Identification and Genetic Division of Fusarium graminearum and Fusarium asiaticum by Species-Specific SCAR Markers

2013 ◽  
Vol 162 (2) ◽  
pp. 81-88 ◽  
Author(s):  
Xu Zhang ◽  
Hong-xiang Ma ◽  
Yong-jin Zhou ◽  
Jin-cheng Xing ◽  
Jian-hua Chen ◽  
...  
Keyword(s):  
Fusarium Graminearum ◽  
Scar Markers ◽  
Fusarium Asiaticum ◽  
Species Specific ◽  
Genetic Division

scholarly journals Trichothecene Genotypes of Fusarium graminearum Populations Isolated from Winter Wheat Crops in Serbia

Toxins ◽  
2018 ◽  
Vol 10 (11) ◽  
pp. 460 ◽  
Author(s):  
Vesna Krnjaja ◽  
Slavica Stanković ◽  
Ana Obradović ◽  
Tanja Petrović ◽  
Violeta Mandić ◽  
...  
Keyword(s):  
Fusarium Graminearum ◽  
Gene Clusters ◽  
Sensu Stricto ◽  
Molecular Techniques ◽  
Weather Data ◽  
Morphological Observation ◽  
Specific Primer ◽  
Head Blight ◽  
As Species ◽  
Species Specific

Fusarium graminearum as the main causal agent of Fusarium head blight (FHB) and its ability to produce trichothecenes was investigated by molecular techniques. A total of 37 strains isolated from the wheat, harvested in Serbia in 2005, 2008 and 2015, and previously designated by morphological observation as F. graminearum, were used for trichothecene genotypes characterization. The strains were identified using the species-specific primer set FG16R/FG16F while genotypic characterization was done using specific TRI13 and TRI3 sequences of the trichothecene gene clusters. The PCR assays identified all strains as species of F. graminearum sensu stricto with the DON/15-ADON genotype. The quantification of the mycotoxin (DON) was performed using the biochemical assay. The high levels of DON (>20,000 µg kg−1) were recorded in all of the strains from 2005, four strains from 2008 and two strains from 2015. Weather data of the investigated seasons, showed that the optimal temperature, frequent rains and high relative humidity (RH) was very favourable for the development of F. graminearum, affecting the DON biosynthesis.

PCR-RFLP for detection of Fusarium graminearum genotypes with resistance to phenamacril

Plant Disease ◽  
2020 ◽  
Author(s):  
Jiao-Sheng Li ◽  
Luo-Yu Wu ◽  
Hui Zhang ◽  
Xiu-Shi Song ◽  
Jian-Xin Wang ◽  
...  
Keyword(s):  
Fusarium Graminearum ◽  
Conventional Method ◽  
Resistance Mutations ◽  
Head Blight ◽  
Fusarium Asiaticum ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Resistant Strains ◽  
Codon Mutation

Phenamacril is a cyanoacrylate fungicide that provides excellent control of Fusarium head blight (FHB) or wheat scab, which is caused predominantly by Fusarium graminearum and Fusarium asiaticum. Previous studies revealed that codon mutations of the myosin-5 gene of Fusarium spp. conferred resistance to phenamacril in vitro lab experiments. In this study, PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) was developed to detect three common mutations (A135T, GCC to ACC at codon 135; S217L, TCA to TTA at codon 217, and E420K, GAA to AAA at codon 420) in F. graminearum induced by fungicide domestication in vitro. PCR products of 841 bp (for mutation of A135T), 802 bp (for mutation of S217L) or 1649 bp (for mutation of E420K) in myosin-5 gene were amplified respectively by appropriate primer pairs. Restriction enzyme KpnⅠ, TasⅠ or DraⅠ was used to distinguish phenamacril-sensitive and -resistant strains with mutation genotypes of A135T, S217L and E420K, respectively. KpnⅠ digested the 841 bp PCR products of phenamacri-resistant strains with codon mutation A135T into two fragments of 256 bp and 585 bp. In contrast, KpnⅠ did not digest the PCR products of sensitive strains. TasⅠ digested the 802 bp PCR products of phenamacril-strains with codon mutation S217L into three fragments of 461 bp, 287bp and 54 bp. In contrast, TasⅠ digestion of the 802 bp PCR products of phenamacril-sensitive strains resulted in only two fragments of 515bp and 287bp. DraⅠ digested the 1649 bp PCR products of phenamacril-resistant strains with codon mutation E420K into two fragments of 932 bp and 717 bp, while the PCR products of phenamacril-sensitive strains was not digested. The three genotypes of resistance mutations were determined by analyzing electrophoresis patterns of the digestion fragments of PCR products. The PCR-RFLP method was evaluated on 48 phenamacril-resistant strains induced by fungicide domestication in vitro and compared with the conventional method (mycelial growth on fungicide-amended agar). The accuracy of the PCR-RFLP method for detecting the three resistant mutation genotypes of F. graminearum to phenamacril was 95.12% compared with conventional method. Bioinformatics analysis revealed that the PCR-RFLP method could also be used to detect the codon mutations of A135T and E420K in F. asiaticum.

Quantification of Fusarium graminearum in infected wheat by species specific real-time PCR applying a TaqMan Probe

2004 ◽  
Vol 59 (1) ◽  
pp. 141-146 ◽  
Author(s):  
G.H. Reischer ◽  
M. Lemmens ◽  
A. Farnleitner ◽  
A. Adler ◽  
R.L. Mach
Keyword(s):  
Real Time ◽  
Fusarium Graminearum ◽  
Real Time Pcr ◽  
Taqman Probe ◽  
Species Specific

scholarly journals Fitness traits of deoxynivalenol and nivalenol-producing Fusarium graminearum species complex strains from wheat

2017 ◽  
Author(s):  
C.P. Nicolli ◽  
F.J. Machado ◽  
P. Spolti ◽  
E.M. Del Ponte
Keyword(s):  
Fusarium Graminearum ◽  
Mycelial Growth ◽  
Species Complex ◽  
Head Blight ◽  
Fitness Traits ◽  
Fusarium Graminearum Species Complex ◽  
Important Trait ◽  
South Of Brazil ◽  
Species Specific ◽  
Moderately Resistant

AbstractFusarium graminearum of the 15-acetyl(A)deoxynivalenol(D0N) chemotype is the main cause of Fusarium head blight (FHB) of wheat in south of Brazil. However, 3-ADON and nivalenol(NIV) chemotypes have been found in other members of the species complex causing FHB in wheat. To improve our understanding of the pathogen ecology, we assessed a range of fitness-related traits in a sample of 30 strains representatives of 15-ADON (F. graminearum), 3-ADON (F. cortaderiae and F. austroamericanum) and NIV (F. meridionale and F. cortaderiae). These included: perithecia formation on three cereal-based substrates, mycelial growth at two suboptimal temperatures, sporulation and germination, pathogenicity towards a susceptible and a moderately resistant cultivar and sensitivity to tebuconazole. The most important trait favoring F. graminearum was its 2x higher sexual fertility (> 40% PPI = perithecia production index) than the other species (< 30% PPI); PPI varied among substrates (maize > rice > wheat). In addition, sensitivity to tebuconazole appeared lower in F. graminearum which had the only strain with EC50 > 1 ppm. In the pathogenicity assays, the DON-producers were generally more aggressive (1.5 to 2x higher final severity) towards the two cultivars, with 3-ADON or 15-ADON leading to higher area under the severity curve than the NIV strains in the susceptible and moderately resistant cv., respectively. There was significant variation among strains of a same species with regards asexual fertility (mycelial growth, macroconidia production and germination), which suggest a strain-rather than a species-specific differences. These results contribute new knowledge to improve our understanding of the pathogen-related traits that may explain the dominance of certain members of the species complex in specific wheat agroecosystems.

scholarly journals First Report of the Nivalenol Chemotype of Fusarium graminearum Causing Head Blight of Wheat in the Grand Duchy of Luxembourg

Plant Disease ◽  
2009 ◽  
Vol 93 (11) ◽  
pp. 1217-1217 ◽  
Author(s):  
M. Pasquali ◽  
F. Giraud ◽  
C. Brochot ◽  
L. Hoffmann ◽  
T. Bohn
Keyword(s):  
Fusarium Graminearum ◽  
Aerial Mycelium ◽  
Triticum Aestivum L ◽  
Negative Control ◽  
Head Blight ◽  
First Report ◽  
Low Levels ◽  
Species Specific ◽  
Control Water ◽  
Positive Controls

Head blight caused by Fusarium graminearum is one of the major diseases of wheat (Triticum aestivum L.) in Luxembourg (2) and there is concern for mycotoxins in diseased grain. Isolates of F. graminearum have been assigned to chemotypes based on the particular toxins produced. Ten wheat fields representing different topoclimatological areas of Luxembourg were surveyed in 2007 and 2008 to determine the frequency and distribution of chemotypes. Partially blighted wheat heads were collected, and diseased grains were plated on Fusarium-selective agar (dichloran-chloramphenicol-peptone) for 12 days at 22 ± 2°C with a 12-h light period. Monoconidial isolates of F. graminearum (79 in 2007 and 85 in 2008) were obtained by conidia dilution on 2% water agar and needle selection under a microscope. F. graminearum isolates showed rapid growth on potato dextrose agar, dense aerial mycelium with red pigment deposits in the plate, macroconidia with five to six defined septa, and a basal cell with the typical foot shape. Microconidia were absent. To confirm species identification, a PCR reaction was carried out using the F. graminearum species-specific primers Fg16F (5′-CTCCGGATATGTTGCGTCAA-3′) and Fg16R (5′-GGTAGGTATCCGACATGGCAA-3′) according to Demeke et al. (1). Chemotype of each isolate was determined according to Ward et al. (4). In particular, PCR primer 12CON (5′ CATGAGCATGGTGATGTC-3′) coupled with primer 12NF (5′-TCTCCTCGTTGTATCTGG-3′) and primer 3CON (5′-TGGCAAAGACTGGTTCAC-3′) coupled with primer 3NA (5′-GTGCACAGAATATACGAGC-3′) identified the nivalenol chemotype, primer 12CON coupled with primer 12-15F (5′-TACAGCGGTCGCAACTTC-3′) and primer 3CON coupled with primer 3D15A (5′-ACTGACCCAAGCTGCCATC-3′) identified the 15-acetylated deoxynivalenol (DON) chemotype, while primer 12CON coupled with primer 12-3F (5′-CTTTGGCAAGCCCGTGCA-3′) and primer 3CON coupled with primer 3D3A (5′-CGCATTGGCTAACACATG-3′) identified 3-acetylated DON chemotype. Reactions were repeated two times and positive controls (provided by Kerry O'Donnell, NRRL collection, Peoria, IL) and a negative control (water) were used in each reaction. Frequency of the nivalenol chemotype was found to be 2.5% in 2007 and 1% in 2008. Interestingly, the nivalenol chemotype was absent in southern Luxembourg. According to this finding, nivalenol was likely to be present at low levels in grain from Reisdorf and Echternach in 2007 (central Luxembourg) and in 2008 from grain of Troisvierges (northern Luxembourg). The remaining isolates in both years belonged to the 15-acetylated DON chemotype and the 3-acetylated DON chemotype was not detected. Compared with a previous report from the Netherlands (3), the nivalenol chemotype in Luxembourg is less frequent and widespread. To our knowledge, this is the first report of the nivalenol chemotype of F. graminearum causing head blight on wheat in Luxembourg. References:(1) T. Demeke et al. Int. J. Food Microbiol. 103:271, 2005. (2) F. Giraud et al. Plant Dis. 92:1587, 2008. (3) C. Waalwijk et al. Eur. J. Plant Pathol. 109:743, 2003. (4) T. J. Ward et al. Fung. Genet. Biol. 45:473, 2008.

scholarly journals Development of species-specific SCAR markers for identification and authentication of three rare Peninsular Malaysian endemic Coelogyne (Orchidaceae) orchids

F1000Research ◽  
2020 ◽  
Vol 9 ◽  
pp. 1161
Author(s):  
Yoh Kok Hon ◽  
Christina Seok-Yien Yong ◽  
Janna Ong Abdullah ◽  
Rusea Go
Keyword(s):  
Scar Marker ◽  
Management Plan ◽  
Peninsular Malaysia ◽  
Single Band ◽  
Scar Markers ◽  
Orchid Species ◽  
Endangered Orchid ◽  
Reproductive Structures ◽  
In The Wild ◽  
Species Specific

Background: Coelogyne kaliana, Coelogyne stenochila and Coelogyne tiomanensis are three valuable rare orchid species endemic to Peninsular Malaysia, currently rampantly traded illegally via the internet and through local nurseries, which label them as hybrids to avoid enforcement detection. Drastic measures to ensure the continued existence of their populations in the wild should be introduced as they are rapidly diminishing into extinction, including the development of rapid and accurate species-specific identification tools. These three orchid species are highly similar morphologically and currently it is impossible to distinguish among them without their reproductive structures. Methods:  RAPD-based species-specific SCAR markers were developed to distinguish and authenticate the identity of these three endemic Peninsular Malaysian Coelogyne species. Results: Three SCAR markers were successfully developed in this study. SCAR marker primer pair, CKL_f / CKL_r was specific to C. kaliana as it produced a unique single band of 271 bp but not in C. stenochila and C. tiomanensis.  SCAR marker primer pair CST_f / CST_r amplified a single band of 854 bp in C. stenochila and two bands of different sizes (372 bp and 858 bp) in C. tiomanensis, but no amplification in C. kaliana. The third SCAR marker primer pair, CTI_f / CTI_r produced a single band (about 500 bp) for both C. stenochila and C. tiomanensis, but showed no amplification in C. kaliana. Conclusions: Although not all these SCAR markers were species amplification specific, they could be used to discriminate among the three Coelogyne species effectively.  Accurate species identification is one of the most important steps to allow a proper management plan to be established in the effort to conserve these three endangered orchid species of Peninsular Malaysia. Besides, it could effectively put a stop to the illegal trading of these rare endangered orchid species worldwide.

Discovery of a species‐specific novel antifungal compound against Fusarium graminearum through an integrated molecular modeling strategy

2020 ◽  
Vol 76 (12) ◽  
pp. 3990-3999
Author(s):  
Weitao Fu ◽  
Ningjie Wu ◽  
Di Ke ◽  
Yun Chen ◽  
Tianming Xu ◽  
...  
Keyword(s):  
Molecular Modeling ◽  
Fusarium Graminearum ◽  
Antifungal Compound ◽  
Species Specific ◽  
Modeling Strategy
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