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Ancient Platycladus orientalis holds significant ecological, landscape, historical, and cultural value. In northern China, leaf blight has significantly impacted the growth and ornamental value of ancient P. orientalis. In this study, 26 blight leaf samples of ancient P. orientalis were collected in Beijing, China. Phylogenetic analysis based on the concatenated internal transcribed spacer (ITS), β-tubulin (tub2), and RNA polymerase second largest subunit (rpb2) DNA sequence data combined with fungal morphological characteristics revealed three taxa of Nothophoma, i.e. N. platycladus, N. spiraeae, and N. juglandis. Of which, N. platycladus is a species new to science. Koch's postulates indicated that all these three species of Nothophoma could cause leaf blight of P. orientalis. [ABSTRACT FROM AUTHOR]

Abstract Ancient Platycladus orientalis holds significant ecological, landscape, historical, and cultural value. In northern China, leaf blight has significantly impacted the growth and ornamental value of ancient P. orientalis. In this study, 26 blight leaf samples of ancient P. orientalis were collected in Beijing, China. Phylogenetic analysis based on the concatenated internal transcribed spacer (ITS), β-tubulin (tub2), and RNA polymerase second largest subunit (rpb2) DNA sequence data combined with fungal morphological characteristics revealed three taxa of Nothophoma, i.e. N. platycladus, N. spiraeae, and N. juglandis. Of which, N. platycladus is a species new to science. Koch’s postulates indicated that all these three species of Nothophoma could cause leaf blight of P. orientalis.

The cause of peanut kernel shrivel (PKS) syndrome, affecting peanut crops in Australia's growing regions, is currently unknown. It is estimated that PKS is costing the peanut industry more than AUD 5 M p.a. and is a potential threat to the industry. Previous investigations have ruled out all abiotic factors and most biotic factors as the cause of PKS. This research aimed at investigating the scope, distribution, and cause of the PKS syndrome. The survey showed PKS symptoms to be present in peanut crops in all the growing regions surveyed. Based on our study of culturable microorganisms, there appears to be no clear-cut involvement of plant pathogenic bacteria and fungi; however, Fusarium spp. were revealed as the most prevalent fungi in affected plants. Moreover, the soil metagenomics study revealed Fusarium spp. as the most abundant fungal communities in the soil microbial profile, and they could contribute to the PKS syndrome. The consistent presence observed of the identified Fusarium oxysporum in PKS-affected samples could indicate a role for this pathogen in the syndrome, especially in conjunction with abiotic stressors. The pathogenicity testing of F. oxysporum resulted in very mild PKS symptoms. A separate report suggesting the involvement of phytoplasma in the PKS syndrome raises the possibility of an interplay of biotic factors in the development of this disease. Further investigation is warranted to determine the true cause or causes of this disease. [ABSTRACT FROM AUTHOR]

Abstract Background Cobweb disease is a fungal disease that commonly affects the cultivation and production of edible mushrooms, leading to serious yield and economic losses. It is considered a major fungal disease in the realm of edible mushrooms. The symptoms of cobweb disease were found during the cultivation of Lyophyllum decastes. This study aimed to identify the causative pathogen of cobweb disease and evaluate effective fungicides, providing valuable insights for field control and management of L. decastes cobweb disease. Results The causal agent of cobweb disease was isolated from samples infected and identified as Cladobotryum mycophilum based on morphological and cultural characteristics, as well as multi-locus phylogeny analysis (ITS, RPB1, RPB2, and TEF1-α). Pathogenicity tests further confirmed C. mycophilum as the responsible pathogen for this condition. Among the selected fungicides, Prochloraz-manganese chloride complex, Trifloxystrobin, tebuconazole, and Difenoconazole exhibited significant inhibitory effects on the pathogen’s mycelium, with EC50 values of 0.076 µg/mL, 0.173 µg/mL, and 0.364 µg/mL, respectively. These fungicides can serve as references for future field control of cobweb disease in L. decastes. Conclusion This study is the first report of C. mycophilum as the causing agent of cobweb disease in L. decastes in China. Notably, Prochloraz-manganese chloride complex demonstrated the strongest inhibitory efficacy against C. mycophilum.

Tsukamurella, a group of multi-drug resistant, Gram-positive, aerobic, and partially acid-fast bacteria, are emerging causes of bacterial conjunctivitis and keratitis. However, the pathogenesis of Tsukamurella keratitis is largely unknown. To address this, we used New Zealand White rabbits to develop the first eye infection model and conducted in vitro tests to study the pathogenesis mechanisms of Tsukamurella. There is increasing evidence that biofilms play a significant role in ocular infections, leading us to hypothesize that biofilm formation is crucial for effective Tsukamurella infection. In order to look for potential candidate genes which are important in biofilm formation and Tsukamurella keratitis. We performed genome sequencing of two ocular isolates, T. pulmonis-PW1004 and T. tyrosinosolvens-PW899, to identify potential virulence factors. Through in vitro and in vivo studies, we characterized their biological roles in mediating Tsukamurella keratitis. Our findings confirmed that Tsukamurella is an ocular pathogen by fulfilling Koch's postulates, and using genome sequence data, we identified tmytC, encoding a mycolyltransferase, as a crucial gene in biofilm formation and causing Tsukamurella keratitis in the rabbit model. This is the first report demonstrating the novel role of mycolyltransferase in causing ocular infections. Overall, our findings contribute to a better understanding of Tsukamurella pathogenesis and provide a potential target for treatment. Specific inhibitors targeting TmytC could serve as an effective treatment option for Tsukamurella infections.

Background: Cobweb disease is a fungal disease that commonly affects the cultivation and production of edible mushrooms, leading to serious yield and economic losses. It is considered a major fungal disease in the realm of edible mushrooms. The symptoms of cobweb disease were found during the cultivation of Lyophyllum decastes. This study aimed to identify the causative pathogen of cobweb disease and evaluate effective fungicides, providing valuable insights for field control and management of L. decastes cobweb disease. Results: The causal agent of cobweb disease was isolated from samples infected and identified as Cladobotryum mycophilum based on morphological and cultural characteristics, as well as multi-locus phylogeny analysis (ITS, RPB1, RPB2, and TEF1-α). Pathogenicity tests further confirmed C. mycophilum as the responsible pathogen for this condition. Among the selected fungicides, Prochloraz-manganese chloride complex, Trifloxystrobin, tebuconazole, and Difenoconazole exhibited significant inhibitory effects on the pathogen's mycelium, with EC50 values of 0.076 µg/mL, 0.173 µg/mL, and 0.364 µg/mL, respectively. These fungicides can serve as references for future field control of cobweb disease in L. decastes. Conclusion: This study is the first report of C. mycophilum as the causing agent of cobweb disease in L. decastes in China. Notably, Prochloraz-manganese chloride complex demonstrated the strongest inhibitory efficacy against C. mycophilum. [ABSTRACT FROM AUTHOR]

The germ theory states that pathogenic microorganisms are responsible for causing infectious diseases. The theory is inherently microbe-centric and does not account for variability in disease severity among individuals and asymptomatic carriership--two phenomena indicating an important role for host variability in infection outcome. The basic tenet of the germ theory was recently challenged, and a radically host-centric paradigm referred to as the "full-blown host theory" was proposed. According to this view, the pathogen is reduced to a passive environmental trigger, and the development of disease is instead due to pre-existing immunodeficiencies of the host. Here, we consider the factors that determine disease severity using established knowledge concerning evolutionary biology, microbial pathogenesis, and host-pathogen interactions. We note that the available data support a noncentric view that recognizes key roles for both the causative microbe and the host in dictating infection outcome. [ABSTRACT FROM AUTHOR]

Leaf-blight disease caused by the Fusarium oxysporum is an emerging problem in Dendrobium chrysotoxum production in China. Symptoms of leaf blight were observed on seedlings of D. chrysotoxum cultivated in a nursery in Ruili City, Yunnan Province, China. In this study, we isolated the Fusarium sp. associated with leaf-blight disease of D. chrysotoxum from the diseased seedlings. A pathogenicity test was performed to fulfill Koch's postulates to confirm the pathogenicity of isolated strains and identified using morphological and molecular techniques. The results revealed that all four isolated Fusarium sp. isolates (DHRL-01~04) produced typical blight symptoms followed by marginal necrosis of leaves on the D. chrysotoxum plants. On the PDA medium, the fungal colony appeared as a white to purplish color with cottony mycelium growth. Microconidia are oval-shaped, whereas macroconidia are sickle-shaped, tapering at both ends with 2–4 septations. The phylogenetic trees were construed based on internal transcribed spacer (ITS), translation elongation factor (EF-1α), and RNA polymerase subunit genes RPB1 and RPB2 genes, respectively, and blasted against the NCBI database for species confirmation. Based on the NCBI database's blast results, the isolates showed that more than 99% identify with Fusarium oxysporum. To our knowledge, this is the first comprehensive report on the identification of Fusarium oxysporum as the causal agent of Dendrobium chrysotoxum leaf blight in Yunnan Province, China, based on morphological and molecular characteristics. [ABSTRACT FROM AUTHOR]

In 2015 and 2016, two Barramundi (Lates calcarifer) farms in Singapore reported a disease outbreak characterized by lethargic behavior, pronounced inappetence, generalized skin lesions, erosions of the fins and tail, and ultimately high mortality in their fish. Next-generation sequencing and PCR confirmed presence of a novel virus belonging to the Alloherpesviridae family, Lates calcarifer herpesvirus (LCHV), which was subsequently isolated and cultured. We characterize, for the first time, the complete genome of two cultured LCHV isolates. The genome contains a long unique region of approximately 105,000 bp flanked by terminal repeats of approximately 24,800 bp, of which the first 8.2 kb do not show any similarity to described genomes in the Alloherpesviridae family. The two cultured isolates share 89% nucleotide identity, and their closest relatives are the viruses belonging to the genus Ictalurivirus. Experimental infections using one of the cultured LCHV isolates resulted in identical clinical signs as originally described in the index farm, both in intraperitoneal-injection infected fish and cohabitant fish, with mortality in both groups. Histopathological analysis showed pronounced abnormalities in the gills. Virus culture and PCR analysis confirmed the replication of LCHV in the infected fish, and thus Koch's postulates were fulfilled. [ABSTRACT FROM AUTHOR]

Background: Leaf spot disease severely impacts Ginkgo biloba (G. biloba) yield and quality. While microbial agents offer effective and non-toxic biological control for plant diseases, research on controlling leaf spot disease in G. biloba is notably scarce. Methods: The pathogenic fungi were isolated and purified from diseased and healthy leaves of G. biloba, Subsequent examinations included morphological observations and molecular identification via PCR techniques. A phylogenetic tree was constructed to facilitate the analysis of these pathogenic fungi, and Koch's postulates were subsequently employed to reaffirm their pathogenic nature. The antagonistic experiment was employed to select biocontrol bacteria, and subsequently, the isolated biocontrol bacteria and pathogenic fungi were inoculated onto healthy leaves to assess the inhibitory effects of the biocontrol bacteria. Results: Two pathologies responsible for the leaf spot disease on G. biloba were identified as Botryosphaeria dothidea and Neofusicoccum parvum via the analysis of phylogenetic tree and the application of Koch's Postulates. Additionally, we isolated two strains of biocontrol bacteria, namely Bacillus velezensis and Bacillus amyloliquefaciens. Their average inhibitory zones were measured at 4.78 cm and 3.46 cm, respectively. The inhibition zone of B. velezensis against N. parvum was 4 cm. B. velezensis showed a stronger inhibitory effect compared to B. amyloliquefaciens on the development of lesions caused by B. dothidea via leaf culture experiment. Conclusion: This research reports, for the first time, the presence of B. dothidea as a pathogenic fungus affecting G. biloba. Moreover, the biocontrol bacteria, B. velezensis and B. amyloliquefaciens, exhibited the capability to effectively inhibit the growth and reproduction of B. dothidea, indicating their promising potential as environmentally friendly biocontrol resources. [ABSTRACT FROM AUTHOR]

Tea (Camellia sinensis (L.) Kuntze) is one of the most important economic plants in China, and has many benefits for human health. Anthracnose is one of the most serious diseases of tea in China, and control of the fungus is important since most Chinese cultivars are susceptible to it. The agent of tea anthracnose was initially described as Gloeosporium theae-sinensis I. Miyake in Japan, which was later transferred to Discula, but this taxonomic position remains problematic. To shed light on these taxonomic and phylogenetic issues, the tea anthracnose pathogens were re-studied. Combining the morphological characteristics and a multigene phylogenetic analysis of nrITS, nrLSU, rpb2, and tef1 sequence data, a new genus Sinodiscula was proposed to accommodate the causal fungi of tea anthracnose, including a new species Sinodiscula camellicola and a new combination Sinodiscula theae-sinensis. Furthermore, the pathogenicity of the pathogens was determined according to Koch's postulates. This study thoroughly resolves the long-standing taxonomic and phylogenetic problems of the tea anthracnose pathogens. [ABSTRACT FROM AUTHOR]

Pink staining in shiitake (Lentinula edodes (Berk.) Pegler) billets (Quercus griffithii Hook.f. and Thomson ex Miq.) causing the inhibition of shiitake mycelium and a reduction in shiitake production was observed throughout Bhutan in 2017-2018. Completion of Koch's postulates confirmed the causal agent to be Scytalidium cuboideum, a first record of this disease in shiitake billets in Bhutan. In failed billets, it was observed that there was no growth of shiitake mycelium from the spawn. Trials suggest that S. cuboideum is responsible for the occurrences of poor colonisation by shiitake spawn--even when pink staining is not evident. Warm temperatures favour S. cuboideum, with shiitake able to sometimes overcome low rates of infection when incubated at low temperatures. [ABSTRACT FROM AUTHOR]

Pink staining in shiitake (Lentinula edodes (Berk.) Pegler) billets (Quercus griffithii Hook.f. and Thomson ex Miq.) causing the inhibition of shiitake mycelium and a reduction in shiitake production was observed throughout Bhutan in 2017–2018. Completion of Koch’s postulates confirmed the causal agent to be Scytalidium cuboideum, a first record of this disease in shiitake billets in Bhutan. In failed billets, it was observed that there was no growth of shiitake mycelium from the spawn. Trials suggest that S. cuboideum is responsible for the occurrences of poor colonisation by shiitake spawn—even when pink staining is not evident. Warm temperatures favour S. cuboideum, with shiitake able to sometimes overcome low rates of infection when incubated at low temperatures.

The cause of peanut kernel shrivel (PKS) syndrome, affecting peanut crops in Australia’s growing regions, is currently unknown. It is estimated that PKS is costing the peanut industry more than AUD 5 M p.a. and is a potential threat to the industry. Previous investigations have ruled out all abiotic factors and most biotic factors as the cause of PKS. This research aimed at investigating the scope, distribution, and cause of the PKS syndrome. The survey showed PKS symptoms to be present in peanut crops in all the growing regions surveyed. Based on our study of culturable microorganisms, there appears to be no clear-cut involvement of plant pathogenic bacteria and fungi; however, Fusarium spp. were revealed as the most prevalent fungi in affected plants. Moreover, the soil metagenomics study revealed Fusarium spp. as the most abundant fungal communities in the soil microbial profile, and they could contribute to the PKS syndrome. The consistent presence observed of the identified Fusarium oxysporum in PKS-affected samples could indicate a role for this pathogen in the syndrome, especially in conjunction with abiotic stressors. The pathogenicity testing of F. oxysporum resulted in very mild PKS symptoms. A separate report suggesting the involvement of phytoplasma in the PKS syndrome raises the possibility of an interplay of biotic factors in the development of this disease. Further investigation is warranted to determine the true cause or causes of this disease.

Abstract Background For more than a century, the Koch’s postulates have been the golden rule for determining the causative agents in diseases. However, in cases of multiple pathogens-one disease, in which different pathogens can cause the same disease, the selection of microorganisms that regress infection is hard when Koch’s postulates are applied. Microbiome approaches can obtain relatively complete information about disease-related microorganisms and can guide the selection of target microorganisms for regression infection. In the present study, whitish muscle syndrome (WMS) of Scylla paramamosain, which has typical symptoms with whitish muscle and blackened hemolymph was used as an example to establish a new research strategy that integrates microbiome approaches and Koch’s postulates to determinate causative agents of multiple pathogens-one disease. Results Microbiome results revealed that Aeromonas, Acinetobacter, Shewanella, Chryseomicrobium, Exiguobacterium, Vibrio and Flavobacterium, and Kurtzmaniella in hemolymph were bacterial and fungal indicators for WMS. A total of 23 bacteria and 14 fungi were isolated from hemolymph and muscle tissues, and among the bacteria, Shewanella chilikensis, S. xiamenensis, Vibrio alginolyticus, S. putrefaciens, V. fluvialis, and V. parahaemolyticus were present in hemolymph and/or muscle tissues in each WMS crab, and the last three species were also present in three Healthy crabs. The target bacteria and fungi were further screened to regression infections based on two criteria: whether they belonged to the indicator genera for WMS, whether they were isolated from both hemolymph and muscle tissues in most WMS crabs. Only S. chilikensis, S. putrefaciens, S. xiamenensis, V. alginolyticus, V. fluvialis, and V. parahaemolyticus met both two criteria. The six bacteria that met both two criteria and six fungi and another bacterium that unmatched any of two criteria were used to perform regression infection experiments based on Koch’s postulates. S. chilikensis, S. putrefaciens, S. xiamenensis, V. alginolyticus, V. fluvialis, and V. parahaemolyticus met both two criteria, and the results indicate that they cause WMS in crabs independently. Conclusions This study fully demonstrated that our research strategy that integrates the microbiome and Koch’s postulates can maximize the ability to catch pathogens in one net for the situation of multiple pathogens-one disease. Video Abstract

Grapevine leafroll disease (GLRD) is the most globally prevalent and destructive disease complex responsible for significant reductions in grape yield and quality as well as wine production. GLRD is associated with several positive-strand RNA viruses of the family Closteroviridae, designated as grapevine leafroll-associated viruses (GLRaVs). However, the specific etiological role of any of these GLRaVs in GLRD has not been demonstrated. Even though GLRaV-3 is considered the chief GLRD agent, little is known about the molecular, cellular, and pathological properties of this virus. Such a knowledge gap is due to multiple factors, including the unavailability of biologically active virus cDNA clones and the lack of reliable experimental systems for launching grapevine infection using such clones. In this work, we tested four methods for inoculating tissue-cultured grapevine plantlets with cDNA clones of GLRaV-3: (i) vacuum agro-infiltration; (ii) agro-pricking; (iii) agro-drenching; and (iv) agro-injection. We showed that vacuum agro-infiltration was the most effective of these methods. Furthermore, we examined the impacts of different experimental conditions on the survival and infectivity rate of grapevines after infiltration. To verify the infectivity rate for different treatments, we used RT-PCR, RT-qPCR, and Western blotting. We found that humidity plays a critical role in the survival of plantlets after agro-infiltration and that the use of RNA silencing suppressor and dormancy treatment both had strong effects on the infection rates. To our knowledge, the experimental protocol reported herein is the most effective system for launching the infection of grapevine using cDNA clones of grapevine viruses featuring up to a 70% infection rate. This system has strong potential to facilitate grapevine virology research including the fulfillment of Koch's postulates for GLRD and other major virus diseases as well as identifying the molecular, cellular, and pathological properties of GLRaVs and, potentially, other important grapevine viruses. [ABSTRACT FROM AUTHOR]

This study aimed to identify fungal species associated with trunk diseases of sweet cherries (Prunus avium) in several commercial cherry orchards in Beijing, Guizhou and Shandong provinces, China. In total, eighteen fungal strains that fitted well into the species concept of Diaporthe were isolated. Based on both morphological and multi-locus phylogenetic analyses of internal transcribed spacer region (ITS), beta-tubulin (tub-2), calmodulin (Cal) and translation elongation factor 1-α (tef1–α) sequencing data, fourteen isolates were identified as Diaporthe eres, while four isolates were classified as D. hongkongensis. Here, we report D. hongkongensis causing sweet cherry branch dieback disease and, further, we confirmed the host association of D. eres with sweet cherries in China. A pathogenicity assay revealed the ability of both D. eres and D. hongkongensis to cause shoot necrosis and stem lesions on Prunus avium cv. 'Brooks' (mean lesion lengths of 1.86 cm and 1.56 cm, respectively). The optimal temperature for the growth of both Diaporthe species was tested. The optimal growth temperature for D. hongkongensis was 30 °C, and the 25–28 °C temperatures were the most favorable for the growth of D. eres strains. This research advances the understanding of fungal trunk diseases in fruit crops, particularly gummosis and branch dieback disease in Chinese cherry orchards, and will aid growers in making decisions about cultural practices and disease management. [ABSTRACT FROM AUTHOR]

Background: For more than a century, the Koch's postulates have been the golden rule for determining the causative agents in diseases. However, in cases of multiple pathogens-one disease, in which different pathogens can cause the same disease, the selection of microorganisms that regress infection is hard when Koch's postulates are applied. Microbiome approaches can obtain relatively complete information about disease-related microorganisms and can guide the selection of target microorganisms for regression infection. In the present study, whitish muscle syndrome (WMS) of Scylla paramamosain, which has typical symptoms with whitish muscle and blackened hemolymph was used as an example to establish a new research strategy that integrates microbiome approaches and Koch's postulates to determinate causative agents of multiple pathogens-one disease. Results: Microbiome results revealed that Aeromonas, Acinetobacter, Shewanella, Chryseomicrobium, Exiguobacterium, Vibrio and Flavobacterium, and Kurtzmaniella in hemolymph were bacterial and fungal indicators for WMS. A total of 23 bacteria and 14 fungi were isolated from hemolymph and muscle tissues, and among the bacteria, Shewanella chilikensis, S. xiamenensis, Vibrio alginolyticus, S. putrefaciens, V. fluvialis, and V. parahaemolyticus were present in hemolymph and/or muscle tissues in each WMS crab, and the last three species were also present in three Healthy crabs. The target bacteria and fungi were further screened to regression infections based on two criteria: whether they belonged to the indicator genera for WMS, whether they were isolated from both hemolymph and muscle tissues in most WMS crabs. Only S. chilikensis, S. putrefaciens, S. xiamenensis, V. alginolyticus, V. fluvialis, and V. parahaemolyticus met both two criteria. The six bacteria that met both two criteria and six fungi and another bacterium that unmatched any of two criteria were used to perform regression infection experiments based on Koch's postulates. S. chilikensis, S. putrefaciens, S. xiamenensis, V. alginolyticus, V. fluvialis, and V. parahaemolyticus met both two criteria, and the results indicate that they cause WMS in crabs independently. Conclusions: This study fully demonstrated that our research strategy that integrates the microbiome and Koch's postulates can maximize the ability to catch pathogens in one net for the situation of multiple pathogens-one disease. EgKJnBsvf5xgzELdpRDt_Q Video Abstract [ABSTRACT FROM AUTHOR]

Symptoms of foliar blight were observed on turfgrass in Oran (Algeria), including yellow chlorotic patches on leaves during the 2020 summer (temperatures between 35 and 40°C). Symptoms extended downward from leaf tips and entire leaves became blighted, leading to irregular discoloured areas that later turned brown. Isolations from infected plants included 214 isolates identified as Curvularia or Bipolaris, based on morphological traits. Other isolates included Fusarium, Myrothecium and Acremonium spp. Three molecular loci, ITS rDNA, gpd and tef1, were amplified and sequenced. Morphological and multi-locus phylogenetic analyses revealed four fungal species viz. B. sorokiniana, C. spicifera, C. verruculosa, C. geniculata, and two additional Curvularia lineages, some of these fungi are reported are first records for Algeria. Koch's postulates were confirmed by inoculating potted turfgrass with spore suspensions of 16 isolates and re-isolating of the inoculated pathogens from symptomatic tissues. Bipolaris sorokiniana was the most virulent pathogen causing numerous foliar necrotic lesions similar to those observed in the field. Other isolates infected basal leaves only, and caused less severe symptoms. The results show that Curvularia species may be secondary pathogens infecting stressed plants, and that simultaneous occurrence of high temperatures and poor water quality have influenced disease progression. Correct identification of these pathogens is important for applying appropriate and timely disease management. [ABSTRACT FROM AUTHOR]

Phoenix dactylifera L. (Date palm) is one of the leading perennial fruit crops regarding its nutritional profile, medicinal significance, excellent yields, long life, and high adaptability to diverse climatic conditions of semi-arid and arid regions globally. Eleven major palm-growing districts of Punjab, Sindh, and Khyber Pakhtunkhwa were surveyed for wilt disease assessment on date palms. The date palm plantation was highly affected with Fusarium wilt diseases with 55%, the incidence in Khairpur (Sindh province), followed by Bahawalpur at 47.4% (Punjab Province) and 45% in Dera Ismail Khan (KPK province). Disease severity was recorded as 65% in Khairpur, 55% in Bahawalpur, and 50% in Dera Ismail Khan (Sindh, Punjab, and KPK, respectively). Sixty-three Fusarium isolates were obtained from three hundred and eighty (380) diseased samples, and thirty isolates randomly selected out of 63 were studied for morpho-cultural and microscopic characterization. Morpho-cultural characters, i.e., colony pigmentation, growth pattern, conidia (macro and microconidia), shape, apical cell shape, and phalidies studied, coinciding with typical characteristics of Fusarium oxysporum. Twenty isolates of the Fusarium population were subjected to the ITS region sequence data-based phylogenetic network. The Neighbor net algorithm computed an unrooted phylogenetic network displayed as four branched genetic network that depicted the division of twenty fungal isolates into four groups. F. oxysporum (FMB-FO-PD-011) inoculated onto healthy symptomless detached date palm leaves, seedlings, and suckers under controlled and field conditions showed typical wilt disease symptoms. Hence this study proved that F. oxysporum is the potential cause of date palm wilt in Pakistan. [ABSTRACT FROM AUTHOR]

Leaf-blight disease caused by the Fusarium oxysporum is an emerging problem in Dendrobium chrysotoxum production in China. Symptoms of leaf blight were observed on seedlings of D. chrysotoxum cultivated in a nursery in Ruili City, Yunnan Province, China. In this study, we isolated the Fusarium sp. associated with leaf-blight disease of D. chrysotoxum from the diseased seedlings. A pathogenicity test was performed to fulfill Koch’s postulates to confirm the pathogenicity of isolated strains and identified using morphological and molecular techniques. The results revealed that all four isolated Fusarium sp. isolates (DHRL-01~04) produced typical blight symptoms followed by marginal necrosis of leaves on the D. chrysotoxum plants. On the PDA medium, the fungal colony appeared as a white to purplish color with cottony mycelium growth. Microconidia are oval-shaped, whereas macroconidia are sickle-shaped, tapering at both ends with 2–4 septations. The phylogenetic trees were construed based on internal transcribed spacer (ITS), translation elongation factor (EF-1α), and RNA polymerase subunit genes RPB1 and RPB2 genes, respectively, and blasted against the NCBI database for species confirmation. Based on the NCBI database’s blast results, the isolates showed that more than 99% identify with Fusarium oxysporum. To our knowledge, this is the first comprehensive report on the identification of Fusarium oxysporum as the causal agent of Dendrobium chrysotoxum leaf blight in Yunnan Province, China, based on morphological and molecular characteristics.

Tea (Camellia sinensis (L.) Kuntze) is one of the most important economic plants in China, and has many benefits for human health. Anthracnose is one of the most serious diseases of tea in China, and control of the fungus is important since most Chinese cultivars are susceptible to it. The agent of tea anthracnose was initially described as Gloeosporium theae-sinensis I. Miyake in Japan, which was later transferred to Discula, but this taxonomic position remains problematic. To shed light on these taxonomic and phylogenetic issues, the tea anthracnose pathogens were re-studied. Combining the morphological characteristics and a multigene phylogenetic analysis of nrITS, nrLSU, rpb2, and tef1 sequence data, a new genus Sinodiscula was proposed to accommodate the causal fungi of tea anthracnose, including a new species Sinodiscula camellicola and a new combination Sinodiscula theae-sinensis. Furthermore, the pathogenicity of the pathogens was determined according to Koch’s postulates. This study thoroughly resolves the long-standing taxonomic and phylogenetic problems of the tea anthracnose pathogens.

White mold of cabbage caused by Sclerotinia sclerotiorum was identified in Bangladesh Agricultural Research Institute (BARI), Bangladesh. The fungus was isolated from the symptomatic leaves of cabbage on potato dextrose agar (PDA) medium. Subsequent inoculation of S. sclerotiorum on detached cabbage leaves and cabbage heads confirmed the pathogenicity of fungus. Based on morphological characteristics and phylogenetic analysis, the pathogen was identified as Sclerotinia sclerotiorum. [ABSTRACT FROM AUTHOR]

This study aimed to identify fungal species associated with trunk diseases of sweet cherries (Prunus avium) in several commercial cherry orchards in Beijing, Guizhou and Shandong provinces, China. In total, eighteen fungal strains that fitted well into the species concept of Diaporthe were isolated. Based on both morphological and multi-locus phylogenetic analyses of internal transcribed spacer region (ITS), beta-tubulin (tub-2), calmodulin (Cal) and translation elongation factor 1-α (tef1–α) sequencing data, fourteen isolates were identified as Diaporthe eres, while four isolates were classified as D. hongkongensis. Here, we report D. hongkongensis causing sweet cherry branch dieback disease and, further, we confirmed the host association of D. eres with sweet cherries in China. A pathogenicity assay revealed the ability of both D. eres and D. hongkongensis to cause shoot necrosis and stem lesions on Prunus avium cv. ‘Brooks’ (mean lesion lengths of 1.86 cm and 1.56 cm, respectively). The optimal temperature for the growth of both Diaporthe species was tested. The optimal growth temperature for D. hongkongensis was 30 °C, and the 25–28 °C temperatures were the most favorable for the growth of D. eres strains. This research advances the understanding of fungal trunk diseases in fruit crops, particularly gummosis and branch dieback disease in Chinese cherry orchards, and will aid growers in making decisions about cultural practices and disease management.

Primary biliary cholangitis (PBC) is an autoimmune liver disease characterized by the production of diagnostic antimitochondrial antibodies (AMA) reactive to the pyruvate dehydrogenase complex. A human betaretrovirus (HBRV) resembling mouse mammary tumor virus has been characterized in patients with PBC. However, linking the viral infection with the disease is not a straight-forward process because PBC is a complex multifactorial disease influenced by genetic, hormonal, autoimmune, environmental, and other factors. Currently, PBC is assumed to have an autoimmune etiology, but the evidence is lacking to support this conjecture. In this review, we describe different approaches connecting HBRV with PBC. Initially, we used co-cultivation of HBRV with biliary epithelial cells to trigger the PBC-specific phenotype with cell surface expression of cryptic mitochondrial autoantigens linked with antimitochondrial antibody expression. Subsequently, we have derived layers of proof to support the role of betaretrovirus infection in mouse models of autoimmune biliary disease with spontaneous AMA production and in patients with PBC. Using Hill's criteria, we provide an overview of how betaretrovirus infection may trigger autoimmunity and propagate biliary disease. Ultimately, the demonstration that disease can be cured with antiviral therapy may sway the argument toward an infectious disease etiology in an analogous fashion that was used to link H. pylori with peptic ulcer disease. [ABSTRACT FROM AUTHOR]

Inflammatory bowel disease (IBD) is a chronic non-specific inflammatory disease that occurs in the intestinal tract. It is mainly divided into two subtypes, i.e., the Crohn’s disease (CD) and ulcerative colitis (UC). At present, its pathogenesis has not been fully elucidated, but it has been generally believed that the environment, immune disorders, genetic susceptibility, and intestinal microbes are the main factors for the disease pathogenesis. With the development of the sequencing technology, microbial factors have received more and more attention. The gut microbiota is in a state of precise balance with the host, in which the host immune system is tolerant to immunogenic antigens produced by gut commensal microbes. In IBD patients, changes in the balance between pathogenic microorganisms and commensal microbes lead to changes in the composition and diversity of gut microbes, and the balance between microorganisms and the host would be disrupted. This new state is defined as dysbiosis. It has been confirmed, in both clinical and experimental settings, that dysbiosis plays an important role in the occurrence and development of IBD, but the causal relationship between dysbiosis and inflammation has not been elucidated. On the other hand, as a classic research method for pathogen identification, the Koch’s postulates sets the standard for verifying the role of pathogens in disease. With the further acknowledgment of the disease pathogenesis, it is realized that the traditional Koch’s postulates is not applicable to the etiology research (determination) of infectious diseases. Thus, many researchers have carried out more comprehensive and complex elaboration of Koch’s postulates to help people better understand and explain disease pathogenesis through the improved Koch’s postulates. Therefore, focusing on the new perspective of the improved Koch’s postulates is of great significance for deeply understanding the relationship between dysbiosis and IBD. This article has reviewed the studies on dysbiosis in IBD, the use of microbial agents in the treatment of IBD, and their relationship to the modified Koch’s postulates.

El encrespamiento amarillo (EA), la enfermedad más severa detectada en Argentina, causa síntomas y daños en cultivos de batata en toda la región productora. Desde 2010/11 se ha detectado la presencia de cuatro virus en plantas sintomáticas del cv. Arapey INIA recolectadas en lotes de Colonia Caroya, provincia de Córdoba. Los virus son sweet potato feathery mottle virus (SPFMV) y sweet potato virus G (SPVG), dos potyvirus transmitidos de forma no persistente por Myzus persicae; un closterovirus: sweet potato chlorotic stunt virus (SPCSV) y un geminivirus: sweet potato leaf curl virus (SPLCV), ambos transmitidos por Bemisia tacabi de manera semipersistente y persistente, respectivamente. Los objetivos de este trabajo fueron aislar e identificar el o los virus involucrados en la enfermedad EA de la batata y determinar la combinación de virus que reproduce la sintomatología de EA Solo la combinación de los cuatro virus permitió reproducir la sintomatología más severa del encrespamiento amarillo observada a campo en dicho genotipo. Los síntomas incluyen clorosis, achaparramiento, mosaico, ampollado, enrulado de la hoja, manchas cloróticas, diseños cloróticos, reducción y distorsión del área foliar, bordes de la hoja curvados hacia arriba. La presencia de cada uno de los virus se detectó mediante pruebas serológicas (DAS, NCM y TAS-ELISA) y moleculares (PCR). Se concluye que la interacción de SPFMV, SPVG, SPCSV y SPLCV es necesaria para el desarrollo de EA. [ABSTRACT FROM AUTHOR]

Abstract Pennisetum plants (Pennisetum alopecuroides L.), displaying a dwarfing phenotype along with delayed flowering and mosaic symptom on leaves, were found in Beijing, China. Flexuous filamentous particles with a size of approximate 15 × 850 nm were observed in symptomatic leaves via transmission electron microscopy. Deep sequencing of small RNAs (sRNA) from symptomatic leaves and analysis of sRNA populations were then conducted to determine the genome sequence of the viral agent in diseased plant tissues. It showed that the viral agent had one positive-sense and single-stranded RNA genome, which consisted of 9717 nucleotides (nts) excluding poly(A) tail. The complete viral genome contained a large open reading frame, encoding a polyprotein of 3131 amino acids (aa). Sequence comparison and phylogenetic analysis demonstrated that the viral agent belonged to the genus Potyvirus in the family Potyviridae. In the cladogram it was most closely related to johnsongrass mosaic virus, sharing 72% nt and 65% aa sequence identity. This viral agent was provisionally named pennisetum alopecuroides mosaic virus (PalMV). Subsequently, it was confirmed that PalMV is the causal agent of this new disease in P. alopecuroides by Koch’s postulates and reverse transcription-polymerase chain reaction analysis. Moreover, maize, millet, wheat, sorghum and rice plants were experimentally infected by PalMV via rub inoculation. Consequently, we proposed that PalMV could be a potentially dangerous virus threating a wide range of cereal crops.

Tsukamurella , a group of multi-drug resistant, Gram-positive, aerobic, and partially acid-fast bacteria, are emerging causes of bacterial conjunctivitis and keratitis. However, the pathogenesis of Tsukamurella keratitis is largely unknown. To address this, we used New Zealand White rabbits to develop the first eye infection model and conducted in vitro tests to study the pathogenesis mechanisms of Tsukamurella . There is increasing evidence that biofilms play a significant role in ocular infections, leading us to hypothesize that biofilm formation is crucial for effective Tsukamurella infection. In order to look for potential candidate genes which are important in biofilm formation and Tsukamurella keratitis. We performed genome sequencing of two ocular isolates, T. pulmonis -PW1004 and T. tyrosinosolvens -PW899, to identify potential virulence factors. Through in vitro and in vivo studies, we characterized their biological roles in mediating Tsukamurella keratitis. Our findings confirmed that Tsukamurella is an ocular pathogen by fulfilling Koch's postulates, and using genome sequence data, we identified tmytC, encoding a mycolyltransferase, as a crucial gene in biofilm formation and causing Tsukamurella keratitis in the rabbit model. This is the first report demonstrating the novel role of mycolyltransferase in causing ocular infections. Overall, our findings contribute to a better understanding of Tsukamurella pathogenesis and provide a potential target for treatment. Specific inhibitors targeting TmytC could serve as an effective treatment option for Tsukamurella infections.

We argue that Koch's postulates are best understood within an interventionist account of causation, in the sense described in Woodward (2003). We show how this treatment helps to resolve interpretive puzzles associated with Koch's work and how it clarifies the different roles the postulates play in providing useful, yet not universal criteria for disease causation. Our paper is an effort at rational reconstruction; we attempt to show how Koch's postulates and reasoning make sense and are normatively justified within an interventionist framework and more difficult to understand within alternative frameworks for thinking about causation.

Citrus wilt is a disease of recent appearance in the northern area of Veracruz that causes economic losses to producers in the region. The present work aimed to identify the causative agent of this disease and evaluate different fungicides to determine its in vitro sensitivity. A fungus was consistently isolated in plants with wilt symptoms; it was morphologically identified in different culture media and molecularly identified by PCR using the EF1-728F/EF1-986R primers. The fungus was inoculated in three varieties of citrus under greenhouse conditions. The sensitivity test was carried out with the fungicides chlorothalonil, benomyl thiabendazole, prochloraz, and a biological agent (Bacillus subtilis) at different concentrations, plus a negative control. Fusarium sp. (Accession No. MW438335) was morphologically and molecularly identified as the causal agent of vascular wilt in citrus fruits, causing growth retardation, decreased number of roots, wilting of the apical bud, and necrosis in the vascular system of the three varieties inoculated. The most effective fungicides in inhibiting mycelial growth were thiabendazole, prochloraz, and the biological agent Bacillus subtilis. [ABSTRACT FROM AUTHOR]

Abstract Purpose Aphis gossypii and A. punicae are the most damaging pests. The emergence of large populations has created concern among farmers because this pest complex is considered critical as it has a significant effect on major crops around the globe. The lack of new technologies for the control of A. gossypii and A. punicae is also worrying due to the indiscriminate use of chemical insecticides. Besides, this leads to the rapid development of resistance, which strangles their control in the field. Hence, there is a dire need to find the effective biocontrol agent for the management of Aphis gossypii and A. punicae. Methods The present investigation emphasizes the isolation and characterization of Aphidicidal Bacillus thuringiensis from the Andaman and Nicobar Islands, Karnataka, Assam, Arunachal Pradesh, and Sikkim soil samples. Phase contrast and scanning electron microscopy analysis used to characterize and identify the crystal morphology. Molecular profiling of Bt cry genes was determined by PCR using aphidicidal cry gene-specific primers, and molecular cloning and sequencing were carried out. Protein profiling by SDS–PAGE analysis was further studied. Finally, a qualitative bioassay of insecticidal activity was carried out against A. gossypii and A. punicae. Results A total of 65 Bacillus-like colonies were screened; 15 putative Aphidicidal B. thuringiensis isolates were identified based on morphological as well as through microbiological studies. Spherical and amorphous crystal inclusion was predominantly present in 34.28% of the Bt isolates. Crystal protein profiling of Bt isolates by SDS–PAGE analysis showed the presence 130, 73, 34, 25, and 13 kDa bands, among which 50−66 kDa bands were present abundantly. The detection of the cry gene of these isolates was done by PCR analysis, which indicated that cry1, cry2A, cry3A, and cry11A were on plasmid DNA. All cry genes were 80–100% homologous when aligned on alignment tool NCBI-BLASTn and BLASTp. All isolates of Bt were tested for their insecticidal activity against aphids. Three of the 15 isolates are Aphidicidal toxin specific by PCR analysis which were observed to be toxic to Aphis gossypii and A. punicae at a concentration of 35 μg/mL. The observed physical changes were induced by B. thuringiensis infection; these strains had been re-isolated from the dead aphids, and the presented results fulfilled Koch’s postulates. Conclusion The present study brought promising Bt isolates, primarily capable of creating an efficient biocontrol agent for Aphis gossypii and A. punicae and various sucking pests soon.

Verticillium spp., including V. nonalfalfae and V. dahliae, are known vascular wilt pathogens of the invasive Ailanthus altissima (tree-of-heaven) in the United States and in Europe. Herein we provide evidence of the presence of a previously unreported wilt disease of A. altissima in Tuscany (Central Italy). Several isolates were collected from two locations and identified as V. dahliae, based on microscopical features of conidiophores, conidia and microsclerotia. Genomic DNA was extracted from the mycelium, the ITS region was amplified and the sequence was deposited in GenBank as VdGL16 (accession no. MK474459). BLASTn analysis showed 100% similarity with V. dahliae. To confirm pathogenicity of VdGL16, inoculations of Ailanthus seedlings were performed with the root dipping technique whereas mature trees were stem-inoculated. All inoculated seedlings exhibited wilt symptoms after 20 days, while mature Ailanthus trees showed wilting and dieback after six months. The pathogen was easily re-isolated from seedlings and re-identified as V. dahliae, thus satisfying Koch’s postulates. Results from intraspecific resistance screening of nine seed sources from across Italy revealed that Ailanthus provenances from all the six sampled regions were susceptible to V. dahliae. Stem inoculated adult plants exhibited abundant production of epicormic sprouts along the stem within six months, and most of these sprouts wilted following initial dieback of the main stem; furthermore, sprouting from the crown was intense. Petioles and rachises tissues of leaves fallen from infected trees were a good source for re-isolation of the pathogen; we proved that such petioles and rachises can effectively transfer the fungus to healthy Ailanthus seedlings via root infections. Host-specificity of the V. dahliae isolate VdGL16 was also determined on 40 non-target species/varieties/cultivars. The isolate caused disease in herbaceous species belonging to five botanical families: Asteraceae, Lamiaceae, Leguminoseae, Linaceae and Solanaceae. Given the difficulties in countering Ailanthus invasion with mechanical and chemical methods, the biological control using Verticillium may provide an efficient, low cost and sustainable control of this invasive species.

Anthracnose is one of the major problems for cultivating many crops, including vegetables, fruits, and trees. It is a continual threat for fruits grower worldwide. Colletotrichum fructicola was isolated from Shine Muscat berries showing typical anthracnose symptom in Korea. It was identified as C. fructicola based on morphology, pathological signs and concatenated sequences of internal transcribed spacer region of rDNA, glyceraldehyde-3-phosphate dehydrogenase, β-tubulin-2, chitin synthase-1, calmodulin, and the Apn2-Mat1-2 intergenic spacer and partial mating type (Mat1-2) gene. To the best of our knowledge, this is the first report first report of anthracnose of Shine Muscat caused by C. fructicola in Korea.

“Hongtuozhusun” (Phallus rubrovolvatus) is an important edible and medicinal mushroom endemic to Southwest China. However, yellow rot disease is a severe disease of P. rubrovolvatus that occurs extensively in Guizhou Province. It has caused major economic losses and hinders the development of the P. rubrovolvatus industry. In this study, 28 microorganism strains were isolated from diseased fruiting bodies of P. rubrovolvatus at various stages, two of which were confirmed to be pathogenic based on Koch’s postulates. These two strains are introduced herein as Saccharomycopsis phalluae sp. nov. based on morphological, physiological, and molecular analysis. We reported a high-quality de novo sequencing and assembly of the S. phalluae genome using singlemolecule real-time sequencing technology. The whole genome was approximately 14.148 Mb with a G+C content of 43.55%. Genome assembly generated 8 contigs with an N50 length of 1,822,654 bp. The genome comprised 5966 annotated protein-coding genes. This is the first report of mushroom disease caused by Saccharomycopsis species. We expect that the information on genome properties, particularly in pathogenicity-related genes, assist in developing effective control measures in order to prevent severe losses and make amendments in management strategies. [ABSTRACT FROM AUTHOR]

Medical practice is increasingly coming under the guidance of statistical-mathematical models that are, undoubtedly, valuable tools but are also only a partial representation of reality. Indeed, given that statistics may be more or less adequate, a model is still a subjective interpretation of the researcher and is also influenced by the historical context in which it operates. From this opinion, I will provide a short historical excursus that retraces the advent of probabilistic medicine as a long process that has a beginning that should be sought in the discovery of the complexity of disease. By supporting the belonging of this evolution to the scientific domain it is also acknowledged that the underlying model can be imperfect or fallible and, therefore, confutable as any product of science. Indeed, it seems non-trivial here to recover these concepts, especially today where clinical decisions are entrusted to practical guidelines, which are a hybrid product resulting from the aggregation of multiple perspectives, including the probabilistic approach, to disease. Finally, before the advent of precision medicine, by limiting the use of guidelines to the original consultative context, an aged approach is supported, namely, a relationship with the individual patient.

Primary biliary cholangitis (PBC) is an autoimmune liver disease characterized by the production of diagnostic antimitochondrial antibodies (AMA) reactive to the pyruvate dehydrogenase complex. A human betaretrovirus (HBRV) resembling mouse mammary tumor virus has been characterized in patients with PBC. However, linking the viral infection with the disease is not a straight-forward process because PBC is a complex multifactorial disease influenced by genetic, hormonal, autoimmune, environmental, and other factors. Currently, PBC is assumed to have an autoimmune etiology, but the evidence is lacking to support this conjecture. In this review, we describe different approaches connecting HBRV with PBC. Initially, we used co-cultivation of HBRV with biliary epithelial cells to trigger the PBC-specific phenotype with cell surface expression of cryptic mitochondrial autoantigens linked with antimitochondrial antibody expression. Subsequently, we have derived layers of proof to support the role of betaretrovirus infection in mouse models of autoimmune biliary disease with spontaneous AMA production and in patients with PBC. Using Hill’s criteria, we provide an overview of how betaretrovirus infection may trigger autoimmunity and propagate biliary disease. Ultimately, the demonstration that disease can be cured with antiviral therapy may sway the argument toward an infectious disease etiology in an analogous fashion that was used to link H. pylori with peptic ulcer disease.

To identify the agent of black scab on prickly pear (Opuntia ficus-indica), the fungi associated with the disease were isolated and morphologically identified. The study was carried out from cladodes with characteristic symptoms of the disease collected in the town of Santa Cecilia Clavijero, in the municipality of San Juan Ixcaquixtla, Puebla. The fungi associated with the disease were identified as: Cladosporium cladosporioides, Aplosporella hesperidica and Didymella glomerata. To demonstrate their pathogenicity, the fungi were inoculated individually and in their combinations in six-month-old cladodes. Symptoms started 50 days after inoculation. The inoculation method that allowed the reproduction of the symptoms was the injection of a suspension at a concentration of 6 × 10³ conidia mL-1 of the mixture of the three fungi; cladodes inoculated with one and two fungi, did not result in the development of the disease. C. cladosporioides, A. hesperidica and D. glomerata are the causative agents of the nopal black scab, this being the first report of these pathogens in the nopal cultivation. [ABSTRACT FROM AUTHOR]

To elucidate the symptoms and pathogens diversity of corn Fusarium sheath rot (CFSR), diseased samples were collected from 21 county-level regions in 12 prefecture-level districts of Sichuan Province from 2015 to 2018 in the present study. In the field, two symptom types appeared including small black spots with a linear distribution and wet blotches with a tawny or brown color. One hundred thirty-seven Fusarium isolates were identified based on morphological characteristics and phylogenetic analysis (EF1-α), and Koch's postulates were also assessed. The results identified the isolates as 8 species in the Fusarium genus, including F. verticillioides, F. proliferatum, F. fujikuroi, F. asiaticum, F. equiseti, F. meridionale, F. graminearum and F. oxysporum, with isolation frequencies of 30.00, 22.67, 15.33, 7.33, 6.00, 5.33, 3.33 and 1.33%, respectively. Fusariumverticillioides and F. proliferatum were the dominant and subdominant species, respectively. Two or more Fusarium species such as F. verticillioides and F. proliferatum were simultaneously identified at a mixed infection rate of 14.67% in the present study. The pathogenicity test results showed that F. proliferatum and F. fujikuroi exhibited the highest virulence, with average disease indices of 30.28 ± 2.87 and 28.06 ± 1.96, followed by F. equiseti and F. verticillioides, with disease indices of 21.48 ± 2.14 and 16.21 ± 1.84, respectively. Fusarium asiaticum, F. graminearum and F. meridonale showed lower virulence, with disease indices of 13.80 ± 2.07, 11.57 ± 2.40 and 13.89 ± 2.49, respectively. Finally, F. orysporum presented the lowest virulence in CFSR, with a disease index of 10.14 ± 1.20. To the best of our knowledge, this is the first report of F. fujikuroi, F. meridionale and F. asiaticum as CFSR pathogens in China. [ABSTRACT FROM AUTHOR]

The present study was conducted to identify and confirm the pathogenicity of the causal agent of a wet finger rot of Cavendish banana, a newly reported pre-harvest disease in commercial plantations in Moneragala and Badulla districts of Sri Lanka. Bacterial species were isolated from banana fingers at three stages of symptom development: initial, middle, and late. Isolated bacterial colonies were inoculated to healthy and mature banana fingers by two methods, namely peel and pulp inoculation, to confirm the pathogenicity by Koch's postulates. Re-isolated bacterial isolates from inoculated banana fingers showing typical symptoms were identified by biochemical tests and subjected to PCR amplification using universal primers of bacteria, B27F/1492R. Subsequent DNA sequencing of the PCR products and homology search identified the pathogen as a strain of Dickeya dadantii. Phylogenetic analysis with selected Dickeya spp. sequences available in databases identified that the D. dadantii strain of the present study groups with D. dadantii 3937 (CP002038.1), of which the complete genome has been sequenced. Cross infection studies revealed that D. dadantii isolate is more virulent on banana variety Anamalu than on Cavendish banana in terms of days taken to develop the symptoms. The findings identified the causal agent of wet finger rot of banana in the present study as a strain of D. dadantii and confirmed its pathogenicity. [ABSTRACT FROM AUTHOR]

Shell (cuticular) disease manifests in various forms and affects many crustaceans, including lobsters. Outbreaks of white leg disease (WLD) with distinct signs of pereiopod tissue whitening and death have been observed in cultured larvae (phyllosomas) of ornate spiny lobster Panulirus ornatus, eastern rock lobster Sagmariasus verreauxi, and slipper lobster Thenus australiensis. This study aimed to characterise and identify the causative agent of WLD through morphological and molecular (16S rRNA gene and whole genome sequencing) analysis, experimental infection of damaged/undamaged P. ornatus and T. australiensis phyllosomas, and bacterial community analysis (16S rRNA gene amplicon sequencing) of P. ornatus phyllosomas presenting with WLD during an outbreak. Bacterial communities of WLD-affected pereiopods showed low bacterial diversity and dominant abundance of Aquimarina spp. compared to healthy pereiopods, which were more diverse and enriched with Sulfitobacter spp. 16S rRNA gene Sanger sequencing of cultures from disease outbreaks identified the dominant bacterial isolate (TRL1) as a Gram-negative, long non-flagellated rod with 100% sequence identity to Aquimarina hainanensis. Aquimarina sp. TRL1 was demonstrated through comparative genome analysis (99.99% OrthoANIu) as the bacterium reisolated from experimentally infected phyllosomas presenting with typical signs of WLD. Pereiopod damage was a major predisposing factor to WLD. Histopathological examination of WLD-affected pereiopods showed masses of internalised bacteria and loss of structural integrity, suggesting that Aquimarina sp. TRL1 could enter the circulatory system and cause death by septicaemia. Aquimarina sp. TRL1 appears to have important genomic traits (e.g., tissue-degrading enzymes, gliding motility, and aggregate-promoting factors) implicated in the pathogenicity of this bacterium. We have shown that Aquimarina sp. TRL1 is the aetiological agent of WLD in cultured Palinurid and Scyllarid phyllosomas and that damaged pereiopods are a predisposing factor to WLD.

ABSTRACT Potomac horse fever (PHF), a severe and frequently fatal febrile diarrheal disease, has been known to be caused only by Neorickettsia risticii, an endosymbiont of digenean trematodes. Here, we report the cell culture isolation of a new Neorickettsia species found in two locations in eastern Ontario, Canada, in 2016 and 2017 (in addition to 10 variable strains of N. risticii) from N. risticii PCR-negative horses with clinical signs of PHF. Gene sequences of 16S rRNA and the major surface antigen P51 of this new Neorickettsia species were distinct from those of all previously characterized N. risticii strains and Neorickettsia species, except for those from an uncharacterized Neorickettsia species culture isolate from a horse with PHF in northern Ohio in 1991. The new Neorickettsia species nonetheless had the characteristic intramolecular repeats within strain-specific antigen 3 (Ssa3), which were found in all sequenced Ssa3s of N. risticii strains. Experimental inoculation of two naive ponies with the new Neorickettsia species produced severe and subclinical PHF, respectively, and the bacteria were reisolated from both of them, fulfilling Koch’s postulates. Serological assay titers against the new Neorickettsia species were higher than those against N. risticii. Whole-genome sequence analysis of the new Neorickettsia species revealed unique features of this bacterium compared with N. risticii. We propose to classify this new bacterium as Neorickettsia finleia sp. nov. This finding will improve the laboratory diagnosis of and vaccine for PHF, environmental risk assessment of PHF, and understanding of PHF pathogenesis and Neorickettsia biology in general. IMPORTANCE Despite the detection of Neorickettsia species DNA sequences in various trematode species and their hosts, only three Neorickettsia species have been cell culture isolated and whole-genome sequenced and are known to infect mammals and/or cause disease. The molecular mechanisms that enable the obligatory intracellular bacterium Neorickettsia to colonize trematodes and to horizontally transmit from trematodes to mammals, as well as the virulence factors associated with specific mammalian hosts, are unknown. Potomac horse fever (PHF) is a severe and acute systemic infectious disease of horses, with clinical signs that include diarrhea. Neorickettsia risticii is the only known bacterial species that causes PHF. Ingestion of insects harboring N. risticii-infected trematodes by horses leads to PHF. Our discovery of a new Neorickettsia species that causes PHF and whole-genome sequence analysis of this bacterium will improve laboratory diagnosis and vaccine development for PHF and will contribute to our understanding of Neorickettsia ecology, pathogenesis, and biology.

Previous studies reported that sagebrush plants near experimentally clipped neighbors experienced less herbivory than did plants near unclipped neighbors. Blocking air flow with plastic bags made this effect undetectable. However, some scientists remained skeptical about the possibility of volatile communication between plants since the existence and identity of a cue that operates in nature have never been demonstrated. We conducted an air transfer experiment that collected air from the headspace of an experimentally clipped donor plant and delivered it to the headspace of an unclipped assay plant. We found that assay plants treated with air from clipped donors were less likely to be damaged by naturally occurring herbivores in a field experiment. This simple air transfer experiment fulfills the most critical of Koch's postulates and provides more definitive evidence for volatile communication between plants. It also provides an inexpensive experimental protocol that can be used to screen plants for interplant communication in the field.

Camellia japonica is a native tree species with high economic value that is widely cultivated in southern China. In recent years, canker disease has been observed in camellia plantations in Zhejiang Province, China, with the disease incidence rate in some plantations exceeding 20%. Canker disease severely affects the trunks and branches of C. japonica in China, but the causal agent has not yet been identified. In this study, the pathogen was isolated from infected C. japonica tissues through a conventional tissue isolation approach. Species identification was conducted using morphological methods combined with multilocus phylogenetic analysis. Pathogenicity was tested based on Koch’s postulates. The results showed that the pathogen could be isolated from the diseased bark of C. japonica ‘Hongluzhen’. The pathogen was identified as Nectria pseudotrichia based on morphological, cultural, and molecular traits. The inoculation of the pathogen into C. japonica ‘Hongluzhen’ caused necrotic lesions on healthy seedlings, and the fungus N. pseudotrichia could be re-isolated from such lesions. Therefore, N. pseudotrichia is the causal agent of canker disease affecting C. japonica in China.

Shell (cuticular) disease manifests in various forms and affects many crustaceans, including lobsters. Outbreaks of white leg disease (WLD) with distinct signs of pereiopod tissue whitening and death have been observed in cultured larvae (phyllosomas) of ornate spiny lobster Panulirus ornatus , eastern rock lobster Sagmariasus verreauxi , and slipper lobster Thenus australiensis. This study aimed to characterise and identify the causative agent of WLD through morphological and molecular (16S rRNA gene and whole genome sequencing) analysis, experimental infection of damaged/undamaged P. ornatus and T. australiensis phyllosomas, and bacterial community analysis (16S rRNA gene amplicon sequencing) of P. ornatus phyllosomas presenting with WLD during an outbreak. Bacterial communities of WLD-affected pereiopods showed low bacterial diversity and dominant abundance of Aquimarina spp. compared to healthy pereiopods, which were more diverse and enriched with Sulfitobacter spp. 16S rRNA gene Sanger sequencing of cultures from disease outbreaks identified the dominant bacterial isolate (TRL1) as a Gram-negative, long non-flagellated rod with 100% sequence identity to Aquimarina hainanensis. Aquimarina sp. TRL1 was demonstrated through comparative genome analysis (99.99% OrthoANIu) as the bacterium reisolated from experimentally infected phyllosomas presenting with typical signs of WLD. Pereiopod damage was a major predisposing factor to WLD. Histopathological examination of WLD-affected pereiopods showed masses of internalised bacteria and loss of structural integrity, suggesting that Aquimarina sp. TRL1 could enter the circulatory system and cause death by septicaemia. Aquimarina sp. TRL1 appears to have important genomic traits (e.g., tissue-degrading enzymes, gliding motility, and aggregate-promoting factors) implicated in the pathogenicity of this bacterium. We have shown that Aquimarina sp. TRL1 is the aetiological agent of WLD in cultured Palinurid and Scyllarid phyllosomas and that damaged pereiopods are a predisposing factor to WLD. [ABSTRACT FROM AUTHOR]

Effective control disease measures require a great deal of knowledge about the pathogen, which is also applied to dieback disease of the nutmeg tree in Aceh Selatan District, Province of Aceh, Indonesia. The objectives of this study were to identify the causal agent of dieback disease on a nutmeg plantation and to assess the effect of drought stress on the development of dieback disease. Pathogen identification was carried out by observing the symptoms of infected plants, morphological characters of the isolated pathogens, Koch's postulates, and molecular analysis and pure pathogen cultures. The samples were taken from an infected part of the nutmeg plant (twigs, stems, and roots) in 6 subdistricts at the center of the nutmeg plantation in Aceh Selatan. Koch's postulates were performed by using the mycelia of pathogens on 1-year-old nutmeg seedlings. Drought stress was simulated everyday, and then once every 2 weeks, to study the effect of drought stress on dieback. The results showed that there were 4 genera of fungal pathogens (Botryodiplodia theobromae, Fusarium solani, Graphium euwallaceae, and Rigidoporus microporus) out of 6 fungi associated with dieback disease. However, B. theobromae Pat. turned out to be the dominant fungi associated with dieback on the nutmeg tree in Aceh Selatan. Drought stress was proven to be a predisposing factor for explaining this problem epidemic in Aceh Selatan. [ABSTRACT FROM AUTHOR]

Curvularia includes plant pathogenic species with a worldwide distribution and a wide host range, particularly cereals and grass (Poaceae). Chinese fir (Cunninghamia lanceolata) is an important afforestation tree species in southern China, with a high economic value. Leaf blight disease on Chinese fir was discovered in Hunan and Zhejiang, China, and two fungal species were found to be associated with the symptoms. Using morphological and phylogenetic approaches, they were identified as Curvularia spicifera and Curvularia muehlenbeckiae. They were determined to be the pathogens by fulfilling Koch's postulates. Both showed a pathogenicity on Citrus reticulata and Citrus tangerina, which have a similar geographic distribution to Chinese fir. The similar geographic distribution of these hosts may increase the risk of disease. Based on a literature review, leaf blight caused by C. spicifera and C. muehlenbeckiae is recorded here on a gymnosperm for the first time. [ABSTRACT FROM AUTHOR]

For the first time, Stagonosporopsis caricae is reported causing leaf spots on the non-conventional fruit crop Vasconcellea monoica (Caricaceae). The fungus was identified in Brazil based on morphological and molecular features and its pathogenicity was demonstrated. This is the first pathogen to be reported on V. monoica. [ABSTRACT FROM AUTHOR]

Anthracnose is one of the major problems for cultivating many crops, including vegetables, fruits, and trees. It is a continual threat for fruits grower worldwide. Colletotrichum fructicola was isolated from Shine Muscat berries showing typical anthracnose symptom in Korea. It was identified as C. fructicola based on morphology, pathological signs and concatenated sequences of internal transcribed spacer region of rDNA, glyceraldehyde-3-phosphate dehydrogenase, β-tubulin-2, chitin synthase-1, calmodulin, and the Apn2-Mat1-2 intergenic spacer and partial mating type (Mat1-2) gene. To the best of our knowledge, this is the first report first report of anthracnose of Shine Muscat caused by C. fructicola in Korea. [ABSTRACT FROM AUTHOR]