Variation in xylem vulnerability to cavitation shapes the photosynthetic legacy of drought.

Increased drought conditions impact tree health, negatively disrupting plant water transport which, in turn, affects plant growth and survival. Persistent drought legacy effects have been documented in many diverse ecosystems, yet we still lack a mechanistic understanding of the physiological processes limiting tree recovery after drought. Tackling this question, we exposed saplings of a common Australian evergreen tree (Eucalyptus viminalis) to a cycle of drought and rewatering, seeking evidence for a link between the spread of xylem cavitation within the crown and the degree of photosynthetic recovery postdrought. Individual leaves experiencing >35% vein cavitation quickly died but this did not translate to a rapid overall canopy damage. Rather, whole canopies showed a gradual decline in mean postdrought gas exchange rates as water stress increased. This gradual loss of canopy function postdrought was due to a significant variation in cavitation vulnerability of leaves within canopies leading to diversity in the capacity of leaves within a single crown to recover function after drought. These results from the evergreen E. viminalis emphasise the importance of within-crown variation in xylem vulnerability as a central character regulating the dynamics of canopy death and the severity of drought legacy through time.

Diaporthe phaseolorum causing dieback disease on Melia dubia cav. in Karnataka state (India).

Melia dubia is an important tree species grown worldwide for its medicinal and timber values. It is widely used in timber and pulp industry and also as an organic pesticide, fertilisers, agro-forestry and herbal formulations. During 2019-2022, a dieback disease in plantations of M. dubia was recorded in Mysore, Mandya, Chamarajanagar, Hassan and Tumkur districts of Karnataka state (India) with disease incidence of 26.25%. The associated pathogen was isolated on PDA medium and its morpho-cultural characteristics were studied. The genomic DNA of the pathogen was isolated, and rDNA was amplified and sequenced using universal primers. Based on the microscopic, morpho-cultural, sequence data and phylogenetic analysis, the pathogen was identified as Diaporthe phaseolorum (Cooke & Ellis) Sacc. Koch's postulates were performed both in vitro and in vivo and the typical symptoms of dieback disease were recorded on post-inoculated saplings. The dieback disease is responsible for the poor growth of Melia species in the region, and hence, there is an urgent need to manage the disease in plantations using integrated management practices. This is the first report of the occurrence of D. phaseolorum on M. dubia plantations in India.

Grapevines under drought do not express esca leaf symptoms.

In the context of climate change, plant mortality is increasing worldwide in both natural and agroecosystems. However, our understanding of the underlying causes is limited by the complex interactions between abiotic and biotic factors and the technical challenges that limit investigations of these interactions. Here, we studied the interaction between two main drivers of mortality, drought and vascular disease (esca), in one of the world's most economically valuable fruit crops, grapevine. We found that drought totally inhibited esca leaf symptom expression. We disentangled the plant physiological response to the two stresses by quantifying whole-plant water relations (i.e., water potential and stomatal conductance) and carbon balance (i.e., CO2 assimilation, chlorophyll, and nonstructural carbohydrates). Our results highlight the distinct physiology behind these two stress responses, indicating that esca (and subsequent stomatal conductance decline) does not result from decreases in water potential and generates different gas exchange and nonstructural carbohydrate seasonal dynamics compared to drought.

Determining the Timing of Spore Release by Botryosphaeriaceae Species in Oregon Vineyards.

Botryosphaeria dieback, caused by a group of pathogens in the Botryosphaeriaceae family, is one of the most common grapevine trunk disease complexes (GTDs) found in Oregon vineyards. To understand the period of spores released by Botryosphaeria spp. in Oregon vineyards, four Burkard 7-day recording volumetric spore traps were placed in vineyard blocks in northern and southern Oregon. Each trap was placed near a younger (<10 years) and older (>30 years) block in both regions. Spore traps were deployed at the beginning of December 2019 and continued until March 2021. Between these timeframes, 475 and 477 days of samples were collected from each spore trap in northern and southern Oregon, respectively. DNA extraction was performed from individual day samples and followed by qPCR analysis of Botryosphaeria spores trapped in each tape sample. Weather data such as temperature, precipitation, relative humidity (RH), and wind speed were collected from nearby weather stations. Association between these data and number of spores detected were analyzed using Pearson correlation analysis. In northern Oregon, the detection occurred between December and February, and the first spore detection occurred when cumulative growing degree day (GDD) totaled to 4,357 and 4,351 units (TBase = 0°C, biofix date = January 1) during the first and second seasons, respectively. Similarly, in southern Oregon, the detection occurred between November and January, and the first spore detection occurred when cumulative GDD was 4,405 units during the second season. Hours of continuous RH >86% was significantly associated with number of spores released (P = 0.026; r = 0.42). During the spore detected dates, the RH was >86% for at least 19 consecutive hours. These findings provide an important implication to manage Botryosphaeria dieback by protecting pruning wounds during the most-spore-released periods. Furthermore, the knowledge of weather variables and their possible association with spore detection provides important information towards developing predictive models in future studies.

A glimmer of hope - ash genotypes with increased resistance to ash dieback pathogen show cross-resistance to emerald ash borer.

Plants rely on cross-resistance traits to defend against multiple, phylogenetically distinct enemies. These traits are often the result of long co-evolutionary histories. Biological invasions can force naïve plants to cope with novel, coincident pests, and pathogens. For example, European ash (Fraxinus excelsior) is substantially threatened by the emerald ash borer (EAB), Agrilus planipennis, a wood-boring beetle, and the ash dieback (ADB) pathogen, Hymenoscyphus fraxineus. Yet, plant cross-resistance traits against novel enemies are poorly explored and it is unknown whether naïve ash trees can defend against novel enemy complexes via cross-resistance mechanisms. To gain mechanistic insights, we quantified EAB performance on grafted replicates of ash genotypes varying in ADB resistance and characterized ash phloem chemistry with targeted and untargeted metabolomics. Emerald ash borer performed better on ADB-susceptible than on ADB-resistant genotypes. Moreover, changes in EAB performance aligned with differences in phloem chemical profiles between ADB-susceptible and ADB-resistant genotypes. We show that intraspecific variation in phloem chemistry in European ash can confer increased cross-resistance to invasive antagonists from different taxonomic kingdoms. Our study suggests that promotion of ADB-resistant ash genotypes may simultaneously help to control the ADB disease and reduce EAB-caused ash losses, which may be critical for the long-term stability of this keystone tree species.

Ferulic acid is a putative surrender signal to stimulate programmed cell death in grapevines after infection with Neofusicoccum parvum.

An apoplectic breakdown from grapevine trunk diseases (GTDs) has become a serious challenge to viticulture as a consequence of drought stress. We hypothesize that fungal aggressiveness is controlled by a chemical communication between the host and colonizing fungus. We introduce the new concept of a 'plant surrender signal' accumulating in host plants under stress and facilitating the aggressive behaviour of the strain Neofusicoccum parvum (Bt-67) causing Botryosphaeriaceae-related dieback in grapevines. Using a cell-based experimental system (Vitis cells) and bioactivity-guided fractionation, we identify trans-ferulic acid, a monolignol precursor, as a 'surrender signal'. We show that this signal specifically activates the secretion of the fungal phytotoxin fusicoccin A aglycone. We show further that this phytotoxin, mediated by 14-3-3 proteins, activates programmed cell death in Vitis cells. We arrive at a model showing a chemical communication facilitating fusicoccin A secretion that drives necrotrophic behaviour during Botryosphaeriaceae-Vitis interaction through trans-ferulic acid. We thus hypothesize that channelling the phenylpropanoid pathway from this lignin precursor to the trans-resveratrol phytoalexin could be a target for future therapy.

Lasiodiplodia regiae sp. nov.: A New Species Causing Canker and Dieback of Fruit Trees in China.

Canker and dieback are serious fungal diseases of woody plants that can cause huge economic losses to orchards. The purpose of this study was to classify and assess the pathogenicity of fungal species associated with canker and dieback on fruit trees growing in Henan Province, China. In total, 150 isolates of Botryosphaeriaceae were obtained from six different fruit trees exhibiting typical symptoms of stem canker, branch dieback, and gummosis. Morphological examinations and phylogenetic analysis of ITS, tef1, tub2, and rpb2 revealed two Botryosphaeriaceae species, which are Botryosphaeria dothidea and a novel species, Lasiodiplodia regiae, respectively. Using Koch's postulates, we confirmed that the different isolates of L. regiae can cause disease in their original hosts. The pathogenicity tests showed that L. regiae can cause canker, dieback, and gummosis symptoms in four different hosts, indicating a relatively wider host range. Moreover, 10 L. regiae isolates exhibited similar symptoms but different levels of virulence on shoots of peach trees under field conditions. This study demonstrated that L. regiae was a new causal agent of canker and dieback of six fruit tree species, which could be a serious risk to the orchard industry in China. Furthermore, the findings provide a foundation for further epidemiological studies and the development of management strategies.

Prevalence, Identity, Pathogenicity, and Infection Dynamics of Botryosphaeriaceae Causing Avocado Branch Canker in California.

Botryosphaeria branch canker and dieback of avocado (Persea americana Mill.) has expanded in avocado-growing areas in recent years. Twenty-one avocado groves in the major producing regions of California were surveyed in 2018 and 2019. Monthly inoculations of wounded, green, and lignified branches of 'Hass' and 'Lamb Hass' were conducted. Botryosphaeriaceae were the predominant fungi recovered from cankered tissues collected across the surveyed traditional and high-density orchards and caused symptoms on all six sampled cultivars. These fungi were also recovered in asymptomatic twigs and other organs and thus exist as a potential reservoir for future infections. Molecular analyses of 173 isolates showed that Neofusicoccum luteum had the greatest incidences across sites and cultivars, with 83 and 29% recovered from Hass and Lamb Hass, respectively. Pathogenicity tests on excised (Hass, GEM, and Hass mutants) and attached shoots from potted (Hass) and mature avocado trees (Hass and Lamb Hass) showed that all species were pathogenic on wounded, green, and mature branches of the specified cultivars. Monthly inoculations of wounded, green, and lignified branches of Hass and Lamb Hass showed that both stem types were susceptible throughout the inoculation periods, regardless of the avocado phenological stage. In temperature-dependent growth and infection studies, growth of three points could vary during the growing season. Botryosphaeriaceae grown was higher between 20 and 30°C, but only Lasiodiplodia theobromae significantly grew and caused external lesions at 35°C. Lasiodiplodia theobromae also grew more on perseitol-amended media, all indicating its adaptation to warmer temperatures and capacity in metabolizing the avocado-produced sugar. Overall, this study extended our knowledge of the prevalence, identity, and pathogenicity of Botryosphaeriaceae on avocado cultivars, which will be useful to tailor management strategies.

Antifungal and Insecticidal Activities of Selected Plant Species from Cloud Forest of Veracruz, Mexico: A Contribution to the Search of Novel Control Agents against Ambrosia Pest Complexes.

The antifungal and insecticidal activities of 34 extracts from 27 plant species were evaluated against fungal phytopathogens of the genus Fusarium and Xyleborus Scolytine ambrosia beetles involved in Fusarium dieback (FD) and laurel wilt (LW) diseases. Sixteen extracts caused mycelial growth inhibition (MGI) above 23 % at 2 mg mL-1 against F. solani, those from S. nudum and M. argyrophylla exhibited the highest MGI (57 % and 49 %, respectively). Thirteen extracts displayed significant antifungal activity against F. kuroshium, those from C. nocturnum and M. argyrophylla exhibited the highest MGI (100 % and 54.9 %, respectively). Additionally, ten plants extracts caused mortality in at least one of the beetle species tested, mainly from Solanaceae species. In the most active species, 39 phenolics were identified that may have contributed to their biological effects. This study is one of the first to report the potential of plant-derived natural products against the causative agents of FD and LW.

A Systematic Survey on Prevalence of Grapevine Trunk Disease Pathogens in Oregon Vineyards.

Grapevine trunk diseases (GTDs) are found in vineyards worldwide and can be caused by different fungal pathogens. To characterize types of GTDs in Oregon vineyards, and how the GTD pathogens' prevalence is affected by two geographical regions, a survey was conducted in which grapevine trunk samples were collected from 15 and 14 wine grape (Vitis vinifera) vineyards in southern and northern Oregon, respectively. Fungal species were identified through culture and PCR-based methods. GTD pathogens that were identified included Botryosphaeriaceae spp. and Phaeoacremonium spp. from 72 and 21% of the surveyed vineyards, respectively; Phaeomoniella chlamydospora, Cryptovalsa ampelina, Truncatella angustata, Seimatosporium lichenicola, Hormonema viticola from 7% of the surveyed vineyards; and Dactylonectria macrodidyma, and Pestaloptiopsis sp. from 3% of the surveyed vineyards. Pathogens were identified in both regions and in young and mature vineyards. The presence of GTD from the Botryosphaeria dieback complex was significantly affected by regions (P = 0.021), with pathogens being significantly more abundant in Willamette Valley (northern region) compared with Rogue Valley (southern region) vineyards. Some differences among other tested variables such as vineyard age, cultivars, rootstocks, and pruning methods were observed for all disease complexes; however, the differences were not statistically significant. Our study summarizes that Botryosphaeria dieback and Esca disease complexes are the most prevalent diseases infecting grapevines in Oregon vineyards and management practices need to be geared toward these economically important diseases. In addition, pathogens from other disease complexes are also present, suggesting a need for regular disease monitoring and following practices to limit the spread of these pathogens.

First Report of Cytospora azerbaijanica Causing Cytospora Canker and Shoot Dieback on Peach (Prunus persica) in California, U.S.A.

Peaches (Prunus persica L.) are an important crop in the United States with California leading the nation in peach production, with approximately 505,000 tons valued at $378.3 million (USDA National Agricultural Statistics Service, 2021, https://www.nass.usda.gov/). From April to July 2022, symptoms of branch and scaffold canker as well as shoot dieback were observed in three peach (cvs. Loadel, Late Ross and Starn) orchards located in San Joaquin County, California. Samples were collected from about 12 trees for each cultivar. Fast-growing, white, flat colonies were consistently isolated from active cankers on acidified potato dextrose agar (APDA) following the method described by (Lawrence et al. 2017). Pure fungal cultures were obtained by transferring single hyphal tips onto new APDA Petri plates. A total of 22 isolates were obtained. Each fungal isolate was recovered from a single diseased branch (40 to 55% recovery). All isolates in this study shared similar morphological characteristics. Fungal colonies were fast-growing with relatively even but slightly dentate margin, flat with white to off-white mycelium that turned vinaceous buff to pale greyish sepia (Rayner 1970) with age. Black, globose, ostiolated pycnidia, 0.8-(1.3)-2.2 mm diameter, with brownish surface hyphae formed on peach wood embedded in PDA after approximately three weeks and exudated buff-colored mucilage. Pycnidia were both solitary and aggregated and had multiple internal locules sharing invaginated walls. Conidiogenous cells were hyaline, smooth-walled, septate, tapering towards the apex, 13-(18.2)-25.1 × 0.8-(1.3)-1.9 µm (n = 40). Conidia were hyaline, allantoid, smooth, aseptate, 5.5-(6.3)-7.1 × 1.4-(1.9)-2.3 µm (n = 40). Genomic DNA was extracted and sequences of the internal transcribed spacer region (ITS) using ITS5/ITS4 universal primers, translation elongation factor 1α gene (TEF) using primers EF1-728F/EF1-986R, second largest subunit of RNA polymerase II (RPB2) using primers RPB2-5F2/fRPB2-7cR, and actin gene region (ACT) using primers ACT-512F/ACT-783R were obtained and compared with sequences available in GenBank (Lawrence et al. 2018; Hanifeh et al. 2022). Isolates were identified as Cytospora azerbaijanica following DNA sequencing and morphological identification. Consensus sequences of the four genes of two representative isolates (SJC-66 and SJC-69) were deposited into GenBank database (ITS: OQ060581 and OQ060582; ACT: OQ082292, OQ082295; TEF: OQ082290 and OQ082293; RPB2: OQ082291 and OQ082294). The Basic Local Alignment Search Tool (BLAST) indicated that the sequenced RPB2 genes of isolates (SJC-66 and SJC-69) were at least 99% identical to that of Cytospora sp. strain shd47 (Accession: MW824360) covering at least 85% of the sequences. The actin genes from our isolates were at least 97.85% identical to that of Cytospora sp. strain shd47 (Accession: MZ014513), covering 100% of the sequences. The translation elongation factor gene from isolates (SJC-66 and SJC-69) was at least 96.4% identical to that of Cytospora sp. strain shd166 (Accession: OM372512), covering 100% of the query. Those top hit strains belong to C. azerbaijanica, recently reported by Hanifeh et al. (2022). Pathogenicity tests were performed by inoculating eight wounded, 2- to 3-year-old healthy branches on each of eight 7-year-old peach trees, cvs. Loadel, Late Ross and Starn, using 5-mm-diameter mycelium plugs collected from the margin of an actively growing fungal colony on APDA. Controls were mock-inoculated with sterile agar plugs. Inoculation sites were covered with petroleum jelly and wrapped with Parafilm to keep moisture. The experiment was performed twice. After four months, inoculation tests resulted in vascular discoloration (canker) above and below the inoculation sites (average necrosis length of 114.1 mm). Cytospora azerbaijanica was re-isolated from all infected branches (70 to 100% recovery) completing Koch's postulates. Controls remained symptomless and no fungi were isolated from the slightly discolored tissue. Cytospora species are destructive canker and dieback pathogens of numerous woody hosts worldwide. Recently, C. azerbaijanica was reported in causing canker disease of apple trees in Iran (Hanifeh et al. 2022). To our knowledge, this is the first report of C. azerbaijanica causing canker and shoot dieback of peach trees in the United States and worldwide. These findings will aid towards a better understanding of genetic diversity and host range of C. azerbaijanica.

First report of canker caused by Cytospora sorbicola on Sweet Cherry (Prunus avium) in Chile.

Chile leads cherry exports in the southern hemisphere with a total of 415.315 t exported in the 2022 to 2023 season (IQonsulting, 2023). Cytospora canker, produced by Cytospora spp., causes destructive infections and limit the productivity of sweet cherry orchards (Luo et al. 2019). This study was focused on isolating Cytospora strains to identify and characterize the species present in sweet cherry. During the period 2019-2022, ten samples of stem or branch presenting canker, dieback, gummosis or dead buds, were collected from sweet cherry cultivars 'Skeena', 'Lapins', 'Santina', 'Sweetheart', and 'Regina', in the regions Ñuble and O'Higgins, Chile. Five mm pieces from the necrotic wood margins of the samples were rinsed with sterile deionized water, placed on potato dextrose agar (PDA, Difco) and incubated at 20±2 ºC for 5 days. One isolate was recovered from each sample, resulting in ten Cytospora-like strains. Single hyphal tips were transferred onto PDA plates and all isolates were deposited in the Chilean Collection of Microbial Genetic Resources (CChRGM). Colonies grown on PDA reached 89 mm in diameter in 10 d at 25 °C, showing irregular margin, lacking aerial mycelium, initially off-white to cream that turned greenish gray in the center, which darkens with age. After 20 days of culturing on pine needle agar (Chen et al. 2015), isolates produced conidiomata pycnidial, semi-immersed, black, and subglobose (362)445-555(681)×(357)528-700(1053) µm (n=10), generating amber slimy conidia masses; Conidiophores were phialidic, cylindrical, aseptate, hyaline (6.77)9-10.04(12.88)×(0.82)1.1-1.28(1.99) µm (n = 30); conidia were abundant, allantoid, hyaline to light brown, aseptate (3.39)4.28-4.57(5.36)×(0.69)0.96-1.09(1.47) µm (n = 30) (Supplementary Figure 1). No sexual morph was observed. With the exception of the strain RGM 3390, all the isolates shared morphological characters to the descriptions of Cytospora sorbicola Norphanph., Bulgakov, T. C. Wen & K. D. Hyde (Norphanphoun et al. 2017). Isolates were identified at species level, by sequencing DNA regions described by Pan et al. (2020): ITS1-5.8S-ITS2, LSU; act, tef-1α, and tub2 with the exception of the RBP2, because this region could not be amplified in seven out of ten isolates. The consensus tree included the concatenated sequences of the ten isolates and those of reference Cytospora species reported by Ilyukhin et al. (2023) using a maximum likelihood analysis with the tool IQ-TREE webserver. MLSA confirmed the taxonomic affiliation of nine of the isolates with C. sorbicola and one isolate with Cytospora sp. (RGM 3390), that might represent a novel species (Supplementary Figure 2). The isolates RGM 3399 and RGM 3400, were selected randomly for pathogenicity tests. Inoculations were performed on 2-year-old sweet cherry cv. 'Lapins' grow in pots in a greenhouse at 26±6°C. Seven plants per isolate were cut to about 6-cm length from the main stem, and inoculated onto fresh cuts with 5-mm mycelium PDA plugs of 5-d-old culture and wrapped in moist sterile cotton and parafilm to keep moisture. Six plants were inoculated with non-colonized PDA agar plugs as control. The average canker length 3 months after inoculation was 3.1 and 0.8 cm, for RGM 3389 and RGM 3400, respectively (Supplementary Figure 1). Symptomatic twigs were incubated in moist chambers at 20±2 ºC for 10 d, resulting in the re-isolation of Cytospora strains that produced pycnidia and conidia structures in agreement with C. sorbicola. Both strains were reidentified to fulfill Koch's postulates, control twigs remained asymptomatic and no fungus was isolated from these twigs. This is the first report of C. sorbicola causing canker on sweet cherry in Chile. Our findings suggest that this species could be the most recurrent in cherry in central Chile, coinciding with it found in California where C. sorbicola has been described as the main causal agent of Cytospora canker of stone fruits in California (Lawrence et al. 2018).

First report of branch dieback caused by Neoscytalidium dimidiatum on Styphnolobium japonicum (L.) Schott in China.

Styphnolobium japonicum (L.) Schott (family Fabaceae Juss.) also called pagoda tree, is widely planted in northern China in landscape plantings, for erosion control and forestry. In recent years, symptoms of branch dieback were observed on S. japonicum in the southern part of Xinjiang province, China. From 2019 to 2022, in total ca. 1000 ha area was surveyed in Korla (41.68°N, 86.06°E), Bohu (41.95°N, 86.53°E) and Alaer (41.15°N, 80.29°E). Typical symptoms were observed in 70% of the surveyed branches. To identify the cause, we collected 50 symptomatic branches. Symptoms were initially observed on green current-year twigs, which turned grayish white in color. In the later stages of disease development, a large number of nacked black conidia formed under epidermis of perennial branches, causing visible black protrusions (pycnidia) on branch surface. The disease occurred throughout the entire growing season of S. japonicum. Symptoms also occurred on the inflorescence, fruit, and twigs. In some cases, infection resulted in tree mortality. Isolations were made from the margin between healthy and diseased tissues. Small pieces were excised, surface disinfested (75% ethanol 30 s, 1% NaClO solution 5 mins), cut into pieces (5 to 10 mm2), and incubated on PDA medium at 28℃ for 3 days. A total of 16 isolates (GH01-GH16) with similar colony morphology were obtained. The colonies were initially white, gradually turning to olive-green on the surface and black on the underside after 7 days. Microscopically, the conidia were aseptate, 1-septate, two-septate, and muriform, 2.6-4.5 × 2.9-27.6 µm (n=50). Pycnidia ranged in size from 120.2 to 135.5 × 112.4 to 118.6 µm (n=20). Those morphological characters matched the descriptions of Neoscytalidium dimidiatum (previously N. novaehollandiae) (Alizadeh et al. 2022; Pavlic et al. 2008). For molecular identification, genomic DNA of GH01-GH16 were extracted from fresh mycelia. The internal transcribed spacer (ITS), large subunit ribosomal RNA gene (LSU), and translation elongation factor 1-alpha (EF1-α) gene were amplified using the primer sets ITS1/ITS4 (White 1990), LRoR/LR5 (Vilgalys and Hester 1990) and EF1-728F/EF1-986R (Carbone and Kohn 1999). The sequences were deposited in GenBank (accession No. OP379832, OQ096643-OQ096657 for ITS, OP389048, OQ127403-OQ127417 for LSU, and OQ136617, OQ586044-OQ586058 for EF1-α). The ITS sequence had 100% identity (505/505 bp) to MT362600. Similarly, the LSU and EF1-α sequences were found to be identical to MW883823 (100%, 821/821 bp) and KX464763(99%, 256/258 bp), respectively. Pathogenicity was tested on one-year-old healthy S. japonicum seedlings. Spores of representative isolate GH01 were produced on PDA by incubating for 7-days at 28℃. Conidia were washed with sterile water. Five trees were inoculated with 1 × 106 conidia/ml conidial suspensions and five trees were sprayed with sterile water. All trees were covered with plastic bags for 24 h and kept at 25°C in a greenhouse. Signs and symptoms were similar to those observed in field collections one month after inoculation, while no symptoms occurred on the controls. The original fungus was successfully reisolated from the inoculated trees and was identified as N. dimidiatum following the methods described above. N. dimidiatum has been reported in many Asian country such as Malaysia, India, Turkey, and Iran(Akgül et al. 2019; Alizadeh et al. 2022; Khoo et al. 2023; Salunkhe et al. 2023). To our knowledge, this is the first report of N. dimidiatum associated with branch dieback of S. japonicum in China. Our findings have expanded the host range of N. dimidiatum in China and provides a theoretical basis for the diagnosis and treatment of the disease.

First Report of Neofusicoccum australe Causing Branch Dieback of English Walnut cv. Chandler in Central Chile.

The English walnut (Juglans regia L.) is the second most important fruit crop of importance in Chile, with 43,700 hectares mainly in the Central Valley (www.odepa.cl, 2022). For several seasons symptoms of a branch dieback have been observed in walnut orchards with 3 to 50% of trees incidence levels. During the 2020 winter season (July) a total of 150 symptomatic spurs of 15 trees were sampled from an 8-year-old walnut cv. Chandler orchard located in Buin (33°42' S, 70° 42' W). The collected spurs showed external and internal brown necroses, starting from the tip with well-defined margins. The symptomatic tissue was cut in to small pieces (5 x 4 x 2 mm), surface disinfected by dipping in a 10% solution made from a commercial bleach solution (4,9% NaOHCl), rinsed twice in sterile water and plated on APDA (PDA Difco laboratories acidified with lactic acid (2,5 ml of 25% (vol/vol) per liter of medium). After five days at 20 °C in darkness, fast-growing, white-grey turning to black colonies were obtained, tentatively classified as a member of the Botryosphaeraceae family and two single-spore isolates (SS1, SS2) were selected for identification. Colony mycelia were first white and turned to light grey, dark grey or black, with tufts of mouse gray aerial mycelia. The pycnidia and conidia production was induced by inoculating autoclaved pine needles placed on APDA an incubation for 25 to 30 days at 20 °C in darkness. Black pycnidia solitary and globose were obtained producing hyaline, aseptate, fusiform to obovoid conidia with truncated ends with dimensions of (22.6-) 19.1 ± 1.4 (-13.3) x (6.7-) 5.5 ± 0.5 (-3.7) µm and 3.5 length/width ratio (n=100). Both isolates were identified using dichotomous keys confirming the description of Crous et al, 2006 as Neofusicoccum australe. The identification was molecularly confirmed by amplifying the nuclear ribosomal gene 5,8S (ITS1-5.8S-ITS2) using the ITS1/ITS4 primers, a partial region of ß-tubulin gene (Bt2a/Bt2b), and the translation elongation factor 1-α gene (TEF1) with TEF1-728F/TEF1-986R primers. The BLASTn search revealed 100% of identity for ITS and TEF according to sequences of N. australe reference strains MT587467.1 and MK759852.1, respectively; and over 99% for ß-tubulin compared to N. australe strain KX464929.1. The DNA sequences were submitted to the GenBank (ITS, OP142414, OP142416; BT, OP209981, OP209978; and TEF OP209979, OP209980) for SS1 and SS2 isolates, respectively, and deposited in the fungal collection of CChRGM - INIA, Chillán, Chile (RGM 3409 and 3410). Pathogenicity of both isolates was tested in 8-year-old asymtomatic English walnut cv. Chandler in the field during 2020 spring season, by cutting transversally 15 twigs of different tress and inoculating with a 5 day-old PDA plug. An equal number of wounded twigs were inoculated with a sterile PDA plug and served as control. After six months, all inoculated twigs developed the same necrotic lesions observed in field of 2.0 to 10.1 cm (SS1) and 1.9 to 10.8 cm (SS2) in length while control twigs showed only a scar without any dieback tissues. The inoculated pathogens of N. australe were recovered from the diseased tissues, thus fulfilling Koch's postulates. A similar dieback of walnut was reported in Chile, which caused Diplodia mutila (Díaz et al, 2018), and N. parvum (Luna et al, 2022) while N. australe has been reported in other hosts (Auger et al, 2013, Besoain et al, 2013). To the best of our knowledge, this is the first report of N. australe associated with walnut branch dieback in Chile.

Traces of Hymenoscyphus fraxineus in Northeastern Europe Extend Further Back in History than Expected.

Herbaria are a promising but still poorly applied information source for retrospective microbiological studies. In order to find any evidence of the virulent European origin of ash dieback agent Hymenoscyphus fraxineus and other fungal pathogens, we analyzed 109 leaf samples from three different Estonian botanical herbaria, sampled during 171 years from 20 ash species and cultivars, using a PacBio third-generation sequencing of the fungal internal transcribed spacer ITS1-5.8S-ITS2 ribosomal DNA region. We identified a large amount of saprotrophic fungi naturally colonizing ash leaves. Hymenoscyphus fraxineus colonized a Fraxinus chinensis subsp. rhynchophylla specimen and a F. chinensis specimen collected from Tallinn Botanic Garden in July 1978 and July 1992, respectively. The samples originated from trees grown in this garden from seeds collected from Shamora, Far-East Russia, in 1961 and from a Beijing botanical garden in eastern China in 1985, respectively. Repeated subsequent DNA extraction, real-time quantitative PCR, and Sanger and Illumina sequencing confirmed our findings of these apparently oldest cases of the ash dieback agent in Europe. These results show that H. fraxineus evidently was present in Estonia 19 years earlier than our previous data from fungal herbaria documented and 14 years before the first visible damage of ash trees was registered in Poland. Because we found no evidence of the saprotrophic H. albidus from earlier mycological and botanical herbarium specimens, the presence of H. albidus in Estonia remains questionable.

Health Status of Ready-to-Plant Grapevine Nursery Material in Canada Regarding Young Vine Decline Fungi.

Young vine decline (YVD), caused by several taxonomically different fungi, results in the decline and death of grapevines within a few years after planting. Infection can occur in nursery mother blocks and/or at several stages in the nursery propagation process, but the final plant material may remain asymptomatic. Four nurseries that sell ready-to-plant grapevines in Canada were sampled to evaluate the health status with regard to YVD fungi, including Botryosphaeriaceae spp., Cadophora luteo-olivacea, Dactylonectria macrodidyma, Dactylonectria torresensis, Phaeoacremonium minimum, and Phaeomoniella chlamydospora. Plants representing three cultivars, 'Chardonnay', 'Merlot', and 'Pinot noir', either grafted onto '3309C' rootstock or self-rooted, were provided by the nurseries. Samples from the roots, base of the rootstock or self-rooted cultivar, graft-union, and scion were collected from each plant. DNA was extracted, and the total abundance of each fungus was quantified using Droplet Digital PCR. Results revealed that 99% of plants harbored at least one of the fungi studied, with a mean of three different fungal species that were present per grapevine. Droplet Digital PCR results showed that the abundance of the different fungi significantly varied between different sections of each plant, individual plants for each cultivar, and cultivars from the same nursery. Necrosis measurements were recorded from the base of the rootstock or self-rooted cultivars and did not correlate with fungal abundance recorded in that section for each grapevine, but necrosis was consistent across cultivars within nurseries. Five different rootstocks were compared from one nursery, and results showed no differences between rootstocks and their health status. Among all nurseries, C. luteo-olivacea was the most prevalent fungus (97% of the plants), while D. macrodidyma was the least commonly found (13% of the plants). This study shows that ready-to-plant nursery material sold in Canada is likely to be infected with several YVD fungi and that presence and abundance of fungi vary significantly among individual grapevines and nurseries.

Spatially explicit modeling of disease surveillance in mixed oak-hardwood forests based on machine-learning algorithms.

Incidences of disease, dieback, decline or mortality, some of which induced or enhanced by climate change, threaten the sustainability of forest stands in many ecosystems. Spatially explicit prediction of disease onset remains challenging, however, due to the involvement of several causative agents. In this paper, we developed a generic framework based on machine-learning algorithms and spatial analyses for landscape-level prediction of oak disease outbreaks caused by the charcoal fungus Biscogniauxia mediterranea in a mixed-oak forest of Mediterranean climate. For prediction, we used a set of fifteen causative factors as a cross-function of soil, site and stand-related predictors. A total of 80 sample plots, including 1134 affected trees, were surveyed and used for the modeling process at the 5600-ha landscape level of the southern Zagros, Iran, where the disease occurs in roughly 25% of forest lands. Ten machine learning algorithms were explored and the performance of each algorithm to predict oak disease outbreak was evaluated. The modeling framework used maximum entropy to remove the least influential variables and build the status-quo management scenario to which the results of the prediction models were compared. Results showed that the random forests algorithm (AUC = 0.96: Precision = 0.71: Accuracy = 0.90: F-Measure = 0.70) achieved significantly better results than the status-quo management (Precision = 0.13: Accuracy = 0.67: F-Measure = 0.12) and any other algorithm. Soil chemical properties (NPK, organic carbon and EC) and landform predictors (slope, distance to roads, and TWI) were major forecasters of oak disease outbreak identified by the random forest algorithm. Geostatistical analysis enabled the creation of a map that identified sites at higher risk of infestation, allowing epidemiologists and forest managers to find sites likely to be infested. Consequently, financial resources can be allocated and management practices such as sanitation felling treatments applied across large forest landscapes to minimize the risk of spread and severity to uninfested high-value trees on nearby or adjacent land zones that are in the early stage of epidemics.

Tree dieback, woody plant diversity, and ecosystem driven by topography in semi-arid mountain forests: Implication for ecosystem management.

Mountain landscapes are highly heterogeneous due to topography, notably positions along slope and slope shapes, which control ecosystem mechanisms. We hypothesized that tree dieback is controlled by topography, selecting productive and less diverse communities in lower slopes, and stress-resistant and more diverse communities on upper slopes. Understanding how this heterogeneity drives vegetation patterns should provide benchmarks for ecosystem management of mountain forest dominated by Quercus brantii. Woody communities were sampled along convex vs concave topography (i.e., ridge vs talweg), and with measurements of tree dieback severity, environmental variables (litter depth, soil quality, rock outcrop), stand structure (canopy cover, mistletoe infestation, tree diameter and height, diameter and height differentiations, oaks' number from sprout-clumps or seed-origin), and biodiversity. Slope position was the most significant driver that affected all variables, excepted evenness. Dieback severity was higher on slope shoulders and summits, and lower in lower slopes where trees were the most productive: taller, larger, more homogeneous, and mostly seed-origin. Catena shape affected the diversity and dieback severity, both higher in talwegs, but had no effect on environmental variables and little on stand structure. Outputs indicate that the higher diversity of woody plants is on upper slopes supporting stress-resistant community associated with more severe dieback and mistletoe infection probably because frugivore birds attracted by the shrubs' fruits. Semi-arid forest management must consider the shaped-slope ecosystem heterogeneity by preserving ridges that are more susceptible to tree dieback, and naturally support biodiversity. Restoration measures on lower fertile slopes could be carried out by oak planting or seedlings under the cover of shrubs to counter dieback effects and environmental stresses. In addition, forestry measures can be taken in lower positions for the conversion of coppice to high oak forest to potentially consider a moderate forestry.

Canopy damage during a natural drought depends on species identity, physiology and stand composition.

Vulnerability to xylem cavitation is a strong predictor of drought-induced damage in forest communities. However, biotic features of the community itself can influence water availability at the individual tree-level, thereby modifying patterns of drought damage. Using an experimental forest in Tasmania, Australia, we determined the vulnerability to cavitation (leaf P50 ) of four tree species and assessed the drought-induced canopy damage of 2944 6-yr-old trees after an extreme natural drought episode. We examined how individual damage was related to their size and the density and species identity of neighbouring trees. The two co-occurring dominant tree species, Eucalyptus delegatensis and Eucalyptus regnans, were the most vulnerable to drought-induced xylem cavitation and both species suffered significantly greater damage than neighbouring, subdominant species Pomaderris apetala and Acacia dealbata. While the two eucalypts had similar leaf P50 values, E. delegatensis suffered significantly greater damage, which was strongly related to the density of neighbouring P. apetala. Damage in E. regnans was less impacted by neighbouring plants and smaller trees of both eucalypts sustained significantly more damage than larger trees. Our findings demonstrate that natural drought damage is influenced by individual plant physiology as well as the composition, physiology and density of the surrounding stand.

Lack of hydraulic recovery as a cause of post-drought foliage reduction and canopy decline in European beech.

European beech (Fagus sylvatica) was among the most affected tree species during the severe 2018 European drought. It not only suffered from instant physiological stress but also showed severe symptoms of defoliation and canopy decline in the following year. To explore the underlying mechanisms, we used the Swiss-Canopy-Crane II site and studied in branches of healthy and symptomatic trees the repair of hydraulic function and concentration of carbohydrates during the 2018 drought and in 2019. We found loss of hydraulic conductance in 2018, which did not recover in 2019 in trees that developed defoliation symptoms in the year after drought. Reduced branch foliation in symptomatic trees was associated with a gradual decline in wood starch concentration throughout summer 2019. Visualization of water transport in healthy and symptomatic branches in the year after the drought confirmed the close relationship between xylem functionality and supported branch leaf area. Our findings showed that embolized xylem does not regain function in the season following a drought and that sustained branch hydraulic dysfunction is counterbalanced by the reduction in supported leaf area. It suggests acclimation of leaf development after drought to mitigate disturbances in canopy hydraulic function.