Occurrence and diversity of black-foot pathogens on asymptomatic nursery-produced grapevines in Turkiye

[EN] Black-foot (BF) disease of grapevines in nurseries and young vineyards is caused by soil-borne Cylindrocarpon-like asexual morphs. They can be found both in symptomatic and asymptomatic vines, being spread to new grape growing areas during vineyard establishment. In this study, 42 grapevine nurseries located in different geographical regions in Turkiye were surveyed in 2021 to determine the presence of BF pathogens on asymptomatic marketable plants. Black-foot fungi were isolated from the roots or basal ends of asymptomatic dormant vines in 39 of the nurseries (92.9%). The percentage of isolation of BF pathogens ranged from 1.4 to 51.4% (average 18.4%). Seven species: Cylindrodendrum alicantinum, Cylindrocladiella peruviana, Dactylonectria macrodidyma, D. novozelandica, D. torresensis, Ilyonectria liriodendri, and I. robusta were identified based on DNA sequencing of histone H3 gene and phylogenetic analyses, D. torresensis being the most frequent. From these species Ca. peruviana, D. novozelandica and I. robusta were detected for the first time on grapevines in Turkiye. Pathogenicity tests on 1103P rootstock cuttings revealed that all species significantly decreased root biomass and increased root disease severity index, when compared with the non-inoculated control, D. novozelandica being the most virulent. Pathogenicity of Cm. alicantinum to grapevine was confirmed for the first time, thus this species should be included as causal agent of BF of grapevines. These findings point out that BF pathogens are highly prevalent in the nurseries and could represent a serious threat for Turkish viticulture.Akgül, DS.; Yildiz, M.; Savas, NG.; Bülbül, I.; Özarslandan, M.; León Santana, M.; Armengol Fortí, J. (2022). Occurrence and diversity of black-foot pathogens on asymptomatic nursery-produced grapevines in Turkiye. European Journal of Plant Pathology. 164(1):21-32. https://doi.org/10.1007/s10658-022-02535-521321641Abreo, E., Martínez, S., Betucci, L., & Lupo, S. (2010). Morphological and molecular characterization of Campylocarpon and Cylindrocarpon spp. associated with black foot disease of grapevines in Uruguay. Australasian Plant Pathology, 39, 446–452. https://doi.org/10.1071/AP10021Agustí-Brisach, C., & Armengol, J. (2013). Black-foot disease of grapevine: An update on taxonomy, epidemiology and management strategies. Phytopathologia Mediterranea, 52, 245–261. https://doi.org/10.14601/Phytopathol_Mediterr-12662.Agustí-Brisach, C., Gramaje, D., García-Jiménez, J., & Armengol, J. (2013). Detection of black-foot disease pathogens in the grapevine nursery propagation process in Spain. European Journal of Plant Pathology, 137, 103–112. https://doi.org/10.1007/s10658-013-0221-8Agustí-Brisach, C., Cabral, A., González-Domínguez, E., Pérez-Sierra, A., León, M., Abad-Campos, P., García-Jiménez, J., Oliveira, H., & Armengol, J. (2016). Characterization of Cylindrodendrum, Dactylonectria and Ilyonectria isolates associated with loquat decline in Spain, with description of Cylindrodendrum alicantinum sp. nov. European Journal of Plant Pathology, 145(1), 103–118. https://doi.org/10.1007/s10658-015-0820-7Aiello, D., Gusella, G., Vitale, A., Guarnaccia, V., & Polizzi, G. (2020). Cylindrocladiella peruviana and Pleiocarpon algeriense causing stem and crown rot on avocado (Persea americana). European Journal of Plant Pathology, 158(2), 419–430. https://doi.org/10.1007/s10658-020-02082-xAigoun-Mouhous, W., Elena, G., Cabral, A., Leon, M., Sabaou, N., Armengol, J., Chaouia, C., & Mahamedi, & Berraf-Tebbal, A. (2019). Characterization and pathogenicity of Cylindrocarpon-like asexual morphs associated with black foot disease in Algerian grapevine nurseries, with the description of Pleiocarpon algeriense sp. nov. European Journal of Plant Pathology, 154, 887–901. https://doi.org/10.1007/s10658-019-01708-zAkgül, D. S., Savaş, N. G., Önder, S., Özben, S., & Kaymak, S. (2014). First report of Campylocarpon fasciculare causing black foot disease of grapevine in Turkey. Plant Disease, 98(9), 1277. https://doi.org/10.1094/PDIS-03-14-0284-PDNAlaniz, S., Leon, M., García-Jiménez, J., Abad, P., & Armengol, J. (2007). Characterization of Cylindrocarpon species associated with black-foot disease of grapevine in Spain. Plant Disease, 91, 1187–1193. https://doi.org/10.1094/PDIS-91-9-1187Alaniz, S., Abad-Campos, P., García-Jiménez, J., & Armengol, J. (2011). Evaluation of fungicides to control Cylindrocarpon liriodendri and Cylindrocarpon macrodidymum in vitro, and their effect during the rooting phase in the grapevine propagation process. Crop Protection, 30, 489–494. https://doi.org/10.1016/j.cropro.2010.12.020Berlanas, C., Ojeda, S., López-Manzanares, B., Andrés-Sodupe, M., Bujanda, R., Martinez-Diz, M. D. P., Diaz-Losada, E., & Gramaje, D. (2020). Occurrence and diversity of black-foot disease fungi in symptomless grapevine nursery stock in Spain. Plant Disease, 104, 94–104. https://doi.org/10.1094/PDIS-03-19-0484-RECabral, A., Groenewald, J. Z., Rego, C., Oliveira, H., & Crous, P. W. (2012). Cylindrocarpon root rot: Multi-gene analysis reveals novel species within the Ilyonectria radicicola species -complex. Mycological Progress, 11, 655–668. https://doi.org/10.1007/s11557-011-0777-7Carlucci, A., Lops, F., Mostert, L., Halleen, F., & Raimondo, M.L. (2017). Occurrence fungi causing black foot on young grapevines and nursery rootstock plants in Italy. Phytopathologia Mediterranea, 56, 10–39. https://doi.org/10.14601/Phytopathol_Mediterr-18769.Crous, P. W., Groenewald, J. Z., Risede, J. M., & Hywel-Jones, N. L. (2004). Calonectria species and their Cylindrocladium anamorphs: Species with sphaeropedunculate vesicles. Studies in Mycology, 50, 415–429.Felsenstein, J. (1985). Confidence limits on phylogenies: An approach using the bootstrap. Evolution, 39, 783–791.Fischer, M., & Kassemeyer, H. H. (2003). Fungi associated with esca disease of grapevine in Germany. Vitis, 42, 109–116. https://doi.org/10.5073/vitis.2003.42.109-116Gatica, M., Césari, C., Magnin, S., & Dupont, J. (2001). Phaeoacremonium species and Phaeomoniella chlamydospora in vines showing “hoja de malvón” and young vine decline symptoms in Argentina. Phytopathologia Mediterranea, 40, 317–324 https://www.jstor.org/stable/44981636Gomez, K. A., & Gomez, A. A. (1984). Statistical procedures for agricultural research (2nd ed.). Wiley 680 pp.Gramaje, D., & Armengol, J. (2011). Fungal trunk pathogens in the grapevine propagation process: Potential inoculum sources, detection, identification, and management strategies. Plant Disease, 95, 1040–1055. https://doi.org/10.1094/PDIS-01-11-0025Gramaje, D., Úrbez-Torres, J. R., & Sosnowski, M. R. (2018). Managing grapevine trunk diseases with respect to etiology and epidemiology: Current strategies and future prospects. Plant Disease, 102, 12–39. https://doi.org/10.1094/PDIS-04-17-0512-FEGrasso, S., & Magnano Di San Lio, G. (1975). Infezioni di Cylindrocarpon obtusisporum su piante di vite in Sicilia. Vitis, 14, 38–39.Halleen, F., Schroers, H. J., Groenewald, J. Z., & Crous, P. W. (2004). Novel species of Cylindrocarpon (Neonectria) and Campylocarpon gen. nov. associated with black-foot disease of grapevines (Vitis spp). Studies in Mycology, 50, 431–455.Halleen, F., Fourie, P. H., & Crous, P. W. (2006). A review of black foot disease of grapevine. Phytopatologia Mediterranea, 45, 55–67 https://www.jstor.org/stable/26463236Jones, E. E., Brown, D. S., Bleach, C. M., Pathrose, B., Barclay, C., Jaspers, M. V., & Ridgway, H. J. (2012). First report of Cylindrocladiella parva as a grapevine pathogen in New Zealand. Plant Disease, 96, 144. https://doi.org/10.1094/PDIS-04-11-0347Kumar, S., Stecher, G., Li, M., Knyaz, C., & Tamura, K. (2018). MEGA X: Molecular evolutionary genetics analysis across computing platforms. Molecular Biology and Evolution, 35, 1547–1549. https://doi.org/10.1093/molbev/msy096Langenhoven, S.D., Halleen F., Spies, C.F.J., Stempien, E., & Mostert, L. (2018). Detection and quantification of black foot and crown and root rot pathogens in grapevine nursery soils in the Western cape of South Africa. Phytopathologia Mediterranea, 57, 519–537. https://doi.org/10.14601/Phytopathol_Mediterr-23921.Lawrence, D. P., Nouri, M. T., & Trouillas, F. P. (2019). Taxonomy and multi-locus phylogeny of Cylindrocarpon-like species associated with diseased roots of grapevine and other fruit and nut crops in California. Fungal Systematics and Evolution, 4, 59–75. https://doi.org/10.3114/fuse.2019.04.06Maluta, D. R., & Larignon, P. (1991). Pied-noir: mieux vaut prévenir. Viticulture, 11, 71–72.Mohammadi, H., Alaniz, S., Banihashemi, Z., & Armengol, J. (2009). Characterization of Cylindrocarpon liriodendri associated with black foot disease of grapevine in Iran. Journal of Phytopathology, 157, 642–645. https://doi.org/10.1111/j.1439-0434.2009.01555.xO’Donnell, K., Cigelnik, E., & Nirenberg, H. I. (1998). Molecular systematics and phylogeography of the Gibberella fujikuroi species complex. Mycologia, 90(3), 465–493. https://doi.org/10.2307/3761407Özben, S. (2020). Asma fidanlıklarında önemli odun dokusu fungal hastalıkların tespiti ve bazı üzüm çeşitlerinin Phaeoacremonium aleophilum’a karşı reaksiyonlarının belirlenmesi. PhD Thesis (in Turkish), Ankara University, Graduate School of Natural and Applied Science, Turkey. 175 pp.Özben, S., Demirci, F., Degirmenci, K., & Uzunok, S. (2012). First report of Cylindrocarpon macrodidymum associated with black foot disease of grapevine in Turkiye. Plant Disease, 96, 762. https://doi.org/10.1094/pdis-05-11-0393-pdnÖzben, S., Demirci, F., & Uzunok, S. (2019). First report of Cylindrocladiella parva associated with black foot disease of grapevine in Turkiye. 1st International Molecular Plant Protection Congress, 96 pp. Adana, .Petit, E., Barriault, E., Baumgartner, K., Wilcox, W. F., & Rolshausen, P. E. (2011). Cylindrocarpon species associated with black-foot of grapevine in northeastern United States and southeastern Canada. American Journal of Enology and Viticulture, 62, 177–183. https://doi.org/10.5344/ajev.2011.10112Pintos, C., Redondo, V., Costas, D., Aguin, O., & Mansilla, P. (2018). Fungi associated with grapevine trunk diseases in nursery-produced Vitis vinifera plants. Phytopathologia Mediterranea, 57, 407–424. https://doi.org/10.14601/Phytopathol_Mediterr-22964.Rego, C., Nascimento, T., & Oliviera, H. (2001). Characterization of Cylindrocarpon destructans isolates from grapevines in Portugal. Phytopathologia Mediterranea, 40, 343-350. https://doi.org/10.14601/Phytopathol_Mediterr-1614.Reis, P., Cabral, A., Nascimento, T., Oliviera, H., & Rego, C. (2013). Diversity of Ilyonectria species in a young vineyard affected by black foot disease. Phytopathologia Mediterranea, 52, 335-346. https://doi.org/10.14601/Phytopathol_Mediterr-12719.Savaş, N., Akgül, D. S., & Albaz, E. A. (2015). First report of Ilyonectria liriodendri associated with black foot disease of grapevine in Turkey. Plant Disease, 99, 1855. https://doi.org/10.1094/PDIS-03-15-0246-PDNSavaş, N., Akgül, D. S., Özarslandan, M., & Yıldız, M. (2020). First report of Dactylonectria alcacerensis and Dactylonectria torresensis associated with black foot disease of grapevine in Turkey. Plant Disease, 104, 2027. https://doi.org/10.1094/PDIS-02-20-0385-PDNStamatakis, A. (2014). RAxML version 8: A tool for phylogenetic analysis and post-analysis of large phylogenies. Bioinformatics, 30, 1312–1313. https://doi.org/10.1093/bioinformatics/btu033Thompson, J. D., Higgins, D. G., & Gibson, T. J. (1994). CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Research, 22, 4673–4680. https://doi.org/10.1007/978-1-4020-6754-9_3188Towsend, G. R., & Heuberger, J. V. (1943). Methods for estimating losses caused by diseases in fungicide experiments. Plant Disease Reporter, 24, 340–343.Whitelaw-Weckerd, M. A., Nair, N. G., Lamont, R., Alonso, M., Priest, M. J., & Huang, R. (2007). Root infection of Vitis vinifera by Cylindrocarpon liriodendri in Australia. Australasian Plant Pathology, 36, 403–406. https://doi.org/10.1071/AP07041Ye, Q., Zhang, W., Jia, J., Li, X., Zhou, Y., Han, C., Wu, X., & Yan, J. (2021) Fungal pathogens associated with black foot of grapevine in China. Phytopathologia Mediterranea, 60(2), 303-319. https://doi.org/10.36253/phyto-12353

Pre-emptive Use of Riboflavin in a Rat Model of Bilateral Cavernous Nerve Injury

Objective: Erectile dysfunction is commonly encountered after radical prostatectomy due to cavernous nerve injury (CNI). We investigated the effects of riboflavin (Rb) on bilateral CNI in a rat model. Materials and Methods: Twenty-four male rats were divided into four groups: control (C), patients with bilateral CNI, those with CNI receiving postoperative Rb treatment (CNI+Rb), and those with CNI receiving pre- and post-operative Rb treatment (Rb+CNI+Rb). Bilateral CNI was performed in all groups except for C. The CNI+Rb group was treated with 30 mg/kg Rb daily after CNI for two weeks; the Rb+CNI+Rb group was treated with 30 mg/kg Rb daily one week before CNI and then for two weeks after injury. Mean arterial pressure (MAP) and intracavernosal pressure (ICP) were measured 14 days after CNI in all groups. Tissue malondialdehyde, cyclic guanosine monophosphate, nerve growth factor, superoxide dismutase and total nitric oxide synthase (NOS) activities, neuronal NOS (nNOS) and inducible NOS (iNOS) were analyzed. Results: ICP/MAP ratio was significantly lower in the CNI (p<0.01) and CNI+Rb groups (p<0.05) compared to the control group, however, the Rb+CNI+Rb group had results comparable to the C group in terms of nNOS and iNOS expression in the Western Blot analysis. Conclusion Rb exerted anti-oxidative and anti-inflammatory effects on CNI in a CNI rat model. Rb can be a potential beneficial agent to improve erectile function in nerve-sparing radical prostatectomy patients as a preemptive penile rehabilitation strategy, although further clinical studies are needed

5-fluorourasil ile oluşturulan ileum ve kolon epitel hasarına karşı Saint John’s Wort’un olası koruyucu rolünün değerlendirilmesi

ÖZET 5-Fluorourasil ile Oluşturulan İleum ve Kolon Epitel Hasarına Karşı Saint John’s Wort’un Olası Koruyucu Rolünün Değerlendirilmesi Öğrenci Adı ve Soyadı: Nurdan BÜLBÜL Danışman: Prof. Dr. Şule ÇETİNEL Anabilim dalı: Histoloji ve Embriyoloji Amaç: 5-Fluorourasil (5-FU) kanser tedavisinde yaygın olarak kullanılan bir kemoterapötik ajandır. Ancak ileit ve kolit gibi yan etkileri vardır. Saint John’s Wort (SJW) antioksidan, antiinflamatuvar ve antibakteriyal özelliklerinin yanında orta düzeyde depresyonların tedavisinde kullanılan bir tıbbi bitkidir. Bu çalışmanın amacı 5-FU uygulanarak oluşturulmuş ileit ve kolite karşı Saint John’s wort’un olası koruyucu etkisinin araştırılmasıdır. Gereç ve Yöntem: Çalışmada dişi Sprague Dawley sıçanlar (220-300 gr) kullanıldı ve üç deney grubuna (n=5) ayrıldı: Grup 1- Serum fizyolojik (SF) grubu, Grup 2- 5-FU uygulama grubu, Grup 3- 5-FU+SJW uygulama grubu. Her gün 150 mg/kg dozda i.p. 5-FU uygulamasının ardından SF ya da 300 mg/kg dozda SJW 4 gün boyunca verildi. Dekapitasyon sonrası ileum ve kolon dokuları histopatoloji için alınıp %10’luk formaldehitte fikse edilerek ışık mikroskopi için rutin olarak takip edildi. Apoptotik hasar TUNEL ve Western Blot yöntemleriyle, proliferasyon PCNA immünohistokimyası ile araştırılıp, sonuçları istatistiki olarak değerlendirildi. Bulgular ve Sonuçlar: Kolon ve ileum dokularında kontrol grupları düzenli epitel ve çok sayıda goblet hücresi içeren glanduler morfoloji sergiledi. Kolon dokusunda 5-FU tedavisi bezlerde dejenerasyon ve lamina propriyada yüksek düzeyde lökosit birikimiyle epitelyal organizasyon bozukluğuna yol açtı. SJW bezlerdeki dejenerasyonu ve lökosit konsantrasyonunu azalttı. İleumda 5-FU tedavisi ile epitelde oldukça fazla dökülmeyle beraber dejenerasyon ve lamina propriyada lökosit birikimi vardı. SJW tedavisi epitelde ve bezlerde orta düzeyde rejenerasyon sağladı. Western Blot metodu, TUNEL metodu ve PCNA immünohistokimyası sonuçları bu bulgularla uyum gösterdi. Anahtar Sözcükler: 5-Fluorourasil, Saint John’s wort, İleit, Kolit, ApoptozSUMMARY Evaluation of Possible Protective Effect of Saint John’s Wort Against 5-Fluorouracil-Induced Ileum and Colon Epithelium Injury Student Name and Surname: Nurdan Bülbül, Advisor: Prof. Dr. Şule Çetinel Department: Histology and Embryology Aim: 5-Fluorouracil (5-FU) is a commonly used chemotherapeutic agent for cancer treatment. However, it has some side effects including ileitis and colitis. St. John's wort (SJW) is an herbal medicine that is used for therapy of mild depression mainly besides it has antioxidant, anti-inflammatory, and anti-bacterial properties. The aim of this study is to investigate the possible preventive effect of Saint John’s wort against ileitis and colitis which are induced by 5-FU. Materials and Methods: Female Sprague Dawley rats (220-300g) were used in the study and three experimental groups (n=5) : Group ( 1) –saline group, Group ( 2) 5-fluorouracil (5-FU) treated control, and Group (3) 5-FU + SJW treated group were set. Following 150 mg/kg dose of 5-FU, either saline or SJW (300 mg/kg) were administered in each day during 4 days. After decapitation, ileum and colon tissues of the intestine were removed, fixed in 10% formaldehyde and processed routinely for light microscopy. Apoptotic injury was shown via TUNEL assay and Western Blot method. Proliferation was shown via PCNA immunohistochemistry. Data were analyzed statistically. Results: In the colon and ileum tissues, the control groups demonstrated regular epithelial and glandular morphology with abundance of goblet cells. In the colon tissue, 5-FU treatment degenerated the glands prominently and had led to epithelial disorganization along with a high degree of accumulation of leukocytes in lamina propria. The SJW treatment appeared to have reduced the degeneration of glands and concentration of leukocytes. In the ileum, there was degeneration with severe desquamation in the epithelium, and accumulation of leukocytes in the lamina propria, SJW treatment led to a moderate regeneration in the epithelium and glands. Western Blot method, TUNEL assay and PCNA immunohistochemistry results confirmed histology. Key Words: 5-Fluorouracil, Saint John’s Wort, İleitis, Colitis, Apoptosis

Evaluation of the oxytocin effect in a rat model with experimental periodontitis

The aim of the present study was to evaluate the inhibitory effects of oxytocin on the development of periodontitis based on its properties against bone loss and resorption. Thirty-two Wistar albino rats were divided into four equal groups: control, periodontitis + saline, periodontitis + 0.5 mg/kg/day oxytocin, and periodontitis + 1 mg/kg/day oxytocin. Periodontitis groups received 4.0 silk ligatures around their cervixes of the right and left mandibular incisors in an 8 shape, kept for 14 days. Animals in oxytocin groups were injected once every day during 14 days with oxytocin. The mandibles were fixed and scanned using microcomputed tomography to quantify bone resorption and volumetric measurements. Blood samples were collected to analyze the concentrations of macrophage colony-stimulating factor (M-CSF), receptor activator of nuclear factor-kappa Beta ligand (RANKL), osteoprotegerin (OPG), matrix metalloproteinase-8 (MMP-8), tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-6, glutathione peroxidase (GPx), superoxide dismutase (SOD), and malondialdehyde (MDA). Histopathological evaluations were conducted to examine the gingiva and alveolar bone. Oxytocin prevented the development of periodontitis by decreasing ligament deteriorations and leukocytes in the gingival connective tissue and promoting reintegration with the alveolar bone. Bone resorption in all regions was less in the periodontitis + 1 mg/kg/day oxytocin group than in the periodontitis + saline group. Although TNF-alpha, IL-6, and RANKL values were lower in the periodontitis + 1 mg/kg/day oxytocin group, OPG was higher than that in the periodontitis + saline group. M-CSF, MMP-8, and MDA were lower in the oxytocin groups than in the periodontitis + saline group. Oxytocin may be an effective agent for periodontal diseases because it decreased bone resorption, oxidative stress, and inflammation in an experimental periodontitis