Effect of Fungicide Spray Cover on Botrytis Cinerea Infection in Grape Bunches

The effect of fungicide spray cover on Botrytis cinerea infection in grape bunches was determined. Bunches were sprayed at pea size and bunch closure with different volumes of a mixture of fenhexamid and a yellow fluorescent pigment, and the percentage fluorescent pigment coverage on pedicels was determined. Bunches were subsequently dusted with dry airborne conidia of B. cinerea in a settling tower and incubated for 24 h at high relative humidity (98%). Infection was determined by estimating the amount of B. cinerea infections on susceptible bunch parts (pedicel, receptacle and rachis) with isolations onto paraquat (herbicide) and Kerssies (B. cinerea selective) mediums.  Linear regressions for the part × stage combinations of percentage B. cinerea incidence on different bunchparts were fitted on mean coverage levels. An increase in spray cover caused linear reductions in levels of B. cinerea on bunch parts. Higher B. cinerea incidences were recorded at pea size. Furthermore, higher B. cinerea incidences were found on paraquat medium for both stages, than on Kerssies medium. The information gathered from this study will be used to facilitate future determination of minimum effective coverage levels for effective B. cinerea control in grape bunches

A fissitunicate ascus mechanism in the Calosphaeriaceae, and novel species of Jattaea and Calosphaeria on Prunus wood

During a survey of Prunus wood from South Africa, isolations were made of three presumably Calosphaerialean fungi that formed hyphomycetous, phialidic anamorphs in culture. In order to reveal the phylogenetic relationship of these fungi, they were characterised on a morphological and molecular (LSU and ITS rDNA) basis. Two isolates that formed a teleomorph in culture are newly described as Calosphaeria africana sp. nov. Although asci of Calosphaeria are characterised by having non-amyloid apical rings, two functional wall layers were observed in asci of C. africana, which has hitherto not been observed in any member of the Calosphaeriaceae. However, Calosphaeriaceae (Calosphaeriales, Sordariomycetes) are not closely related to other bitunicate fungi like Dothideomycetes, Chaetothyriales and bitunicate lichens. Possession of two separating wall layers is considered to be a result of both inherited abilities and convergent evolution under a strong selection pressure of the environmental conditions that favour an extension of the ascus. The other two species represented a separate lineage within Calosphaeriaceae, and formed phialophora-like anamorphs. By obtaining the teleomorph in culture, one of them could be identified as a species of Jattaea, described here as Jattaea prunicola sp. nov., while the second, which only produced the anamorph, is named as Jattaea mookgoponga sp. nov. These findings suggest that some species of Jattaea are true members of the Calosphaeriaceae, though the phylogenetic relation of the type, J. algeriensis, remains unknown. Furthermore, it also represents the first report of Jattaea on Prunus wood, and from South Africa

Coniochaeta (Lecythophora), Collophora gen. nov. and Phaeomoniella species associated with wood necroses of Prunus trees

Species of the genus Coniochaeta (anamorph: Lecythophora) are known as pathogens of woody hosts, but can also cause opportunistic human infections. Several fungi with conidial stages resembling Lecythophora were isolated from necrotic wood samples of Prunus trees in South Africa. In order to reveal their phylogenetic relationships, these fungi were studied on a morphological and molecular (5.8S nrDNA, ITS-1, ITS-2, GAPDH, EF-1α, 28S nrDNA, 18S nrDNA) basis. Some of the isolates were identified as Coniochaeta (Sordariomycetes), including C. velutina and two new species, C. africana and C. prunicola. The majority of the isolates, however, formed pycnidial or pseudopycnidial synanamorphs and were not closely related to Coniochaeta. According to their 28S nrDNA phylogeny, they formed two distinct groups, one of which was closely related to Helotiales (Leotiomycetes). The new genus Collophora is proposed, comprising five species that frequently occur in necrotic peach and nectarine wood, namely Co. africana, Co. capensis, Co. paarla, Co. pallida and Co. rubra. The second group was closely related to Phaeomoniella chlamydospora (Eurotiomycetes), occurring mainly in plum wood. Besides P. zymoides occurring on Prunus salicina, four new species are described, namely P. dura, P. effusa, P. prunicola and P. tardicola. In a preliminary inoculation study, pathogenicity was confirmed for some of the new species on apricot, peach or plum wood

Protection of Grapevine Pruning Wounds against Eutypa lata by Biological and Chemical Methods

Eutypa dieback, caused by the fungus Eutypa lata, is a serious disease of grapevines that infects mainly through pruningwounds. The aim of this study was to evaluate the in vitro efficacy of fungicides from various chemical groups againstE. lata, as well as the in vivo efficacy of the most effective fungicides and selected bacterial and fungal antagonists ofE. lata, in grapevine pruning wound protection trials. In vitro studies revealed that flusilazole, tebuconazole, benomyl,fenarimol and myclobutanil were the most effective fungicides to inhibit mycelial growth of E. lata. Two field trialswere conducted, one subjected to artificial inoculation and the second to natural infection only. In the first, benomyl,flusilazole and commercially available Trichoderma harzianum-containing products and an experimental Bacillussubtilis strain were applied to fresh pruning wounds. Two Cabernet Sauvignon vineyards were pruned in August2001 and 2002 and immediately treated and inoculated with a spore suspension of E. lata one day later. Isolationswere made from the treated pruning wounds after 12 months to assess the effectiveness of the treatments. Thefungicides benomyl and flusilazole were the most effective treatments, although the Trichoderma treatments T77 andTrichoseal spray caused a significant reduction in E. lata infection. In a second trial, pruning wounds of CabernetSauvignon, Sauvignon blanc, Red Globe and Bonheur were treated with the Trichoderma products Vinevax (=Trichoseal spray) and Eco77 (= T77) in August 2005 and 2006, subjected to natural infection only and evaluatedafter seven months. Vinevax and Eco77 not only reduced E. lata, but they also reduced the incidence of othergrapevine trunk disease pathogens

Aplosporella prunicola, a novel species of anamorphic Botryosphaeriaceae

Aplosporella prunicola is a newly described species associated with a dead branch of Prunus persica var. nucipersica from South Africa. Based on its phylogeny of the 28S rDNA (LSU) gene, the genus Aplosporella represents yet another anamorph lineage within the Botryosphaeriaceae (Botryosphaeriales). Aplosporella is characterized by having multilocular conidiomata opening by a single ostiole, verrucose, brown conidia, and the presence of prominent paraphyses. Details pertaining to the pathogenicity and host specificity of Aplosporella spp., remain to be elucidated

Novel Paraconiothyrium species on stone fruit trees and other woody hosts

Coniothyrium-like fungi are common wood and soil inhabitants and hyperparasites on other fungi. They belong to different fungal genera within the Pleosporales. Several isolates were obtained on wood of different Prunus species (plum, peach and nectarine) from South Africa, on Actinidia species from Italy and on Laurus nobilis from Turkey. Morphological and cultural characteristics as well as DNA sequence data (5.8S nrDNA, ITS1, ITS2, partial SSU nrDNA) were used to characterise them. The isolates belonged to three species of the recently established genus Paraconiothyrium. This is the first report of Paraconiothyrium brasiliense on Prunus spp. from South Africa. Two new species are described, namely Paraconiothyrium variabile sp. nov. on Prunus persica and Prunus salicina from South Africa, on Actinidia spp. from Italy and on Laurus nobilis from Turkey, and Paraconiothyrium africanum sp. nov. on Prunus persica from South Africa. Although other known species of Paraconiothyrium commonly produce aseptate conidia, those of P. africanum and P. hawaiiense comb. nov. are predominantly two-celled

Botryosphaeria spp. as Grapevine Trunk Disease Pathogens

Several species of Botryosphaeria, including Botryosphaeria dothidea, B. obtusa, B. parva and B. australis, were isolated and/or described from declining grapevines in association with a wide range of decline and dieback symptoms. Internal wood symptoms most commonly included black streaking, wedge-shaped necrosis and brown internal necrosis. Several diseases under names such as excoriose, grapevine decline syndrome and black dead arm have been described for decline and dieback symptoms associated with Botryosphaeria. This has led to the confusing situation where the same symptoms and causal species are often associated with more than one disease. This review summarises the research on Botryosphaeria on grapevine by focusing on the causal species, their associated symptoms, known epidemiology and possible effective control and management strategies

DNA phylogeny, morphology and pathogenicity of Botryosphaeria species on grapevines

Several species of Botr yosphaeria are known to occur on grapevines, causing a wide range of disorders including bud mortality, dieback, brown wood streaking and bunch rot. In this study the 11 Botryosphaeria spp. associated with grapevines growing in various parts of the world, but primarily in South Africa, are distinguished based on morphology, DNA sequences (ITS-1, 5.8S, ITS-2 and EF1-) and pathological data. Botryosphaeria australis, B. lutea, B. obtusa, B. parva, B. rhodina and a Diplodia sp. are confirmed from grapevines in South Africa, while Diplodia porosum, Fusicoccum viticlavatum and F. vitifusiforme are described as new. Although isolates of B. dothidea and B. stevensii are confirmed from grapevines in Portugal, neither of these species occurred in South Africa, nor were any isolates of B. ribis confirmed from grapevines. All grapevine isolates from Portugal, formerly presumed to be B. ribis, are identified as B. parva based on their EF1- equence data. From artificial inoculations on grapevine shoots, we conclude that B. australis, B. parva, B. ribis and B. stevensii are more virulent than the other species studied. The Diplodia sp. collected from grapevine canes is morphologically similar but phylogenetically distinct from D. sarmentorum. Diplodia sarmentorum is confirmed as anamorph of Otthia spiraeae, the type species of the genus Otthia (Botryosphaeriaceae). A culture identified as O. spiraeae clustered within Botryosphaeria and thus is regarded as probable synonym. These findings confirm earlier suggestions that the generic concept of Botryosphaeria should be expanded to include genera with septate ascospores and Diplodia anamorphs

Systematic reappraisal of Coniella and Pilidiella, with specific reference to species occurring on Eucalyptus and Vitis in South Africa

The genus Pilidiella, including its teleomorphs in Schizoparme, has a cosmopolitan distribution and is associated with disease symptoms on many plants. In the past, conidial pigmentation has been used as a character to separate Pilidiella (hyaline to pale brown conidia) from Coniella (dark brown conidia). In recent years, however, the two genera have been regarded as synonymous, the older name Coniella having priority. To address the generic question, sequences of the internal transcribed spacer region (ITS1, ITS2), 5.8S gene, large subunit (LSU) and elongation factor 1-[alpha] gene (EF 1-[alpha]) were analysed to compare the type species of Pilidiella and Coniella. All three gene regions supported the separation of Coniella from Pilidiella, with the majority of taxa residing in Pilidiella. Pilidiella is characterised by having species with hyaline to pale brown conidia (avg. length[ratio]width >1.5), in contrast to the dark brown conidia of Coniella (avg. length[ratio]width [less-than-or-eq, slant]1.5). Pilidiella diplodiella, which is a pathogen associated with white rot of grapevines, was shown to be an older name for C. petrakii. To delineate species in the P. diplodiella species complex, isolates were also compared based on histone (H3) gene sequences. Analyses derived from these sequence data separated P. diplodiella from a newly described species, P. diplodiopsis. The new species P. eucalyptorum sp. nov. is proposed for isolates formerly treated as C. fragariae and associated with leaf spots of Eucalyptus spp. This species clustered basal to Pilidiella, and may represent yet a third genus within this complex. Pilidiella destruens sp. nov. is newly described as anamorph of Schizoparme destruens, which is associated with twig dieback of Eucalyptus spp. in Hawaii. A key based on morphological characteristics is provided to separate the taxa treated in this stud

Laboratory evaluation of a specimen transport medium for downstream molecular processing of sputum samples to detect Mycobacterium tuberculosis

BACKGROUND : Modern molecular-based approaches for the detection of Mycobacterium tuberculosis in sputum samples promise quicker and more accurate detection of cases. However, processing sputum samples at central diagnostic facilities provides a diagnostic approach, but requires a safe and efficient system that is not affected by transport delays and ambient temperature to be feasible. We evaluated the technical properties of PrimeStore®-Molecular Transport Medium(PS-MTM) for its ability to inactivate mycobacteria, ensuring stability of DNA over time at ambient temperatures and to assess the compatibility of the transport medium with DNA extraction systems. METHODS : Assessment of the transport medium for application of sputum samples processed for the detection of M. tuberculosis included the inactivation of M. tuberculosis in spiked sputum samples, compatibility of the medium with three commercial nucleic extraction systems and stability of DNA in the medium at ambient temperature over 28 days. We further performed a clinical laboratory evaluation on 256 sputum specimens sent for tuberculosis investigation. RESULTS : Complete inactivation ofM. tuberculosis occurredwithin 30 min of exposure at a ratio of 1:3 for sputumto PS-MTM. Sputum specimen in PS-MTMshowed very good compatibility with automated bead-based extraction systems, producing high DNA output (estimated lower limits of detection: ~170 CFU/ml). Furthermore, PS-MTM samples remained stable over 28 days at ambient temperature displaying no significant change over time in Ctvalues (b5% on a mean starting value of 22.47). Of the 256 clinical sputumspecimens, 10.2%were culture positive and 11.0% were positive by real-time PCR of PS-MTM samples. CONCLUSIONS : Collecting and transporting sputum from TB suspects in PS-MTM offer safe transport at ambient temperature, DNA stability for extended periods without cooling and specimens directly suitable for molecular testing. This novel approach may support introduction and further scale-up of molecular diagnostics for TB in resource-limited settings.http://www.elsevier.com/locate/jmicmeth2016-10-31hb201