Search

Field surveys conducted during 2021 and 2022 in Western Sicily, Italy, revealed the presence of common fig trees severely affected by trunk and crown root canker and bark cracking. Moreover, in conjunction with the symptomatic tissues, the same surveyed plants showed the presence of bark beetle holes and internal wood galleries. The predominant beetle Criphalus dilutus was previously reported attacking figs in Sicily. Phylogenetic analyses based on multilocus DNA data showed the presence of different fungal taxa associated with disease symptoms, including Botryosphaeria dothidea , Ceratocystis ficicola , Diaporthe foeniculina , Neocosmospora bostrycoides , N. perseae , and Neofusicoccum luteum . Pathogenicity tests conducted on potted fig plants showed that all the species were pathogenic to fig, with C. ficicola and Neocosmospora spp. as the most aggressive fungal species. Moreover, isolations conducted from the bodies of emerging adult insects recovered from disease samples confirmed the presence of C. ficicola and Neocosmospora spp., suggesting the potential involvement of C. dilutus in their dissemination., Competing Interests: The author(s) declare no conflict of interest.

Italian ryegrass is widely cultivated for the production of forage, hay, and silage because of its high nutritional value and good palatability. Leaf spots caused by fungi pose a serious threat to forage crops. In order to expand the knowledge of fungi causing leaf spots in ryegrass ( Lolium multiflorum ) in Sichuan, Yunnan, Chongqing, and Guizhou of southwestern China, a comprehensive survey was undertaken from 2015 to 2022. The survey discovered that Epicoccum leaf spot (ELS) was a common and widespread disease, more serious at the late stage of growth (after late May). Symptomatic leaf samples collected from the four different provinces were analyzed, and a total of 202 Epicoccum isolates were obtained. Based on both multilocus phylogeny (ITS, LSU, TUB2 , and RPB2 ) and morphology, 10 Epicoccum species were finally identified, including three novel species ( E. endololii sp. nov., E. lolii sp. nov., and E. loliicola sp. nov.), six new host records ( E. draconis , E. endophyticum , E. oryzae , E. plurivorum , E. thailandicum , and E. tobaicum ), and an unknown species ( Epicoccum sp.1). Pathogenicity tests showed that E. endophyticum , E. endololii , and Epicoccum sp.1 were nonpathogenic to Italian ryegrass, which were confirmed as endophytes in this study; the other six species could infect Italian ryegrass and cause leaf lesions to different degrees, of which E. draconis was more aggressive ( P ≤ 0.05). Coupled with the isolation rates and geographical distributions of these species, it was found that E. plurivorum was the predominant pathogen in Yunnan while E. oryzae and E. tobaicum were the predominant pathogens in the other three provinces. This work provides an initial understanding of the taxonomy, virulence, and distribution of Epicoccum species associated with ELS in southwestern China and lays a solid foundation for the diagnosis in the field and scientific control of ELS on Italian ryegrass., Competing Interests: The author(s) declare no conflict of interest.

Bananas ( Musa spp.) are among the world's most economically important staple food crops. The most important fungal leaf diseases of Musa spp. worldwide are caused by the Sigatoka disease complex, which comprises black Sigatoka ( Pseudocercospora fijiensis ), yellow Sigatoka ( P. musae ), and Eumusae leaf spot ( P. eumusae ). Considering the rapid spreading rate of black Sigatoka in Puerto Rico since its first observation in 2004, a disease survey was conducted from 2018 to 2020 to evaluate the Sigatoka disease complex on the island. Sixty-one leaf samples showing Sigatoka-like symptoms were collected throughout the island for diagnosis by molecular approaches and fungal isolation. Molecular analysis using species-specific primers for P. fijiensis , P. musae , and P. eumusae detected the presence of P. fijiensis in 50 leaf samples. Thirty-eight fungal isolates were collected and identified by morphology and genomic sequencing from various nuclear genes. The analysis identified 24 isolates as P. fijiensis , while the rest of the isolates belonged to the genus Cladosporium spp. and Cladosporium -like spp. ( n = 5) , Neocordana musae ( n = 2), Zasmidium spp. ( n = 6), and Z. musigenum ( n = 1). The high frequency of P. fijiensis found in leaf samples and collected isolates suggests that black Sigatoka has displaced the yellow Sigatoka ( P. musae ) in Puerto Rico. Accurate identification of fungal species causing foliar diseases in Musa spp. will allow the establishment of quarantine regulations and specific management approaches in Puerto Rico., Competing Interests: The author(s) declare no conflict of interest.

Hazelnut ( Corylus avellana ), a nut crop that is rapidly expanding worldwide, is endangered by a rot. Nut rot results in hazelnut defects. A survey was conducted in northwestern Italy during 2020 and 2021 to identify the causal agents of hazelnut rots. Typical symptoms of black rot, mold, and necrotic spots were observed on hazelnuts. The prevalent fungi isolated from symptomatic hazelnut kernels were Diaporthe spp. (38%), Botryosphaeria dothidea (26%), Diplodia seriata (14%), and other fungal genera with less frequent occurrences. Among 161 isolated Diaporthe spp., 40 were selected for further analysis. Based on morphological characterization and multilocus phylogenetic analysis of the ITS, tef-1α , and tub2 , seven Diaporthe species were identified as D. eres , D. foeniculina , D. novem , D . oncostoma , D. ravennica , D. rudis , and D. sojae . D. eres was the main species isolated from hazelnut rots, in particular from moldy nuts. The pathogenicity test performed on hazelnuts 'Tonda Gentile del Piemonte' using a mycelium plug showed that all the Diaporthe isolates were pathogenic on their original host. To our knowledge, this work is the first report of D. novem , D . oncostoma , and D. ravennica on hazelnuts worldwide. D. foeniculina , D. rudis , and D. sojae were reported for the first time as agents of hazelnut rot in Italy. Future studies should focus on the comprehension of epidemiology and climatic conditions favoring the development of Diaporthe spp. on hazelnut. Prevention and control measures should target D. eres , representing the main causal agents responsible for defects and nut rot of hazelnuts in Italy., Competing Interests: The author(s) declare no conflict of interest.

Fungi in the Botryosphaeriaceae family cause dieback, fruit rots, and stem cankers in many tropical fruit trees. To identify which species of Botryosphaeriaceae were present in tropical fruit in Puerto Rico and the symptoms they cause in rambutan and longan, a disease survey was conducted throughout the island from 2008 to 2016. Diseased organs of rambutan, longan, mango, and tangerine were collected and 39 isolates belonging to the Botryosphaeriaceae family were isolated and identified. Phylogenetic analysis of three nuclear genes identified nine species: six Lasiodiplodia spp. and three Neofusicoccum spp. All 39 isolates were inoculated on healthy 1-year-old rambutan and longan seedlings to confirm their pathogenicity. Dieback on both rambutan and longan was observed at 14 days after inoculation (DAI). Fourteen isolates from seven Botryosphaeriaceae species ( Lasiodiplodia brasiliensis, L. hormozganensis, L. iraniensis, L. pseudotheobromae, L. theobromae, Neofusicoccum batangarum , and N. parvum ) caused dieback in rambutan. Five of these pathogenic isolates were collected from rambutan, four from longan, two from mango, and three from tangerine. Ten isolates of four Lasiodiplodia spp. caused dieback in longan: L. hormozganensis , L. iraniensis, L. pseudotheobromae and L. theobromae,. Of these, three were collected from rambutan, three from longan, one from mango, and three from tangerine. Abundant development of pycnidia on branches, called corky bark, were observed on both rambutan and longan 60 DAI. Cross-inoculations showed that pathogenicity is wide in spectrum, indicating that different planting alternatives should be considered for better crop management.

California has been invaded by two distinct Euwallacea spp. that vector unique plant pathogenic symbiotic fungi on multiple hosts and cause Fusarium dieback. The objective of this study was to develop multiplex real-time quantitative PCR assays using hydrolysis probes targeting the β-tubulin gene to detect, distinguish, and quantify fungi associated with the polyphagous shot hole borer (PSHB; Euwallacea whitfordiodendrus , Fusarium euwallaceae , Graphium euwallaceae , and Paracremonium pembeum ) as well as the Kuroshio shot hole borer (KSHB; Euwallacea kuroshio , Fusarium kuroshium , and Graphium kuroshium ) from various sample types. Absolute quantification reaction efficiencies ranged from 88.2 to 104.3%, with a coefficient of determination >0.992 and a limit of detection of 100 copies µl -1 for all targets across both assays. Qualitative detection using the real-time assays on artificially inoculated avocado shoot extracts showed more sensitivity compared with conventional fungal isolation from wood. All symbiotic fungi, except P. pembeum , from PSHB and KSHB female heads were detectable and quantified. Field samples from symptomatic Platanus racemosa , Populus spp., and Salix spp. across 17 of 26 city parks were positively identified as PSHB and KSHB through detection of their symbiotic fungi, and both were found occurring together on five trees from three different park locations. The molecular assays presented here can be utilized to accurately identify fungi associated with these invasive pests in California.

Canker and wood rot pathogens cause dieback and, in severe cases, the death of young apple trees. Recently, a higher occurrence of cankers was observed on 1-year-old apple trees in the Western Cape Province of South Africa. This study aimed to assess the phytosanitary status of nursery trees and propagation material as possible inoculum sources for canker pathogens. Thirteen 1-year-old apple orchards showing canker or dieback symptoms were sampled. Certified nursery apple trees were collected from four nurseries as well as scion and rootstock mother plant material. Isolations were made from the discoloration observed in the vascular tissue of the plant parts and from asymptomatic material. Possible canker and wood rot species were identified with PCR and sequence comparisons of the relevant gene regions and phylogenetic analyses. Similar canker and wood rot species were isolated from 1-year-old diseased apple trees, nursery apple trees, and the propagation material. Forty-five fungal species associated with canker or wood rot symptoms were identified. The top five most abundant fungal species found causing disease on commercial 1-year-old trees were also found in high numbers causing latent infection in certified apple nursery trees. These species were Didymosphaeria rubi-ulmifolii sensu lato, Diplodia seriata , Schizophyllum commune , Didymella pomorum , and Coniochaeta fasciculata , with D. rubi-ulmifolii sensu lato being the dominant species in both sampling materials. In all, 65% of certified nursery apple trees, 5% of scion shoots used for budding, and 21% of rooted rootstock cuttings from layer blocks had latent infections of canker and wood rot pathogens. Pathogenicity trials were conducted with isolates of 39 species, inoculated onto 2-year-old branches of 14-year-old Golden Delicious trees. All species caused lesions that were significantly longer than the control. This study confirmed the presence of canker and wood rot pathogens in apple propagation material as well as certified nursery apple trees, which will aid the improvement of management practices in nurseries.

Gray blight disease, caused by Pestalotiopsis -like fungi, is one of the deadliest threats to tea ( Camellia sinensis ) production. However, little information is known about the traits and characteristics of this pathogen. Here, a systematic survey was performed, and a total of 20 representative isolates were obtained from the leaves of tea plants affected by gray blight in two main tea plantations located in Anhui Province, China. Further analyses showed that two isolates were identified as Neopestalotiopsis ellipsospora , three isolates were regarded as Pseudopestalotiopsis chinensis , one isolate was considered as Pseudopestalotiopsis camelliae-sinensis , and the remaining isolates belonged to Pseudopestalotiopsis spp., on the basis of morphological characteristics and multigene phylogenetic analyses of the internal transcribed spacer, β-tubulin, and translation elongation factor 1-α. Pathogenicity tests indicated that there were significant differences in virulence among the Neopestalotiopsis and Pseudopestalotiopsis isolates when inoculated on the leaves of the tea plant ( C. sinensis 'Shuchazao'). Furthermore, varied pathogenicity was also observed for the same isolate when inoculated on different varieties of tea plants. To our knowledge, this is the first record of Neopestalotiopsis ellipsospora and Pseudopestalotiopsis chinensis causing gray blight disease of tea plants in China.

Cranberry fruit rot (CFR) is arguably one of the most limiting factors of cranberry ( Vaccinium macrocarpon ) production throughout its growing areas. The disease is caused by a group of closely related fungi that require identification using long and cumbersome steps of isolation and microscopic observations of structural features. The objective of this study was to develop a molecular assay to simultaneously detect and discriminate 12 of the most important fungal species reported to be pathogenic on cranberry fruit to facilitate the diagnosis of CFR. As the first approach, internal transcribed spacers and large subunit regions of all fungi were sequenced and confirmed with sequences available in the NCBI database. These data were used to develop primers able to differentiate seven of the 12 species. The five remaining species, including three in the Phacidiaceae family and two in the Glomerellaceae family, were differentiated on the basis of a more discriminant marker, the translation elongation factor 1-α. Two PCR reactions were optimized to clearly delineate the 12 species. The multiplex test was first validated using pure fungal cultures; it was subsequently validated using fruit collected in cranberry beds in eastern Canada. In the latter case, the test was rigorous enough to clearly discriminate the fungal pathogens from contaminants. Within the tested samples, Physalospora vaccinii and Coleophoma empetri were most commonly found. This molecular test offers scientists, diagnosticians, and growers a powerful tool that can rapidly and precisely identify fungi causing CFR so they can implement appropriate control methods.

Several Pestalotiopsis -like species cause gray blight disease in tea plants, resulting in severe tea production losses. However, systematic and comprehensive research on the diversity, geographical distribution, and pathogenicity of pathogenic species associated with tea plants in China is limited. In this study, 168 Pestalotiopsis -like isolates were obtained from diseased tea plant leaves from 13 primary tea-producing provinces and cities in China. Based on a multilocus (internal transcribed spacer, translation elongation factor 1-α, and β-tubulin gene region) phylogenetic analysis coupled with an assessment of conidial characteristics, 20 Neopestalotiopsis unclassified isolates, seven Pestalotiopsis species, including two novel ( Pestalotiopsis menhaiensis and Pestalotiopsis sichuanensis ), four known ( Pestalotiopsis camelliae , Pestalotiopsis chamaeropis , Pestalotiopsis kenyana , and Pestalotiopsis rhodomyrtus ) and one indistinguishable species, and three Pseudopestalotiopsis species, including two known ( Pseudopestalotiopsis camelliae-sinensis and Pseudopestalotiopsis chinensis ) and one indistinguishable species, were identified. This study is the first to evaluate Pestalotiopsis chamaeropis on tea plants in China. The geographical distribution and pathogenicity tests showed Pseudopestalotiopsis camelliae-sinensis to be the dominant cause of gray blight of tea plants in China. In vitro antifungal assays demonstrated that theobromine not only derepressed mycelial growth of the 29 representative isolates but also increased their growth. Correlation analysis revealed a linear positive relationship between the mycelial growth rate and pathogenicity ( P = 0.0148).

The Botryosphaeriaceae family is considered a fungal family that includes pathogens causing latent infection of woody plants, and a number of species were identified as causal pathogens of canker and shoot blight diseases. To better understand the process of latent infection of major canker-causing pathogens in woody tissues in different tree crops important in California, shoot and bud samples were randomly collected from four tree crops: almond, dried plum, pistachio, and walnut. The previously developed DNA primers and quantitative real-time PCR (qPCR) assay systems were applied to detect six canker-causing pathogen groups, including Botryosphaeria dothidea , and species of Cytospora , Diplodia , Lasiodiplodia , Neofusicoccum , and Phomopsis. The concepts of molecular severity (MS) and latent infection index (LII) were introduced and applied to quantify the latent infection levels for these samples. Variation in incidence of latent infection among pathogen groups was observed, whereas the incidences were relatively low among species of Phomopsis and Diplodia. High incidences of Cytospora spp. were observed in two dried plum (prune) orchards. Most orchards showed high incidences of B. dothidea and Lasiodiplodia spp. and moderate incidences of Neofusicoccum spp. Variations in MS were observed among samples of the studied orchards, ranging from 4 to 8. The overall results of LII demonstrated that species of Diplodia and Phomopsis were less important in population development of canker-causing pathogens at the latent phase. Lasiodiplodia spp. were the most aggressive and had been well developed in populations among the studied tree crops. Cytospora spp. became predominant in two of the three dried plum orchards, whereas B. dothidea and Neofusicoccum spp. showed trends of increase in incidence across various tree crops. This study also demonstrated the usefulness of this sensitive qPCR approach in providing evidence of the latent phase of major canker-causing pathogens of stone fruit and nut crops at an early stage of latent infection in woody plant tissues.

Blackleg, caused by the fungus Leptosphaeria maculans , is a widespread disease of winter canola ( Brassica napus ) in Oklahoma and Kansas. Deployment of genetic resistance is the primary strategy for managing blackleg. Resistance genes ( Rlm ) in canola interact with avirulence genes in the fungus ( AvrLm ) in a gene-for-gene manner. Little is known about the diversity and frequency of avirulence genes and the race structure in the region. Isolates of Leptosphaeria spp. were collected from diseased leaves in nine counties in Oklahoma and one county in Kansas from 2009 to 2013. Based on pathogenicity and PCR amplification of mating type and species-specific internal transcribed spacer loci, most isolates ( n = 90) were L. maculans. The presence of avirulence genes was evaluated using phenotypic interactions on cotyledons of differential cultivars with Rlm1 , Rlm2 , Rlm3 , and Rlm4 and amplification of AvrLm1 , AvrLm4-7 , and AvrLm6 by PCR. The avirulence alleles AvrLm6 and AvrLm7 were present in the entire L. maculans population. AvrLm1 was found in 34% of the population, AvrLm2 in 4%, and AvrLm4 in only 1%. A total of five races, defined as combinations of avirulence alleles, were identified that included AvrLm1-2-6-7 , AvrLm2-6-7 , AvrLm4-6-7 , AvrLm1-6-7 , and AvrLm6-7 . Races virulent on the most Rlm genes, AvrLm1-6-7 at 32% and AvrLm6-7 at 62%, were predominant. Defining the avirulence allele frequency and race structure of L. maculans should be useful for the identification and development of resistant cultivars and hybrids for blackleg management in the region. The results suggest that Rlm6 and Rlm7 would be effective, although their deployment should be integrated with quantitative resistance and cultural practices, such as crop rotation, that limit selection pressure on Rlm genes.

Spot blotch, caused by the fungal pathogen Cochliobolus sativus , is a limiting factor for barley ( Hordeum vulgare ) production in northeast China, which causes significant grain yield losses and kernel quality degradation. It is critical to determine the virulence diversity of C . sativus populations for barley resistance breeding and the judicious grouping of available resistance varieties according to the predominant pathotypes in disease epidemic regions. With little information on the barley pathogen in China, this study selected 12 typical barley genotypes to differentiate the pathotypes of C . sativus isolates collected in China. Seventy-one isolates were grouped into 19 Chinese pathotypes based on infection responses. Seventeen isolates were classified as pathotype 3, which has only been identified in China, whereas most (52 of 71) were classified as pathotype 1. All of the tested isolates had low virulence on the North Dakota (ND) durable, resistant line ND B112. Using 22 selected amplified fragment-length polymorphism (AFLP) primer combinations, genetic polymorphism was used to analyze 68 isolates, which clustered into three distinct groups using the unweighted pair group method average with the genetic distance coefficient. No relationship was found between the virulence of isolates and their origins. Isolates of the same pathotype or those collected from the same location did not group into clusters based on the AFLP analysis.

California produces 99.1% of pistachios grown in the United States, and diseases affecting pistachio rootstocks represent a constant challenge to the industry. Field surveys of fungi associated with pistachio rootstocks with symptoms of crown rot and stem canker in three central California counties followed by phylogenetic analyses of translation elongation factor 1-α and second largest subunit of RNA polymerase II gene fragments identified three Fusarium species ( Fusarium equiseti , Fusarium oxysporum , and Fusarium proliferatum ) and two Neocosmospora species ( Neocosmospora falciformis and Neocosmospora solani ). F. oxysporum and N. falciformis were the fungal species most frequently recovered from symptomatic pistachio trees. Inoculations of detached twigs of cultivar Kerman pistachio Pioneer Gold I and clonal University of California, Berkeley I (UCBI) rootstocks showed that all five species could colonize pistachio wood and cause vascular discolorations. Pathogenicity tests in potted pistachio trees completed Koch's postulates and confirmed that F. oxysporum , F. proliferatum , N. falciformis , and N. solani were capable of producing rot and discoloration in stems of clonal UCBI rootstocks, the most widely planted pistachio rootstock in California. To our knowledge, this study is the first to present insights into the biodiversity and biology of Fusarium and Neocosmospora species associated with pistachio trees in California.

Cercospora beticola , the cause of Cercospora leaf spot (CLS) of sugar beet and table beet, has a broad range of potential alternative hosts. The role of these hosts as inoculum sources in the field is unclear and has had limited investigation since the advent of DNA-based pathogen identification. The presence of C. beticola on alternative hosts associated with table beet fields of New York was assessed in field surveys during 2016. Lesions were collected, and 71 cercosporoid conidia were isolated for phylogenetic comparison. C. beticola was identified from Solanum ptycanthum ( n = 4), Chenopodium album ( n = 2), and Spinacia oleracea ( n = 1), whereas C. chenopodii was identified on Chenopodium album ( n = 51). Artificial inoculation of 21 plants species demonstrated that C. beticola was pathogenic to Brassica kaber , Chenopodium album , Carthamus tinctorius , Rumex obtusifolius , and Spinacia oleracea . These results indicate that although C. beticola may be pathogenic to a range of plant species, the role of symptomatic tissue for inoculum production on alternative hosts in the field appears limited. Observations of C. beticola on necrotic and naturally senescent tissue suggest saprophytic survival on plant debris of a range of species, which has implications for CLS epidemics and disease management.

Apple fruit spot disease has caused serious economic losses for years in China since the widespread application of fruit bagging in production. Although the three genera Trichothecium , Alternaria , and Acremonium have been reported to be the causal agents, studies on the disease etiology and pathogen biology are still sparse. Here, we report characterization of eight fungal isolates from lesions on 126 symptomatic fruit samples collected in Shaanxi Province, China. Pathogenicity of the isolates was assessed. DNA sequences were obtained at four loci, including D1/D2 domains of the large-subunit nrRNA gene, internal transcribed spacer regions 1 and 2, 5.8S nrDAN gene, a fragment of the actin gene, and a fragment of the β-tubulin. Based on phylogenetic analysis and morphological features, three new species were found: Acremonium mali , Sarocladium liquanensis , and Sarocladium mali . In addition, we made the first report of Sarocladium terricola as a plant pathogen. Temperature and moisture significantly affected in vitro conidial germination of five Acremonium -like species, and their impact on infection of apple fruit was tested using Acremonium sclerotigenum . Conidia of five species germinated from 15 to 35°C in free water; four of the species had optimum temperature around 25°C, whereas conidia of S. terricola had an optimum temperature of 30°C. Conidial germination rate increased as relative humidity (RH) increased. The five isolates had relatively high conidial germination rates at RH > 97%, with a significant decline at 95% RH. Incidence of infection also increased in proportion to RH. In free water, conidial germination was relatively unaffected by temperature.

Blueberries ( Vaccinium spp.) are largely cultivated in China because of their nutritional benefits and economic value. Blueberry stem blight caused by members of the Botryosphaeriaceae has become one of the most severe diseases affecting blueberry cultivation in China. In this study, we examined the causal agent of blueberry stem blight at commercial greenhouse farms in the suburban area of Beijing, China. In total, 37 isolates of Botryosphaeriaceae were obtained from 100 stem blight samples of blueberry. Twelve of 37 strains were morphologically consistent with the genus Lasiodiplodia , showing ellipsoid to ovoid, one-celled, hyaline conidia that sometimes turned brown, with median septa and longitudinal striations when mature. These 12 strains were identified as belonging to a novel fungal species, Lasiodiplodia vaccinii , based on phylogenetic analysis of the concatenated internal transcribed spacer, RNA polymerase II gene, β-tubulin gene, and translation elongation factor-1α gene sequences as well as morphological characteristics. Pathogenicity tests indicated that L. vaccinii can cause twig blight on blueberry seedlings in the greenhouse. Mycelial growth of L . vaccinii occurred at pH values ranging from 3.0 to 10.0, with an optimum at 6.2, and at temperatures from 15 to 40°C, with an optimum at 30.3°C. Of the seven carbon sources tested, sucrose, fructose, and glucose were all highly efficient in supporting the mycelial growth of L. vaccinii , and xylose was the least effective. Of six nitrogen sources tested, yeast extract and tryptone best promoted mycelial growth of L. vaccinii . The ability of L. vaccinii to grow at high temperatures may help to explain its occurrence in Beijing greenhouses in this study.

Northern corn leaf blight (NCLB) caused by Exserohilum turcicum is the most common and economically significant fungal leaf disease of corn in Ontario, Canada. During the past 10 years in Ontario, severity and incidence of NCLB have increased, possibly owing to the appearance of new races. Several races have been identified in various parts of the world; however, information on occurrence and distribution of races in Ontario is lacking. In the current study, 677 single conidial isolates of E. turcicum were isolated from 687 symptomatic leaf samples collected between 2012 and 2016. These isolates were evaluated for pathogenicity on six corn differential inbreds (A619, A619 Ht1 , A619 Ht2 , A619 Ht3 , A632 Htn1 , and H102 Htm1 ) under controlled environmental conditions and then grouped into 17 physiological races (0, 1, 2, 3, M, N, 12, 1M, 1N, 3M, 13M, 12N, 13N, 1MN, 12MN, 13MN, 123MN) based on the reaction of the inbreds to infection (resistant or susceptible). Four races (0, 1M, 1N, and 1MN) were most frequent, with an isolation frequency of 13, 10, 12, and 41%, respectively. Seventy-six percent of the isolates were virulent on more than one Ht resistance gene, with 2.4% (16 isolates) virulent on all five Ht resistance genes used in this study. Further analysis of the distribution of races in four regions over the years revealed that the occurrence and distribution of the races changed with time in Ontario. Overall, the frequency of virulence of the 677 isolates screened on the differentials with resistance genes Ht1 , Ht2 , Ht3 , Htm1 , and Htn1 varied from 6 to 81% ( Ht1 81%, Ht2 6%, Ht3 12%, Htm1 64%, and Htn1 64%). Virulent isolates produced fewer lesions on the Htm1 differential, and smaller lesions that were slower and having less sporulation on the Htn1 differential, compared with infection of the differentials with Ht1 , Ht2 , and Ht3 resistance genes. Virulence frequency also changed within the four geographical regions of Ontario, with fewer isolates virulent on all resistance genes in eastern Ontario compared with southern and western Ontario. Isolates from southern Ontario had greater virulence frequency against Ht1 and Htm1 , whereas isolates from western Ontario were more frequently virulent on Ht1 and Htn1. The information generated in this study on the distribution of E. turcicum races in Ontario corn will help growers to select appropriate hybrids with required resistance genes and will assist seed companies in deploying resistance genes in corn hybrids across the province or within a particular region.

Persian lime ( Citrus latifolia Tan.) is an important and widely cultivated fruit crop in several regions of Mexico. In recent years, severe symptoms of gummosis, stem cankers, and dieback were detected in the Persian lime-producing region in the states of Veracruz and Puebla, Mexico. The aims of this study were to identify the species of Lasiodiplodia associated with these symptoms, determine the distribution of these species, and test their pathogenicity and virulence on Persian lime plants. In 2015, symptomatic samples were collected from 12 commercial Persian lime orchards, and 60 Lasiodiplodia isolates were obtained. Fungal identification of 32 representative isolates was performed using a phylogenetic analysis based on DNA sequence data of the internal transcribed spacer region and part of the translation elongation factor 1-α and β-tubulin genes. Sequence analyses were carried out using the Maximum Likelihood and Bayesian Inference methods. Six Lasiodiplodia species were identified as Lasiodiplodia pseudotheobromae , Lasiodiplodia theobromae , Lasiodiplodia brasiliense , Lasiodiplodia subglobosa , Lasiodiplodia citricola , and Lasiodiplodia iraniensis . All Lasiodiplodia species of this study are reported for the first time in association with Persian lime in Mexico and worldwide. L. pseudotheobromae (46.9% of isolates) was the most frequently isolated species followed by L. theobromae (28.1%) and L. brasiliense (12.5%). Pathogenicity on Persian lime young plants using a mycelial plug inoculation method showed that all identified Lasiodiplodia species were able to cause necrotic lesions and gummosis, but L. subglobosa , L. iraniensis , and L. pseudotheobromae were the most virulent.

Several species of the Botryosphaeriaceae family have been associated with branch canker, dieback, and stem end rot in avocado ( Persea americana Mill.). In Chile, the incidence of diseases affecting the avocado tree increased from 2011 to 2016, which coincided with a severe drought that affected avocado production. Moreover, distant countries importing avocados from Chile also reported an increase of stem end rot of ripe avocados. Therefore, the aims of this study were to identify the pathogen species associated with branch canker, dieback, and stem end rot of avocado in Chile and to study their pathogenicity. This study was conducted between 2015 and 2016 in 'Hass' avocado orchards located in the main avocado-producing regions in Chile. A diverse collection of fungal species was recovered from both necrotic woody tissue and necrotic tissue on harvested ripe fruit. On the basis of morphology and phylogenetic analyses of the internal transcribed spacer region (ITS1-5.8S-ITS2) and the translation elongation factor 1-α (TEF1-α) gene, eight species in the Botryosphaeriaceae family were identified: Diplodia mutila , D. pseudoseriata , D. seriata , Dothiorella iberica , Lasiodiplodia theobromae , Neofusicoccum australe , N. nonquaesitum , and N. parvum . For each of these species, pathogenicity studies were conducted on 1-year-old healthy Hass avocado plants. All isolates produced brown gum exudate and caused necrosis in the vascular system 3 weeks after inoculation. N. nonquaesitum , N. parvum , and D. pseudoseriata were the most virulent species. Necrotic lesions and cavities with white mycelia near the peduncle union were observed on Hass avocado fruit inoculated postharvest. L. theobromae , N. australe , and N. parvum were significantly more virulent than the other tested species in the Botryosphaeriaceae family. This study identified and characterized the pathogenicity of Botryosphaeriaceae species in Chile, which will prove useful to future research on these pathogens directed at establishing effective control strategies in avocado.

Blackberry cane diseases with the symptoms of necrosis, canker, and wilting are caused by several fungi worldwide. Surveys conducted from 2013 to 2016 in Serbia revealed the occurrence of Gnomoniopsis idaeicola, the causal agent of cane canker and wilting, which was found to be distributed in almost half of the surveyed orchards, in three blackberry cultivars, and with disease incidence of up to 80%. Wide distribution and high disease incidence suggest that G. idaeicola has been present in Serbia for some time. Out of 427 samples, a total of 65 G. idaeicola isolates were obtained (isolation rate of 34.19%). Reference isolates, originating from different localities, were conventionally and molecularly identified and characterized. G. idaeicola was detected in single and mixed infections with fungi from genera Paraconiothyrium, Colletotrichum, Diaporthe, Botryosphaeria, Botrytis, Septoria, Neofusicoccum, and Discostroma, and no diagnostically specific symptoms could be related directly to the G. idaeicola infection. In orchards solely infected with G. idaeicola, blackberry plant mortality was up to 40%, and yield loses were estimated at 50%. G. idaeicola isolates included in this study demonstrated intraspecies diversity in morphological, biological, pathogenic, and molecular features, which indicates that population in Serbia may be of different origin. This is the first record of a massive outbreak of G. idaeicola infection, illustrating its capability of harmful influence on blackberry production. This study represents the initial step in studying G. idaeicola as a new blackberry pathogen in Serbia, aiming at developing efficient control measures.

Persimmon (Diospyros kaki) fruit production is severely affected by circular leaf spot worldwide. Mycosphaerella nawae causes circular leaf spot of persimmon (CLSP) and can result in leaf spot, defoliation, early fruit maturation, and subsequent softening and abscission. The morphology and phylogenetic position of M. nawae within the family Mycosphaerellaceae is, therefore, of utmost importance given its impact on persimmon production. Based on previous morphological and molecular studies, the phylogenetic position of the anamorphic genera associated with M. nawae remain in confusion. In the present study, 15 isolates of M. nawae were collected from the tissue of living leaves exhibiting leaf spot symptoms. A subsample of three isolates was characterized phylogenetically and morphologically. Isolates were compared based on DNA sequence data for the internal transcribed spacer region (ITS1-5.8S ITS2), part of the 28S nrDNA including domains D1-D3 (LSU), actin (Act), translation elongation factor 1-alpha (EF-1α), and RNA polymerase II second largest subunit (rpb2). The anamorph and teleomorph structures, ascospore germination patterns, as well as host specificity were used to describe the isolates. The phylogenetic and morphological analyses revealed that M. nawae requires a new holomorphic genus within Mycosphaerellaceae, described herein as Plurivorosphaerella gen. nov. A host specificity test revealed that Plurivorosphaerella nawae comb. nov. (M. nawae) can superficially colonize, but not infect, apple, peach, cherry, and plum.

Cut branches of deciduous holly (Ilex spp. L.) harboring colorful berries are traditionally used as ornaments in holiday decorations. Since 2012, a fruit rot of unspecified cause has resulted in significant yield reduction and economic losses across Midwestern and Eastern U.S. nurseries. In this study, symptomatic fruit samples collected from nine different locations over five years were analyzed, and several fungal species were isolated. A combination of morphological characterization, multilocus phylogenetic analyses, and pathogenicity assays revealed that Alternaria alternata and Diaporthe ilicicola sp. nov. were the primary pathogens associated with symptomatic fruit. Other fungi including A. arborescens, Colletotrichum fioriniae, C. nymphaeae, Epicoccum nigrum, and species in the D. eres species complex appeared to be minor pathogens in this disease complex. In detached fruit pathogenicity assays testing the role of wounding and inoculum concentration on disease development, disease incidence and severity increased when fruit was wounded and inoculated with a higher inoculum concentration. These findings indicate that management strategies that can protect fruit from injury or reduce inoculum may lower disease levels in the field. This research established the basis for further studies on this emerging disease and the design of research-based management strategies. To our knowledge, it also represents the first report of species of Alternaria, Colletotrichum, Diaporthe, and Epicoccum causing fruit rot of deciduous holly.

In the early 2000s, spruce trees in Michigan began displaying basal needle drop and branch death that slowly progressed upward, symptoms of what we call spruce decline. A survey in 2013 revealed that spruce decline was common throughout Michigan's Lower Peninsula, and Diaporthe was the most likely pathogen causing the cankers associated with these symptoms. Greenhouse inoculation studies completed Koch's postulates, confirming that Diaporthe could cause cankers that cause needle loss and branch death. The five different Diaporthe haplotypes isolated from symptomatic branches during the survey differed in virulence. Haplotypes 2, 4, and 5 were more virulent, and differed from each other by only one or two base pairs using the internal transcribed spacer (ITS) region and did not differ using the β-tubulin (TUB) gene. These haplotypes were unresolved phylogenetically. Haplotypes 1 and 3 were weakly virulent to avirulent on multiple spruce taxa, and fell into a resolved Diaporthe eres clade. Spruce taxa varied in susceptibility, with Colorado blue spruce (Picea pungens) the most susceptible, followed by Norway (P. abies), then white spruce (P. glauca). Spruce taxa that were much less susceptible were Black Hills (P. glauca var. densata), Serbian (P. omorika), and Meyer spruce (P. meyeri). We demonstrate that one or more Diaporthe species is causing cankers on declining spruce in Michigan, and these cankers elicit symptoms similar to the branch death expressed by declining spruce in the landscape. Future work will focus on further characterizing Diaporthe to species, and determining biotic and abiotic stressors that may predispose spruce trees to express decline symptoms.

Monilinia spp. are responsible for brown rot decay of stone and pome fruit in the field as well as in postharvest. Monilinia laxa and M. fructigena are considered indigenous to Europe, while M. fructicola is a quarantine pathogen in the European and Mediterranean Plant Protection Organization area included in the A2 List. In Italy, it was first reported in 2009 in Piedmont (northern Italy) and rapidly spread to central Italy. We carried out a monitoring program on the occurrence of Monilinia spp. in southern Italy and a comparative characterization of the three main fungal pathogens. Molecular assays based on direct polymerase chain reaction (PCR) and real-time quantitative PCR for molecular identification of Monilinia spp. from rotted fruit were set up, validated, and applied in a monitoring program. Of the tested 519 isolates from 26 orchards, 388 (74.8%) were identified as M. fructicola, 118 (22.7%) as M. laxa, 10 (1.9%) as M. fructigena, and 3 (0.6%) were M. polystroma. M. fructicola colonies grew faster and had a higher optimal temperature for growth (26°C) than M. laxa (23°C) and M. fructigena (20°C). No relevant difference in virulence could be observed on artificially inoculated apricot, cherry, and peach fruit. The fungal species showed different responses to fungicides, because M. fructicola was more sensitive than M. laxa, especially to cyflufenamid, and M. fructigena revealed a lower sensitivity to succinate dehydrogenase inhibitors (boscalid, fluopyram, and fluxapyroxad) and quinone outside inhibitors (mandestrobin). In summary, the two species M. fructicola and M. polystroma were first detected in southern Italy where M. fructicola has largely displaced the two indigenous pathogens M. laxa and M. fructigena; the relative proportions of the three pathogens in orchards should be considered when defining the management of brown rot of stone fruit due to differences in their responses to fungicides.

Almond trees with trunk and branch cankers were observed in several orchards across almond-producing counties in California. Symptoms of cankers included bark lesions, discoloration of xylem tissues, longitudinal wood necrosis, and extensive gumming. Spur and shoot blight associated with rotted fruit were detected in two orchards in Kern County. The fungus Neoscytalidium dimidiatum was consistently recovered from the various cankers, infected fruit, and blighted shoots and its identity was confirmed based on phylogenetic and morphological studies. Phylogenetic analyses of the internal transcribed spacer, translation elongation factor 1-α, and β-tubulin genes comparing 47 strains from California with reference specimens within the family Botryosphaeriaceae and coupled with detailed morphological observations validated the identity of the pathogenic fungus. Pathogenicity tests conducted in the field using 1- to 2-year-old branches inoculated with mycelium plugs or conidial suspensions and attached fruit inoculated with conidial suspensions fulfilled Koch's postulates. N. dimidiatum appeared highly virulent in almond-producing cankers of up to 22 cm in length within 4 weeks using mycelium plug inoculations as well as severe fruit rot combined with spur blight on the fruit-bearing spurs. This study reports, for the first time, the fungus N. dimidiatum as a pathogen of almond in California causing canker, shoot blight, and fruit rot. Disease symptoms are described and illustrated.

Botryosphaeria dieback, caused by species of Botryosphaeriaceae, is an important grapevine trunk disease in Australia. Inocula produced by the pathogens are primarily dispersed by rain splash and wind and infect pruning wounds leading to cankers, dieback, and eventually death of vines. The objective of this study was to develop molecular tools to detect and quantify Botryosphaeriaceae inocula from the environment. These tools are essential for investigating spore dispersal patterns of Botryosphaeriaceae pathogens in Australian vineyards. DNA extraction protocols were evaluated and one modified protocol was found suitable for extracting Botryosphaeriaceae DNA from artificially and naturally inoculated Burkard volumetric spore sampler tapes. Multispecies primers and a hydrolysis probe for quantitative PCR (qPCR) were further developed to detect and quantify Botryosphaeriaceae inocula from environmental samples. Specificity tests showed that the multispecies primers were able to amplify the DNA of 10 Botryosphaeriaceae species (58 isolates) found in Australia while none of the 27 nontarget fungal species (90 isolates) tested were amplified. The qPCR assay was suitable for amplifying purified DNA, synthetic DNA fragments (gBlocks), and mixed DNA from spore trap tapes. The qPCR method developed in this study was shown to be rapid and sensitive in detecting Botryosphaeriaceae inocula from the environment using spore traps.

Brown rot is a severe disease affecting stone and pome fruit. This disease was recently confirmed to be caused by the following six closely related species: Monilinia fructicola, M. laxa, M. fructigena, Monilia polystroma, M. mumecola, and M. yunnanensis. Because of differences in geographic distributions, some of these species are important quarantine pathogens in certain countries. In this study, we developed TaqMan real-time polymerase chain reaction (PCR) assays to detect and identify the six species. Primer pairs and probes were designed for Monilinia fructicola, M. fructigena, M. laxa, and Monilia polystroma based on sequence differences in the laccase-2 genes. Additionally, based on sequence differences in the elongation factor genes, primer pairs and probes were designed for Monilia mumecola and M. yunnanensis. The real-time PCR assays were able to specifically identify the target pathogens, with detection limits of 10 to 100 fg of DNA, which is equivalent to one to seven conidia. The assays were also able to detect the target pathogens in a mixed DNA sample comprising all six Monilinia spp. and related species. The real-time PCR assays accurately detected target fungi from infected apple fruit. Furthermore, the identification results were consistent with those of traditional morphological methods.

Citrus black spot, caused by Phyllosticta citricarpa, is one of the most important fungal diseases in many citrus-growing regions with hot and humid summers. Ascospores and conidia are known to contribute to epidemic development of the disease. However, pathogenicity testing has never been done for pure ascospores produced from fully characterized P. citricarpa isolates, due to the inability to induce the sexual state in vitro. Recently, an in vitro mating technique was developed to readily produce pure P. citricarpa ascospores for use in host inoculation studies. To test the pathogenicity of P. citricarpa ascospores, we inoculated Troyer citrange leaves and Murcott tangor fruit with ascospores produced in vitro from characterized P. citricarpa isolates. Typical symptoms of citrus black spot occurred. Recovery of P. citricarpa isolates from symptomatic lesions and their characterization using genetic markers enabled us to identify recombinant genotypes among the isolates recovered from ascospore inoculations and, as such, fulfill Koch's postulates for ascospores. We have also identified Troyer citrange seedlings as a potential model system for citrus black spot inoculation studies, because it allows typical symptoms of citrus black spot to be expressed with a much shorter latent period than on fruit. This will facilitate future studies of epidemiological aspects of P. citricarpa ascospores relative to conidia and improve our understanding of the citrus black spot pathosystem. The susceptibility of Troyer citrange seedlings will also facilitate experimenting with disease management methods, aimed at reducing the impact of citrus black spot.

Grapevine trunk diseases (GTDs) represent one of the most important problems for viticulture worldwide. Beyond the original causes of this outbreak in some countries like France, the lack of efficient control protocols and the prohibition of using active ingredients such as sodium arsenite and benzimidazoles, until recently used to reduce the impact of some GTDs but deleterious for humans and the environment, have probably worsened the impact of the diseases, leading to increasing economic losses. Since 1990, searches have been made to find efficient tools to control GTDs, testing a wide range of active ingredients and biocontrol agents. This review provides readers with an overview of the results reported in the scientific literature over the last 15 years. In particular, the review focuses on the trials carried out applying chemicals or microorganisms to control Esca complex diseases, Botryosphaeria dieback, and Eutypa dieback, the most widespread GTDs.

Seimatosporium spp. and closely related "pestalotioid fungi" have been isolated from vineyards worldwide, but their ecological status in grapevine wood is unclear. To determine their involvement in the grapevine trunk-disease complex, we tested the pathogenicity of Californian isolates obtained from vines with general symptoms of Botryosphaeria, Eutypa, and Phomopsis diebacks. Multilocus phylogenetic analyses revealed three species: Seimatosporium vitis and two newly described and typified species, S. luteosporum sp. nov. and S. vitifusiforme sp. nov. Inoculations to woody stems of potted grapevines of both isolates of S. vitis and one isolate of S. vitifusiforme, but not S. luteosporum, were associated with significantly larger lesions than those of noninoculated controls. Coinoculations with trunk pathogens (Cryptovalsa ampelina, Diaporthe ambigua, Diatrypella verruciformis, Diplodia seriata, and Eutypa lata), coisolated from the same wood cankers in the field, brought about increased lesion lengths for S. vitifusiforme paired with D. seriata, and S. luteosporum paired with Diaporthe ambigua. In contrast, there were no differences in lesion lengths of S. vitis and Diatrypella verruciformis or S. vitis and E. lata, inoculated alone or together. Our findings suggest that Seimatosporium spp. are involved in the grapevine trunk-disease complex, and their virulence may depend on or affect that of trunk pathogens.

Boxwood blight caused by Calonectria pseudonaviculata (Cps) is an emerging disease in the United States. Understanding the biology and survival of microsclerotia, the resting propagules of Cps, is important to managing the disease and restoring Cps-infested sites. In this study, an image-analyzing software (OpenCFU) was adapted to enumerate microsclerotia produced in culture plates. Microsclerotial counts increased with time up to 42 days while their size continued to grow for 70 days. Additionally, younger microsclerotia were less sensitive to extreme temperatures of -10 and 40°C than older ones. For instance, 21-day-old microsclerotia survived at almost twice the rate of 70-day-old ones after a 24-h exposure to -10°C. Likewise, the 21-day-old microsclerotia survived at much greater rates than 70-day-old ones (50 to 70% versus <10%) after a 24-h exposure to 40°C. The practical implications of these results are discussed.

Brown rot, caused by Monilinia spp., is an economically important pre- and postharvest disease of pome and stone fruits worldwide. In Serbia, apple is the most widely grown pome fruit, and the distribution of economically important Monilinia spp. responsible for apple brown rot is unknown. Hence, we conducted a three year survey, from 2010 to 2012, where 349 isolates were obtained from six orchards and four storage facilities from five different apple cultivars with brown rot symptoms. Morphological characterization of the isolates, multiplex PCR, and phylogenetic analysis revealed four species: M. fructigena, M. laxa, M. fructicola, and Monilia polystroma. All species were found in the orchard and in storage, with M. fructigena predominating, followed by M. polystroma. Representative isolates were analyzed in vitro and in vivo where differences in growth rate, sporulation, and virulence on apple fruit were observed. Findings from this investigation demonstrate diversity in the species responsible for pre- and postharvest apple brown rot, which has significant implications for pathogen detection and for developing disease-specific management strategies.

Grape white rot is a common disease and causes considerable yield losses in many grape-growing regions when environmental conditions are favorable. We surveyed grape white rot in five provinces in China and collected 27 isolates from diseased grape tissues. Multigene phylogenetic analyses of the internal transcribed spacer region (ITS1-5.8S-ITS2), the 28S large subunit of nuclear ribosomal RNA (LSU), partial translation elongation factor 1-alpha gene (TEF 1-α), and partial histone 3 gene (HIS), coupled with genealogical concordance phylogenetic species recognition and morphological observations, revealed that Coniella vitis sp. nov. and C. diplodiella are the causal agents of grape white rot in China. Koch's postulates were performed on Vitis vinifera cv. Summer Black in a greenhouse. These results confirmed the pathogenicity on grapes, as symptoms were reproduced, and also indicated significant variations in the virulence among C. vitis isolates. This work provides evidence that C. vitis is the main pathogen of grape white rot in China and also provides an optimized multigene backbone for resolving Coniella species.

A novel disease characterized by small brown-black spots (1 to 2 mm in diameter) on tender tea leaves (Camellia sinensis) has been observed in many regions of Hubei Province, China, which severely affects the yield and quality of tea. Tea leaf samples with typical symptoms were collected from three major tea-cultivation regions of Hubei, and were subjected to pathogen isolation for etiological analysis. As a result, 34 Pestalotiopsis isolates were obtained from 20 samples, and they were identified as Pestalotiopsis theae (14 isolates), P. camelliae (12), and P. clavispora (8), determined by morphologies and phylogenetic analysis based on internal transcribed spacer, and partial β-tubulin and translation elongation factor 1-alpha genes. Pathogenicity tests on detached tea leaves showed that no matter what mycelial discs or conidium suspensions were used, inoculation of the Pestalotiopsis fungi could result in small brown-black spots (1 to 2 mm in diameter) on wounded leaves, similar to those observed in the field in the sizes and colors. It also revealed that only P. theae had pathogenicity on unwounded tea leaves, and P. theae and P. clavispora showed significantly higher virulence than P. camelliae. Inoculation test with conidium suspension on intact tea leaves in the field further confirmed that P. theae as the pathogen of brown-black spots. Reisolation of the pathogens from diseased leaves confirmed that the symptom was caused by the inoculation of Pestalotiopsis fungi. The P. theae isolates responsible for brown-black spots were also compared with those for tea gray blight disease in growth rate, pathogenicity, and molecular characteristics in parallel. To our knowledge, this is the first report that the Pestalotiopsis fungi cause brown-black spot disease on tender tea leaves. The results provide important implications for the prevention and management of this economically important disease.

Diaporthe spp. are important plant pathogens causing wood cankers, blight, dieback, and fruit rot in a wide range of hosts. During surveys conducted during the 2013 and 2014 seasons, a postharvest rot in Hayward kiwifruit (Actinidia deliciosa) was observed in Chile. In order to identify the species of Diaporthe associated with this fruit rot, symptomatic fruit were collected from seven kiwifruit packinghouses located between San Francisco de Mostazal and Curicó (central Chile). Twenty-four isolates of Diaporthe spp. were identified from infected fruit based on morphological and cultural characters and analyses of nucleotides sequences of three loci, including the internal transcribed spacer (ITS) region (ITS1-5.8S-ITS2), a partial sequences of the β-tubulin, and translation elongation factor 1-α genes. The Diaporthe spp. identified were Diaporthe ambigua, D. australafricana, D. novem, and D. rudis. Multilocus phylogenetic analysis revealed that Chilean isolates were grouped in separate clades with their correspondent ex-types species. All species of Diaporthe were pathogenic on wounded kiwifruit after 30 days at 0°C under normal and controlled-atmosphere (2% O 2 and 5% CO 2 ) storage and they were sensitive to benomyl, pyraclostrobin, and tebuconazole fungicides. D. ambigua isolates were the most virulent based on the lesion length measured in inoculated Hayward and Jintao kiwifruit. These findings confirm D. ambigua, D. australafricana, D. novem, and D. rudis as the causal agents of kiwifruit rot during cold storage in Chile. The specie D. actinidiae, a common of Diaporthe sp. found associated with kiwifruit rot, was not identified in the present study.