Multi-locus phylogenetic analyses of Colletotrichum gloeosporioides species complex causing crown rot on strawberry in Florida

Phylogenetic Analyses ◽

Management Strategies ◽

Crown Rot ◽

Cultivated Plants ◽

Complex Species ◽

Colletotrichum Gloeosporioides Species Complex ◽

Genomic Regions

Colletotrichum gloeosporioides is the causal agent of Colletotrichum crown rot of strawberry in the southern United States. Recent multi-gene studies defined C. gloeosporioides as a complex species comprised of 37 species. In our study, we phylogenetically characterized C. gloeosporioides isolates from strawberry and other non-cultivated plants around strawberry fields. One hundred and fifteen strawberry isolates and 38 isolates from non-cultivated hosts were sequenced for five genomic regions: internal transcribed spacer (ITS), actin (ACT), calmodulin (CAL), chitin synthase (CHS-1), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Phylogenetic analysis using the maximum likelihood and Bayesian inference methods, based on partition-specific models, revealed that most of the isolates in Florida (86%) were closely related to C. siamense, whereas 14 isolates were closely related to C. theobromicola (syn. C. fragariae), four isolates were C. fructicola, and three were C. clidemiae. However, only the first three species were pathogenic to strawberry. Morphological characteristics evaluated show that mycelial growth of all species is about 5 mm/day but colony morphology varies by species and incubation conditions. In vitro mating of the isolates demonstrated that C. fructicola is homothallic whereas C. siamense and C. theobromicola isolates are heterothallic. The biological importance of these different Colletotrichum species is currently being investigated to determine whether different management strategies are needed in strawberry production fields.

Two new species of Undifilum, fungal endophytes of Astragalus (locoweeds) in the United States

Botany ◽

10.1139/b2012-056 ◽

2012 ◽

Vol 90 (9) ◽

pp. 866-875 ◽

Cited By ~ 33

Author(s):

Deana L. Baucom ◽

Marie Romero ◽

Robert Belfon ◽

Rebecca Creamer

Keyword(s):

New Species ◽

New Mexico ◽

Random Amplified Polymorphic Dna ◽

Phylogenetic Analyses ◽

Fungal Endophytes ◽

Its Region ◽

The United States ◽

Neighbor Joining

New species of Undifilum , from locoweeds Astragalus lentiginosus Vitman and Astragalus mollissimus Torr., are described using morphological characteristics and molecular phylogenetic analyses as Undifilum fulvum Baucom & Creamer sp. nov. and Undifilum cinereum Baucom & Creamer sp. nov. Fungi were isolated from dried plants of A. lentiginosus var. araneosus , diphysus , lentiginosus , and wahweapensis collected from Arizona, Oregon, and Utah, USA, and A. mollissimus var. biglovii , earleii , and mollissimus collected from New Mexico, Oklahoma, and Texas, USA. Endophytic fungi from Astragalus locoweeds were compared to Undifilum oxytropis isolates obtained from dried plant material of Oxytropis lamberteii from New Mexico and Oxytropis sericea from Arizona, Colorado, New Mexico, Utah, and Wyoming. Extremely slow growth in vitro was observed for all, and conidia, if present, were ellipsoid with transverse septa. However, in vitro color, growth on four different media, and conidium size differed between fungi from Astragalus spp. and U. oxytropis. Neighbor-joining analyses of internal transcribed spacer (ITS) region and glyceraldehyde-3-phosphate dehydrogenase (GPD) gene sequences revealed that U. fulvum and U. cinereum formed a clade distinct from U. oxytropis. This was supported by neighbor-joining analyses of results generated from random amplified polymorphic DNA (RAPD) fragments using two different primers.

First Report of Leaf Spot Disease Caused by Colletotrichum gloeosporioides on Chinese Bean Tree in China

Plant Disease ◽

10.1094/pdis-03-12-0261-pdn ◽

2013 ◽

Vol 97 (1) ◽

pp. 138-138 ◽

Cited By ~ 1

Author(s):

B. Z. Fu ◽

M. Yang ◽

G. Y. Li ◽

J. R. Wu ◽

J. Z. Zhang ◽

...

Keyword(s):

Leaf Spot ◽

Disease Incidence ◽

Phylogenetic Analyses ◽

Leaf Spot Disease ◽

First Report ◽

Spot Disease ◽

Rdna Its ◽

Its Sequence Analysis

Chinese bean tree, Catalpa fargesii f. duciouxii (Dode) Gilmour, is an ornamental arbor plant. Its roots, leaves, and flowers have long been used for medicinal purposes in China. During July 2010, severe outbreaks of leaf spot disease on this plant occurred in Kunming, Yunnan Province. The disease incidence was greater than 90%. The symptoms on leaves began as dark brown lesions surrounded by chlorotic halos, and later became larger, round or irregular spots with gray to off-white centers surrounded by dark brown margins. Leaf tissues (3 × 3 mm), cut from the margins of lesions, were surface disinfected in 0.1% HgCl2 solution for 3 min, rinsed three times in sterile water, plated on potato dextrose agar (PDA), and incubated at 28°C. The same fungus was consistently isolated from the diseased leaves. Colonies of white-to-dark gray mycelia formed on PDA, and were slightly brown on the underside of the colony. The hyphae were achromatic, branching, septate, and 4.59 (±1.38) μm in diameter on average. Perithecia were brown to black, globose in shape, and 275.9 to 379.3 × 245.3 to 344.8 μm. Asci that formed after 3 to 4 weeks in culture were eight-spored, clavate to cylindrical. The ascospores were fusiform, slightly curved, unicellular and hyaline, and 13.05 to 24.03 × 10.68 to 16.02 μm. PCR amplification was carried out by utilizing universal rDNA-ITS primer pair ITS4/ITS5 (2). Sequencing of the PCR products of DQ1 (GenBank Accession No. JN165746) revealed 99% similarity (100% coverage) with Colletotrichum gloeosporioides isolates (GenBank Accession No. FJ456938.1, No. EU326190.1, No. DQ682572.1, and No. AY423474.1). Phylogenetic analyses (MEGA 4.1) using the neighbor-joining (NJ) algorithm placed the isolate in a well-supported cluster (>90% bootstrap value based on 1,000 replicates) with other C. gloeosporioides isolates. The pathogen was identified as C. gloeosporioides (Penz.) Penz. & Sacc. (teleomorph Glomerella cingulata (Stoneman) Spauld & H. Schrenk) based on the morphological characteristics and rDNA-ITS sequence analysis (1). To confirm pathogenicity, Koch's postulates were performed on detached leaves of C. fargesii f. duciouxii, inoculated with a solution of 1.0 × 106 conidia per ml. Symptoms similar to the original ones started to appear after 10 days, while untreated leaves remained healthy. The inoculation assay used three leaves for untreated and six leaves for treated. The experiments were repeated once. C. gloeosporioides was consistently reisolated from the diseased tissue. C. gloeosporioides is distributed worldwide causing anthracnose on a wide variety of plants (3). To the best of our knowledge, this is the first report of C. gloeosporioides causing leaf spots on C. fargesii f. duciouxii in China. References: (1) B. C. Sutton. Page 1 in: Colletotrichum: Biology, Pathology and Control. CAB International. Wallingford, UK, 1992. (2) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990. (3) J. Yan et al. Plant Dis. 95:880, 2011.

Evaluation of an EpitypifiedOphiocordyceps formosana(Cordyceps s.l.) for Its Pharmacological Potential

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2015/189891 ◽

2015 ◽

Vol 2015 ◽

pp. 1-13 ◽

Cited By ~ 6

Author(s):

Yen-Wen Wang ◽

Tzu-Wen Hong ◽

Yu-Ling Tai ◽

Ying-Jing Wang ◽

Sheng-Hong Tsai ◽

...

Keyword(s):

Phylogenetic Analyses ◽

Water Extract ◽

Hot Water ◽

Clinical Complications ◽

Molecular Phylogenetic ◽

Potential Applicability ◽

Pharmacological Potential

The substantial merit ofCordyceps s.l.spp. in terms of medicinal benefits is largely appreciated. Nevertheless, only few studies have characterized and examined the clinical complications of the use of health tonics containing these species. Here, we epitypifiedC. formosanaisolates that were collected and characterized asOphiocordyceps formosanabased on morphological characteristics, molecular phylogenetic analyses, and metabolite profiling. Thus, we renamed and transferredC. formosanato the new protologueOphiocordyceps formosana(Kobayasi & Shimizu) Wang, Tsai, Tzean & Shencomb. nov.Additionally, the pharmacological potential ofO. formosanawas evaluated based on the hot-water extract from its mycelium. The relative amounts of the known bioactive ingredients that are unique toCordyceps s.l.species inO. formosanawere found to be similar to the amounts inO. sinensisandC. militaris, indicating the potential applicability ofO. formosanafor pharmacological uses. Additionally, we found thatO. formosanaexhibited antioxidation activitiesin vitroandin vivothat were similar to those ofO. sinensisandC. militaris. Furthermore,O. formosanaalso displayed conspicuously effective antitumor activity compared with the testedCordyceps s.l.species. Intrinsically,O. formosanaexhibited less toxicity than the otherCordycepsspecies. Together, our data suggest that the metabolites ofO. formosanamay play active roles in complementary medicine.

Molecular Identification Based on Chloroplast Sequences and Anti-complementary Activity Comparison of Juniperus Samples from the Qinghai-Tibet Plateau

Planta Medica ◽

10.1055/a-1194-0471 ◽

2020 ◽

Vol 86 (16) ◽

pp. 1176-1184

Author(s):

Yi-Ting Yang ◽

Ji De ◽

Ze-Long Fu ◽

Xie-Li Wang ◽

Dao-Feng Chen ◽

...

Keyword(s):

Phylogenetic Trees ◽

Tibetan Medicine ◽

Tibet Plateau ◽

Efficacy Evaluation ◽

Original Species ◽

Genomic Regions ◽

Qinghai Tibet Plateau

Abstract Juniperus (Cupressaceae, Pinales) plants are widely distributed in the Qinghai-Tibet Plateau of China. The leaves and twigs of at least 8 Juniperus species (J. pingii, J. pingii var. wilsonii, J. squamata, J. recurva var. coxii, J. saltuaria, J. indica, J. tibetica and J. convallium var. microsperma) have been used as the Tibetan medicine Xuba. At present, it is difficult to distinguish among the original species of Xuba based only on their similar morphological characteristics. However, in our previous studies, 4 Xuba samples from different Juniperus species exhibited significant differences in both anticomplementary activity in vitro and anti-inflammatory effects on acute lung injury in vivo. To identify the effective original species of Xuba reliably, in this study, we developed a sequencing-based DNA molecular technology to distinguish 14 populations of 8 Juniperus species collected from Tibet region, using trnS-G, trnD – T, and petN-psbM genomic regions to build phylogenetic trees. In addition, their anticomplementary activities were evaluated. The results showed that combined sequence of these 3 genomic regions could identify 8 Juniperus species clearly and clustered individuals of one species but from different locations, whichever phylogenetic tree was constructed. Moreover, the anticomplementary activities of the 8 species were clustered into 2 groups. Among them, J. saltuaria and J. recurva var. coxii, which formed an independent branch apart from the other 6 species in phylogenetic trees, were the most potent (CH50: 0.029 – 0.032 mg/mL). Consequently, DNA identification of Juniperus using the combined sequence could provide beneficial guidance for further efficacy evaluation and quality control of Xuba.

Efficacy and Baseline Sensitivity of Succinate-Dehydrogenase-Inhibitor Fungicides for Management of Colletotrichum Crown Rot of Strawberry

Plant Disease ◽

10.1094/pdis-01-20-0083-re ◽

2020 ◽

Vol 104 (11) ◽

pp. 2860-2865

Author(s):

Michelle S. Oliveira ◽

Leandro G. Cordova ◽

Natalia A. Peres

Keyword(s):

Succinate Dehydrogenase ◽

Fungal Growth ◽

Crown Rot ◽

Baseline Sensitivity ◽

Current Management ◽

Qoi Fungicides ◽

Dilution Assay ◽

Succinate Dehydrogenase Inhibitor

The current management of Colletotrichum crown rot (CCR) of strawberry, caused by Colletotrichum gloeosporioides sensu lato, relies on the use of a few fungicide classes, particularly QoI fungicides. Since resistance to QoI fungicides has recently been detected, alternative fungicide groups are needed to control this disease. Our objective was to evaluate the efficacy of succinate-dehydrogenase-inhibitor (SDHI) fungicides in managing CCR. Five SDHI fungicides, fluopyram, isofetamid, penthiopyrad, fluxapyroxad, and benzovindiflupyr, were applied 2 days before or 1 day after inoculation of cultivar Strawberry Festival. SDHI treatments were compared with the most common fungicides used for CCR management, i.e., thiophanate-methyl, pyraclostrobin, and captan. Benzovindiflupyr applied 1 day after inoculation was effective in reducing plant mortality and disease development. The baseline sensitivity of C. gloeosporioides isolates was determined in vitro using a spiral gradient dilution assay. The EC50 for benzovindiflupyr and penthiopyrad varied from 0.08 to 1.11 and 0.45 to 3.17 µg/ml, respectively, whereas the other SDHI fungicides did not inhibit fungal growth. If registered, benzovindiflupyr could serve as an alternative to manage CCR in Florida.

Investigation of the Colletotrichum gloeosporioides Species Complex Causing Peach Anthracnose in South Carolina

Plant Disease ◽

10.1094/pdis-10-14-1076-re ◽

2015 ◽

Vol 99 (6) ◽

pp. 797-805 ◽

Cited By ~ 24

Author(s):

Meng-Jun Hu ◽

Anja Grabke ◽

Guido Schnabel

Keyword(s):

South Carolina ◽

Species Complex ◽

Colletotrichum Gloeosporioides Species Complex

Colony Growth ◽

Pcr Assay ◽

Peach Fruit ◽

Multiplex Pcr Assay ◽

Colletotrichum Spp ◽

Colletotrichum gloeosporioides, C. acutatum, and C. truncatum are causal agents of anthracnose disease of peach in South Carolina, but more recent investigations show that C. gloeosporioides is a species complex that has not yet been investigated among peach isolates. A total of 28 Colletotrichum spp. isolates associated with peach fruit anthracnose were collected in 2012 from Chesnee (10 isolates), McBee (10 isolates), Monetta (2 isolates), and Ridge Spring (6 isolates), South Carolina. Morphological characteristics indicated that all 28 isolates belonged to the C. gloeosporioides species complex. Phylogenetic analysis of the combined calmodulin (CAL), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and β-tubulin (TUB2) gene sequences identified two species, C. siamense and C. fructicola. Cultural characteristics such as colony growth rate, shape and size of conidia, and appressoria from representative isolates of the two species largely matched previous descriptions for C. siamense and C. fructicola. Koch’s postulates for C. siamense and C. fructicola were fulfilled, confirming pathogenicity of the two species on peach. A new, two-step multiplex PCR assay was developed to facilitate differentiation of the four known Colletotrichum spp. causing anthracnose of peach in South Carolina, including C. acutatum, C. truncatum, C. siamense, and C. fructicola. The first step distinguished C. acutatum from C. truncatum and the two members of the C. gloeosporioides species complex. The second assay distinguished C. siamense from C. fructicola isolates.

The fungal endophyte Epicoccum dendrobii as a potential biocontrol agent against Colletotrichum gloeosporioides

Phytopathology ◽

10.1094/phyto-05-20-0170-r ◽

2020 ◽

Cited By ~ 1

Author(s):

Jin-Yue Bian ◽

Yu-Lan Fang ◽

Qing Song ◽

Mei-Ling Sun ◽

Ji-Yun Yang ◽

...

Keyword(s):

Biocontrol Agent ◽

Phylogenetic Analyses ◽

Fungal Endophytes ◽

Fungal Endophyte ◽

Chinese Fir ◽

Dual Culture ◽

Potential Biocontrol Agent ◽

Fungal Phytopathogen

Anthracnose caused by Colletotrichum gloeosporioides is one of most serious fungal diseases on Chinese fir (Cunninghamia lanceolata). Eight fungal endophytes were isolated from a young heathy branch of Chinese fir and screened against the pathogen in vitro. One isolate, designated as SMEL1 and subsequently identified as Epicoccum dendrobii based on morphological and phylogenetic analyses, suppressed mycelial growth of C. gloeosporioides on dual culture plates. Additionally, the metabolites of E. dendrobii significantly decreased the biomass of C. gloeosporioides. E. dendrobii was able to enter the internal tissues of the host plant via stomatal cells. The metabolites of E. dendrobii significantly inhibited conidial germination and appressorium formation, which at least partly explained why the endophyte significantly inhibited lesion development caused by C. gloeosporioides on various host plants. We further confirmed that some components with antifungal activity could be extracted from E. dendrobii using ethyl acetate as an organic solvent. To our knowledge, this is the first report of E. dendrobii as a potential biocontrol agent against a fungal phytopathogen.

Developing Actinobacterial Endophytes as Biocontrol Products for Fusarium pseudograminearum in Wheat

Frontiers in Bioengineering and Biotechnology ◽

10.3389/fbioe.2021.691770 ◽

2021 ◽

Vol 9 ◽

Author(s):

Cathryn A. O’Sullivan ◽

Margaret M. Roper ◽

Cindy A. Myers ◽

Louise F. Thatcher

Keyword(s):

Management Strategies ◽

Agar Plate ◽

Treatment Strategies ◽

Crown Rot ◽

Yield Losses ◽

Disease Symptoms ◽

Fusarium Pseudograminearum ◽

Unsterilized Soil ◽

Novel Treatment Strategies

Crown rot of wheat, caused by Fusarium pseudograminearum, results in millions of dollars of yield losses globally each year. Management strategies to control crown rot are limited and there are concerns about development of fungicide resistance so novel treatment strategies are desirable. A collection of endophytic Actinobacteria was screened for their ability to suppress the growth of F. pseudograminearum and the development of crown rot symptoms in wheat with the aim of identifying candidates that can be developed into biocontrol products. The ability of the Actinobacteria isolates to suppress the growth of three different F. pseudograminearum strains in vitro was assessed using agar-plate competition assays. Soil-free seedling assays were used to screen for suppression of development of early disease symptoms in the susceptible wheat (Triticum aestivum) cv. Tamaroi. Four of the isolates were tested in a glasshouse pot experiment to assess their ability to decrease disease symptoms and prevent yield losses in wheat cv. Tamaroi grown to maturity in an unsterilized soil. The screening of 53 isolates identified two Streptomyces isolates, MH71 and MH243, with very strong antifungal activity against F. pseudograminearum strains in agar-plate competition and seedling assays. In the glasshouse pot trial, plants treated with seed coatings of either MH71 or MH243 had > 24% lower disease severity than control plants infected with F. pseudograminearum. These two cultures show potential for development as biocontrol products because they are easy to culture, grow on relatively inexpensive media, produce highly durable spores and can be delivered to plants as a seed coat.

Colletotrichum Gloeosporioides Species Complex

Characterization and Fungicide Sensitivity of Colletotrichum Species Causing Strawberry Anthracnose in Eastern China

Plant Disease ◽

10.1094/pdis-10-19-2241-re ◽

2020 ◽

Vol 104 (7) ◽

pp. 1960-1968

Author(s):

Liqing Zhang ◽

Lili Song ◽

Xiangming Xu ◽

Xiaohua Zou ◽

Ke Duan ◽

...

Keyword(s):

Species Complex ◽

Phylogenetic Analyses ◽

Eastern China ◽

Intergenic Spacer ◽

Fungicide Sensitivity ◽

Strawberry Anthracnose ◽

Dmi Fungicides ◽

Colletotrichum Spp ◽

Strawberry anthracnose caused by Colletotrichum spp. is one of the most serious diseases in the strawberry fields of China. In total, 196 isolates of Colletotrichum were obtained from leaves, stolons, and crowns of strawberry plants with anthracnose symptoms in eastern China and were characterized based on morphology, internal transcribed spacer (ITS), and β-tubulin (TUB2) gene sequences. All 196 isolates were identified as the Colletotrichum gloeosporioides species complex. In total, 62 strains were further identified at the species level by phylogenetic analyses of multilocus sequences of ITS, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), actin (ACT), Apn2-Mat1-2 intergenic spacer and partial mating type (ApMat), calmodulin (CAL), and TUB2. Three species from the C. gloeosporioides species complex were identified: Colletotrichum siamense, C. fructicola, and C. aenigma. Isolates of C. siamense were tolerant to high temperatures, with a significantly larger colony diameter than the other two species when grown above 36°C. The inoculation of strawberry plants confirmed the pathogenicity of all three species. C. siamense isolates resulted in the highest disease severity. The in vitro sensitivities of C. siamense and C. fructicola isolates to azoxystrobin and three demethylation-inhibitor (DMI) fungicides (difenoconazole, tebuconazole, and prochloraz) were determined. Both species were sensitive to DMI fungicides but not to azoxystrobin. C. siamense isolates were more sensitive to prochloraz, while C. fructicola isolates were more sensitive to difenoconazole and tebuconazole. The present study provides valuable information for the effective management of strawberry anthracnose.

Ochraceocephala foeniculi gen. et sp. nov., a new pathogen causing crown rot of fennel in Italy

MycoKeys ◽

10.3897/mycokeys.66.48389 ◽

2020 ◽

Vol 66 ◽

pp. 1-22

Author(s):

Dalia Aiello ◽

Alessandro Vitale ◽

Giancarlo Polizzi ◽

Hermann Voglmayr

Keyword(s):

New Genus ◽

Southern Italy ◽

Phylogenetic Analyses ◽

Large Subunit ◽

Management Strategies ◽

Small Subunit ◽

Ssu Rdna ◽

Crown Rot ◽

New Genus And Species ◽

Pathogenicity Tests

A new disease of fennel is described from Sicily (southern Italy). Surveys of the disease and sampling were conducted during spring 2017 and 2018 in Adrano and Bronte municipalities (Catania province) where this crop is widely cultivated. Isolations from the margin of symptomatic tissues resulted in fungal colonies with the same morphology. Pathogenicity tests with one isolate of the fungus on 6-month-old plants of fennel reproduced similar symptoms to those observed in nature. Inoculation experiments to assess the susceptibility of six different fennel cultivars to infection by the pathogen showed that the cultivars ‘Narciso’, ‘Apollo’, and ‘Pompeo’ were more susceptible than ‘Aurelio’, ‘Archimede’, and ‘Pegaso’. Phylogenetic analyses based on a matrix of the internal transcribed spacer (ITS), the large subunit (LSU), and the small subunit (SSU) rDNA regions revealed that the isolates represent a new genus and species within the Leptosphaeriaceae, which is here described as Ochraceocephala foeniculi gen. et sp. nov. This study improves the understanding of this new fennel disease, but further studies are needed for planning effective disease management strategies. According to the results of the phylogenetic analyses, Subplenodomus iridicola is transferred to the genus Alloleptosphaeria and Acicuseptoria rumicis to Paraleptosphaeria.