Browsing by Author "Halleen, Francois"
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- ItemCharacterisation of Cylindrocarpon spp. associated with black foot disease of grapevine(Stellenbosch : Stellenbosch University, 2005-12) Halleen, Francois; Crous, Pedro W.; Fourie, Paul H.; Stellenbosch University. Faculty of Agrisciences. Dept. of Plant Pathology.ENGLISH ABSTRACT: During the past few years a drastic reduction has been noted in the survival rate of grafted grapevines in nurseries, as well as in young vineyards in the Western Cape Province of South Africa. Circumstantial evidence suggested that Cylindrocarpon spp., which cause black foot disease of grapevine, were associated with this decline. Black foot disease of grapevine is a relatively new, and as yet poorly known disease affecting vines in various countries where grapevines are cultivated. Primary aims of this research have been (1) to conduct nursery surveys in order to determine which fungi are involved in the decline phenomenon, with special reference to the involvement of Cylindrocarpon spp., (2) to identify and characterise the organisms believed to be the causal organisms of black foot disease, and (3) the development of control and/or management strategies to prevent or eradicate Cylindrocarpon infections. Nursery grapevines were sampled at different stages from three commercial nurseries in the Wellington area of the Western Cape Province and were investigated during the 19992000 season by means of destructive sampling. The first samples were taken in September from callused cuttings prior to planting in nurseries. After planting, asymptomatic rooted cuttings were selected from nurseries after 3, 6 and 9 months. Isolation studies clearly demonstrated that different “Cylindrocarpon spp.” infected cuttings from nursery soils. These species rarely occurred in rootstock propagation material prior to planting. At the time of planting, the susceptible basal ends (especially the pith area) of most of the nursery cuttings are partly or even fully exposed. Callus roots also break during the planting process, resulting in small wounds susceptible to infection by soilborne pathogens. The isolation studies revealed that the first infections occurred in the roots, followed by infections of the rootstocks. These infections increased progressively during the course of the growing season. Substantial variation in cultural and morphological characters was observed among the Cylindrocarpon isolates obtained from the nursery survey, as well as from isolations that were made from diseased grapevines. Morphological and phylogenetic studies were conducted to identify these “Cylindrocarpon spp.” and to establish their association with black foot disease. Sequences of the partial nuclear large subunit ribosomal DNA (LSU rDNA), internal transcribed spacers 1 and 2 of the rDNA including the 5.8S rDNA gene (ITS), and partial β-tubulin gene introns and exons were used for phylogenetic inference. Phylogenetic analyses confirmed the diversity observed among the isolates and four Cylindrocarpon-like species were identified. One of these species was initially identified as Cylindrocarpon destructans. However, further research revealed C. destructans to represent a species complex. Grapevine isolates of “C. destructans” proved to be identical to the ex-type strain of Cylindrocarpon liriodendri, which also produced a teleomorph, Neonectria liriodendri in culture. A second species was newly described in this study as Cylindrocarpon macrodidymum (Neonectria macrodidyma). The two remaining Cylindrocarpon-like species were placed in a new genus, Campylocarpon. The two species were named Campylocarpon fasciculare and Campylocarpon pseudofasciculare. Pathogenicity studies confirmed that all four species were able to reduce root and shoot mass significantly. Knowledge obtained pertaining to the disease cycle of black foot disease suggest that suitable management strategies should focus on prevention of primary infection in nurseries. However, at present, no fungicides are registered for control of this disease in South African vineyards or nurseries. Thirteen fungicides were screened in vitro for mycelial inhibition of these pathogens. Prochloraz manganese chloride, benomyl, flusilazole and imazalil were the most effective fungicides tested, and were subsequently included in semi-commercial field trials. Basal ends of grafted cuttings were dipped (1 min) in various chemical and biological treatments prior to planting in open-rooted nurseries. Black foot pathogens were not isolated from grafted cuttings prior to planting in nurseries. Additional treatments involved soil amendments with Trichoderma formulations and hot water treatment (50°C for 30 min) of dormant nursery grapevines. Field trials were evaluated after a growing season of eight months. The incidence of black foot pathogens was not significantly and/or consistently reduced by the majority of chemical or biological treatments. However, these pathogens were not isolated from uprooted plants that were subjected to hot water treatment. It is therefore recommended that hot water treatment of dormant nursery plants be included in an integrated strategy for the proactive management of black foot disease in grapevine nurseries.
- ItemDetection and quantification of black foot and crown and root rot pathogens in grapevine nursery soils in the Western Cape of South Africa(Firenze University Press, 2018) Langenhoven, Shaun; Halleen, Francois; Spies, Christoffel F. J.; Stempien, Elodie; Mostert, LizelBlack foot disease (BFD) and crown and root rot (CRR) are important soilborne diseases that affect young grapevines in nurseries and vineyards. A 3-year survey (2013–2015) of five open-field grapevine nurseries was conducted in the Western Cape Province of South Africa. The survey involved the isolation of BFD and CRR pathogens from grafted rootstocks (ten plants per nursery, per year) that were rooted in soil for 1 year. In 2013 and 2015, grapevines were sampled, while in 2014, sampling was focused on rotation crops and weeds (ten plants each). The rotation crops included white mustard, lupins, canola, triticale and forage radish. The weed species sampled included Johnson grass, ryegrass, winter grass, Cape marigold and corn spurry. Soil samples from ten sites per nursery were also collected in close proximity to the sampled plants, at depths of 0–30 cm and 30–60 cm (ten samples per depth). Isolations were made from the grapevines, rotation crops and weeds. Pathogen detection and quantification in the soil were determined using quantitative real-time polymerase chain reaction technology. The predominant BFD pathogens isolated from grapevines were Campylocarpon fasciculare, Ca. pseudofasciculare and Dactylonectria macrodidyma. The predominant CRR pathogens were Pythium irregulare and Phytopythium vexans. Dactylonectria macrodidyma, D. novozelandica, D. pauciseptata, Py. irregulare, Py. ultimum var. ultimum and Py. heterothallicum were isolated from triticale roots. Dactylonectria spp. were also isolated from corn spurry, while Py. irregulare and Py. ultimum var. ultimum were isolated from numerous weeds and rotation crops. Mean soil DNA concentrations of Ilyonectria and Dactylonectria were from 0.04 to 37.14 pg μL-1, and for Py. irregulare were between 0.01 and 3.77 pg μL-1. The Phytophthora mean soil DNA concentrations ranged from 0.01 to 33.48 pg μL-1. The qPCR protocols successfully detected and quantified BFD and CRR pathogens in grapevine nursery soil. This is the first report of D. pauciseptata and D. alcacerensis in South African grapevine nurseries.
- ItemHymenochaetales associated with esca-related wood rots on grapevine with a special emphasis on the status of esca in South African vineyards(Firenze University Press, 2015) Cloete, Mia; Fischer, Michael; Mostert, Lizel; Halleen, FrancoisEsca disease is a problem on grapevines worldwide. This disease complex is characterised by several external and internal symptoms including foliar tiger-stripe chlorosis and necrosis, dieback, wood necrosis and white rot. The causal organisms of esca are primarily Phaeomoniella chlamydospora, several Phaeoacremonium species and basidiomycete species from the order Hymenochaetales, the latter ones responsible for causing the white rot symptom. Basidiomycete species causing the wood rot symptom of esca differ among grapevine-growing areas worldwide. South African vineyards are unique in having a minimum of ten different basidiomycete taxa from five different genera associated with the esca complex. In general, Hymenochaetales species are associated with white rot on woody plants and there are several species that are economically important to the agricultural and forestry industries. Few Hymenochaetales species have been described from the African continent, though this review is an indication of the previously unknown diversity of these fungi in Southern Africa.
- ItemOccurrence fungi causing black foot on young grapevines and nursery rootstock plants in Italy(Firenze University Press, 2017) Carlucci, Antonia; Lops, Francesco; Mostert, Lizel; Halleen, Francois; Raimondo, Maria LuisaYoung grapevine plants with decline and wood necrosis symptoms were collected from vineyards and nurseries in the Apulia and Molise regions, Italy, from 2013 to 2015. Isolations of fungi were prepared from 45 diseased grapevine plants, and the cultures were identified. Several species commonly associated with Petri disease, Botryosphaeria dieback, and black foot disease were isolated. A detailed study was carried out, and 182 isolates resembling Cylindrocarpon-like asexual forms were identified through morphological characterisation and DNA analysis of internal transcribed spacer regions 1 and 2 of the rRNA gene and the partial β-tubulin gene. Dactylonectria torrensensis and Ilyonectria liriodendri were identified based on morphological features and the partial histone 3 gene, so these fungi can be defined as the causal agents of black foot on grapevine for the first time in Italy. Thelonectria blackeriella is also described as a new species, through morphological characterisation and multigenic analysis using sequence data for five loci (large subunit RNA, internal transcribed spacers, β-tubulin, actin, RNA polymerase II subunit 1). This new species was associated with black foot symptoms according to preliminary pathogenicity tests, with representative isolates of each of the three species. Pathogenicity tests showed that these species can cause black streaking in the wood of 1-year-old grapevine rootstock shoots. The identification of D. torresensis, I. liriodendri and T. blackeriella from young grapevine plants and rooted rootstock highlights the importance of black foot disease in Italy, which has previously been overlooked.
- ItemPathogenicity of South African Hymenochaetales taxa isolated from esca-infected grapevines(Firenze University Press, 2015) Cloete, Mia; Mostert, Lizel; Fischer, Michael; Halleen, FrancoisLittle is known about the pathogenicity and etiology of Hymenochaetales taxa associated with esca in South Africa. Ten South African Hymenochaetales taxa associated with esca in grapevine were subjected to basic enzyme assays to determine which ligninolytic enzymes were secreted by each taxon. In addition, a field trial was undertaken to determine the pathogenicity of these taxa. Twenty-seven fungal isolates and two negative controls were inoculated into wounds made on mature grapevines of the cultivars Shiraz and Mourvédre. Inoculated vines were evaluated for white rot symptoms after 24 months. The results of the enzyme assays indicated a difference in enzyme secretion among taxa and also between isolates of the same taxa. All isolates secreted cellulase and laccase, but there was a difference in isolates’ ability to secrete manganese peroxidase and lignin peroxidase. The results of the pathogenicity trial showed that all of the isolates used were capable of causing the characteristic white rot symptom in the wood. There were clear differences in susceptibility to white rot between the two cultivars tested, namely Shiraz and Mourvédre. The cultivars also differed in which taxa proved to be more virulent. On Shiraz a specific isolate of Taxon 6 (an Inonotus sp.), Phellinus sp. and Inonotus setuloso-croceus were significantly virulent. On Mourvédre, Taxon 3 (an Inocutis sp.) was significantly virulent.
- ItemPathogenicity of ten Phaeoacremonium species associated with esca and Petri disease of grapevine(Firenze University Press, 2018) Baloyi, Mahlatse A.; Mostert, Lizel; Halleen, FrancoisNineteen species of Phaeoacremonium have been associated with grapevines in South Africa, of which only six species have been confirmed as pathogens through pathogenicity tests conducted on field-grown grapevines. This study determined the pathogenic status of ten Phaeoacremonium spp. recently found for the first time on South African grapevines. These were: Pm. australiense, Pm. austroafricanum, Pm. fraxinopennsylvanicum, Pm. griseo-olivaceum, Pm. griseorubrum, Pm. iranianum, Pm. italicum, Pm. prunicolum, Pm. scolyti and Pm. sicilianum. In the pathogenicity tests, Ph. parasiticum was used as the positive control, and sterile water as the negative control. Up to three isolates were used per species, depending on isolate availability. Freshly cut pruning wounds in a 9-yearold Cabernet Sauvignon vineyard in Stellenbosch, South Africa, were inoculated with 200 conidia of each fungus per wound. Inoculated pruning wounds were removed after 18 months, cut longitudinally and lesion lengths were measured. Re-isolation proportions were determined by conducting isolations from inoculated spurs. All the inoculated isolates successfully colonized pruning wounds, and caused lesions that were significantly different from the negative control. All isolates were re-isolated at proportions varying from 28.6 to 85.7%. Phaeoacremonium griseo-olivaceum STE-U 7859 produced the longest lesions (mean = 79.5 mm) and Pm. iranianum STE-U 6998 the shortest (62.0 mm). No statistically significant differences in mean lesion lengths were observed between the inoculated species. There were also no significant differences between isolates of the same species, except in Pm. prunicolum where isolate STE-U 5968 produced longer lesions (mean = 77.3 mm) than STE-U 7857 (62.3 mm). This study confirmed the capabilities of all the tested Phaeoacremonium spp. to infect grapevine pruning wounds and cause lesions. The study also confirmed the importance of pruning wounds as ports of entry by these pathogens into host plants.
- ItemA protocol for the management of grapevine rootstock mother vines to reduce latent infections by grapevine trunk pathogens in cuttings(Firenze University Press, 2018) Waite, Helen; Armengol, Josep; Baaijens, Regina Billones; Gramaje, David; Halleen, Francois; Marco, Stefano Di; Smart, RichardA protocol is offered as a guideline for managers of rootstock mother grapevines, and as a potential research framework for to reduce infections by Grapevine Trunk Disease (GTD) pathogens in rootstock mother vines and cuttings. Latent infections by GTD pathogens in rootstock cuttings are a major source of the pathogens in grafted nursery vines and subsequently in new vineyards. The many pruning cuts made at the crowns of mother vines predispose them to infection which is transmitted to the new shoots via the xylem. Direct penetration by epiphytic inoculum on the bark of the shoots/canes can also occur. Mother vines with unprotected pruning wounds are typically heavily infected, particularly if they are not trellised. Availability of pruning wound treatments is limited in many countries. The spread of GTD pathogen inoculum can be reduced by avoiding sprinkler and flood irrigation, by trellising mother vines so that canopies are off the soil, and by spraying fungicides or painting wounds immediately after canes are harvested. Frequent trunk renewal aids in reducing inoculum. Cuts should be made to retain long internodes on the mother vines, cuttings should not contact the soil and pruning debris should be promptly destroyed. Cutting implements should be disinfested regularly and cuttings should be dipped in a registered fungicide or sterilant. Soaking cuttings increases fungal populations in the basal wounds and softens the bark, favouring penetration by pathogen inoculum. Dormant bench grafting in nurseries produces more GTD-symptomatic vines than field chip budding, so improved management for rootstock mother vines is more important where dormant cuttings are bench grafted. GTD epidemiology in source blocks is summarised, and best practice protocols for mother vine management and pre-grafting stages of propagation are suggested. Similar principles could be applied to scion mother vine management.
- ItemResistance in Oidium tuckeri to triazole fungicides(Stellenbosch : Stellenbosch University, 1999) Halleen, Francois; Stellenbosch University. Faculty of . Dept. of .