Species composition,toxigenic potential and pathogenicity of Fusarium graminearum species complex isolates from southern Brazilian rice

This study aimed to assess the extent and distribution of Fusarium graminearum species complex (FGSC) diversity in rice seeds produced in southern Brazil. Four species and two trichothecene genotypes were detected among 89 FGSC isolates, based on a multilocus genotyping assay: F. asiaticum (69·6%) with the nivalenol (NIV) genotype, F. graminearum (14·6%) with the 15‐acetyldeoxynivalenol (ADON) genotype, and F. cortaderiae (14·6%) and F. meridionale (1·1%), both with the NIV genotype. Seven selected F. asiaticum isolates from rice produced NIV in rice‐based substrate in vitro, at levels ranging from 4·7 to 84·1 μg g^?1. Similarly, two F. graminearum isolates from rice produced mainly 15‐ADON (c. 15–41 μg g^?1) and a smaller amount of 3‐ADON (c. 6–12 μg g^?1). One F. meridionale and two F. cortaderiae isolates did not produce detectable levels of trichothecenes. Two F. asiaticum isolates from rice and two from wheat (from a previous study), and one F. graminearum isolate from wheat, were pathogenic to both crops at various levels of aggressiveness based on measures of disease severity in wheat spikes and rice kernel infection in a greenhouse assay. Fusarium asiaticum and the reference F. graminearum isolate from wheat produced NIV, and deoxynivalenol and acetylates, respectively, in the kernels of inoculated wheat heads. No trichothecene was produced in kernels from inoculated rice panicles by any of the isolates. These findings constitute the first report of FGSC composition in rice outside Asia, and confirm the dominance of F. asiaticum in rice agroecosystems.     

Identification of Neofabraea species causing bull's eye rot of apple in Poland and their direct detection in apple fruit using multiplex PCR

Based on partial sequence analysis of the β‐tubulin gene, 19 isolates of fungi causing bull's eye rot on apple in Poland were classified into species: Neofabraea alba, N. perennans and N. kienholzii. To the authors’ knowledge, the detection of N. kienholzii is the second in Europe and the first in Poland. Species affiliation of these fungi was confirmed by a new species‐specific multiplex PCR assay developed on the basis of previously published methods. The new protocol allowed for the specific identification of bull's eye rot‐causing species, both from pure cultures and directly from the skin of diseased or apparently healthy apples. In 550 samples of diseased fruits collected from nine cold storage rooms located in three regions of Poland, in 2011 and 2012, N. alba was detected as the predominant species causing bull's eye rot, occurring on average in 94% of the tested samples. Neofabraea perennans was found in a minority of apple samples, N. kienholzii was found only in two apple samples, while N. malicorticis was not detected in any sample tested. In tests on 120 apparently healthy fruits, only N. perennans was detected in a single sample. The results of genetic diversity analyses of bull's eye rot‐causing fungi based on the β‐tubulin gene sequence and an ISSR (inter‐simple sequence repeat) PCR assay with two primers were consistent, showing the expected segregation of tested isolates with respect to their species boundaries. However, the genetic distance between N. perennans and N. malicorticis was very low, as reported previously.     

Fusarium species and chemotypes associated with fusarium head blight and fusarium root rot on wheat in Sardinia

Environmental conditions in Sardinia (Tyrrhenian Islands) are conducive to fusarium root rot (FRR) and fusarium head blight (FHB). A monitoring survey on wheat was carried out from 2001 to 2013, investigating relations among these diseases and their causal agents. FHB was more frequently encountered in the most recent years while FRR was constantly present throughout the monitored period. By assessing the population composition of the causal agents as well as their genetic chemotypes and EF‐1α polymorphisms, the study examined whether the two diseases could be differentially associated to a species or a population. Fusarium culmorum chemotypes caused both diseases and were detected at different abundances (88% 3‐ADON, 12% NIV). Fusarium graminearum (15‐ADON genetic chemotype) appeared only recently (2013) and in few areas as the causal agent of FHB. In F. culmorum, two haplotypes were identified based on an SNP mutation located 34 bp after the first exon of the EF‐1α partial sequence (60% adenine, 40% thymine); the two populations did not segregate with the chemotype but the A‐haplotype was significantly associated with FRR in the Sardinian data set (P = 0·001), suggesting a possible fitness advantage of the A‐haplotype in the establishment of FRR that was neither dependent on the sampling location nor the sampling year. The SNP determining the Sardinian haplotype is distributed worldwide. The question whether the A‐haplotype segregates with characters facilitating FRR establishment will require further validation on a specifically sampled international data set.     

Detection of quantitative resistance loci associated with resistance to rice false smut (Ustilaginoidea virens) using introgression lines

False smut, caused by Ustilaginoidea virens, is a rice disease of increasing importance worldwide, with no source of high‐level resistance in the existing rice germplasm. To facilitate breeding varieties with good levels of field resistance to false smut, quantitative resistance loci (QRL) were identified using 213 introgression lines (ILs) from a cross between Teqing (recipient) and Lemont (donor) evaluated using natural infection at two hotspots of false smut in northeast China. Ten QRL affecting percentages of diseased hills, diseased panicles and diseased spikelets were detected and mapped to rice chromosomes 2, 3, 4, 6, 8, 10, 11 and 12. The Lemont alleles at all QRL increased false smut resistance. Four QRL (qFSR‐6‐7, qFSR‐10‐5, qFSR‐10‐2 and qFSR‐11‐2) had relatively larger and consistent effects across the two testing sites. Promising resistant ILs were identified, most of which had multiple QRL, suggesting that pyramiding multiple QRL by marker‐assisted selection would be an effective strategy for improving rice resistance to false smut. The identified QRL and their linked DNA markers will facilitate this breeding effort in the future.     

Biological control of Pythium,Rhizoctonia and Sclerotinia in lettuce: association of the plant protective activity of the bacterium Paenibacillus alvei K165 with the induction of systemic resistance

The soilborne fungi Sclerotinia sclerotiorum, Rhizoctonia solani and the oomycete Pythium ultimum are among the most destructive pathogens for lettuce production. The application of the biocontrol agent Paenibacillus alvei K165 to the transplant soil plug of lettuce resulted in reduced S. sclerotiorum, R. solani and P. ultimum foliar symptoms and incidence compared to untreated controls, despite the suppressive effect of the pathogens on the rhizosphere population of K165. In vitro, K165 inhibited the growth of S. sclerotiorum and R. solani but not P. ultimum. Furthermore, the expression of the pathogenesis‐related (PR) gene PR1, a marker gene of salicylic acid (SA)‐dependent plant defence, and of the Lipoxygenase (LOX) and Ethylene response factor 1 (ERF1) genes, markers of ethylene/jasmonate (ET/JA)‐dependent plant defence was recorded. K165‐treated plants challenged with P. ultimum showed up‐regulation of PR1, whereas challenge with R. solani resulted in up‐regulation of LOX and ERF1, and challenge with S. sclerotiorum resulted in up‐regulation of PR1, LOX and ERF1. This suggests that K165 triggers the SA‐ and the ET/JA‐mediated induced systemic resistance against P. ultimum and R. solani, respectively, while the simultaneous activation of the SA and ET/JA signalling pathways is proposed for S. sclerotiorum.     

Investigation of rice blast development in susceptible and resistant rice cultivars using a gfp‐expressing Magnaporthe oryzae isolate

In this study, an isolate of Magnaporthe oryzae expressing the green fluorescent protein gene (gfp) was used to monitor early events in the interaction of M. oryzae with resistant rice cultivars harbouring a blast resistance (R) gene. In the resistant cultivars Saber and TeQing (Pib gene), M. oryzae spores germinated normally on the leaf surface but produced morphologically abnormal germ tubes. Germling growth and development were markedly and adversely affected in leaves of these resistant cultivars. Penetration of host cells was never seen, supporting the idea that disruption of germling development on the leaf surface might be one of the resistance mechanisms associated with Pib function. Thus, this particular R gene appeared to function in the absence of host penetration by the fungal pathogen. Confocal laser scanning microscopy of M. oryzae‐infected susceptible rice cultivars showed the dimorphic growth pattern that is typically observed during the biotrophic and necrotrophic stages of leaf colonization in susceptible cultivars. The suitability of the gfp‐expressing M. oryzae isolate for further research on R‐gene function and identification of resistant genotypes in rice germplasm collections is discussed.     

On the interaction between weed and bird damage in rice

Weeds and granivorous birds are the two major biotic constraints to rice production in sub‐Saharan Africa. Anecdotal evidence advances the hypothesis that weedy fields attract granivorous birds to the extent that the joint weed × bird impact on crops is synergistic. We develop a theoretical model, based on experimental data, of the interaction between weed and bird damage to formally test this hypothesis. We observe that the response of bird‐inflicted yield loss to weeds is unimodal and inverse‐U‐shaped. Bird‐inflicted yield loss increases from 44% in weed‐free conditions to a maximum of 55% at a critical weed infestation level of 20 g dry weight m^?2. We conclude that the 11% increase in yield loss can be attributed to the synergistic interaction between weed and bird damage in rice. Our theory provides new insights into the analysis of the interaction between weeds and pests in general and supports a revision of currently accepted recommendations for weed management in rice. In areas where birds constitute an important production risk, it would be beneficial for rice farmers to remove those weeds that are tall enough to attract birds, even beyond the critical weed period.     

Detection and quantification of Ilyonectria spp. associated with black‐foot disease of grapevine in nursery soils using multiplex nested PCR and quantitative PCR

Three nursery fields and three rootstock mother fields from commercial nurseries located in Comunidad Valenciana region (central‐eastern Spain) were surveyed in July 2011 to detect the presence and to quantify Ilyonectria spp. in the soil. In each field, ten soil samples were taken randomly with a soil probe at a depth of 10–30 cm, and 10–20 cm from the base of the plant. Three replicate subsamples (10 g each) were taken from each soil sample. DNA was extracted and a multiplex nested PCR with species‐specific primer pairs (Mac1/MaPa2, Lir1/Lir2 and Pau1/MaPa2) was used to identify the species present. Among the 180 soil DNA samples analysed, Ilyonectria spp. were detected in 172 of them. Ilyonectria macrodidyma complex was the most frequently detected, being identified in 141 samples from all the fields evaluated. However, I. liriodendri was detected in only 16 samples, but was present in all open‐root field nurseries and in two rootstock mother fields. In addition, quantitative PCR (qPCR) assays were done to assess the levels of I. liriodendri and I. macrodidyma‐complex DNA in the soil samples. Detection of Ilyonectria spp. DNA using qPCR correlated with the fields found positive with the nested multiplex PCR. DNA concentrations of Ilyonectria spp. ranged from 0·004 to 1904·8 pg μL^?1. In general, samples from rootstock mother fields showed the highest DNA concentrations. The ability to detect and quantify Ilyonectria spp. genomic DNA in soil samples from nursery fields and rootstock mother fields confirms soils from both field types as important inoculum sources for black‐foot pathogens.     

Photosynthesis and sugar concentration are impaired by the defective active silicon uptake in rice plants infected with Bipolaris oryzae

The effect of soluble silicon (Si) on photosynthetic parameters and soluble sugar concentrations was determined in leaves of rice cv. Oochikara and mutant plants of Oochikara defective in active Si uptake [low silicon 1 (lsi1)]. Plants were grown in hydroponic culture amended with 0 (?Si) or 2 mm Si (+Si), under either low or high photon flux density (PFD) and with or without inoculation with Bipolaris oryzae, the causal agent of brown spot of rice. Leaf Si concentration increased by 141 and 435% in +Si cv. Oochikara and by 119 and 251% in +Si lsi1 mutant plants under high and low PFD, respectively, compared with ?Si plants. Plant biomass accumulation was improved by Si regardless of PFD, especially plants for cv. Oochikara. Brown spot severity was highest in ?Si plants for cv. Oochikara and lsi1 mutant plants under low PFD. In the presence of Si, disease severity in plants grown under both low and high PFD was reduced, except for lsi1 mutant plants under high PFD. Plant inoculation reduced the photosynthetic parameters measured regardless of plant material or Si supply. A decrease of net carbon assimilation rate (A) of inoculated plants under low PFD compared with non‐inoculated plants was associated with damage in the photosynthetic apparatus, except for +Si cv. Oochikara in which stomatal restriction [low water vapour conductance (g_s)] contributed to A reduction. Under high PFD, damage to the photosynthetic apparatus of inoculated plants was the main reason for the reduction in A for +Si and ?Si lsi1 mutant plants. In addition, for ?Si cv. Oochikara, a reduction in g_s contributed to reduced A. However, for +Si cv. Oochikara, g_s was the limiting factor for A. Inoculated plants of +Si cv. Oochikara had higher A values than +Si lsi1 mutant plants, regardless of environmental conditions. Soluble sugars were not detected in leaf tissues of plants under low PFD. For high PFD, Si improved the hexose concentration in non‐inoculated plants at 144 h after inoculation (hai) for lsi1 mutant plants and from 96 hai onwards for cv. Oochikara compared with ?Si plants. However, plant inoculation reduced hexose concentration compared with non‐inoculated plants, mainly in +Si plants, regardless of plant material. Sucrose concentration increased in leaves of cv. Oochikara in the presence of Si whether inoculated or not. For +Si lsi1 mutant plants, sucrose concentration increased only at 48 hai compared with ?Si plants, whether inoculated or not. The results of this study show that a minimum Si concentration is needed in leaf tissues of rice plants to avoid the negative impact of B. oryzae infection on photosynthesis and sugar concentration. High leaf Si concentration resulted in an increased soluble sugar concentration and together, but in independent ways, soluble sugar and Si reduced brown spot severity of rice.     

Phylogenetic relationships,pathogenicity and fungicide sensitivity of Greeneria uvicola isolates from Vitis vinifera and Muscadinia rotundifolia grapevines

Greeneria uvicola causes bitter rot on Vitis vinifera (bunch grapes) and Muscadinia rotundifolia (muscadine grapes) in warm moist temperate and subtropical regions. This study investigated the phylogenetic relationship of G. uvicola representatives from Australia (67 isolates), the USA (31 isolates), India (1 isolate) and Costa Rica (1 isolate) and compared their pathogenicity and fungicide sensitivity. Differences in cultural and conidial morphology were observed between the isolates from Australia and the USA. Phylogenetic relationships were determined based on three gene regions: the ribosomal DNA (rDNA) internal transcribed spacer 1 (ITS1–5?8S–ITS2), 28S large subunit (LSU) nuclear rDNA and β‐tubulin‐2. Greeneria uvicola isolates were clearly differentiated into four groups: isolates from Australia and India; USA isolates from V. vinifera; USA isolates from M. rotundifolia; and the isolate from Costa Rica. All isolates were pathogenic on V. vinifera (cv. Chardonnay) berries although those originating from M. rotundifolia were not as aggressive as isolates from V. vinifera, irrespective of geographical origin. Sensitivity to pyraclostrobin and salicylhydroxamic acid (SHAM) was studied. Despite differences in fungicide applications, hyphal growth inhibition was not significantly different for geographical location, cultivar, tissue, year of collection or different spray regimes. For the Australian and USA isolates, fungal growth inhibition was significantly greater for pyraclostrobin than for SHAM, and was significantly greater for the combined treatment than for each of the fungicides applied singly. The aetiological and epidemiological knowledge of bitter rot collected through this study will aid better prediction and management strategies of this pathogen.     

Evidence for Lettuce big‐vein associated virus as the causal agent of a syndrome of necrotic rings and spots in lettuce

Lettuce big‐vein associated virus (LBVaV, genus Varicosavirus) was shown to be responsible for characteristic necrotic symptoms observed in combination with big‐vein symptoms in lettuce breeding lines when tested for their susceptibility to lettuce big‐vein disease (BVD) using viruliferous Olpidium virulentus spores in a nutrient film technique (NFT) system. Lettuce plants showing BVD are generally infected by two viruses: Mirafiori lettuce big‐vein virus (MiLBVV, genus Ophiovirus) and LBVaV. New mechanical inoculation methods were developed to separate the two viruses from each other and to transfer both viruses to indicator plants and lettuce. After mechanical inoculation onto lettuce plants MiLBVV induced vein‐band chlorosis, which is the characteristic symptom of BVD. LBVaV caused a syndrome of necrotic spots and rings which was also observed earlier in lettuce plants inoculated in the NFT system, resembling symptoms described for lettuce ring necrosis disease (RND). This observation is in contrast with the idea that LBVaV only causes latent infections in lettuce. De novo next‐generation sequencing demonstrated that LBVaV was the only pathogen present in a mechanically inoculated lettuce plant with symptoms, providing evidence that LBVaV was the causal agent of the observed necrotic syndrome and thus fulfilling Koch’s postulates for this virus. The necrotic syndrome caused by LBVaV in lettuce is referred to as LBVaV‐associated necrosis (LAN).     

Elucidation of the infection process of Ustilaginoidea virens (teleomorph: Villosiclava virens) in rice spikelets

This study reports an efficient inoculation protocol that allowed cytological analysis of the infection process of the rice false smut pathogen Ustilaginoidea virens. Examination of serial semithin and ultrathin sections of infected spikelets showed that the primary infection sites for the pathogen were the upper parts of the three stamen filaments located between the ovary and the lodicules. The stigma and lodicules were also occasionally infected to a limited extent. The pathogen infected the filaments intercellularly and extended intercellularly along the filament base. The host cells were degraded gradually. The pathogen did not penetrate host cell walls directly and did not form haustoria. In the balls the ovary remained alive and was never infected. This suggests that the pathogen is a biotrophic parasite that grows intercellularly in vivo.     

The impact of management on weeds and aquatic plant communities in Hungarian rice crops

This study assessed the cultural and weed management factors influencing the weed communities of Hungarian rice fields. Hungary is situated at the northern limit of rice production with a history of about 300 years of rice culture. We surveyed the weed flora and 25 background variables in 100 active rice fields. Using a minimal adequate model containing 11 terms, 48.5% of the total variation in weed species data could be explained. The net effects of nine variables on species composition were significant. Crop cover was found to be the most important explanatory variable, which was followed by the herbicides penoxsulam and azimsulfuron, tillage depth, phosphorous and potassium fertilisers, years after last rotation, water depth in May, sowing type, pendimethalin and water conductivity. Filamentous algae, as the most abundant group of weeds, were positively associated with deep tillage, deep water and surface sowing. Echinochloa crus‐galli, one of the most troublesome grass weeds, was associated with low rice cover, shallow water and later years after crop rotation, while weedy rice favoured high crop cover, deep water and soil sowing. These findings can be used to design improved weed management strategies. The occurrence of red list species and charophytes in diverse micro‐mosaic patterns deserves attention from a conservation perspective, as well. The maintenance of these unique charophyte communities can be facilitated by shallow tillage without soil inversion.     

Characterization of Alternaria species associated with potato foliar diseases in China

The population structure of Alternaria species associated with potato foliar diseases in China has not been previously examined thoroughly. Between 2010 and 2013, a total of 511 Alternaria isolates were obtained from diseased potato leaves sampled in 16 provinces, autonomous regions or municipalities of China. Based on morphological traits and molecular characteristics, all the isolates were identified as Alternaria tenuissima, A. alternata or A. solani. Of the three species, A. tenuissima was the most prevalent (75·5%), followed by A. alternata (18·6%) and A. solani (5·9%). Phylogenetic analysis based on sequences of the internal transcribed spacer (ITS) region of ribosomal DNA (rDNA) of representative Alternaria isolates showed that A. solani was distinct from the two small‐spored Alternaria species. Phylogenetic analysis of the partial coding sequence of the histone 3 gene divided the same collection of isolates into three main clades representing A. tenuissima, A. alternata and A. solani, respectively. The pathogenicity of the isolates on detached leaves of potato cv. Favorite did not differ significantly between the three species or between isolates from different geographical origins. The results indicate that the population structure of Alternaria species associated with potato foliar diseases differs from that reported previously in China. This is the first report of A. tenuissima causing potato foliar diseases in China.     

Most Colletotrichum species associated with tree tomato (Solanum betaceum) and mango (Mangifera indica) crops are not host‐specific

An important constraint for crop production in Colombia is the high incidence of anthracnose caused by Colletotrichum species. Although several studies have focused on these fungi, the relationship between the different fungal species within the genus and their hosts and whether they display any host preference or host specificity has yet to be examined. In Colombia, diseases caused by Colletotrichum species are particularly severe in mango (Mangifera indica) and tree tomato (Solanum betaceum) crops. In a previous investigation, the Colletotrichum phylogenetic species attacking these crops were identified. The present study aimed to determine whether isolates collected from tree tomato and mango showed host preference or host specificity by assessing aggressiveness, spore density, latent period, and fitness of each strain on the two hosts. In the departments of Cundinamarca and Tolima, Colombia, isolates were collected from plants that presented typical anthracnose symptoms and were identified as C. acutatum, C. asianum, C. boninense, C. gloeosporioides, C. tamarilloi and C. theobromicola. Inoculation of conidia of each isolate onto both hosts showed isolates had no host preference and only the C. gloeosporioides isolate showed host specificity. However, in general, isolates produced a higher spore density when inoculated on the alternate host, which may indicate a difference in the degree of adaptation to each host. Statistical analyses of the assessed parameter values revealed that isolates use different infection strategies when infecting each host. In light of these results, the implications of using quantitative estimations of fitness when studying fungal pathogens are discussed.     

Absence of detectable yield penalty associated with insensitivity to Pleosporales necrotrophic effectors in wheat grown in the West Australian wheat belt

Genetic disease resistance is widely assumed, and occasionally proven, to cause host yield or fitness penalties due to inappropriate activation of defence response mechanisms or diversion of resources to surplus preformed defences. The study of resistance gene trade‐offs has so far been restricted to biotrophic pathogens. In some Pleosporales necrotrophic interactions, quantitative resistance is positively associated with insensitivity to effectors. Host lines that differ in sensitivity can easily be identified amongst current cultivars and advanced breeding lines. Large wheat cultivar trials were used to test whether lines sensitive or insensitive to three necrotrophic effectors from Pyrenophora tritici‐repentis and Parastagonospora nodorum differed in yield when subjected to natural disease and stress pressures in the West Australian wheat belt. There was no significant yield penalty associated with insensitivity to the fungal effectors ToxA, SnTox1 and SnTox3. Some yield gains were associated with insensitivity and some of these gains could be attributed to increased disease resistance. It is concluded that insensitivity to these effectors does not render such plants more vulnerable to any relevant biotic or abiotic stress present in these trials. These results suggest that the elimination of sensitivity alleles for necrotrophic effectors is a safe and facile strategy for improving disease resistance whilst maintaining or improving other desirable traits.     

Modelling the effects of herbicide dose and weed density on rice‐weed competition

The effects of herbicide dose on rice‐weed competition were investigated to develop a combined model, which can be utilised to estimate an optimum herbicide dose for a given weed density in paddy rice cultivation. Field studies were conducted in Suwon for rice‐Echinochloa crus‐galli competition and Iksan for rice‐Eleocharis kuroguwai during 2007. The competitive effect of the weeds E. crus‐galli and E. kuroguwai decreased with increasing doses of flucetosulfuron and azimsulfuron, respectively, in the same manner as the standard dose–response curve. The combination of the rectangular hyperbolic model and the standard dose–response curve adequately described the complex effects of herbicide dose and weed competition on rice yield. Parameter estimates were used with the model to predict rice yield and estimate the doses of flucetosulfuron and azimsulfuron required to restrict rice yield loss caused by E. crus‐galli and E. kuroguwai, respectively, to an acceptable level. For a rice yield of 5.0 t ha^?1, the model recommended flucetosulfuron doses of 8.7, 13.4 and 20.1 g a.i. ha^?1 when infested with E. crus‐galli at 12, 24 and 48 plants m^?2 respectively. For a rice yield of 5.2 t ha^?1, the model recommended azimsulfuron doses of 3.9, 7.5 and 12.6 g a.i. ha^?1 when infested with E. kuroguwai at 24, 48 and 96 plants m^?2 respectively. The theoretical outputs of the combined model appear robust and indicate there are opportunities for reduced herbicide use in the field. These now require evaluation under field conditions.     

Diversity and pathogenicity of the Ceratocystidaceae associated with cacao agroforests in Cameroon

Knowledge of the diversity and ecology of plant pathogenic fungi in cacao agroforests and surrounding natural ecosystems can inform the development of sustainable management strategies for new cacao disease outbreaks. This study investigated the occurrence of fungi related to the Ceratocystidaceae and their nitidulid beetle vectors in cacao agroforests in Cameroon, under diverse agroecological conditions. The fungi and their vectors were collected from artificially induced stem wounds on cacao and associated shade trees. Collections were also made from abandoned cacao pod husks and other tree wounds within and around plantations. Fungal isolates were identified using DNA sequence‐based phylogenies and morphological comparisons, and two representatives of each species were evaluated for pathogenicity on cacao. Five species of Ceratocystidaceae were recovered, including Huntiella chlamydoformis sp. nov., H. pycnanthi sp. nov. and H. moniliformis, as well as Thielaviopsis cerberus and T. ethacetica. The incidence of these fungi appeared to be influenced by the prevailing agroecological conditions. Nitidulid beetles in the genus Brachypeplus were found to be their most common insect associates on cacao. Both T. ethacetica and H. pycnanthi produced extensive lesions after inoculation on branches of mature cacao trees, while T. ethacetica also caused pod rot. Although their impact remains unknown, fungi in the Ceratocystidaceae and their nitidulid beetle vectors are common and probably contribute to the parasitic pressure in Cameroonian cacao agrosystems.     

Application of cycleave PCR to the detection of a point mutation (F167Y) in the β2‐tubulin gene of Fusarium graminearum

BACKGROUND: Resistance of Fusarium graminearum to the benzimidazole fungicide carbendazim is caused by point mutations in the β₂‐tubulin gene (FGSG_06611.3). The point mutation at codon 167 (TTT → TAT, F167Y) occurs in more than 90% of field isolates in China. It is important to find a suitable method for rapid detection and quantification of this point mutation in the F. graminearum populations. RESULTS: A pair of primers, Codon167F/Codon167R, were designed to amplify a fragment containing the mutation site, and two cycling probes labelled with different fluorescent reporters were used to detect whether the mutation was present. A cycleave real‐time PCR method was developed for rapid determination of the frequency of this point mutation in 282 F. graminearum perithecia collected from different fields in Jiangsu Province, China. The mutation frequency in ascospores from the perithecia to carbendazim by a spore germination assay was 6.0%, while the frequency of the point mutation at codon 167 by the cycleave real‐time PCR assay was 3.9%. CONCLUSION: The cycleave real‐time PCR method is suitable for accurate detection of the codon 167 point mutation. The frequency of this mutation in the β₂‐tubulin gene represents the resistance frequency in F. graminearum populations to carbendazim. Copyright © 2011 Society of Chemical Industry