Resistance risk assessment for Fusarium graminearum to pydiflumetofen, a new succinate dehydrogenase inhibitor.

BACKGROUND: Pydiflumetofen is a new generation succinate dehydrogenase inhibitor currently undergoing the process of registration in China for the control of Fusarium head blight in wheat. A resistance risk assessment of Fusarium graminearum to pydiflumetofen was undertaken in this study. RESULTS: A total of 75 pydiflumetofen-resistant mutants were generated through spontaneous selection and displayed high resistance with an average resistance factor (RF) value of 78. Four mutants were generated through UV mutagenesis and displayed very high resistance with an RF value >1000. The sequence analysis results for Sdh genes and fitness studies revealed the existence of four types of mutations. In particular, 32 spontaneous selection mutants (SP mutants) had an arginine (R) to histidine (H) transition at position 86 in FGSdhC, resulting in seriously reduced fitness. Seven SP mutants had an R to cysteine (C) transition at position 86 in FGSdhC, resulting in reduced fitness. Thirty-six SP mutants had an alanine (A) to valine (V) transition at position 83 in FGSdhC and had no fitness penalties. The efficacy of pydiflumetofen towards a mutant carrying A83V in FGSdhC in vivo was significantly decreased at 42.7%. Four UV mutants had no mutations on all Sdh genes and no fitness penalties. Cross-resistance among boscalid, fluopyram and pydiflumetofen was observed. CONCLUSION: Sdhc mutations were found and other target site resistance may be present in laboratory PR mutants of F. graminearum. An overall moderate risk of resistance development in F. graminearum was recommended for pydiflumetofen. © 2019 Society of Chemical Industry.

Fitness of three chemotypes of Fusarium graminearum species complex in major winter wheat-producing areas of China.

In China, Fusarium head blight is caused mainly by the Fusarium graminearum species complex (FGSC), which produces trichothecene toxins. The FGSC is divided into three chemotypes: 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), and nivalenol (NIV). In order to predict the geographical changes in the distribution of these chemotype populations in major winter wheat-producing areas in China, the biological characteristics of twenty randomly selected isolates from each of the three chemotypes were studied. No significant difference was exhibited in the growth rate of 3-ADON, 15-ADON, and NIV isolates at 15°C. At 20°C and 25°C, the growth rate of 15-ADON isolates was the highest. At 30°C, the growth rate of NIV and 3-ADON isolates was significantly higher than that of 15-ADON isolates. The 15-ADON isolates produced the highest quantities of perithecia and two to three days earlier than the other two populations at each temperature, and released more ascospores at 18°C. The aggressiveness test on wheat seedlings and ears indicated there was no significant difference between the 3-ADON and 15-ADON isolates. However, the aggressiveness of NIV isolates was significantly lower than that of the 3-ADON and 15-ADON isolates. The DON content in grains from heads inoculated with the 3-ADON isolates was higher than the content of 15-ADON and NIV isolates. The results showed that 15-ADON population had the advantage in perithecia formation and ascospore release, and the 3-ADON population produced more DON in wheat grains. We suggested that distribution of these three chemotype populations may be related to these biological characteristics.

Homozygous and heterozygous point mutations in succinate dehydrogenase subunits b, c and d of Rhizoctonia cerealis conferring resistance to thifluzamide.

BACKGROUND: Thifluzamide, a succinate dehydrogenase inhibitor (SDHI) fungicide, is a promising fungicide for controlling wheat sharp eyespot (WSE). WSE is caused by Rhizoctonia cerealis. Information on the resistance mechanism of this pathogen to thifluzamide remains unavailable. RESULTS: We used selective reculturing and UV mutagenesis to generate thifluzamide-resistant mutants. Thifluzamide-resistant mutants were only generated through UV mutagenesis. Sequence analysis of succinate dehydrogenase (Sdh) genes revealed that two mutants had no mutation in RCSdhB, RCSdhC and RCSdhD, and the other 18 mutants all had at least one mutation in RCSdhB, RCSdhC or RCSdhD, either in a homozygous or heterozygous state. The majority of mutants included either RCSdhD-H116Y or RCSdhC-H139Y. They showed slight resistance to boscalid, bixafen and penflufen. Only one mutant possessed RCSdhB-H246Y, and it showed medium resistance to boscalid and penflufen and a slight resistance to bixafen. All the thifluzamide mutants were sensitive to flutolanil. Compared with their parental isolates, these mutants present no or minor fitness penalties. CONCLUSION: Homozygous and heterozygous point mutations in the succinate dehydrogenase subunits b, c and d of R. cerealis may be involved in thifluzamide resistance. © 2016 Society of Chemical Industry.

Conjunctively screening of biocontrol agents (BCAs) against fusarium root rot and fusarium head blight caused by Fusarium graminearum.

BACKGROUND AND AIMS: Fusarium root-rot and fusarium head blight are plant diseases caused by Fusarium sp. in different growth periods of wheat, bring heavy losses to crop production in China. This research is aiming to screen biocontrol agents conjunctively for controlling these two diseases at the same time, as well as evaluate our previous BCAs (Biological Control Agents) screening strategies in more complex situation, considering biocontrol is well concerned as an environmental-friendly plant disease controlling method. METHODS: Totally 966 bacterial isolates were screened from different parts of wheat tissues, of which potential biocontrol values were detected according to their abilities in antagonism inhibition and secreting extracellular hydrolytic enzyme. Biocontrol tests against fusarium root rot and fusarium head blight were carried out on 37 bacterial isolates with potential biocontrol capacity after pre-selection through ARDRA- and BOX-PCR analysis on strains with high assessment points. RESULTS: We acquired 10 BCAs with obvious biocontrol efficacy (more than 40%) in greenhouse and field tests. Pseudomonas fluorescens LY1-8 performed well in both two tests (biocontrol efficacy: 44.62% and 58.31%), respectively. Overall, correlation coefficient is 0.720 between assessment values of 37 tested BCA strains and their biocontrol efficacy in trails against fusarium root rot; correlation coefficient is 0.806 between their assessment values and biocontrol efficacy in trails against fusarium head blight. CONCLUSION: We acquired 10 well-performed potential BCAs, especially P. fluorescens LY1-8 displayed good biocontrol capacity against two different diseases on wheat. Biocontrol efficacies results in both greenhouse and field tests showed high positive correlation with assessment values (0.720 and 0.806), suggesting that the BCAs screening and assessing strategy previously developed in our lab is also adaptable for conjunctively screening BCAs for controlling both root and shoot diseases on wheat caused by same fungal pathogen.

Survey of Fusarium spp. Causing Wheat Crown Rot in Major Winter Wheat Growing Regions of China.

Fusarium crown rot of wheat has become more prevalent in China. To investigate the phylogenetic structure of Fusarium causing wheat crown rot in China, wheat basal stems with symptoms of the disease were collected from 2009 to 2013 in Jiangsu, Anhui, Henan, Hebei, and Shandong provinces. In total, 175 Fusarium isolates were collected and their mycotoxin chemotypes and distribution were identified. Among the 175 isolates, 123 were Fusarium asiaticum; 95 of these were the chemotype 3-acetyl-deoxynivalenol (3-AcDON) and 28 were nivalenol (NIV). Thirty-seven isolates belonged to F. graminearum, which were all 15-AcDON. Smaller numbers of isolates consisted of F. acuminatum, F. pseudograminearum, and F. avenaceum. The virulence of F. asiaticum and F. graminearum isolates on wheat crowns and heads was comparable. The virulence of isolates of the DON and NIV chemotype were statistically similar, but DON tended to be more aggressive. The DON concentrations in grains from wheat heads inoculated with isolates causing either Fusarium head blight or crown rot were similar. In the five provinces, F. asiaticum of the 3-AcDON chemotype was the predominant pathogen causing crown rot, followed by F. graminearum. Recent changes in causal Fusarium species, chemotypes, and distribution in China are discussed.

Geographic Distribution of Trichothecene Chemotypes of the Fusarium graminearum Species Complex in Major Winter Wheat Production Areas of China.

Fusarium head blight, caused by members of the Fusarium graminearum species complex (FGSC), is among the most destructive and economically important diseases of small grain crops, including wheat. To determine the phylogenetic species and mycotoxin (trichothecene) chemotypes of the FGSC in the major winter-wheat-producing areas of China, 530 isolates were collected from diseased wheat during the years 2008, 2009, and 2010, and typed using a polymerase chain reaction-based trichothecene genotype assay. Virulence of isolates with different chemotypes was also compared. Of the 530 isolates typed, 348 were F. asiaticum and 182 were F. graminearum. Subdividing the 530 isolates by the trichothecene predicted to be expressed, 482 were of the deoxynivalenol (DON) chemotype and 48 were nivalenol (NIV). Acetylated derivatives of DON included 3-acetyldeoxynivalenol (3-AcDON; 300 isolates), and 15-acetyldeoxynivalenol (15-AcDON; 182 isolates). Chemotypes of the F. asiaticum isolates were either 3-AcDON or NIV, with 3-AcDON being predominant. F. graminearum isolates were all of the 15-AcDON chemotype. F. asiaticum was the predominant phylogenetic species in the Yangtze River Basin and F. graminearum was dominant in the north of China. Two areas of co-occurrence of trichothecene chemotypes were found. The 3-AcDON and 15-AcDON isolates had similar levels of virulence. The DON isolates were significantly more virulent than those of the NIV. The 3-AcDON and 15-AcDON chemotypes were predominant in the Yangtze River Basin and areas north of the Yangtze River Basin, respectively, and it is suggested that geographic distribution is associated with differences in temperature as well as crop rotation systems.

[Analysis of simple sequence repeats in genomes of Sclerotinia sclerotiorum and Botrytis cinerea].

Simple sequence repeats or microsatellites have been used as genetic markers in population genetics because of their abundance and length variation between different individuals. This study examined the SSRs in the completely sequenced Sclerotinia sclerotiorum and Botrytis cinerea genomes. The occurrences, relative abundance, relative density, most common motifs, and the longest SSRs in the two species were analyzed, and compared with other plant pathogenic fungal species, such as Fusarium graminearum, Magnaporthe grisea and Ustilago maydis. The results demonstrated that the SSRs are abundant in S. sclerotiorum and B. cinerea genomes, and 6 539 and 8 627 SSRs were obtained from these species. The types and distributions of SSRs have similarities between the two species. In the genomes of S. sclerotiorum and B. cinerea, tetra-, penta- and hexa-nucleotide repeats were more abundant than other species, indicating high mutation rates in these species. Furthermore, the abundance and relative density of SSRs were not influenced by the genome sizes and GC content. The analysis in this study provided useful information on applications of microsatellites in population genetics of S. sclerotiorum and B. cinerea.