scholarly journals Three-Locus Sequence Identification and Differential Tebuconazole Sensitivity Suggest Novel Fusarium equiseti Haplotype from Trinidad

Pathogens ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 175 ◽  
Author(s):  
Ria T. Villafana ◽  
Sephra N. Rampersad

The Fusarium incarnatum-equiseti species complex (FIESC) consists of 33 phylogenetic species according to multi-locus sequence typing (MLST) and Genealogical Concordance Phylogenetic Species Recognition (GCPSR). A multi-locus dataset consisting of nucleotide sequences of the translation elongation factor (EF-1α), calmodulin (CAM), partial RNA polymerase largest subunit (RPB1), and partial RNA polymerase second largest subunit (RPB2), was generated to distinguish among phylogenetic species within the FIESC isolates infecting bell pepper in Trinidad. Three phylogenetic species belonged to the Incarnatum clade (FIESC-15, FIESC-16, and FIESC-26), and one species belonged to the Equiseti clade (FIESC-14). Specific MLST types were sensitive to 10 µg/mL of tebuconazole fungicide as a discriminatory dose. The EC50 values were significantly different among the four MLST groups, which were separated into two homogeneous groups: FIESC-26a and FIESC-14a, demonstrating the “sensitive” azole phenotype and FIESC-15a and FIESC-16a as the “less sensitive” azole phenotype. CYP51C sequences of the Trinidad isolates, although under positive selection, were without any signatures of recombination, were highly conserved, and were not correlated with these azole phenotypes. CYP51C sequences were unable to resolve the FIESC isolates as phylogenetic inference indicated polytomic branching for these sequences. This data is important to different research communities, including those studying Fusarium phytopathology, mycotoxins, and public health impacts.

2021 ◽  
Vol 58 (04) ◽  
pp. 1301-1313
Author(s):  
Imran Ul Haq

Neopestalotiopsis species are known to be involved in plant diseases as associated pathogens. The taxonomic identification of the fungal group Neopestalotiopsisis little bit difficult due to its complex evolutionary history. In the present study, seven fungal isolates were investigated from canker-affected guava plants. The phylogeny for generic placement of these isolates was analyzed to validate them as Neopestalotiopsis genus by phylogenetic signals from the 28S nrRNA region (LSU). Generated morphological data was segregated as new morpho-species of the Neopestalotiopsis genus. Hence, the internal transcribed spacer (ITS), Translation elongation factor 1-α (TEF1-α) and Tubulin (TUB) genic region of these isolates were studied in juxtaposition with morphological data to resolve species limits. Both phylogenetic and morphological data revealed four novel species of the Neopestalotiopsis genus out of seven isolates studied. These Neopestalotiopsis species could be of great significance for further investigation as putative pathogens associated with canker or scabby canker disease in guava.


Plant Disease ◽  
2021 ◽  
Author(s):  
Ana Obradović ◽  
Jelena Stepanovic ◽  
Vesna Krnjaja ◽  
Aleksandra Bulajic ◽  
Goran Stanković ◽  
...  

The cosmopolitan species Fusarium graminearum Schwabe directly reduces yield, as well as grain quality of cereals, due to its ability to synthesize mycotoxins. Previously it was considered to be one species occurring on all continents. However, phylogenetic analysis employing the GCPSR method (Genealogical Concordance Phylogenetic Species Recognition) revealed the existence of 15 phylogenetic species within what is now recognised as the Fusarium graminearum Species Complex (FGSC) (Sarver et al. 2011). During 1996-2008, a MRIZP collection of FGSC isolates was established and isolates originating from wheat (5), maize (3) and barely (2) were selected for further study. Morphological features including the appearance of colonies and macroconidia (average size 38.5-53.1 × 4.6-5.4 µm, No 50) of all 10 isolates on PDA were consistent with descriptions of F. graminearum (O’Donnell et al. 2004, Leslie and Summerell 2006). Total DNA was isolated from mycelium removed from 7-day old colonies of single-spore isolates grown on PDA using the DNeasy Plant Mini Kit (Qiagen, Hilden). Further identification was based on amplification and sequencing of elongation factor TEF−1α, histone H3 and β−tubulin in both directions, with primers ef1/ef2, H3-1a/H3-1b and T1/T22, respectively (Jacobs et al. 2010). The sequences were deposited in NCBI under accession numbers MF974399 - MF974408 (TEF−1α), MG063783 - MG063792 (β−tubulin) and MF999139 - MF999148 (histone H3). Sequence analysis was performed using BLAST while genetic similarity was calculated using MEGA 6.0 software. Isolate 1339 originating from wheat (collected at the locality of Kikinda in 2006), shared 100% nucleotide identity with TEF−1α (DQ459745), histone H3 (DQ459728) and β−tubulin (DQ459643) of F. vorosii isolate NRRL37605 (Starkey et al. 2007). The remaining nine isolates were identified as F. graminearum as they shared 99% to 100% nucleotide similarity with F. graminearum NRRL 28439 (O’Donnell et al. 2004). Pathogenicity was tested using artificial inoculations of spikes during wheat flowering (Mesterhazy et al. 1999). Thirty classes were inoculated with each isolate, in three replicates. Inoculum was prepared from 7-day colonies on PDA, and 30 ml of a conidia suspension (1x105 conidia/ml) was used. Control plants were inoculated with sterile water. Three weeks after inoculation, typical Fusarium head blight symptoms were visible on inoculated plants, from which all 10 isolates were successfully reisolated. Control spikes remained symptomless. Disease severity was estimated on the 1-7 scale (Blandino et al. 2012). Average pathogenicity of the F. vorosii isolate 1339 was 1.9, and 2.4 -5.1 of F. graminearum isolates. Toxin production was determined using gas chromatography-tandem mass spectrometry. Kernels inoculated with the 10 isolates were ground and tested for the presence of deoxynivalenol (DON) and its acetyl derivatives 3ADON, 15ADON and NIV. F. vorosii isolate 1339 possessed the 15ADON chemotype, as well as eight F. graminearum isolates, while only one F. graminearum isolate was 3ADON chemotype. To date, F. vorosii has only been detected in Hungary on wheat (Toth et al. 2005) and Korea on barley, corn and rice (Lee et al. 2016). This is the first report of F. vorosii in Serbia, which is of great importance, because it indicates the spread of this toxigenic species. Further studies should be focused on determining the distribution, aggressiveness and toxicological profile of F. vorosii.


Plant Disease ◽  
2017 ◽  
Vol 101 (12) ◽  
pp. 2123-2136 ◽  
Author(s):  
K. W. T. Chethana ◽  
Y. Zhou ◽  
W. Zhang ◽  
M. Liu ◽  
Q. K. Xing ◽  
...  

Grape white rot is a common disease and causes considerable yield losses in many grape-growing regions when environmental conditions are favorable. We surveyed grape white rot in five provinces in China and collected 27 isolates from diseased grape tissues. Multigene phylogenetic analyses of the internal transcribed spacer region (ITS1-5.8S-ITS2), the 28S large subunit of nuclear ribosomal RNA (LSU), partial translation elongation factor 1-alpha gene (TEF 1-α), and partial histone 3 gene (HIS), coupled with genealogical concordance phylogenetic species recognition and morphological observations, revealed that Coniella vitis sp. nov. and C. diplodiella are the causal agents of grape white rot in China. Koch’s postulates were performed on Vitis vinifera cv. Summer Black in a greenhouse. These results confirmed the pathogenicity on grapes, as symptoms were reproduced, and also indicated significant variations in the virulence among C. vitis isolates. This work provides evidence that C. vitis is the main pathogen of grape white rot in China and also provides an optimized multigene backbone for resolving Coniella species.


2020 ◽  
Vol 44 (1) ◽  
pp. 113-139
Author(s):  
L.H. Han ◽  
G. Wu ◽  
E. Horak ◽  
R.E. Halling ◽  
J. Xu ◽  
...  

Strobilomyces is broadly distributed geographically and serves an important ecological function. However, it has been difficult to delimit species within the genus, primarily due to developmental variations and phenotypic plasticity. To elucidate phylogenetic relationships among species within the genus and to understand its species diversity, especially in Asia, materials of the genus collected from five continents (Africa, Asia, Australia, Europe, and North/Central America) were investigated. The phylogeny of Strobilomyces was reconstructed based on nucleotide sequences of four genes coding for: the largest and the second largest subunits of the RNA polymerase II (RPB1 and RPB2); the translation elongation factor subunit 1-α (TEF1); and the mitochondrial cytochrome oxidase subunit 3 (COX3). The combined results based on molecular phylogenetics, morphological characters, host tree associations, and geographical distribution patterns support a new classification consisting of two sections, sect. Strobilomyces and sect. Echinati. Using the genealogical concordance phylogenetic species recognition (GCPSR) approach, at least 33 phylogenetic species in Asia can be delimited, all of which are supported by morphological features, and five phylogenetic species remain to be described. The mountainous region of Southwest China is especially special, containing at least 21 species and likely represents a centre of diversification. We further compared our specimens with the type specimens of 25 species of Strobilomyces. Our comparisons suggest that, there are a total of 31 distinct species, while S. sanmingensis, S. verruculosus, S. subnigricans, and S. zangii/S. areolatus, are synonyms of S. mirandus, S. giganteus, S. alpinus and S. seminudus, respectively. Eight new species, namely, S. albidus, S. anthracinus, S. calidus, S. cingulatus, S. densisquamosus, S. douformis, S. microreticulatus and S. pinophilus, are described. A dichotomous key to the Asian Strobilomyces species is provided.


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