Muscat Hamburg’ grapevine (Vitis vinifera). Real-time polymerase chain reaction (PCR) analysis revealed that the expression of VvHOS1 could be induced by the application of exogenous abscisic acid and various abiotic environmental conditions such as low temperature, drought, and salinity. Moreover, VvHOS1 expression could also be induced by cold plus drought conditions (4 °C, 10% polyethylene glycol 6000). In addition, overexpression of VvHOS1 in arabidopsis (Arabidopsis thaliana) decreased the plants’ tolerance to cold, drought, and salt as well as negatively regulated the expression level of two stress-responsive genes, AtRD29A and AtCOR47. The results obtained in this study should help us to elucidate the function of VvHOS1 and understand the cold-responsive pathway in grapevine.

scholarly journals Characterization of a Gene Identified in Pathotype 5 of the Clubroot Pathogen Plasmodiophora brassicae

2015 ◽  
Vol 105 (6) ◽  
pp. 764-770 ◽  
Author(s):  
H. Zhang ◽  
J. Feng ◽  
V. P. Manolii ◽  
S. E. Strelkov ◽  
S.-F. Hwang
Keyword(s):  
Plasmodiophora Brassicae ◽  
Blot Hybridization ◽  
Field Isolate ◽  
Pcr Analysis ◽  
In Planta ◽  
Dot Blot ◽  
Reverse Transcription Quantitative Pcr ◽  
Polymerase Chain ◽  
Differential Hosts

Clubroot caused by Plasmodiophora brassicae is an important disease of crucifers worldwide. Isolates of the pathogen can be classified into pathotypes according to their pathogenicity on differential hosts. In this study, the presence or absence of all database-available nonhousekeeping P. brassicae genes (118 in total) were assessed by polymerase chain reaction (PCR) analysis in isolates belonging to five P. brassicae pathotypes (2, 3, 5, 6, and 8 according to Williams’ differential set). One gene, designated Cr811, was present exclusively in the isolate of pathotype 5. This was further confirmed by dot blot hybridization and by PCR using alternative DNA preparations and primers. Reverse transcription quantitative PCR analysis indicated that in planta expression of Cr811 was up-regulated during canola infection, especially in the stage of secondary plasmodia. Primers specific to Cr811 could distinguish a field isolate of P. brassicae belonging to pathotype 5 from two other field isolates representing pathotypes 3 and 8. These findings suggest that Cr811 is a gene that is potentially involved in clubroot pathogenesis and that it also might serve as a molecular marker for differentiation of pathotype 5 from other pathotypes.

scholarly journals Detection and Quantification of Resistance of Venturia inaequalis Populations to Sterol Demethylation Inhibitors

1997 ◽  
Vol 87 (2) ◽  
pp. 184-190 ◽  
Author(s):  
Wolfram Köller ◽  
W. F. Wilcox ◽  
J. Barnard ◽  
A. L. Jones ◽  
P. G. Braun
Keyword(s):  
New York ◽  
Nova Scotia ◽  
Venturia Inaequalis ◽  
Apple Scab ◽  
Resistant Isolate ◽  
Cross Resistance ◽  
Sensitive Isolate ◽  
Sterol Demethylation Inhibitors ◽  
Sensitivity Data ◽  
Dmi Resistance

Monoconidial isolates of Venturia inaequalis were collected in 1990 and 1991 from orchards in New York, Michigan, and Nova Scotia that had never or only sporadically been treated with fungicides acting as sterol demethylation inhibitors (DMIs). Sensitivities of isolates to two representative DMIs (fenarimol and myclobutanil) were determined by a sensitivity test based on the relative growth (RG) of mycelial colonies at one discriminatory dose. Mean isolate sensitivities were not significantly different (P > 0.2) for the majority of the populations tested, and all sensitivity data obtained from these sites were combined to provide a baseline distribution of isolate sensitivities for both fenarimol and myclobutanil. The baseline distributions were compared with isolate sensitivities determined for an experimental orchard in Nova Scotia with a documented case of DMI resistance and for a commercial orchard in Michigan with a long history of DMI use and first evidence of practical DMI resistance. For both DMIs tested and in both treated orchards, frequencies of isolates with RG values <80 had decreased or only slightly increased compared to the baseline population. In contrast, frequencies of isolates with RG values >80 had increased more than 20-fold over baseline levels. Thus, isolates with RG values >80 were rated DMI resistant. The validity of a qualitative isolate classification was tested in controlled infection studies. At doses of fenarimol and myclobutanil recommended for commercial control of apple scab, reproduction of a typical sensitive isolate on treated apple leaves was suppressed completely. Lesions caused by a resistant isolate continued to expand and produced abundant conidia. Statistical analysis of orchard sensitivities revealed that the analysis of isolate counts grouped into the categories DMI sensitive or resistant was most indicative in comparisons of orchard sensitivities aimed at detection of practical DMI resistance. A high degree of cross-resistance between fenarimol and myclobutanil indicated that sensitivity tests with one of the DMIs employed as the diagnostic tool in this study can serve as a test for other DMIs.

scholarly journals Integrated Use of Pyraclostrobin and Epoxiconazole for the Control of Fusarium Head Blight of Wheat in Anhui Province of China

Plant Disease ◽  
2012 ◽  
Vol 96 (10) ◽  
pp. 1495-1500 ◽  
Author(s):  
Yu Chen ◽  
Ai-Fang Zhang ◽  
Tong-Chun Gao ◽  
Yong Zhang ◽  
Wen-Xiang Wang ◽  
...  
Keyword(s):  
Fusarium Head Blight ◽  
Mycelial Growth ◽  
Field Trials ◽  
Anhui Province ◽  
Cross Resistance ◽  
Head Blight ◽  
Baseline Sensitivity ◽  
Quinone Outside Inhibitor ◽  
Fusarium Asiaticum ◽  
Excellent Activity

Fusarium asiaticum and F. graminearum are the primary causal agents of Fusarium head blight (FHB) of wheat in China. Carbendazim (a benzimadazole fungicide, MBC), has been extensively used for the control of FHB, resulting in severe MBC resistance in China. This article presents the baseline sensitivity of F. asiaticum and F. graminearum isolates from Anhui Province of China to fungicides pyraclostrobin (a quinone outside inhibitor) and epoxiconazole (a sterol demethylation inhibitor). In the presence of salicylhydroxamic acid, the 50% effective concentration (EC50) values for pyraclostrobin in inhibiting mycelial growth of the 126 F. asiaticum isolates and 63 F. graminearum isolates were 0.012 to 0.135 μg/ml and 0.010 to 0.105 μg/ml, and the EC50 values for pyraclostrobin in inhibiting conidium germination of the F. asiaticum and F. graminearum populations were 0.047 to 0.291 and 0.042 to 0.255 μg/ml, respectively. The EC50 values for epoxiconazole in inhibiting mycelial growth of the F. asiaticum and F. graminearum populations were 0.12 to 0.95 and 0.16 to 0.93 μg/ml, respectively. All of the baseline sensitivity curves were unimodal. This study also suggested that there was no cross-resistance between MBC and pyraclostrobin or epoxiconazole. In the protective and curative tests, pyraclostrobin and epoxiconazole applied at 200 and 300 μg/ml exhibited over 75% protective and curative control efficacy in all treatments. In field trials, both pyraclostrobin and epoxiconazole at 225 g a.i./ha provided over 80% efficacy in 2010 and 2011 at both sites where MBC resistance occurred, suggesting excellent activity against FHB. Interestingly, integrated use of pyraclostrobin + epoxiconazole applied at 150 + 150 g a.i./ha provided over 85% efficacy at both sites in 2010 and 2011. Pyraclostrobin and epoxiconazole should be good alternatives to MBC for the control of FHB, and integrated use of these two fungicides might achieve greater efficacy.

scholarly journals Identification and characterization of Edwardsiella ictaluri from diseased Pangasius pangasius, cultured in Cirata Lake, Indonesia

2018 ◽  
Vol 19 (3) ◽  
pp. 816-822 ◽  
Author(s):  
MIRA MAWARDI ◽  
JAELANI JAELANI ◽  
ZAKKI ZAINUN ◽  
YUANI MUNDAYANA ◽  
BAIRD SAM CHILORA ◽  
...  
Keyword(s):  
Edwardsiella Ictaluri ◽  
Pcr Analysis ◽  
Nucleotide Sequencing ◽  
Infection Prevalence ◽  
Chain Reaction ◽  
Prevalence Level ◽  
Conventional Biochemical Test ◽  
Polymerase Chain ◽  
Identification And Characterization

Mawardi M, Jaelani, Zainun Z, Mundayana Y, Chilora BS, Hardi EH. 2018. Identification and characterization of Edwardsiella ictaluri from diseased Pangasius pangasius, cultured in Cirata Lake, Indonesia. Biodiversitas 19: 766-772. In January 2016, there were reported extensive mortalities of Pangasius pangasius cultured on cage, in Cirata Lake. Edwardsiella ictaluri is primarily recognized as a disease-causing pathogen in catfish species, causing an economically important bacterial infection hence affecting productivity of aquaculture enterprises in many regions across the E. ictaluri. The samples were 17 fishes collected from organs containing spleen, kidney, and liver. The total samples were 51 organs of fishes. The high bacteria infection prevalence level was found in kidney (29.41%). The gross pathological signs of sample of organs were abnormal sizes and abnormal color including white nodules. Bacteria were identified by conventional biochemical test, polymerase chain reaction (PCR) analysis and nucleotide sequencing. The results showed that P. pangasius cultured in cages in Cirata Lake is positively infected by E. ictaluri strain (accession number KF9071291). Other infection cases by Aeromonas sp. and Plesiomonas shigelloides as co-infection bacteria in P. pangasius were also found.

scholarly journals Genotypes and Phenotypic Characterization of Field Fusarium asiaticum Isolates Resistant to Carbendazim in Anhui Province of China

Plant Disease ◽  
2015 ◽  
Vol 99 (3) ◽  
pp. 342-346 ◽  
Author(s):  
Yu Chen ◽  
Xue Yang ◽  
Chun-Yan Gu ◽  
Ai-Fang Zhang ◽  
Tong-Chun Gao ◽  
...  
Keyword(s):  
Natural Populations ◽  
Point Mutations ◽  
Anhui Province ◽  
Phenotypic Characterization ◽  
Head Blight ◽  
Single Spore ◽  
Tubulin Gene ◽  
Fusarium Asiaticum ◽  
Moderate Resistance

Fusarium asiaticum is a causal agent of Fusarium head blight (FHB) of wheat in the southern part of China. Carbendazim has been extensively used for controlling FHB for more than 30 years, leading to the widespread carbendazim-resistant isolates in all major wheat-producing provinces in China, especially in Anhui Province. F. asiaticum isolates were collected throughout Anhui Province between 2010 and 2012 to monitor their sensitivity to carbendazim. In total, 74 of 899 single-spore isolates F. asiaticum were found to be resistant to carbendazim. Resistant isolates were collected from all of the sampled sites except Hefei of Anhui Province. The overall frequency of carbendazim resistance was shown to be 8.2%. Of the 74 isolates, 1, 68, and 5 had low resistance (LR), moderate resistance (MR) ,and high resistance (HR), respectively, to carbendazim. Five types of point mutations (F167Y, E198L, E198K, F200Y, and E198Q) in the β2-tubulin gene conferring resistance to carbendazim were detected in the field-resistant isolates with frequencies of 89.2, 2.7, 4.1, 2.7, and 1.4%, respectively. The point mutations at codon 167, 198, or 200 of the β2-tubulin gene were correlated with different levels of carbendazim resistance. Some of the sensitive and resistant isolates appeared to possess different biological characteristics; however, these might not be due to resistance. Because carbendazim resistance was generally widespread throughout Anhui Province, the sensitivity of F. asiaticum populations to carbendazim should be constantly monitored for the development of carbendazim resistance in natural populations.

scholarly journals The Rice Cation/H+ Exchanger Family Involved in Cd Tolerance and Transport

2021 ◽  
Vol 22 (15) ◽  
pp. 8186
Author(s):  
Wenli Zou ◽  
Jingguang Chen ◽  
Lijun Meng ◽  
Dandan Chen ◽  
Haohua He ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Pcr Analysis ◽  
Dependent Manner ◽  
Rice Varieties ◽  
Yeast Saccharomyces Cerevisiae ◽  
Cd Uptake ◽  
Homologous Genes ◽  
Dose Dependent

Cadmium (Cd), a heavy metal toxic to humans, easily accumulates in rice grains. Rice with unacceptable Cd content has become a serious food safety problem in many rice production regions due to contaminations by industrialization and inappropriate waste management. The development of rice varieties with low grain Cd content is seen as an economic and long-term solution of this problem. The cation/H+ exchanger (CAX) family has been shown to play important roles in Cd uptake, transport and accumulation in plants. Here, we report the characterization of the rice CAX family. The six rice CAX genes all have homologous genes in Arabidopsis thaliana. Phylogenetic analysis identified two subfamilies with three rice and three Arabidopsis thaliana genes in both of them. All rice CAX genes have trans-member structures. OsCAX1a and OsCAX1c were localized in the vacuolar while OsCAX4 were localized in the plasma membrane in rice cell. The consequences of qRT-PCR analysis showed that all the six genes strongly expressed in the leaves under the different Cd treatments. Their expression in roots increased in a Cd dose-dependent manner. GUS staining assay showed that all the six rice CAX genes strongly expressed in roots, whereas OsCAX1c and OsCAX4 also strongly expressed in rice leaves. The yeast (Saccharomyces cerevisiae) cells expressing OsCAX1a, OsCAX1c and OsCAX4 grew better than those expressing the vector control on SD-Gal medium containing CdCl2. OsCAX1a and OsCAX1c enhanced while OsCAX4 reduced Cd accumulation in yeast. No auto-inhibition was found for all the rice CAX genes. Therefore, OsCAX1a, OsCAX1c and OsCAX4 are likely to involve in Cd uptake and translocation in rice, which need to be further validated.

Characterization of a cDNA for Rhesus Monkey Protein C Inhibitor – Evidence for N-Terminal Involvement in Heparin Stimulation

1995 ◽  
Vol 74 (04) ◽  
pp. 1079-1087 ◽  
Author(s):  
Klaus-P Radtke ◽  
José A Fernández ◽  
Bruno O Villoutreix ◽  
Judith S Greengard ◽  
John H Griffin
Keyword(s):  
Rhesus Monkey ◽  
Protein C ◽  
Activated Protein C ◽  
Pcr Amplification ◽  
Amino Acid Sequences ◽  
Heparin Binding ◽  
Reactive Center ◽  
Protein C Inhibitor ◽  
Polymerase Chain

SummarycDNAs for protein C inhibitor (PCI) were cloned from human and rhesus monkey 1 liver RNAs by reverse transcription and polymerase chain reaction (PCR) amplification. Sequencing showed that rhesus monkey and human PCI cDNAs were 93% identical. Predicted amino acid sequences differed at 26 of 387 residues. Pour of these differences (T352M, N359S, R362K, L3631) were in the reactive center loop that is important for inhibitory specificity, and two were in the N-terminal helix (M8T, E13K) that is implicated in glycosaminoglycan binding. PCI in human or rhesus monkey plasma showed comparable inhibitory activity towards human activated protein C in the presence of 10 U/ml heparin. However, maximal acceleration of the inhibition of activated protein C required 5-fold lower heparin concentration for rhesus monkey than for human plasma, consistent with the interpretation that the additional positive charge (E13K) in a putative-heparin binding region increased the affinity for heparin.

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