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Chilean avocado orchards have been affected by branch canker and dieback, mainly associated with pathogens species of the family Botryosphaeriaceae. Producers often relate water stress to these phenomena; however, predisposing factors for these diseases in Chilean orchards have not been reported. Therefore, the objective of this study was to identify the main climate, planting, and management variables associated with branch canker and dieback in Chilean avocado orchards. Multivariable statistical analyses (PCA and PLS-DA) were performed to analyze 76 variables possibly associated with the incidence of these diseases during two consecutive growing seasons. Our findings indicated that branch canker and dieback are mainly conditioned by orchard variables such as plant age, canopy volume, trunk diameter, leaf area index, and planting density. Variables associated with aged orchards, such as high age, high canopy volume, and high trunk diameter, appeared to be more associated with high incidence levels than were younger orchards with higher planting densities and high tree vigor levels. In addition, abiotic stresses, such as frost and overirrigation, were associated as secondary factors affecting these diseases' development in avocado orchards. Moreover, this study allowed us to determine that branch canker and dieback can develop indistinctly in the different Chilean agroclimatic zones. [ABSTRACT FROM AUTHOR]

Grapevine grafting is an essential practice in viticulture and over the years, various bench grafting techniques have been developed to mechanize the nursery process and to increase the yield in number of viable cuttings. Bench grafting is a fundamental nursery practice that can potentially affect the quality of propagation material also in young decline associated to grapevine trunk diseases and has been recently reported to influence leaf symptoms development associated with diseases of Esca complex. The study aimed to investigate how three bench grafting methods [i.e., (i) Omega graft as mechanical technique, (ii) Whip and Tongue graft as manual technique and (iii) Full Cleft graft as semi-mechanical technique] can influence these phenomena. Specifically, the different methods were compared for their effect on the anatomical development of the grafting point and the functionality of the xylem, also considering two factors: the grapevine cultivar (Cabernet Sauvignon, Glera and Teroldego) and the scion/rootstock diameter (thin and large). Observations by light microscopy on the anatomical evolution and measurements on the xylem morphology and hydraulic traits were correlated with the grafting methods and the investigated varieties. The anatomical observations revealed that the mechanical (Omega) and semimechanical (Full Cleft) grafting methods have a faster callusing response while the manual technique (Whip and Tongue) has a slower but greater vascularization of the differentiated callus. Significant differences between cultivars and/or grafting types were also detected in necrotic area on the grafted tissues. Statistical analysis of the grapevine vessels suggested differences in xylem parameters between cultivars, while grafting type had no significant effects. On the other hand, the grafting type significantly affected the intrinsic growth rate. The study confirms the potential incidence of lesions and dysfunctionalities correlated with the grafting method applied, which can potentially induce grafted vine declines in vineyards due to the necrotic area detected on the grafted tissues. [ABSTRACT FROM AUTHOR]

Grapevine grafting is an essential practice in viticulture and over the years, various bench grafting techniques have been developed to mechanize the nursery process and to increase the yield in number of viable cuttings. Bench grafting is a fundamental nursery practice that can potentially affect the quality of propagation material also in young decline associated to grapevine trunk diseases and has been recently reported to influence leaf symptoms development associated with diseases of Esca complex. The study aimed to investigate how three bench grafting methods [i.e., (i) Omega graft as mechanical technique, (ii) Whip and Tongue graft as manual technique and (iii) Full Cleft graft as semi-mechanical technique] can influence these phenomena. Specifically, the different methods were compared for their effect on the anatomical development of the grafting point and the functionality of the xylem, also considering two factors: the grapevine cultivar (Cabernet Sauvignon, Glera and Teroldego) and the scion/rootstock diameter (thin and large). Observations by light microscopy on the anatomical evolution and measurements on the xylem morphology and hydraulic traits were correlated with the grafting methods and the investigated varieties. The anatomical observations revealed that the mechanical (Omega) and semi-mechanical (Full Cleft) grafting methods have a faster callusing response while the manual technique (Whip and Tongue) has a slower but greater vascularization of the differentiated callus. Significant differences between cultivars and/or grafting types were also detected in necrotic area on the grafted tissues. Statistical analysis of the grapevine vessels suggested differences in xylem parameters between cultivars, while grafting type had no significant effects. On the other hand, the grafting type significantly affected the intrinsic growth rate. The study confirms the potential incidence of lesions and dysfunctionalities correlated with the grafting method applied, which can potentially induce grafted vine declines in vineyards due to the necrotic area detected on the grafted tissues.

Hazelnut is among the most important nut crops in Chile, currently covering 46,000 ha. In 2023, the country exported 30,000-ton. In recent years the incidence of plants with internal discoloration, cankers and dieback has been increasing. In some cases, the trees died and had to be removed and, after a year, purple resupinate fruiting bodies were observed growing from the stumps. To determine the etiology of the symptoms and signs, wood samples (n=318) were collected since 2020, from 38 symptomatic orchards from Maule to La Araucanía Regions, primarily from the cvs. Tonda di Giffoni and Lewis. Wood sections 0.5 cm diameter were cut from the symptomatic tissues, disinfected using a sodium hypochlorite (10%) solution, and plated on a quarter-strength acidified potato dextrose agar (aPDA1/4). The plates were incubated and purified on PDA. Subsequently, isolates were identified by morphological and molecular means. Almost half of the isolates (47%) were preliminarily identified as basidiomycetes, based on mycelial features such as the presence of clamp connections, with 45% of them exhibiting abundant whitish cottony fast-growth mycelia, resembling Chondrostereum purpureum (Grinbergs et al ., 2020). DNA was extracted and the 500-bp fragment, located between 5S and 18S ribosomal regions, was amplified using APN1 specific primers (Becker et al . 1999), identifying the isolates as C. purpureum . In addition, 5.8S gene of RGM1 (35°13'40.9"S 71°25'14.1"W), RGM2 (36°31'27.95"S 71°46'58.31"W), RGM3 (37°10'54.8"S 72°03'39.6"W), RGM4 (35°19'25.2"S 71°19'54.7"W) and RGM5 (36°35'30.8"S 72°05'18.8"W) isolates, representing different locations within the hazelnut growing area, was amplified using ITS1/ITS4 primers (White et al ., 1990). The PCR product was sequenced, and the analysis showed 100% homology among isolates (Genebank codes: PP839283, PP839284, PP839285, PP839286 and PP839287, respectively). To determine the pathogenicity of the isolates, 30-cm healthy cuttings cv. Lewis were inoculated with mycelial plugs, while control shoots were inoculated with sterile agar plugs. Cuttings were vertically arranged in pots with 3-cm water and incubated for 60-d at 22°C. In addition, fresh cuts of 3-y potted plants cv. Lewis were inoculated with mycelial plugs and incubated for 137-d in a shadehouse. After incubation, bark was removed from inoculated cuttings and the length of necrotic lesions was measured. Although discoloration was reproduced by all the isolates in both pathogenicity tests, RGM1 isolate was the most aggressive, causing the complete discoloration of the cuttings and the death of the inoculated plants. To our knowledge this is the first report of C. purpureum causing wood disease in hazelnut. These findings are significant because the disease may not only reduce orchard longevity but also decrease fruit yield and quality, as observed in other fruit crops (Grinbergs et al. , 2021).

Chilean avocado orchards have been affected by branch canker and dieback, mainly associated with pathogens species of the family Botryosphaeriaceae. Producers often relate water stress to these phenomena; however, predisposing factors for these diseases in Chilean orchards have not been reported. Therefore, the objective of this study was to identify the main climate, planting, and management variables associated with branch canker and dieback in Chilean avocado orchards. Multivariable statistical analyses (PCA and PLS-DA) were performed to analyze 76 variables possibly associated with the incidence of these diseases during two consecutive growing seasons. Our findings indicated that branch canker and dieback are mainly conditioned by orchard variables such as plant age, canopy volume, trunk diameter, leaf area index, and planting density. Variables associated with aged orchards, such as high age, high canopy volume, and high trunk diameter, appeared to be more associated with high incidence levels than were younger orchards with higher planting densities and high tree vigor levels. In addition, abiotic stresses, such as frost and overirrigation, were associated as secondary factors affecting these diseases’ development in avocado orchards. Moreover, this study allowed us to determine that branch canker and dieback can develop indistinctly in the different Chilean agroclimatic zones.

Neofusicoccum parvum and Rhizoctonia solani are fungal pathogens with an increasing incidence in young grapevine plants. In this study, the antagonistic potential of some strains of the genus Trichoderma isolated from grapevine against these pathogens was investigated at the laboratory and greenhouse levels. In-plate confrontation assays showed that the selected Trichoderma strains could inhibit the mycelial growth of both taxa, being more effective against N. parvum. In the in vivo assays, the biocontrol activity of the mentioned strains against the pathogens, when applied either simultaneously or successively, was tested on both grafted plants and seedlings germinated from seed. The effectiveness of the treatments was evaluated by comparing biomass weight and vascular rot lengths data. In seedling trials, successive treatments resulted in higher root development and a lower colonization rate of the pathogens, especially against R. solani. In grafted plants, some disparity was observed against N. parvum: simultaneous treatments resulted in higher aerial biomass, but successive treatments resulted in higher root biomass and lower necrosis. Against R. solani, simultaneous treatments were clearly more effective, with higher root and aerial length values and lower necrosis. The obtained data suggest that the use of Trichoderma spp. isolates can constitute an alternative to conventional fungicides to control certain grapevine wood diseases.

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Phaeomoniella chlamydospora (Pc) and Phaeoacremonium minimum (Pm) are among the main grapevine trunk disease pathogens affecting vines in the nursery. These pathogens can cause infections of the propagation material often appearing as dark wood streaking, and are associated with decline occurrence in young vineyards. The infection may occur during the grafting process, particularly during hydration. Promising preliminary results were obtained in the nursery using electrolyzed acid water (EAW) for the hydration of cuttings; EAW is characterized by low values of pH (2.5), high oxidation reduction potential (ORP>1000) and a certain amount of free chlorine. No differences in absorption kinetics were recorded between EAW and tap water, nor among 1103P, K5BB and SO4 rootstocks. In all combinations no significant additional absorption was observed after 7 h of immersion. In order to assess the characteristics of EAW after the contact with the inner plant tissues ORP and pH were measured from the liquid extracted from rootstocks after immersion in EAW. The values of the liquid, named "weakened" EAW (wEAW) (625–650 mV ORP and 3.1–3.3 pH), were significantly different from the values of both EAW and the liquid extracted after immersion in tap water. When the propagation material was hydrated for 7 h in EAW no significant effect on vegetating growth performance and percentage of certifiable plants was observed for SO4 or 1103P, whereas a reduction in plant vegetative growth was observed in K5BB hydrated vines at 12hr hydration. EAW and wEAW gave no significant reduction of in vitro mycelial growth, whereas a consistent decrease in conidial germination was observed in Pc and Pm. The three-year experiments carried out in the nursery on cuttings inoculated with Pc and hydrated in EAW showed a remarkable reduction of the infection level in the treated plants. • Phaeomoniella chlamydospora or Phaeoacremonium spp. infections in nursery are reduced. • Electrolyzed acid water (EAW) reduces fungal wood infections on grapevine. • EAW has effect against conidia. • EAW is able to control systemic fungal infection due to hydration along the trunk. [ABSTRACT FROM AUTHOR]

Abstract Wood degrading diseases are escalating in vineyards all over the world, inducing heavy economic losses. The diseases are caused by fungal pathogens that invade the innermost xylem part of the trunk and trigger many metabolic disorders of vines leading to hindered productivity. In particular, esca disease in which Phaemoniella chlamydospora and Phaeacremonium minimum are implicated, is very frequently observed. The aim of this work was to establish the basis of a preventive/curative treatment to control these types of diseases, since no efficient treatment is yet available. Therefore, we proposed to design a treatment based on a combination of active chemicals that exhibit antimicrobial properties per se and/or are able to activate the defense reactions of the plant. An essential property of the chemicals should reside in their capacity to be transported to remote areas within the plant. To this end, the chemicals chosen in our experimental model were cysteine, iron sulfate, salicylic acid and fosetyl aluminium. The effective conditions of application were determined and the efficiency of the treatments was verified on cuttings using an immunological assay. The non-toxicity of the treatments was tested on cuttings and vinestocks following a series of sprayings on leaves at various periods of the vine culture process. Highlights • A preventive/curative treatment of grapevine towards fungal induced esca is proposed. • A mixture of antimicrobial molecules inhibit mycelial growth and spore germination. • The efficiency of the treatment can be monitored by an immunological assay. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate the in vitro antifungal activity (AA) of the essential oil (EO) of lemon ( Citrus limon L.) against three pathogenic fungi attacking grapevine wood. The composition of the EO was also studied. Ten volatile components were identified by gas chromatography–mass spectrometry. The results showed that the EO consists of volatile components where monoterpene hydrocarbons are the most abundant ones. Four major components were identified, which represent 99.9% of the total EO (limonene, neral, β-pinene, and γ-terpinene). The AA of the EO was evaluated against three pathogenic fungi attacking grapevine wood ( Eutypa sp., Botryosphaeria dothidea , and Fomitiporia mediterranea ). The results showed that the EO exerts AA against all tested fungi and significantly inhibits their growth. Eutypa sp. is the most sensitive fungus. These results show, for the first time, a new use for the EO of lemon ( C. limon L.) to control fungal diseases of grapevine wood. [ABSTRACT FROM AUTHOR]

Wood-degrading diseases, in particular esca, Botryosphaeria dieback and Eutypa dieback, affect grapevines in vineyards around the world. The wood-inhabiting fungal pathogens degrade wood tissues in the colonization areas and induce, at distance, characteristic foliar symptoms. These effects have been previously partly correlated with the secretion of polypeptides (PFpal) presenting toxic effects on plant cells at various metabolic levels. These diseases are insidious and the symptoms can occur in mild or severe forms, showing differences from one year to another. Observations allowed to assume that variations in climatic conditions may intervene in this behavior. We showed here that a modification in the temperature regime modified the fungal development of Phaeoacremonium aleophilum and induced modifications of the intracellular structures, observed in particular in the nucleus, the vacuolar apparatus, the Golgi, the endoplasmic reticulum and also in the hyphal coat. Furthermore, changes in the pattern of the secreted polypeptides were observed as well quantitatively as qualitatively by electrophoretic analysis and immunolabeling by anti-PFpal antibodies. Subsequently, the different polypeptide fractions obtained from fungal cultures, either maintained at 20 °C or respectively transferred to colder (5 °C) and warmer (30 °C) temperatures, differentially affected the grapevine physiology as shown by the different effects induced on leaf anthocyanin synthesis, modification of proton secretion and lethality of cells in culture. [ABSTRACT FROM AUTHOR]

Phaeoacremonium aleophilum is a fungal pathogen implicated in esca, a devastating disease affecting grapevines around the world. We have previously reported that the fungus secreted in its culture medium a variety of polypeptides, which allowed us to develop a serological method of detection. In this method, rabbit antibodies raised against the polypeptide fraction recognized secreted fungal proteins with high sensitivity (commonly 1 ng). These antibodies were specific since they cross-reacted with polypeptides secreted by various strains of Pm. aleophilum, but not with the proteins secreted by many other fungal pathogens implicated in other grapevine infections. Importantly, as shown by ELISA tests and immunolocalization, they did not cross-react with the secreted polypeptides of Eutypa lata (agent of Eutypa dieback), Neofusicoccum parvum (found in Black Dead Arm), and Phaeomoniella chlamydospora (also found in the fungal complex inducing esca). This serological approach permitted the development of a reliable dot-blot method to detect the presence in woody members of Pm. aleophilum in selectively infected trunk and canes. This method of diagnosis is rapid, easy to perform and non destructive for grapevines. Advantages and drawbacks of the method are discussed. [ABSTRACT FROM AUTHOR]

Botryosphaeria parva, Eutypa lata and Phomopsis viticola are ascomyceteous fungi responsible for severe canker and dieback in numerous woody plants. In grapevine, these pathogens colonise the wood mainly through pruning wounds, and the diseases gradually develop, leading to partial or total vine death. In the present study, the three fungal species were grown in Czapek Dox modified medium. Under these conditions, fungal colonies are able to distinguish self from non-self. The production of reactive oxygen species (ROS) was analysed by specifically staining for superoxide (O2.-) or peroxide (O22-) radicals. The presence of ROS in both isolated cultures and fungal interactions was confi rmed. All fungi produced both radicals, in every interaction. However, the patterns of ROS production depend on the fungus itself and on the fungal species with which it is interacting, being also dependent on the presence of antioxidant compounds in the surrounding medium. It is as though a fungal species hierarchy could be established for every interaction under each set of conditions (i.e. habitat). The results obtained suggest that fungi display more complex behaviours than generally acknowledged. They are able to recognize potential contestants and built up defence reactions, as well as weaken plant defences and structures to induce infection. [ABSTRACT FROM AUTHOR]

Neofusicoccum parvum and Rhizoctonia solani are fungal pathogens with an increasing incidence in young grapevine plants. In this study, the antagonistic potential of some strains of the genus Trichoderma isolated from grapevine against these pathogens was investigated at the laboratory and greenhouse levels. In-plate confrontation assays showed that the selected Trichoderma strains could inhibit the mycelial growth of both taxa, being more effective against N. parvum. In the in vivo assays, the biocontrol activity of the mentioned strains against the pathogens, when applied either simultaneously or successively, was tested on both grafted plants and seedlings germinated from seed. The effectiveness of the treatments was evaluated by comparing biomass weight and vascular rot lengths data. In seedling trials, successive treatments resulted in higher root development and a lower colonization rate of the pathogens, especially against R. solani. In grafted plants, some disparity was observed against N. parvum: simultaneous treatments resulted in higher aerial biomass, but successive treatments resulted in higher root biomass and lower necrosis. Against R. solani, simultaneous treatments were clearly more effective, with higher root and aerial length values and lower necrosis. The obtained data suggest that the use of Trichoderma spp. isolates can constitute an alternative to conventional fungicides to control certain grapevine wood diseases. [ABSTRACT FROM AUTHOR]

Silverleaf caused by the basidiomycete Chondrostereum purpureum affects numerous woody species, including fruit tree crops like apple, resulting in wood necrosis and foliar silvering. There are no curative alternatives for this disease, and its management is by prevention methods. Therefore, the aim of this study was to develop a rapid diagnostic tool for the detection and identification of C. purpureum directly from woody tissues to help distinguish the pathogen from other basidiomycetes that are commonly found on apple. The silverleaf pathogen was isolated from different hosts and locations, and Koch's postulates were performed by inoculating the isolates on apple cuttings and measuring internal necrosis. A previously described APN 1 pair of primers specificity was also tested against 25 C. purpureum isolates in this study, using other wood rotting species as negative controls. Seven virulent isolates were inoculated on apple cuttings, and DNA was extracted from the cuttings' sawdust and amplified using APN 1, after 22 days of incubation. To prove the efficiency of the method in the field, DNA from healthy nursery plants inoculated with two virulent isolates, and naturally infected plants showing different levels of foliar symptoms, were tested. Presence of the fungus was verified by reisolation on APDA in all assays. Koch's postulates indicated that all C. purpureum isolates were pathogenic, showing different virulence levels, and APN 1 primers were able to discriminate them from other basidiomycetes. The method was also able to detect C. purpureum from artificially inoculated plants as well as naturally infected ones, demonstrating that the protocol may become a rapid minimally destructive diagnostic tool to detect the pathogen without the need to isolate it from tissues, and thus taking measures to prevent its dissemination.