scholarly journals First report of Fusarium commune causing root and crown rot on maize in Italy

Plant Disease ◽  
2021 ◽  
Author(s):  
Monica Mezzalama ◽  
Vladimiro Guarnaccia ◽  
Ilaria Martino ◽  
Giulia Tabome ◽  
Maria Lodovica GULLINO

Maize (Zea mays L.) is a cereal crop of great economic importance in Italy; production is currently of 62,587,469 t, with an area that covers 628,801 ha, concentrated in northern Italy (ISTAT 2020). Fusarium species are associated with root and crown rot causing failures in crop establishment under high soil moisture. In 2019 maize seedlings collected in a farm located in San Zenone degli Ezzelini (VI, Italy) showed root and crown rot symptoms with browning of the stem tissues, wilting of the seedling, and collapsing due to the rotting tissues at the base of the stem. The incidence of diseased plants was approximately 15%. Seedlings were cleaned thoroughly from soil residues under tap water. Portions (about 3-5 mm) of tissue from roots and crowns of the diseased plants were cut and surface disinfected with a water solution of NaClO at 0.5% for 2 minutes and rinsed in sterile H20. The tissue fragments were plated on Potato Dextrose Agar (PDA) amended with 50 mg/l of streptomycin sulfate and incubated for 48-72 hours at 25oC. Over the 80 tissue fragments plated, 5% were identified as Fusarium verticillioides, 60% as Fusarium spp., 35% developed saprophytes. Fusarium spp. isolates that showed morphological characteristics not belonging to known pathogenic species on maize were selected and used for further investigation while species belonging to F. oxysporum were discarded. Single conidia of the Fusarium spp. colonies were cultured on PDA and Carnation Leaf Agar (CLA) for pathogenicity tests, morphological and molecular identification. The colonies showed white to pink, abundant, densely floccose to fluffy aerial mycelium. Colony reverse showed light violet pigmentation, in rings on PDA. On CLA the isolates produced slightly curved macronidia with 3 septa 28.1 - 65.5 µm long and 2.8-6.3 µm wide (n=50). Microconidia were cylindrical, aseptate, 4.5 -14.0 µm long and 1.5-3.9 µm wide (n=50). Spherical clamydospores were 8.8 ± 2.5 µm size (n=30), produced singly or in pairs on the mycelium, according to the description by Skovgaard et al. (2003) for F. commune. The identity of two single-conidia strains was confirmed by sequence comparison of the translation elongation factor-1α (tef-1α), and RNA polymerase II subunit (rpb2) gene fragments (O’Donnell et al. 2010). BLASTn searches of GenBank, and Fusarium-ID database, using the partial tef-1α (MW419921, MW419922) and rpb2 (MW419923, MW419924) sequences of representative isolate DB19lug07 and DB19lug20, revealed 99% identity for tef-1α and 100% identity to F. commune NRRL 28387(AF246832, AF250560). Pathogenicity tests were carried out by suspending conidia from a 10-days old culture on PDA in sterile H2O to 5×104 CFU/ml. Fifty seeds were immersed in 50 ml of the conidial suspension of each isolate for 24 hours and in sterile water (Koch et al. 2020). The seeds were drained, dried at room temperature, and sown in trays filled with a steamed mix of white peat and perlite, 80:20 v/v, and maintained at 25°C and RH of 80-85% for 14 days with 12 hours photoperiod. Seedlings were extracted from the substrate, washed under tap water, and observed for the presence of root and crown rots like the symptoms observed on the seedlings collected in the field. Control seedlings were healthy and F. commune was reisolated from the symptomatic ones and identified by resequencing of tef-1α gene. F. commune has been already reported on maize (Xi et al. 2019) and other plant species, like soybean (Ellis et al. 2013), sugarcane (Wang et al. 2018), potato (Osawa et al. 2020), indicating that some attention must be paid in crop rotation and residue management strategies. To our knowledge this is the first report of F. commune as a pathogen of maize in Italy. References Ellis M L et al. 2013. Plant Disease, 97, doi: 10.1094/PDIS-07-12-0644-PDN. ISTAT. 2020. http://dati.istat.it/Index.aspx?QueryId=33702. Accessed December 28, 2020. Koch, E. et al. 2020. Journal of Plant Diseases and Protection. 127, 883–893 doi: 10.1007/s41348-020-00350-w O’Donnell K et al. 2010. J. Clin. Microbiol. 48:3708. https://doi.org/10.1128/JCM.00989-10 Osawa H et al. 2020. Journal of General Plant Pathology, doi.org/10.1007/s10327-020-00969-5. Skovgaard K 2003. Mycologia, 95:4, 630-636, DOI: 10.1080/15572536.2004.11833067. Wang J et al. 2018. Plant Disease, 102, doi/10.1094/PDIS-07-17-1011-PDN Xi K et al. 2019. Plant Disease, 103, doi/10.1094/PDIS-09-18-1674-PDN

Plant Disease ◽  
2010 ◽  
Vol 94 (10) ◽  
pp. 1261-1261 ◽  
Author(s):  
İ. Kurbetli ◽  
K. Değirmenci

Almond (Prunus dulcis) production is currently increasing in Turkey. Losses of approximately 1% associated with root and crown rot of almond seedlings were observed in two commercial nurseries in Ankara and Düzce provinces in 2009. Aboveground symptoms were leaf chlorosis and wilt. Feeder roots were decayed, necrosis occurred on taproots and basal stems, and plants collapsed within several weeks. Roots were washed in tap water and 9 to 10 pieces (3 to 5 mm long) of root tissue taken from the margins of canker lesions, without surface disinfection, were placed on selective medium P5ARPH-CMA (2). Plates were incubated for 3 to 5 days at 20°C in darkness and a number of Phytophthora spp. were recovered. Actively growing mycelium was transferred to carrot piece agar containing β-sitosterol (per liter: carrot piece, 40 g; agar, 20 g; β-sitosterol, 20 mg). Isolates were identified as Phytophthora cactorum and P. citrophthora on the basis of morphological characteristics (1). P. cactorum produced abundant sporangia, oogonia, and paragynous antheridia on carrot piece agar plus β-sitosterol. It had conspicuously papillate and caducous sporangia with short pedicel. Sporangia were usually ovoid but sometimes nearly spherical. P. citrophthora did not produce sexual structures in single culture. It produced papillate, noncaducous sporangia, which were usually ovoid and obpyriform, often asymmetrically shaped and rarely possessed more than one apex. P. citrophthora did not grow at 35°C but it grew well at 30°C. Isolate identities were confirmed by sequence analysis of the ribosomal DNA internal transcribed spacers 1 and 2 (GenBank Accession Nos. HM357622, HM357623, HM357624, HM357625) using primers ITS1 and ITS2 (3). One representative isolate of each species was used to inoculate eight 2-year-old almond plants with an agar plug with actively growing mycelium that was attached to exposed cambium of basal stems. Agar plugs without mycelium were used for eight control plants. All plants inoculated with Phytophthora spp. collapsed within 3 to 4 weeks. Control plants remained healthy. Phytophthora spp. were reisolated from necrotic basal stems. To our knowledge, this is the first report of P. cactorum and P. citrophthora of almond in Turkey. References: (1) M. E. Gallegly and C. Hong. Phytophthora, Identifying Species by Morphology and DNA Fingerprints. The American Phytopathological Society, St. Paul, MN, 2008. (2) S. N. Jeffers and S. B. Martin. Plant Dis. 70:1038, 1986. (3) S. G. Roy et al. J. Phytopathol. 157:666, 2009.


Plant Disease ◽  
2006 ◽  
Vol 90 (8) ◽  
pp. 1109-1109 ◽  
Author(s):  
A. Garibaldi ◽  
G. Gilardi ◽  
M. L. Gullino

Lamb's lettuce or corn salad (Valerianella olitoria) is increasingly grown in Italy and used primarily in the preparation of mixed processed salad. In the fall of 2005, plants of lamb's lettuce, cv Trophy, exhibiting a basal rot were observed in some commercial greenhouses near Bergamo in northern Italy. The crown of diseased plants showed extensive necrosis, progressing to the basal leaves, with plants eventually dying. The first symptoms, consisting of water-soaked zonate lesions on basal leaves, were observed on 30-day-old plants during the month of October when temperatures ranged between 15 and 22°C. Disease was uniformly distributed in the greenhouses, progressed rapidly in circles, and 50% of the plants were affected. Diseased tissue was disinfested for 1 min in 1% NaOCl and plated on potato dextrose agar amended with 100 μg/liter of streptomycin sulfate. A fungus with the morphological characteristics of Rhizoctonia solani was consistently and readily isolated and maintained in pure culture after single-hyphal tipping (3). The five isolates of R. solani, obtained from affected plants successfully anastomosed with tester isolate AG 4, no. RT 31, received from R. Nicoletti of the Istituto Sperimentale per il Tabacco, Scafati, Italy (2). The hyphal diameter at the point of anastomosis was reduced, and cell death of adjacent cells occurred (1). Pairings were also made with AG 1, 2, 3, 5, 7, and 11 with no anastomoses observed between the five isolates and testers. For pathogenicity tests, the inoculum of R. solani (no. Rh. Vale 1) was grown on autoclaved wheat kernels at 25°C for 10 days. Plants of cv. Trophy were grown in 10-liter containers (20 × 50 cm, 15 plants per container) on a steam disinfested substrate (equal volume of peat and sand). Inoculations were made on 20-day-old plants by placing 2 g of infected wheat kernels at each corner of the container with 3 cm as the distance to the nearest plant. Plants inoculated with clean wheat kernels served as controls. Three replicates (containers) were used. Plants were maintained at 25°C in a growth chamber programmed for 12 h of irradiation at a relative humidity of 80%. The first symptoms, consisting of water-soaked lesions on the basal leaves, developed 5 days after inoculation with crown rot and plant kill in 2 weeks. Control plants remained healthy. R. solani was consistently reisolated from infected plants. The pathogenicity test was carried out twice with similar results. This is, to our knowledge, the first report of R. solani on lamb's lettuce in Italy as well as worldwide. The isolates were deposited at the AGROINNOVA fungal collection. The disease continues to spread in other greenhouses in northern Italy. References: (1) D. Carling. Rhizoctonia Species: Pages 37–47 in: Taxonomy, Molecular Biology, Ecology, Pathology and Disease Control. B. Sneh et al., eds. Kluwer Academic Publishers, the Netherlands, 1996. (2) J. Parmeter et al. Phytopathology, 59:1270, 1969. (3) B. Sneh et al. Identification of Rhizoctonia Species. The American Phytopathological Society, St. Paul, MN, 1996.


Plant Disease ◽  
2013 ◽  
Vol 97 (1) ◽  
pp. 147-147
Author(s):  
J. H. Park ◽  
S. E. Cho ◽  
K. S. Han ◽  
H. D. Shin

Garlic chives, Allium tuberosum Roth., are widely cultivated in Asia and are the fourth most important Allium crop in Korea. In June 2011, a leaf blight of garlic chives associated with a Septoria spp. was observed on an organic farm in Hongcheon County, Korea. Similar symptoms were also found in fields within Samcheok City and Yangku County of Korea during the 2011 and 2012 seasons. Disease incidence (percentage of plants affected) was 5 to 10% in organic farms surveyed. Diseased voucher specimens (n = 5) were deposited at the Korea University Herbarium (KUS). The disease first appeared as yellowish specks on leaves, expanding to cause a leaf tip dieback. Half of the leaves may be diseased within a week, especially during wet weather. Pycnidia were directly observed in leaf lesions. Pycnidia were amphigenous, but mostly epigenous, scattered, dark brown to rusty brown, globose, embedded in host tissue or partly erumpent, separate, unilocular, 50 to 150 μm in diameter, with ostioles of 20 to 40 μm in diameter. Conidia were acicular, straight to sub-straight, truncate at the base, obtuse at the apex, hyaline, aguttulate, 22 to 44 × 1.8 to 3 μm, mostly 3-septate, occasionally 1- or 2-septate. These morphological characteristics matched those of Septoria allii Moesz, which is differentiated from S. alliacea on conidial dimensions (50 to 60 μm long) (1,2). A monoconidial isolate was cultured on potato dextrose agar (PDA). Two isolates have been deposited in the Korean Agricultural Culture Collection (Accession Nos. KACC46119 and 46688). Genomic DNA was extracted using the DNeasy Plant Mini DNA Extraction Kit (Qiagen Inc., Valencia, CA). The internal transcribed spacer (ITS) region of rDNA was amplified using the ITS1/ITS4 primers and sequenced. The resulting sequence of 482-bp was deposited in GenBank (JX531648 and JX531649). ITS sequence information was at least 99% similar to those of many Septoria species, however no information was available for S. allii. Pathogenicity was tested by spraying leaves of three potted young plants with a conidial suspension (2 × 105 conidia/ml), which was harvested from a 4-week-old culture on PDA. Control leaves were sprayed with sterile water. The plants were placed in humid chambers (relative humidity 100%) for the first 48 h. After 7 days, typical leaf blight symptoms started to develop on the leaves of inoculated plants. S. allii was reisolated from the lesions of inoculated plants, confirming Koch's postulates. No symptoms were observed on control plants. The host-parasite association of A. tuberosum and S. allii has been known only from China (1). S. alliacea has been recorded on several species of Allium, e.g. A. cepa, A. chinense, A. fistulosum, and A. tuberosum from Japan (4) and A. cepa from Korea (3). To the best of our knowledge, this is the first report of S. allii on garlic chives. No diseased plants were observed in commercial fields of garlic chives which involved regular application of fungicides. The disease therefore seems to be limited to organic garlic chive production. References: (1) P. K. Chi et al. Fungous Diseases on Cultivated Plants of Jilin Province, Science Press, Beijing, China, 1966. (2) P. A. Saccardo. Sylloge Fungorum Omnium Hucusque Congnitorum. XXV. Berlin, 1931. (3) The Korean Society of Plant Pathology. List of Plant Diseases in Korea, Suwon, Korea, 2009. (4) The Phytopathological Society of Japan. Common Names of Plant Diseases in Japan, Tokyo, Japan, 2000.


Plant Disease ◽  
2006 ◽  
Vol 90 (9) ◽  
pp. 1260-1260 ◽  
Author(s):  
A. Garibaldi ◽  
D. Bertetti ◽  
D. Minerdi ◽  
M. L. Gullino

Penstemon barbatus (Cav.) Roth (synonym Chelone barbata), used in parks and gardens and sometimes grown in pots, is a plant belonging to the Scrophulariaceae family. During the summers of 2004 and 2005, symptoms of a root rot were observed in some private gardens located in Biella Province (northern Italy). The first symptoms resulted in stunting, leaf discoloration followed by wilt, root and crown rot, and eventually, plant death. The diseased tissue was disinfested for 1 min in 1% NaOCl and plated on a semiselective medium for Oomycetes (4). The microorganism consistently isolated from infected tissues, grown on V8 agar at 22°C, produced hyphae with a diameter ranging from 4.7 to 5.2 μm. Sporangia were papillate, hyaline, measuring 43.3 to 54.4 × 26.7 to 27.7 μm (average 47.8 × 27.4 μm). The papilla measured from 8.8 to 10.9 μm. These characteristics were indicative of a Phytophthora species. The ITS region (internal transcribed spacer) of rDNA was amplified using primers ITS4/ITS6 (3) and sequenced. BLASTn analysis (1) of the 800 bp obtained showed a 100% homology with Phytophthora citrophthora (R. & E. Sm.) Leonian. The nucleotide sequence has been assigned GenBank Accession No. DQ384611. For pathogenicity tests, the inoculum of P. citrophthora was prepared by growing the pathogen on autoclaved wheat and hemp kernels (2:1) at 25°C for 20 days. Healthy plants of P. barbatus cv. Nano Rondo, 6 months old, were grown in 3-liter pots (one plant per pot) using a steam disinfested substrate (peat/pomix/pine bark/clay 5:2:2:1) in which 200 g of kernels per liter of substrate were mixed. Noninoculated plants served as control treatments. Three replicates were used. Plants were maintained at 15 to 20°C in a glasshouse. The first symptoms, similar to those observed in the gardens, developed 21 days after inoculation, and P. citrophthora was consistently reisolated from infected plants. Noninoculated plants remained healthy. The pathogenicity test was carried out twice with similar results. A nonspecified root and crown rot of Penstemon spp. has been reported in the United States. (2). To our knowledge, this is the first report of P. citrophthora on P. barbatus in Italy as well as in Europe. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997 (2) F. E. Brooks and D. M. Ferrin. Plant Dis. 79:212, 1995. (3) D. E. L. Cooke and J. M. Duncan. Mycol. Res. 101:667, 1997. (4) H. Masago et al. Phytopathology 67:425, 1977.


Plant Disease ◽  
2008 ◽  
Vol 92 (3) ◽  
pp. 482-482 ◽  
Author(s):  
J. E. Woodward ◽  
M. A. Batla ◽  
P. A. Dotray ◽  
T. A. Wheeler ◽  
T. A. Baughman

Sclerotinia blight, caused by the soilborne fungus Sclerotinia minor Jagger, is a major disease of peanut (Arachis hypogaea L.) in parts of west Texas. Previous reports have indicated that annual weed species may serve as collateral hosts for S. minor (2). Several Ipomoea spp. are commonly found in peanut fields throughout the region. In September of 2007, Ipomoea hederacea and I. coccinea plants with bleached, shredded stems, and signs of black sclerotia were collected from a field known to be infested with S. minor. Symptomatic stem sections were rinsed in tap water, surface disinfested in 0.5% sodium hypochlorite for 1 min, air dried, and plated on potato dextrose agar (PDA). Pure cultures of S. minor consisting of white, fluffy mycelia and small (<2 mm), black, irregular sclerotia were consistently recovered. Pathogenicity tests were conducted by wound-inoculating healthy I. hederacea and I. coccinea transplants (n = 3) with agar plugs obtained from the edges of actively growing S. minor cultures. Plants were incubated in a dew chamber at 20°C and 95% relative humidity for 5 days. Plants inoculated with sterile PDA plugs served as controls (n = 3). A similar test was conducted using the susceptible peanut cultivar Flavorunner 458. Characteristic symptoms of Sclerotinia blight (3) were observed on all inoculated weed and peanut plants; whereas, the controls remained healthy. Pathogenicity tests were repeated with similar results. Cultures of S. minor were obtained from all symptomatic tissues, fulfilling Koch's postulates. These results indicate that I. hederacea and I. coccinea are additional hosts of S. minor and that sclerotia produced on infected plants can significantly augment soil inoculum. S. minor has been observed to infect I. batatas seedlings in New Jersey (1); however, this to our knowledge is the first report of S. minor infecting Ipomoea spp. in Texas. Therefore, weed management should inevitability be a part of disease management strategies for the control of Sclerotinia blight in peanut. References: (1) Anonymous. Index of Plant Diseases in the United States. USDA Handb. No. 165, 1960. (2) J. E. Hollowell et al. Plant Dis. 87:197, 2003. (3) D. M. Porter and H. A. Melouk. Sclerotinia blight. Page 34 in: Compendium of Peanut Diseases. 2nd ed. N. Kokalis-Burelle et al., eds. The American Phytopathologicial Society, St. Paul, MN, 1997.


Plant Disease ◽  
2012 ◽  
Vol 96 (5) ◽  
pp. 766-766 ◽  
Author(s):  
W. Cheon ◽  
Y. S. Kim ◽  
Y. H. Jeon

In 2010 and 2011, crab apples in Andong Province, Korea were found with dark brown spots on the fruit and mummified fruit on a tree. The fruit surface had red, circular spots that contained smaller, white spots; the color of the inner spots later changed to brown or black. Eventually, the rotten fruit dried and became mummified. Microscopic examination revealed the presence of acervuli and dark brown-to-black, needle-shaped setae. To isolate potential pathogens from infected fruit, small sections (5 to 10 mm2) were excised from the margins of lesions. These sections were surface sterilized with 70% ethanol and 1% NaOCl for 1 min and then rinsed three times with sterile distilled water. The fungus that was isolated produced whitish mycelia when grown on potato dextrose agar (PDA); the mycelia later became gray to dark gray with aerial mycelia in tufts and numerous conidia were produced. The conidia were straight, cylindrical with an obtuse apex and a truncated base, and measured 11.4 to 17.5 × 4.2 to 7.1 μm. The measurements and taxonomic characteristics coincide with those of Colletotrichum gloeosporioides (Penz.) (1). The isolated fungus was tested for pathogenicity on crab apples and cv. Fuji apples by inoculation with a conidial suspension (105 conidia/ml) prepared from 20-day-old PDA cultures. A 20-μl drop of the conidial suspension was placed onto crab apple and apple fruits that had been wounded by piercing them 1 to 2 mm deep with a pin. Small, dark lesions were observed on the artificially inoculated fruit 3 days after inoculation. Nine days after inoculation, dark lesions with salmon-colored masses of conidia were observed on fruit, which were also soft and sunken. Abundant masses of conidia were produced in the decayed tissues. The fungus was reisolated from the parts of the fruits showing the symptoms. The internal transcribed spacer (ITS) rDNA of the isolated fungus was amplified and sequenced by PCR as described by White et al. (2). The resulting 582-bp of ITS rDNA sequence was deposited in GenBank (Accession No. JQ405742). A BLAST analysis for sequence similarity of the ITS region revealed 100% identity with nucleotide sequences for C. gloeosporioides isolates (Accession Nos. HQ645080 and AB458667). The results obtained on morphological characteristics, pathogenicity, and molecular data corresponded with those of C. gloeosporioides described by Sutton (1). To our knowledge, this is the first report of the presence of C. gloeosporioides on crab apple in Korea (3). Crab apple is used as a pollinator for single-cultivar apple orchards and may become a possible source of inoculum for cultivated apple. References: (1) T. B. Sutton. Compendium of Apple and Pear Diseases. The American Phytopathological Society, St. Paul, MN, 1990. (2) T. J. White et al. PCR Protocols: A Guide to Methods and Applications, Academic Press, Inc., New York, 1990. (3) S. H. Yu. List of Plant Diseases in Korea. 5th ed. (in Korean). The Korean Society of Plant Pathology, 2009.


Plant Disease ◽  
2021 ◽  
Author(s):  
Sumyya Waliullah ◽  
Greg E. Fonsah ◽  
Jason Brock ◽  
Yonggang Li ◽  
Emran Ali

Crown rot is one of the most damaging disease of banana fruit characterized by rot and necrosis of crown tissues. In severe cases, the disease can spread to the pedicel and banana pulp. Crown rot can be infected by several common fungi, including Lasiodiplodia theobromae, Musicillium theobromae, Colletotrichum musae, and a complex of Fusarium spp. and lead to softening and blackening of tissues (Lassois et al., 2010; Kamel et al., 2016; Triest et al., 2016; Snowdon, 1990). In November 2020, typical crown rot of banana fruits (cv. Pisang Awak, belonging to the tetraploid AABB genome) were observed from UGA Banana Research 12 Plots, Tifton, GA, with incidence rates of 15%. Initial symptoms appeared in the infected crown of green banana fruits. As the infection progressed, the crown tissues became blackened and softened, followed by an internal development of infection affecting the peduncle and the fruit, triggered early ripening of bananas. At last, the development of necrosis on the pedicels and fruits appeared and caused the fingers to fall off. To identify the pathogen, tissue pieces (~0.25 cm2) from the infected crown and pedicles were surface-sterilized in a 10% bleach solution for 1 min, followed by 30 s in 70% EtOH. The disinfected tissues were rinsed in sterile water 3 times and cultured on potato dextrose agar (PDA) amended with 50 µg/ml streptomycin at 25°C in the dark for 5–10 days. Isolates of the pathogen were purified using the single-spore isolation method (Leslie and Summerell 2006). Colonies on PDA produced fluffy aerial mycelium and developed an intense purple pigment when viewed from the underside. A range of colony pigmentation and growth rates were observed among the isolates. The microconidia were ovoid, hyaline, or ellipse in shape. The morphological features of the isolates were identified as Fusarium proliferatum (Leslie and Summerell, 2006). To further identify the isolates, genomic DNA was extracted from a representative isolate. And the internal transcribed spacer (ITS) region, the partial elongation factor (TEF1-α) gene and the β-tubulin gene (TUB2)were amplified and sequenced using the primers ITS1/ITS4 (Yin et al. 2012), EF-1 /EF-2 (O’Donnell et al. 1998) and B-tub1 /B-tub2 (O’Donnell and Cigelnik, 1997), respectively. The amplicons were sequenced and deposited in NCBI (accessions no. MZ292989, MZ293071 for ITS: MZ346602, MZ346603 for TEF1-α and MZ346600 and MZ346601 for B-tub). The ITS, TEF1-α, and B-tub sequences of the isolates showed 100% sequence similarity with Fusarium proliferatum isolates (accessions no. MT560212, LS42312, and LT575130, respectively) using BLASTn in Genbank. For pathogenicity testing, three whole bunched bananas sterilized with 10% bleach solutions and washed by sterilized water, were cut into 5 bananas per brunch. The cut surface of the banana crown was inoculated with conidial suspension (1.0 × 107 cfu/ml) of the pathogen with pipette tips. Equal number of bananas were treated with sterilized water in the same volume as a control. All bananas were sealed in a plastic bag and incubated at 25°C. After 7 days post inoculation, all inoculated bananas showed initial crown rot symptoms while no symptoms were observed on the control bananas. The fungus was re-isolated from the symptomatic tissues of infected bananas and confirmed to be genetically identical to F. proliferatum of the original inoculated strains according to morphological characteristics and molecular identification, fulfilling Koch’s postulates. To the best of our knowledge, this is the first report of F. proliferatum causing crown rot on bananas in Georgia, USA.


Plant Disease ◽  
2001 ◽  
Vol 85 (4) ◽  
pp. 448-448 ◽  
Author(s):  
S. T. Koike ◽  
P. W. Crous

Myrtle (Myrtus communis) is a woody, evergreen plant used in California as a landscape shrub or potted plant. In 2000, a new root and crown disease was found in commercial nursery myrtle being grown as potted plants. Roots were necrotic and crown tissue was brown. Affected plants became gray-green in color, withered, and died. A Cylindrocladium sp. was consistently isolated from roots, crowns, and lower stems of symptomatic plants. Isolates were characterized by having penicillate conidiophores terminating in obpyriform to broadly ellipsoidal vesicles. Conidia were hyaline, 1-septate, straight with rounded ends, (50-) 53 to 56 (-58) × (3.5-) 4 to 6 μm, placing it in the Cylindrocladium candelabrum Viégas species complex. Single-conidial isolates (STE-U 4012 to 4018) produced perithecia with viable progeny of Calonectria pauciramosa C.L. Schoch & Crous when mated on carnation leaf agar with tester strains of Cylindrocladium pauciramosum C.L. Schoch & Crous (2). Matings with tester strains of all other species in this complex proved unsuccessful. Only one mating type of C. pauciramosum has thus far been found in the United States. Pathogenicity of representative isolates was confirmed by applying 5 ml of a conidial suspension (1.0 × 106 conidia/ml) to the crowns of potted, 5-month-old, rooted mytle cuttings that were subsequently maintained in a greenhouse (23 to 25°C). After 4 weeks, plant crowns and roots developed symptoms similar to those observed in the nursery, and plants later wilted and died. C. pauciramosum was re-isolated from all plants. Control plants, which were treated with water, did not develop any symptoms. The tests were repeated and the results were similar. This is the first report of C. pauciramosum as a pathogen of myrtle in California. The disease has been reported on myrtle in Europe (1). References: (1) G. Polizzi and P. W. Crous. Eur. J. Plant Pathol. 105:407, 1999. (2) C. L. Schoch et al. Mycologia 91:286, 1999.


Plant Disease ◽  
2021 ◽  
Author(s):  
Wenhao Zhang ◽  
Dan Su ◽  
Rui Sun

Rubus chingii is used as an important traditional Chinese medicine, and belongs to the family Rosaceae. The fruit has multiple pharmacological activities, including antioxidant, anti-inflammatory, and improving cognitive impairment (Na Han et al. 2012). In June 2019, a new fungal infection was observed on the leaves of R. chingii in Qiandongnan Miao and Dong Autonomous Prefecture, Guizhou Province, China, forming small lesions with reddish-brown edges along leaf veins. Over 500 plants were surveyed, and nearly 20% of the plants were symptomatic. The diseased plants grew poorly and appeared stunted, and severely affected plants died. Five symptomatic leaves were randomly collected from the field and washed with tap water and distilled water successively. The edges of infected leaf tissue were cut into small pieces (4 to 5 mm2), surface sterilized with 70% ethanol for 30 s and 0.1% HgCl2 for 1 minute, and then rinsed three times in sterile distilled water (Chen et al. 2016). The same fungus was isolated from 41 pieces. The hyphae of a representative isolate were gray, the colony surface was granular, the edges were uneven and white, and the culture turned black over time with black spherical conidia. Conidia were nearly elliptical, unicellular, and each with a hyaline, unstable apical appendage, 3 to 10 µm long. The size of conidia was 10 to 18 μm in length and 4 to 8 μm in width. These morphological characteristics are consistent with those described for the fungus Phyllosticta capitalensis. (Wikee et al. 2013). For an accurate identification, genomic DNA of a representative isolate of the pathogen was extracted to amplify the internal transcribed spacer (ITS) region, the transcription elongation factor (tefa-1), and actin (ACT) genes with the ITS1/ITS4, EF1-728F/EF1-986R, and ACT-512F/ACT-783R (Cheng, L. L. ,et al. 2019), respectively. The ITS, tefa-1 and actin gene sequences were deposited in GenBank and assigned accession numbers MW308365, MW714380 and MW714381, respectively. BLAST search analysis of GenBank (NCBI) showed that the sequences had 100% similarity with those of Phyllosticta capitalensis (GenBank accession no. ITS, MN548091; tefa-1, MN958711; and ACT, MN565575). The pathogenicity of Phyllosticta capitalensis was verified using six healthy detached leaves from healthy R. chingii plants around 40 cm tall. A total of nine plants were used, and three leaves from each plant were artificially inoculated. Each wound was inoculated with conidial suspension (106 mL-1), while the control leaves were coated by sterile water. All the treated plants were covered with plastic bags for 2 days, incubated at 28ºC and 85% relative humidity, with a 12-hour photoperiod. After 15 days following inoculation, the injured leaves showed similar symptoms to the above-mentioned lesions, while the control and uninjured leaves were still healthy. P. capitalensis were reisolated from inoculated leaves, fulfilling Koch’s postulates. P. capitalensis is an endophyte, widely distributed in various host plants in China. (Lu, J. M, et al. 2016). To the best of our known, this is the first report of black freckle disease caused by P. capitalensis on Rubus chingii in China. P. capitalensis is a destructive plant pathogen with an unusually broad host range and our findings will be useful for its management and for further research. The author(s) declare no conflict of interest.


Plant Disease ◽  
2021 ◽  
Author(s):  
Rui Qiu ◽  
Qi Li ◽  
Juan Li ◽  
Ningyu Dong ◽  
Shujun Li ◽  
...  

Tobacco (Nicotiana tabacum L.) is an economically important crop in China, with an estimated production of 2.2 million tons every year. In June 2018, tobacco plants within the municipality of Sanmenxia (Henan, China) showed symptoms of wilting with leaf yellowing and stunting. Diseased plants exhibited severe necrosis that advanced through the main root (Figure 1 A). The symptoms were observed in nineteen surveyed tobacco fields, 60 ha in total, and approximately 25% of the plants were symptomatic. The disease resulted in a severe loss in tobacco leaf production. Five symptomatic tobacco plants were sampled. Diseased tissues from roots were surface sterilized in 75% ethanol and placed on potato dextrose agar (PDA) medium. Eighteen of the 25 diseased tissues had cultures growing from them, and all the cultures were white colonies with abundant aerial mycelium produced scarlet pigmentation on PDA. One pure culture was obtained by single-spore culturing (SL1). A 10-day-old culture grown on CLA (carnation leaf agar) produced macroconidia that were falcate, straight or slightly curved, 3-septate, 25-35×3.5-4.5 μm (average 26.8×3.7 μm) (n=50). Two types of microconidia (napiform and fusiform) were formed on CLA that were hyaline, with one to two cells. Napiform conidia were 4.5-9.3×3.8-5.9 (average 7.3×5.0 μm) (n=50); fusiform conidia were 6.9-15.8×1.8-3.1 (average 9.9×2.5 μm). Spherical chlamydospores (7-12.5 μm) (n=50) were terminal or intercalary and produced in clumps or in chains (Figure1 B-D). Morphological characteristics of the isolate were similar to the features of Fusarium sinensis previously described by Zhao and Lu (2008). Molecular identification was performed using partial sequences of EF1-α gene (primers EF1/EF2, O’Donnell et al. 1998). Maximum parsimony and maximum likelihood-based methods were fitted using MEGA 7 (Moreira et al. 2019,Figure 2). The isolate was also sequenced for β-tubulin (primers T1/Bt-2b, O’Donnell & Cigelnik 1997),ribosomal RNA gene (LSU, LROR/LR5 primers, Vu et al. 2019) and rDNA-ITS (ITS 1/ ITS 4 primers, White et al. 1990). Sequences were deposited in GenBank under accession numbers MT947797 (EF1-α), MW484999 (β-tubulin), MW486649 (LSU) and MT907471 (ITS). The obtained EF1-α sequence was 98.10% identity with those of F. sinensis (MG670388.1) in the GenBank database, whereas the β-tubulin, LSU and ITS sequences showed 100% identities to the corresponding DNA sequences in F. sinensis (GenBank Acc. Nos. KX880370.1, NG_067454.1 and MH863232.1, respectively). Morphological and molecular results confirmed this species as F. sinensis (Zhao and Lu 2008). Pathogenicity tests were performed on tobacco seedlings grown on an autoclaved matrix (YC/T310-2009). Healthy 6-leaf stage tobacco seedlings were inoculated by pouring a 20 mL conidial suspension (1×106 conidia/mL-1) around the stem base of each plant, 30 plant were inoculated. Thirty control seedlings received sterilized water. All treatments were maintained for 30 days under greenhouse conditions with a 12-h light/dark photoperiod at 25±0.5℃ and 70% relative humidity. The assay was conducted three times. Root rot and foliage chlorosis similar to the ones observed on infected plants in the field were observed on the inoculated tobacco seedlings, whereas the control seedlings remained asymptomatic after 30 days (Figure1 E). The pathogen isolated from the inoculated plant exhibited morphological characteristics identical to F. sinensis and was identified by a partial EF1-α gene sequence. This disease has previously been reported as the causal agent of root and crown rot of wheat in China (Zhao and Lu 2008; Xu et al. 2018). To our knowledge, this is the first report of F. sinensis causing root rot on tobacco in China. Funding: Funding was provided by the Science and Technology Project of Henan Provincial Tobacco Company (2020410000270012), Independent Innovation Project of Hennan Academy of Agricultural Sciences (2020ZC18) and Research and Development project of Henan Academy of Agricultural Sciences (2020CY010). References: Moreira, G.M., et al. 2019 Plant Dis. O’Donnell, K., et al. 1998. Proc. Natl. Acad. Sci. USA 95:2011. O'Donnell, K., et al. 2008. J. Clin. Microbiol. 46:2477. Xu, F., et al. 2018. Front Microbiol. 9:1054. Zhao, Z.H., and Lu, G. Z., 2008. Mycologia, 100:746. The author(s) declare no conflict of interest. Keywords: tobacco root rot, Henan Province, Fusarium sinensis


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