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

An integrated strategy for the proactive management of grapevine trunk disease pathogen infections in grapevine nurseries

CITATION: Halleen, F. & Fourie, P. H. 2016. An integrated strategy for the proactive management of grapevine trunk disease pathogen infections in grapevine nurseries. South African Journal of Enology & Viticulture, 37(2):104-114, doi:10.21548/37-2-825.The original publication is available at http://www.journals.ac.za/index.php/sajevThe aim of this study was to compare 11 treatment regimes (TR) in grapevine nurseries in order to eradicate trunk pathogens. Grapevine propagation material was subjected to treatments before cold storage, before and after grafting, before planting and after uprooting. Isolations were made from roots, rootstocks and graft unions of uprooted vines. None of the treatments consistently affected the number of certifiable vines produced. TR 1 (benomyl), TR 3 (Sporekill) and TR 4 (Trichoflow) increased root mass. Phaeomoniella chlamydospora was the most frequently isolated pathogen from all plant parts. In general, TR 2, 7, 9 and 11 caused the lowest occurrence of Phaeomoniella. TR 9 consistently reduced the incidence of Phaeoacremonium. Pleurostomophora richardsiae occurred most frequently in graft unions, but treatments were too inconsistent to draw meaningful conclusions. TR 7 (hot water treatment (HWT) of uprooted grapevines) consistently reduced incidences of black foot disease (BFD) pathogens. The highest incidence of Botryosphaeriaceae occurred in graft unions, but all treatments significantly reduced infections. The colonisation of rootstocks and graft unions by Trichoderma was significantly better with TR 4 (Trichoflow). The other Trichoderma treatments (TR 9, 10 and 11) differed from the control treatment only in the graft unions. This is a first report of an integrated strategy covering all the phases of the propagation process, from the moment the nursery receives the propagation material until the dormant vines are removed. TR 9 is recommended for use in nurseries, although HWT of dormant vines is recommended to eradicate BFD pathogens. Benomyl can be replaced by carbendazim when benomyl is no longer available.http://www.journals.ac.za/index.php/sajev/article/view/825Publisher's versio

Advances in propagation of grapevine in the world.

Grapevine production by classical grafting methods and in commercial scale emerged over 130 years. This system remained handmade until the mid-1950s, when emerged the first international certification programs aimed at obtaining mother lants with high viral sanity. The necessity to increase the scale of production on industrial model and plant material production based on minimum morphological standards appeared at the end of the 1960s. Along the 1970s, research unlocked knowledge on emi-automated grafting, process hygiene, use of plant growth regulators and understanding of physiological events of rootstock-scion compatibility, callus formation and rooting. So, until the mid-2000s, certification schemes and propagation processes advanced little in technical standard. However, grapevine growing areas were expanded and demands for plant material increased, and new diseases emerged from contaminated nurseries. These new diseases (new viral complexes, phytoplasmas, bacteria and grapevine trunk diseases) were discovered by high-sensitivity diagnostic methods. Today, there is a new discussion on the nursery segment worldwide. The propagation techniques have been reviewed from the perspective of reducing the incidence of new diseases and minimum physiological damage of nursery plants during the production stages. Therefore, technological innovations regarding equipment, practices and production inputs have been incorporated in new certification schemes. However, despite these dvantages, these schemes have become more complex and multidisciplinary than previous ones, bringing difficulties in adaptation of nurserymen. Index-terms: Vitis grafting, nursery, certification, grapevine cuttings. Propagação de videiras a partir dos métodos clássicos de enxertia e em escala comercial teve origem há mais de 130 anos. Este sistema permaneceu artesanal até meados da década de 1950, quando se iniciaram os primeiros programas internacionais de certificação com foco na obtenção de plantas básicas com elevada sanidade para vírus. No fim da década de 1960, surgiu a necessidade de aumentar a escala para produção em um modelo industrial em que a muda apresentasse um padrão morfológico mínimo. Ao longo da década de 1970, aprofundaram-se as pesquisas relacionadas à automatização da prática de enxertia, à higienização do processo, ao uso de reguladores de crescimento e ao entendimento dos eventos fisiológicos da compatibilidade entre enxerto e porta-enxerto, formação de calos e enraizamentos. Assim, até meados dos anos 2000, os esquemas de certificação e o processo de propagação pouco evoluíram em termos técnicos. Porém, a medida que a área vitícola foi expandindo e a demanda por mudas aumentando, verificou-se que novas doenças se alastravam em escala global a partir de viveiros contaminados. Estas novas doenças (complexos virais, fitoplasmas, bactérias e fungos causadores de podridões vasculares) foram sendo descobertas à medida que os métodos de diagnose avançaram em sensibilidade de detecção. Hoje, surge nova discussão no segmento viveirista mundial fundamentada no fato de que o processo de propagação está sendo revisto sob o foco da redução de incidência das novas pragas e mínimo dano à muda ao longo das etapas da produção. Surgem, assim, inovações tecnológicas, tanto em equipamentos quanto em práticas e insumos, sendo incorporadas aos novos modelos de certificação. Mas, se por um lado, estes esquemas tornam-se cada vez mais multidisciplinares, por outro, a complexidade gerada pode trazer dificuldades para a adesão pelos viveiristas. Termos para indexação: Vitis enxertia, viveiro, certificação, mudas de videira

Cross pathogenicity of Neofusicoccum australe and Neofusicoccum stellenboschiana on grapevine and selected fruit and ornamental trees

[EN] Neofusicoccum australe is one of the most important Botryosphaeriaceae pathogens occurring on fruit and vine crops. This fungus was recently taxonomically reassessed, identifying N. stellenboschiana as a separate species. Previous pathogenicity studies used N. stellenboschiana and N. australe isolates as N. australe, so assessment of the pathogenicity of these two species on grapevine and other hosts was required. A pathogenicity trial was conducted on detached shoots of grapevine, plum, apple, olive and Peruvian pepper tree. Shoots were individually inoculated with 11 N. australe and eight N. stellenboschiana isolates originally isolated from grapevine, plum, apple, olive, Peruvian pepper and fig. Both species formed lesions on all five hosts and were reisolated 5 weeks post-inoculation. In general, the largest lesions were formed on plum and smallest on Peruvian pepper. Isolate host origin did not influence ability to cause lesions on other hosts. Isolates of N. australe and N. stellenboschiana differed in virulence on the various hosts, ranging from those that caused the largest lesions, a group causing intermediate lesions, and another causing lesions similar to uninoculated controls. The study demonstrates that N. australe and N. stellenboschiana isolates originating from various fruit hosts can infect alternative hosts including grapevine and other major fruit crops.This research developed from a mobility sojourn funded by the Erasmus Mundus Joint Master Degrees Programme of the European Commission under the PLANT HEALTH Project. The authors also acknowledge financial support from ARC Infruitec-Nietvoorbij and the National Research Foundation of South Africa (NRF UID88771). Technical Assistance was provided by Carine Vermeulen, Danie Marais, Julia Marais, Muriel Knipe, Lydia Maart, Christopher Paulse, Bongiwe Sokwaliwa, Nadeen van Kervel, and Levocia Williams (Plant Protection Division, ARC Infruitec-Nietvoorbij).Mojeremane, K.; Lebenya, P.; Du Plessis, IL.; Van Der Rijst, M.; Mostert, L.; Armengol Fortí, J.; Halleen, F. (2020). Cross pathogenicity of Neofusicoccum australe and Neofusicoccum stellenboschiana on grapevine and selected fruit and ornamental trees. PHYTOPATHOLOGIA MEDITERRANEA. 59(3):581-593. https://doi.org/10.14601/Phyto-11609S58159359

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

Increased tartrate-resistant acid phosphatase (TRAP) expression in malignant breast, ovarian and melanoma tissue: an investigational study

BACKGROUND: Tartrate-resistant acid phosphatase (TRAP) is a metalloprotein enzyme that belongs to the acid phosphatases and is known to be expressed by osteoclasts. It has already been investigated as a marker of bone metastases in cancer patients. In this study, which examined the value of serum TRAP concentrations as a marker of bone disease in breast cancer patients, we observed high concentrations of TRAP even in patients without bone metastases. To elucidate this phenomenon, we examined the expression of TRAP in breast cancer cells and the cells of several other malignancies. METHODS: TRAP concentrations in the serum of tumor patients were determined by ELISA. The expression of TRAP in breast, ovarian, and cervical cancer and malignant melanoma was analyzed by immunohistochemistry. RT-PCR and immunocytology were used to evaluate TRAP expression in cultured tumor cells. RESULTS: A marked increase in serum TRAP concentrations was observed in patients with breast and ovarian cancer, regardless of the presence or absence of bone disease. TRAP expression was found in breast and ovarian cancers and malignant melanoma, while cervical cancer showed only minimal expression of TRAP. Expression of TRAP was absent in benign tissue or was much less marked than in the corresponding malignant tissue. TRAP expression was also demonstrated in cultured primary cancer cells and in commercially available cell lines. CONCLUSION: Overexpression of TRAP was detected in the cells of various different tumors. TRAP might be useful as a marker of progression of malignant disease. It could also be a potential target for future cancer therapies

Osteoclasts Are Active in Bone Forming Metastases of Prostate Cancer Patients

BACKGROUND: Bone forming metastases are a common and disabling consequence of prostate cancer (CaP). The potential role of osteoclast activity in CaP bone metastases is not completely explained. In this study, we investigated ex vivo whether the osteolytic activity is present and how it is ruled in CaP patients with bone forming metastases. METHODOLOGY: Forty-six patients affected by newly diagnosed CaP and healthy controls were enrolled. At diagnosis, 37 patients had a primary tumour only, while 9 had primary tumour and concomitant bone forming metastases. In all patients there was no evidence of metastasis to other non-bone sites. For all patients and controls we collected blood and urinary samples. We evaluated patients' bone homeostasis; we made peripheral blood mononuclear cell (PBMC) cultures to detect in vitro osteoclastogenesis; we dosed serum expression of molecules involved in cancer induced osteoclatogenesis, such as RANKL, OPG, TNF-alpha, DKK-1 and IL-7. By Real-Time PCR, we quantified DKK-1 and IL-7 gene expression on micro-dissected tumour and healthy tissue sections. PRINCIPAL FINDINGS: CaP bone metastatic patients showed bone metabolism disruption with increased bone resorption and formation compared to non-bone metastatic patients and healthy controls. The CaP PBMC cultures showed an enhanced osteoclastogenesis in bone metastatic patients, due to an increase of RANKL/OPG ratio. We detected increased DKK-1 serum levels and tissue gene expression in patients compared to controls. IL-7 resulted high in patients' sera, but its tissue gene expression was comparable in patients and controls. CONCLUSIONS: We demonstrated ex vivo that osteoclastogenesis is an active mechanism in tumour nesting of bone forming metastatic cancer and that serum DKK-1 levels are increased in CaP patients, suggesting to deeply investigate its role as tumour marker