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- ItemA cost-effective protocol for molecular detection of fungal pathogens in soil(Academy of Science for South Africa, 2005) Damm, Ulrike; Fourie, PaulDNA EXTRACTION FROM FUNGI IN SOIL often fails because of humic substances that are co-extracted with the DNA and subsequently inhibit PCR analyses. Moreover, it is difficult to release the fungal DNA because of the diverse fungal structures residing in soil. Since available DNA extraction protocols and commercial kits are expensive or time-consuming, we have devised a superior method by testing different components of these procedures on grapevine nursery soils. The best DNA yield and sensitivity were obtained by a short and easy extraction method with sodium dodecyl sulphate buffer using the FastPrep homogenizer. An easy-to-prepare spin column with polyvinylpoly-pyrrolidone was developed to remove PCR inhibitors. In the presence of bovine serum albumin, PCR reactions were possible without further dilutions of the DNA. Our method was more sensitive for detecting Phaeomoniella chlamydospora, the organism responsible for Petri grapevine decline, and Cylindrocarpon black-foot pathogens in grapevine nursery soils than the FastDNA SPIN Kit for Soil and enabled us to perform 25 extractions for the price of one with the kit. This is the first report of molecular detection of Cylindrocarpon macrodidymum from soil and the first account of Pa. chlamydospora from soils in South Africa.
- ItemAgricultural practices and their potential role in mycotoxin contamination of maize and groundnut subsistence farming(Academy of Science of South Africa, 2019-09-26) Phokane, Sylvia; Flett, Bradley C.; Ncube, Edson; Rheeder, John P.; Rose, Lindy J.Mycotoxigenic fungi are common pathogens of maize and groundnuts; they produce mycotoxins which reduce the yield and quality of these grain crops. Numerous agricultural practices including crop rotation and storage methods have been shown to impact mycotoxin accumulation. Therefore, the farming and storage practices in maize and groundnut subsistence farming systems in Pongola, Vryheid, Jozini, Manguzi and Mbazwana Districts of northern KwaZulu-Natal (South Africa) were surveyed to determine their potential role in promoting or mitigating mycotoxin contamination. A questionnaire about agricultural farming practices and storage facilities was presented to 65 subsistence maize and/or groundnut farmers. At least 90% of the farmers surveyed were not aware of mycotoxins and their consequences to animal and human health. The majority of the farmers did not practise crop rotation. However, they practised intercropping and sorted damaged and mouldy grain (maize and groundnuts) before storage. The damaged or mouldy grain was largely used as animal feed, thereby exposing animals to an increased risk of mycotoxicoses. Metal tanks and inqolobane (a type of wooden structure) were identified as the most common storage structures. Harvested homegrown maize was mostly used for the farmers’ own consumption but also sometimes sold to the local community. The implementation of mycotoxin awareness campaigns is necessary, particularly in these districts. The storage facilities used by the subsistence farmers allowed increased moisture and insect invasion. The need for the surveillance of mycotoxins in subsistence-farmed food crops is vital.
- ItemApplication of fungicides against postharvest botrytis bunch rot of table grapes in the Western Cape(South African Society for Enology and Viticulture, 1994) De Kock, P. J.; Holz, G.Fungicide programmes for the control of postharvest Botrytis bunch rot on table grapes were evaluated in six trials from 1984/85 to 1991/92 in the Western Cape. The study demonstrated the ineffectiveness of dicarboximide applications during bloom to early pea size in well managed vineyards. Dicarboximides were most effective when applied from bunch closure to ripening. lprodioue/sulphur treatments at veraison and before harvest reduced Botrytis bunch rot, but they were ineffective in inhibiting infection during storage. Control was only achieved when grapes were exposed to S02 during storage. Although bunch dip treatments reduced infection in the vineyard, this control was not commercially acceptable. Therefore no real advantage was found when bunches were dipped in fungicide at veraison to ensure better coverage. The fact that berries became infected primarily during harvest, package operations and storage, emphasised the necessity for reducing B. cinerea inoculum on harvested grapes. It is suggested that the results of this investigation may lay the foundation for incorporating biological control in Botrytis bunch rot control.
- ItemAssessing genotype-by-environment interactions in aspergillus ear rot and pre-harvest aflatoxin accumulation in maize inbred lines(MDPI, 2017) Okoth, Sheila; Rose, Lindy J.; Ouko, Abigael; Netshifhefhe, Nakisani E. I.; Sila, Henry; Viljoen, AltusAspergillus flavus, causal agent of the Aspergillus ear rot (AER) of maize, also produces aflatoxins that cause aflatoxicosis in humans and livestock. Ten maize inbred lines were evaluated in replicated trials in two aflatoxicosis outbreak hot spots in Kenya and in three maize-growing areas in South Africa for resistance to AER, A. flavus colonization, and pre-harvest aflatoxin accumulation during the 2012/13 growing season. AER severity was measured by visual assessment, while A. flavus colonization and aflatoxin content were quantified by real-time polymerase chain reaction (PCR) and liquid chromatography tandem mass spectrometry, respectively. Genotype by environment interaction (GEI) was determined using analysis of variance (ANOVA), additive main effects and multiplicative models (AMMI), and genotype plus by environment (GGE) biplot analyses. Stability of genotypes was evaluated using AMMI analysis. AER severity and fungal colonization significantly (p < 0.001) varied between genotypes. GEI influenced the severity of AER symptoms and aflatoxin accumulation significantly (p < 0.001), while fungal colonization was not affected. The inbred lines response was consistent for this trait in the test environments and was thus considered a desirable measure to indicate maize lines with a high risk of aflatoxin accumulation. CML495, CKL05019, LaPosta, and MIRTC5 were the least diseased lines, with the lowest aflatoxin contamination and a stable phenotypic response across the environments. Kiboko was determined as the ideal representative test environment, with discriminative ability of the genotypes for selection of the desired stable responses of the three traits.
- ItemCharacterisation of South African isolates of Fusarium oxysporum f.sp. cubense from Cavendish bananas(Academy of Science of South Africa (ASSAf), 2010-04) Visser, Marinda; Gordon, Tom; Fourie, Gerda; Viljoen, AltusFusarium wilt, caused by the soil-borne fungus Fusarium oxysporum f.sp. cubense (Foc), is a serious vascular disease of bananas in most subtropical and tropical regions of the world. Twenty-four vegetative compatibility groups (VCGs) and three pathogenic races have been identified in Foc, reflecting a relatively high genetic diversity for an asexual fungus. To characterise a South African population of Foc, a collection of 128 isolates from diverse geographic origins were isolated from diseased Cavendish bananas and subjected to VCG analysis and sequencing of the translation elongation factor 1-α (TEF) gene region. The presence of mating type genes was also determined using MAT-1 and MAT-2 specific primers. VCG 0120 was established as the only VCG of Foc present in the South African population studied. Only the MAT-2 idiomorph was present in all the local isolates of Foc. A phylogenetic analysis of DNA sequences of the TEF gene region revealed that the South African isolates grouped closely with VCG 0120 isolates from Australia and Asia. These results suggest that the South African population of Foc was most likely introduced in a limited number of events and that it had spread with infected planting material within the country. The presence of only one mating type and the limited diversity in this pathogen render it unlikely to rapidly overcome disease management strategies involving host resistance. © 2010. The Authors.
- ItemCitrus black spot is absent in the Western Cape, Northern Cape and Free State Provinces(Academy of Science of South Africa, 2012) Carstens, Elma; Le Roux, Hendrik F.; Holtzhausen, Michael A.; Van Rooyen, Liezl; Coetzee, Joey; Wentzel, Ria; Laubscher, Wilhelm; Dawood, Zorina; Venter, Elrita; Schutte, Gerhardus C.; Hattingh, Vaughan; Fourie, Paul H.The South African citrus industry is strongly focused on exports and South Africa is a signatory member of both the World Trade Organisation Agreement on the application of Sanitary and Phytosanitary Measures and the International Plant Protection Convention. Citrus black spot, caused by Guignardia citricarpa, does not occur in all the South African citrus production areas and, therefore, South Africa has a responsibility to provide those trading partners that have identified G. citricarpa as a regulated pest with reliable information about the distribution of citrus black spot within South Africa. Detection surveys were conducted in citrus production areas in the Western Cape, Northern Cape and Free State Provinces and appropriate diagnostic protocols were used to ensure reliable detection of G. citricarpa. Trees in commercial orchards and home gardens on farms and in towns of 17, 9 and 5 magisterial districts in the Western Cape, Northern Cape and Free State Provinces, respectively, were sampled between 1995 and 2010. Fruit samples were taken during June and July, and leaf samples from November to January. None of the 3060 fruit and leaf samples collected during these surveys tested positive for G. citricarpa. Phyllosticta capitalensis, a non-pathogenic, ubiquitous, endophytic species was, however, detected during these surveys. In compliance with relevant International Standards for Phytosanitary Measures and based on the outcome of these official surveys, these three provinces in South Africa can be recognised as citrus black spot pest free areas.
- ItemCleistothecia and flag shoots : sources of primary inoculum for grape powdery mildew in the Western Cape Province, South Africa(South African Society for Enology and Viticulture, 2000) Halleen, F.; Holz, G.Little is known about the mode of survival and sources of primary inoculum of Uncinula necator, the causal pathogen of grapevine powdery mildew, in vineyards in the Western Cape province. A study was therefore undertaken to determine whether cleistothecia and flag shoots are formed on vines in local vineyards. Flag shoots were found shortly after budbreak in September 1997 in a Carignane vineyard near Somerset West. Cleistothecia were first observed during April to May 1996 on severely infected leaves from three vineyards in the main grapegrowing areas of Stellenbosch. This was the first report of cleistothecia and flag shoot formation in vineyards in the Western Cape province. Cleistothecia occurred in small numbers on leaves (1 - 10 per leaf) and all were immature. Cleistothecia were dispersed by late summer and autumn rains from leaves to bark of grapevines, where they overwinter. No conclusion could be made regarding the viability of cleistothecia. However, the characteristics of the first symptoms that developed on leaves, namely separate, individual lesions formed at random on first-formed leaves growing in close proximity to the bark, provided circumstantial evidence that cleistothecia are dispersed to the bark. Weather conditions suitable for release of ascospores from overwintered cleistothecia occurred frequently between budbreak and bloom in all the areas.
- ItemColonization of table grape bunches by alternaria alternata and rot of cold-stored grapes(South African Society for Enology and Viticulture, 1994) Swart, A. E.; Holz, G.Assessment of latent Alternaria alternata infections in table grapes indicated that infections occurred in commercial vineyards during the entire period of bunch development. Mature bunches were asymptomatic despite high levels of A. alternata recovered from triple-sterilized bunch tissue. Inoculation studies showed no shift in disease susceptibility of ripening grape berries, and postharvest rot was not related to the level of natural infection. Late season fungicidal sprays, or dip treatments that ensured better penetration and coverage of inner parts, resulted in no meaningful reduction in postharvest rot. Based on the behaviour of the pathogen, it is suggested that additional fungicide programmes for the control of the disease should not be followed in commercial vineyards. Instead, attention should be given to physiological and stress factors, such as mechanical and sulphur dioxide damage that might predispose cold-stored bunches to A. alternata decay.
- ItemContamination of bananas with beauvericin and fusaric acid produced by Fusarium oxysporum f. sp. cubense(Public Library of Science, 2013) Li, Chunyu; Zuo, Cunwu; Deng, Guiming; Kuang, Ruibin; Yang, Qiaosong; Hu, Chunhua; Sheng, Ou; Zhang, Sheng; Ma, Lijun; Wei, Yuerong; Yang, Jing; Liu, Siwen; Biswas, Manosh Kumar; Viljoen, Altus; Yi, GanjunBackground Fusarium wilt, caused by the fungal pathogen Fusarium oxysporum f. sp. cubense (Foc), is one of the most destructive diseases of banana. Toxins produced by Foc have been proposed to play an important role during the pathogenic process. The objectives of this study were to investigate the contamination of banana with toxins produced by Foc, and to elucidate their role in pathogenesis. Methodology/Principal Findings Twenty isolates of Foc representing races 1 and 4 were isolated from diseased bananas in five Chinese provinces. Two toxins were consistently associated with Foc, fusaric acid (FA) and beauvericin (BEA). Cytotoxicity of the two toxins on banana protoplast was determined using the Alamar Blue assay. The virulence of 20 Foc isolates was further tested by inoculating tissue culture banana plantlets, and the contents of toxins determined in banana roots, pseudostems and leaves. Virulence of Foc isolates correlated well with toxin deposition in the host plant. To determine the natural occurrence of the two toxins in banana plants with Fusarium wilt symptoms, samples were collected before harvest from the pseudostems, fruit and leaves from 10 Pisang Awak ‘Guangfen #1’ and 10 Cavendish ‘Brazilian’ plants. Fusaric acid and BEA were detected in all the tissues, including the fruits. Conclusions/Signficance The current study provides the first investigation of toxins produced by Foc in banana. The toxins produced by Foc, and their levels of contamination of banana fruits, however, were too low to be of concern to human and animal health. Rather, these toxins appear to contribute to the pathogenicity of the fungus during infection of banana plants.
- 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.
- ItemDevelopment of a hydrolysis probe-based real-time assay for the detection of tropical strains of Fusarium oxysporum f. sp. cubense race 4(Public Library of Science, 2017-02-08) Aguayo, Jaime; Mostert, Diane; Fourrier-Jeandel, Celine; Cerf-Wendling, Isabelle; Hostachy, Bruno; Viljoen, Altus; Ioos, RenaudFusarium oxysporum f. sp. cubense (Foc) is one of the most important threats to global banana production. Strategies to control the pathogen are lacking, with plant resistance offering the only long-term solution, if sources of resistance are available. Prevention of introduction of Foc into disease-free areas thus remains a key strategy to continue sustainable banana production. In recent years, strains of Foc affecting Cavendish bananas have destroyed plantations in a number of countries in Asia and in the Middle East, and one African country. One vegetative compatibility group (VCG), 01213/16, is considered the major threat to bananas in tropical and subtropical climatic conditions. However, other genetically related VCGs, such as 0121, may potentially jeopardize banana cultures if they were introduced into disease-free areas. To prevent the introduction of these VCGs into disease-free Cavendish banana-growing countries, a real-time PCR test was developed to accurately detect both VCGs. A previously described putative virulence gene was used to develop a specific combination of hydrolysis probe/primers for the detection of tropical Foc race 4 strains. The real-time PCR parameters were optimized by following a statistical approach relying on orthogonal arrays and the Taguchi method in an attempt to enhance sensitivity and ensure high specificity of the assay. This study also assessed critical performance criteria, such as repeatability, reproducibility, robustness, and specificity, with a large including set of 136 F. oxysporum isolates, including 73 Foc pathogenic strains representing 24 VCGs. The validation data demonstrated that the new assay could be used for regulatory testing applications on banana plant material and can contribute to preventing the introduction and spread of Foc strains affecting Cavendish bananas in the tropics.
- ItemDiaporthaceae associated with root and crown rot of maize(International Mycological Association, 2011-03-10) Lamprecht, Sandra C.; Crous, Pedro W.; Groenewald, Johannes Z.; Tewoldemedhin, Yared T.; Marasas, Walter F. O.Several isolates of coelomycetous fungi with pigmented conidia were consistently isolated from diseased roots of Zea mays in irrigated plots monitored in the KwaZulu-Natal Province of South Africa. Based on their morphology, these isolates could be identified as representative of Stenocarpella macrospora, S. maydis, and Phaeocytostroma ambiguum. Although species of Stenocarpella are well-known as causal agents of cob and stalk rot and leaf blight of maize in South Africa, the occurrence and importance of P. ambiguum is less well documented and understood. To determine the role of P. ambiguum as a root pathogen of maize, pathogenicity tests were conducted under glasshouse conditions at 18 °C night and 28 °C day temperatures using a pasteurised soil, river sand and perlite medium and a 0.5 % sand-bran inoculum. Based on these results, P. ambiguum was shown to be a primary pathogen of maize, but to be less virulent than the positive control, S. maydis. Furthermore, to clarify the higher-level phylogeny of these fungal genera, isolates were subjected to DNA sequencing of the nuclear ribosomal DNA(ITS & LSU). Partial gene sequences of the translation elongation factor 1-alpha gene were added to confirm the species monophyly. To resolve the generic placement of Phaeocytostroma, additional species such as P. sacchari, P. plurivorum and P. megalosporum were also added to the analysis. Based on these results, Stenocarpella and Phaeocytostroma were shown to be two well defined genera, belonging to Diaporthales, Diaporthaceae, being closely allied to Phomopsis (Diaporthe). All three genera were also observed to form alpha as well as beta conidia, and although this phenomenon is well documented for Phomopsis and Phaeocytostroma, it is a new observation for Stenocarpella. In spite of the differences in conidial pigmentation, no support could be obtained for polyphyly in Diaporthaceae, suggesting that as observed in Botryosphaeriaceae (Botryosphaeriales), conidial pigmentation is not informative at the family level in Diaporthales.
- ItemThe distribution and host range of the banana fusarium wilt fungus, fusarium oxysporum f. sp. cubense , in Asia(Public Library of Science, 2017) Mostert, Diane; Molina, Agustin B.; Daniells, Jeff; Fourie, Gerda; Hermanto, Catur; Chao, Chih- Ping; Fabregar, Emily; Sinohin, Vida G.; Masdek, Nik; Thangavelu, Raman; Li, Chunyu; Yi, Ganyun; Mostert, Lizel; Viljoen, AltusFusarium oxysporum formae specialis cubense (Foc) is a soil-borne fungus that causes Fusarium wilt, which is considered to be the most destructive disease of bananas. The fungus is believed to have evolved with its host in the Indo-Malayan region, and from there it was spread to other banana-growing areas with infected planting material. The diversity and distribution of Foc in Asia was investigated. A total of 594 F. oxysporum isolates collected in ten Asian countries were identified by vegetative compatibility groups (VCGs) analysis. To simplify the identification process, the isolates were first divided into DNA lineages using PCR-RFLP analysis. Six lineages and 14 VCGs, representing three Foc races, were identified in this study. The VCG complex 0124/5 was most common in the Indian subcontinent, Vietnam and Cambodia; whereas the VCG complex 01213/16 dominated in the rest of Asia. Sixty-nine F. oxysporum isolates in this study did not match any of the known VCG tester strains. In this study, Foc VCG diversity in Bangladesh, Cambodia and Sri Lanka was determined for the first time and VCGs 01221 and 01222 were first reported from Cambodia and Vietnam. New associations of Foc VCGs and banana cultivars were recorded in all the countries where the fungus was collected. Information obtained in this study could help Asian countries to develop and implement regulatory measures to prevent the incursion of Foc into areas where it does not yet occur. It could also facilitate the deployment of disease resistant banana varieties in infested areas.
- ItemThe effects of postharvest treatments and sunlight exposure on the reproductive capability and viability of Phyllosticta citricarpa in citrus black spot fruit lesions(MDPI, 2020-12-21) Moyo, Providence; Fourie, Paul H.; Masikane, Siyethemba L.; Fialho, Regis de Oliveira; Mamba, Lindokuhle C.; Du Plooy, Wilma; Hattingh, VaughanCitrus black spot (CBS) is caused by Phyllosticta citricarpa, which is classified as a quarantine organism in certain countries whose concerns are that CBS-infected fruit may be a pathway for introduction of the pathogen. This study evaluated the reproductive capability and viability of P. citricarpa under simulated conditions in which the whole fruit, peel segments, or citrus pulp with CBS lesions were discarded. Naturally infected ‘Midknight’ Valencia orange and ‘Eureka’ lemon fruit, either treated using standard postharvest sanitation, fungicide, and wax coating treatments or untreated, were placed into cold storage for 5 weeks (oranges at 4 ◦C and lemons at 7 ◦C). Thereafter, treated and untreated fruit were incubated for a further 2 weeks at conditions conducive for CBS symptom expression and formation of pycnidia. The ability of pycnidia to secrete viable pycnidiospores after whole fruit and peel segments or peel pieces from citrus pulp were exposed to sunlight at warm temperatures (±28 ◦C) and ±75% relative humidity levels was then investigated. The combination of postharvest treatments and cold storage effectively controlled CBS latent infections (>83.6% control) and pycnidium formation (<1.4% of lesions formed pycnidia), and the wax coating completely inhibited pycnidiospore release in fruit and peel segments. Pycnidiospores were secreted only from lesions on untreated fruit and peel segments and at low levels (4.3–8.6%) from peel pieces from pulped treated fruit. However, spore release rapidly declined when exposed to sunlight conditions (1.4% and 0% after 2 and 3 days, respectively). The generally poor reproductive ability and viability of CBS fruit lesions on harvested fruit, particularly when exposed to sunlight conditions, supports the conclusion that citrus fruit without leaves is not an epidemiologically significant pathway for the entry, establishment, and spread of P. citricarpa.
- ItemEpidemiology of botrytis cinerea on grape : wound infection by dry, airborne conidia(South African Society for Enology and Viticulture, 2002) Coertze, S.; Holz, G.This study describes the infection of fresh wounds on berries exposed to freshly deposited airborne Botrytis cinerea conidia, and on berries carrying previously deposited conidia and germlings (latent infections). Grapes at bunch closure and at the mature (harvest) stage, as well as mature, cold-stored grapes, were used. The grapes were dusted with dry conidia in a settling tower. The inocula were subjected to conditions commonly encountered by the pathogen in grape bunches: dry conidia on dry berries under dry conditions, dry conidia on dry berries under high relative humidity, and dry conidia exposed to a film of water on the berry surface. The mean number of wounds that yielded B. cinerea decay 14 days after inoculation was calculated. Fluorescence microscopy revealed that conidia occurred evenly as single cells on the grape berry surface and seldom landed at the wound periphery. They remained dormant on dry berries, but germinated freely on the unbroken skin and at the wound periphery on moist and wet berries. In the case of berries inoculated at bunch closure and harvest stages, wounds were not infected by conidia deposited on berries four days prior to wounding. This finding indicated that, following adhesion and the first stages of growth, the pathogen did not survive for extended periods on surfaces of immature and mature grape berries. Freshly deposited dry conidia were needed to infect the wounds. The freshly deposited conidia furthermore needed free water, and not high humidity or wound exudates, to infect the fresh wounds. Proportions of wounds infected were extremely low. According to these findings, this mode of infection should not contribute to a gradual build-up of secondary inoculum and to B. cinerea epiphytotics in the vineyard. The previously and freshly deposited conidia both infected wounds made on cold-stored mature berries. Of the two inocula, freshly deposited conidia were more successful in causing wound infection. Therefore, in the event of wounding, postharvest decay would be caused primarily by conidia occurring on the grape berry surface, and not by mycelia in the berry skin.
- ItemThe essential symbiosis of mycology and plant pathology : present and future needs(Academy of Science of South Africa, 1995) Crous, P. W.Correct disease diagnosis is essential for plant pathologists striving to combat plant disease. To support them, mycologists are integrating traditional and molecular techniques. Attention must therefore be given to harnessing the power of molecular genetics to provide rapid methods of identification, and to study the adaptability of plant pathogens in terms of virulence and sensitivity to fungicides. To combat African plant diseases effectively, better co-operation bem'een mycologists and plant pathologists is essential.
- ItemFirst report of Phyllosticta citricarpa and description of two new species, P. paracapitalensis and P. paracitricarpa, from citrus in Europe(Elsevier, 2017) Guarnaccia, V.; Groenewald, J. Z.; Li, H.; Glienke, C.; Carstens, E.; Hattingh, V.; Fourie, P. H.; Crous, P. W.The genus Phyllosticta occurs worldwide, and contains numerous plant pathogenic, endophytic and saprobic species. Phyllosticta citricarpa is the causal agent of Citrus Black Spot disease (CBS), affecting fruits and leaves of several citrus hosts (Rutaceae), and can also be isolated from asymptomatic citrus tissues. Citrus Black Spot occurs in citrus-growing regions with warm summer rainfall climates, but is absent in countries of the European Union (EU). Phyllosticta capitalensis is morphologically similar to P. citricarpa, but is a non-pathogenic endophyte, commonly isolated from citrus leaves and fruits and a wide range of other hosts, and is known to occur in Europe. To determine which Phyllosticta spp. occur within citrus growing regions of EU countries, several surveys were conducted (2015–2017) in the major citrus production areas of Greece, Italy, Malta, Portugal and Spain to collect both living plant material and leaf litter in commercial nurseries, orchards, gardens, backyards and plant collections. A total of 64 Phyllosticta isolates were obtained from citrus in Europe, of which 52 were included in a multi-locus (ITS, actA, tef1, gapdh, LSU and rpb2 genes) DNA dataset. Two isolates from Florida (USA), three isolates from China, and several reference strains from Australia, South Africa and South America were included in the overall 99 isolate dataset. Based on the data obtained, two known species were identified, namely P. capitalensis (from asymptomatic living leaves of Citrus spp.) in Greece, Italy, Malta, Portugal and Spain, and P. citricarpa (from leaf litter of C. sinensis and C. limon) in Italy, Malta and Portugal. Moreover, two new species were described, namely P. paracapitalensis (from asymptomatic living leaves of Citrus spp.) in Italy and Spain, and P. paracitricarpa (from leaf litter of C. limon) in Greece. On a genotypic level, isolates of P. citricarpa populations from Italy and Malta (MAT1-2-1) represented a single clone, and those from Portugal (MAT1-1- 1) another. Isolates of P. citricarpa and P. paracitricarpa were able to induce atypical lesions (necrosis) in artificially inoculated mature sweet orange fruit, while P. capitalensis and P. paracapitalensis induced no lesions. The Phyllosticta species recovered were not found to be widespread, and were not associated with disease symptoms, indicating that the fungi persisted over time, but did not cause disease.
- ItemFitness on grape berries of botrytis cinerea isolates belonging to different dicarboximide sensitivity classes(South African Society for Enology and Viticulture, 2003) Fourie, P. H.; Holz, G.Seasonal fluctuations in the frequency of dicarboximide-resistant Botrytis cinerea isolates in Western Cape vineyards suggest a reduced fitness of these isolates. In this study fitness of sensitive, ultra-low-level and low-level dicarboximide-resistant isolates of B. cinerea were compared on grape berries. Conidia were dispersed as single cells on berry surfaces from airborne inoculum in a settling tower or deposited as clusters in water droplets. Investigations were conducted at two potential infection courts on dicarboximide treated and untreated berries, namely the intact cuticle and wounds. Surface sterilisation, isolation and freezing techniques were used to determine surface colonisation and penetration by airborne inoculum. Decay incidence, severity and sporulation incidence were determined following inoculation with conidial clusters. The different tests indicated that germlings of dicarboximide-sensitive and -resistant isolates had similar surface-colonising abilities of dicarboximide-free berries. However, sensitive strains penetrated significantly more often. Fitness decreased with an increase in the level of dicarboximide resistance. Iprodione caused a drastic disturbance in the ratio of different dicarboximide sensitivity classes that occupied the berry surface and allowed the development of germlings of predominantly resistant isolates, but with few successful infections. Significantly higher levels of infection and proliferation of dicarboximide-resistant isolates on sprayed or unsprayed berries were facilitated by wounding or the termination of host resistance (freezing). According to these findings, these modes of infection should not contribute to a gradual build-up of inoculum of either dicarboximide-sensitive or -resistant isolates. Trends by airborne conidia described here suggest that another primary infection event in the vineyard, most likely floral infection and subsequent debris colonisation, should largely regulate the dynamics between dicarboximide-sensitive and -resistant isolates in B. cinerea populations on grapevine.
- ItemFrequency of dicarboximide resistant strains of botrytis cinerea in South African table grape vineyards and influence of spray schedules on resistant sub-populations(South African Society for Enology and Viticulture, 1998) Fourie, P. H.; Holz, G.Between 1993 and 1995, 1139 isolates of Botrytis cinerea were collected from table grape vineyards in the Western and Northern Cape provinces and tested for sensitivity to dicarboximides (vinclozolin) and benzimidazoles (carbendazim). Overall the frequency of resistance to vinclozolin and carbendazim averaged 9,4 and 11,9%, respectively. Ninety-seven percent of the vinclozolin-resistant isolates were dual resistant to carbendazim. Fifty-nine percent of the isolates were from the Hexriver region; of these 7,7% were vinclozolin, and 12,6% carbendazim resistant. The Paarl region had the highest resistance incidence (37,7% vinclozolin and 41,5% carbendazim resistant). Only two of the 191 isolates tested from the Orange River region were vinclozolin resistant. ED50 values of ultra-low-level and low-level dicarboximide-resistant isolates ranged from 0,843 to 1,834 and 2,053 to 5,129 μg vinclozolin/ml, respectively. Vinclozolin-resistant strains were not abnormally osmotically sensitive. Monitoring of changes in the frequency of resistance in 10 commercial vineyards under high-, medium- and low-schedule dicarboximide programmes from 1993-1995 showed that the resistance frequency fluctuated from low (average 12,7% ) during the winter to high (average 55,8 % ) during the growing season. Maximum levels of dicarboximide resistance were recorded during bunch closure. Distinct differences were, however, observed in the resistance frequency in individual vineyards. In four high-schedule dicarboximide vineyards resistance frequencies increased early in the season, prior to any dicarboximide applications. This finding suggests dual resistance between broad-spectrum fungicides and dicarboximides.
- ItemFungicide efficacy against botrytis cinerea at different positions on grape shoots(South African Society for Enology and Viticulture, 2003) Van Rooi, C.; Holz, G.Fungicide efficacy at different positions on grape shoots was determined on vinelets inoculated with Botrytis cinerea. To ensure proper fungicide coverage, vinelets (table grape cultivar Dauphine, wine grape cultivar Merlot noir), prepared from cuttings, were sprayed in a spray chamber at the recommended dosages with iprodione, pyrimethanil, cyprodinil/fludioxonil and fenhexamid. Vinelets for the control treatment were left unsprayed. After 24 h the vinelets were dusted with dry, airborne conidia in a settling ower to reduce the sporadic occurrence of the pathogen on shoots and incubated for 24 h at high relative humidity (±93 % ) to promote infection. Fungicide efficacy was determined by observing intact vinelets for symptom expression at nodes, internodes, leaf blades, petioles and inflorescences, and by determining surface colonisation (shoots left unsterile) and penetration (surfacedisinfested shoots) by isolation studies. Symptoms of B. cinerea decay did not develop on sprayed and unsprayed vinelets that were kept in dry chambers during the 2-week observation period. The pathogen, however, developed from the isolated parts. On the unsprayed vinelets for both cultivars B. cinerea incidences recorded on two media and the two sterility regimes were significantly higher on leaf blades than on the petioles, shoot pieces and inflorescences. The isolation studies showed that the different fungicides were highly and nearly equally efficient in reducing superficial B. cinerea and latent infection at the various positions. In the case of leaf blades B. cinerea incidence was significantly reduced by each fungicide on both cultivars. This was not the case for the other parts, which yielded B. cinerea at low incidences under the two sterility regimes. The study showed that the fungicides used in this study, if applied properly to shoots at the pre-bloom stage, should effectively reduce the amount of B. cinerea in leaves and completely prevent the infection of nodes, internodes and inflorescences.