- Pyrethrum (Tanacetum cinerariifolium) is produced for extraction of insecticidal compounds from the flower achenes. In 2004 and 2006, isolations from necrotic lesions on stems and leaves in three fields in northern Tasmania, Australia yielded 4 unidentified fungal isolates. Leaf lesions have been mediumĀ brownĀ and round (2 to Four mm in diameter) or irregular in form (2 to five mm lengthy). Stem lesions have been irregular, necrotic spots, 5 to 15 mm beneath the flower peduncle, mediumĀ brown, 2 to Four mm lengthy, and 1 to 2 mm huge.
- Isolations have been carried out on water agar following floor sterilization. Isolates have been recognized by colony traits and the presence of metabolite ‘E’ (1). On oatmeal agar (OA), colonies had irregular margins, have been greenish olivaceous-to-olivaceous grey with sparse, white, floccose, aerial mycelia.
- On malt extract agar (MEA), cultures have been variable in shade with olivaceous black facilities with mushy, dense, aerial mycelia. Conidia have been hyaline, ellipsoidal to rectangular, primarily aseptate, however often 1-septate with dimensions ranging from 2.5 to 7.5 à 1.Eight to three.Eight μm (size/width ratio = 1.7 to 2.1).
- All isolates had reasonable reactions to the NaOH check for metabolite ‘E’. DNA was extracted from all 4 isolates with a DNeasy Plant Mini Kit (QIAGEN Inc., Valencia, CA). For identification, the interior transcribed spacer area (ITS1, 5.8s, and ITS2) and half of the interpretation elongation issue (TEF) area have been amplified and sequenced.
- Primers ITS1 and ITS4 (2) have been used for the ITS area and primers EFCF1 (5′-AGTGCGGTGGTATCGACAAG) and EFCF6 (3′-CATGTCACGGACGGCGAAAC) have been used for the TEF.
- Amplicons have been sequenced in each instructions and consensus sequences assembled. The ITS sequence was 100% an identical to Boeremia exigua var. exigua (GenFinancial institution Accession No. GU237715).
Base pairs 413 to 1,214 of the TEF sequence from the pyrethrum isolates matched base pairs 1 to 802 (799 of 802 identities) of B. exigua var. exigua (GenFinancial institution Accession No. GU349080). All isolates have been confirmed Transparent Plastic Bottles as B. exigua var. exigua utilizing morphology and sequencing.
Pathogenicity exams have been carried out thrice in separate glasshouse trials for 2 of the 4 isolates.
- For every isolate, conidial suspensions in water (Three ml/plant) from MEA, adjusted to five Ć 105/ml have been utilized with Tween 20 (1 drop per 100 ml of water) to 8-week-old pyrethrum vegetation (5 pots per isolate with 4 vegetation per pot) utilizing a hand-held sprayĀ bottle.
- Twenty vegetation have been sprayed with water and Tween 20 as nontreated controls. Plants have been lined with plastic luggage for 48 h after inoculation and examined for signs after 15 days at 20°C.
- Disease incidence (quantity of symptomatic leaves affected per whole quantity of leaves) of the inoculated vegetation diverse from 7.5 to 9.4%. Noninoculated vegetation didn’t develop signs. Isolations resulted in cultures morphologically an identical on MEA and OA to these inoculated.
- To our information, that is the primary report of B. exigua var. exigua inflicting illness in pyrethrum. Cultures have been deposited in the New South Wales Department of Agriculture assortment (DAR79101 to 79104) and TEF and ITS sequences for DAR79101 in GenFinancial institution (Accession Nos. JF925328 and JF925329, respectively).
- Boeremia blight is more likely to contribute to the fungal illness complicated inflicting reductions in inexperienced leaf space in Australian pyrethrum manufacturing. References: (1) M. M. Aveskamp et al. Stud. Mycol. 65:1, 2010. (2) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

First Report of Cobweb Disease Caused by Cladobotryum mycophilum on the Edible Mushroom Pleurotus eryngii in Korea.
- Pleurotus eryngii is one of essentially the most commercially necessary mushrooms in Korea. In May 2009, uncommon signs have been noticed in P. eryngii grown in mushroom farms in Changnyeong and Hapcheon, in Gyeong-nam Province, Korea. One of the principle signs was cobweb-like progress of fungal mycelia over the mushroom floor. Colonies on the floor quickly overwhelmed the mushrooms, which turned paleĀ brownĀ or yellow. Mushrooms finally turned darkishĀ brownĀ and have become rotten.
- Colonies of the isolates on potato dextrose agar (PDA) have been yellowish, and a reddish or orange shade was evident in the agar. The colonies grew 20 to 30 mm per day on PDA. Large spores with a single septum have been produced on vertically branched conidiophores bearing two to 4, largely three to 4, sporogenous cells, starting from 17.2 to 20.5 μm lengthy and eight.zero to 10.2 μm thick.
- The form of the conidia was ellipsoid and obovoid. These morphological traits are according to descriptions of Cladobotryum mycophilum, a causal agent of cobweb illness in Agaricus bisporus (1,4). To establish the remoted fungal pathogen, the ITS area was amplified with ITS1 and ITS4 primers and sequenced.
- The sequence information from the isolate was deposited in GenFinancial institution (Accession No. JF693809). A BLAST search confirmed that the remoted pressure belonged to a species of Cladobotryum. The highest similarity (99.5%) was to the ITS sequence of C. mycophilum (teleomorph Hypomyces odoratus) (GenFinancial institution Accession Nos. JF505112 and Y17096) (3,4). The pressure that was examined for pathogenicity was grown on PDA at 25°C for 72 h.
The inoculum was ready by flooding the agar floor with 10 ml of sterilized double distilled water and scraping it with a spatula.
The ensuing spore suspension was filtered by way of three layers of cheesecloth. Conidial focus was adjusted with a hemacytometer to 1 Ć 106Ā conidia ml-1. A conidia suspension was inoculated onto every of a number of phases of mushroom cultivation with a pipette. The management was noticed with double distilled water. In the case of an infection through the inoculation and spawn operating phases, the fungal mycelia colonized the media and hampered growth of the mycelium of P. eryngii.
In the regeneration and primordia formation phases of the host, the mycelium of the pathogen lined the floor of theĀ plasticĀ bottleĀ containing the substrates and developed many spores. In the rising and harvesting phases, the floor of mushroom was overwhelmed by the mycelium of the fungal pathogen and turned pale or darkishĀ brown, accompanied by cracking of the stipe floor and at last rotting with a foul odor. These signs have been much like the commentary from pure an infection.
The signs of the cobweb-like illness in A. bisporus (1,2) have been noticed inside 5 to 7 days of inoculation with conidia suspensions of C. mycophilum. Fungi remoted from inoculated mushrooms have been proven to be an identical, primarily based on phenotypic attribute, to the inoculated pressure used in these pathogenicity exams. No signs have been noticed on controls.
Disposable Meat Sample Plastic tubes with caps (5 ml) (1000/pk) |
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MST5-1000 | Alpha Diagnostics | 1 pk |
LCP EZ-Cut films (50 pieces, with liners) |
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MLCP-EZCTF-50 | MiTeGen | 50 FILMS |
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P3030 | MTC Bio | 1000/pack |
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TruStrip Sample Transfer Strips, 50-ul, 50/Pk (with sample tracking dye) |
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GP102 | GenDepot | 1,000 tubes |
0.5 ml Individual PCR Tubes with attached Flat Caps |
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GP104 | GenDepot | 1,000 tubes |
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Human Calcyphosine (CAPS) ELISA Kit |
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ARS8-50 | Alpha Diagnostics | 5 L |
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Antigen Retrieval Solution, Tris-EDTA pH 9.0 (50 ml, 100X) |
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ARS9-50 | Alpha Diagnostics | 5 L |
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28957502 | Scientific Laboratory Supplies | EACH |
To our information, that is the primary report on the incidence of C. mycophilum on the edible mushroom P. eryngii in Korea. Based on the pathogenicity check outcomes, the pathogen might assault P. eryngii in any cultivation stage, making it a doubtlessly severe fungal pathogen in P. eryngii. References: (1) C. G. Back et al. J. Gen. Plant Pathol. 76:232, 2010. (2) R. H. Gaze. Mushroom J. 546:23, 1995. (3) F. J. Gea et al. Plant Dis. 95:1030, 2011. (4) H. M. Grogan and R. H. Gaze. Mycol. Res. 104:357, 2000.