Light Microscopic Observation of Nuclei and Mitotic Chromosomes of<i>Botrytis</i>Species

Shirane, Noboru; Masuko, Michio; Hayashi, Yoshihiko

doi:10.1094/phyto-79-728

Cited by 47 publications

(24 citation statements)

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“…b Observed quantity of Botrytis DNA/seed detected using a real-time PCR assay. c Estimated B. allii genome weight based on the assumption of one nucleus per conidium and an average ascomycete genome size of 36 Mb, which is approximately 38 fg (22,51,58). d Estimated DNA extraction and real-time PCR efficiency, calculated per extract as: [(observed quantity of DNA)/(expected quantity of DNA)]*100. viously been reported.…”

Section: Discussionmentioning

confidence: 99%

“…Nonetheless, Owen et al (42) demonstrated that B. byssoidea and B. aclada are valid species. Two subgroups within B. aclada (AI and AII) can be distinguished based on chromosome number and conidial dimensions (20,58). Polymorphic polymerase chain reaction (PCR) alleles and internal transcribed spacer restriction fragment length polymorphisms (ITS-RFLPs) have been used to demonstrate that isolates in subgroups AI and AII are distinct from B. byssoidea (36,39).…”

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confidence: 99%

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A Real-Time, Quantitative PCR Seed Assay for Botrytis spp. that Cause Neck Rot of Onion

Chilvers

Toit²,

Akamatsu

et al. 2007

Plant Disease

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A real-time fluorescent polymerase chain reaction (PCR) assay was developed using SYBR Green chemistry to quantify the Botrytis spp. associated with onion (Allium cepa) seed that are also able to induce neck rot of onion bulbs, i.e., B. aclada, B. allii, and B. byssoidea. The nuclear ribosomal intergenic spacer (IGS) regions of target and nontarget Botrytis spp. were sequenced, aligned, and used to design a primer pair specific to B. aclada, B. allii, and B. byssoidea. Primers and amplification parameters were optimized to avoid amplifying the related species B. cinerea, B. porri, and B. squamosa, as well as Sclerotinia sclerotiorum and isolates of 15 other fungal species commonly found associated with onion seed. The primers reliably detected 10 fg of genomic DNA per PCR reaction extracted from pure cultures of B. aclada and B. allii. Conventional assays of surface-disinfested and nondisinfested seed on an agar medium were used to determine the incidence of neck rot Botrytis spp. associated with each of 23 commercial onion seed lots, and the real-time PCR assay was used to determine the quantity of DNA of neck rot Botrytis spp. in each seed lot. A linear relationship could not be found between the incidence of seed infected with the neck rot Botrytis spp. using the conventional agar seed assays and the quantity of DNA of the neck rot Botrytis spp. detected by the real-time PCR assay. However, the real-time PCR assay appeared to be more sensitive than the conventional agar assay, allowing detection of neck rot Botrytis spp. in 5 of the 23 seed lots that tested negative using the conventional agar seed assay.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

A Real-Time, Quantitative PCR Seed Assay for Botrytis spp. that Cause Neck Rot of Onion

Chilvers

Toit²,

Akamatsu

et al. 2007

Plant Disease

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“…Mitotic chromosomes of cytological specimens were prepared by modifying protocols of the germ tube burst method (GTBM) used for other fungi (Shirane et al, 1989;Taga and Murata, 1994;Taga et al, 1998). Eighty to one-hundred fifty microliters of a conidial suspension of C. parasitica in PDB (5 Â 10 5 -1 Â 10 6 conidia/ml) was placed on a slide glass and incubated at 23°C in a humid chamber for 36-40 h. One hour before the end of incubation, thiabendazole (Sigma, St. Louis, MO; stock solution: 10 mg/ml dimethyl sulfoxide) was added to the conidial suspension at a final concentration of 50 lg/ml.…”

Section: Cytological Observationmentioning

confidence: 99%

Cytological and electrophoretic karyotyping of the chestnut blight fungus Cryphonectria parasitica

Eusebio-Cope

Suzuki

Garmaroodi

et al. 2009

Fungal Genetics and Biology

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“…Three replicates were prepared for each isolate and then incubated for one week at 23°C. Length, width and volume of 200 conidia from each isolate were measured according to Shirane (1989). The conidial volume was determined by measuring conidial dimensions using an objective (40X) of a light microscope, Olympus microscope (BH2), using the following formula:…”

Section: Introductionmentioning

confidence: 99%

Molecular and Pathological Variability Associated with Transposable Elements of Botrytis cinerea Isolates Infecting Grape and Strawberry in Egypt

Wagih

Wahab

Shehata

et al. 2019

Int. J. Phytopathol.

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Grey mold caused by Botrytis cinerea, is known to cause great losses in most vegetable and fruit crops. Fifty-one isolates of B. cinerea were collected from grape (BCG) and strawberry (BCS) grown in different Egyptian locations. Variation among isolates was demonstrated using fenhexamid resistance and genetic approaches. Isolates were classified into various pathogenic groups depending on their reactions towards lettuce leaves. Genetic variability was identified in all isolates using transposable elements (TEs) analysis which revealed either the presence or absence of boty and flipper transposons. Furthermore, TEs typing of B. cinerea isolates demonstrated four TE types, on the basis of TE distribution in B. cinerea populations, namely, transposa (having both boty and flipper), flipper (possessing only flipper), boty (having only boty), and vacuma (lacking both boty and flipper elements). Transposa type was predominant (43.1%) and both transposa and vacuma isolate types showed no specialization with respect to host plant or plant location, while flipper type revealed a geographical preference in (BCG) isolates. Pathogenicity was also correlated to TE type as isolates containing transposa type revealed some degree of correlation with virulence behaviour, suggesting that transposa populations have higher pathogenic potential as compared to vacuma ones. The sensitivity of sampled isolates was tested against fenhexamid as one of the most important botryticides. Sensitivity to fenhexamid was shown in all isolates from strawberry and grape, grown in different locations, with low EC50 values between 0.012-0.084 μg/ml. This finding provided a cue for effective usage of fenhexamid for grey mold management. The present work demonstrated a correlation between the distribution of TEs and some fungal features such as isolate source and virulence, but no correlation was found between morphological characteristics, TE type, and sensitivity to fenhexamid. Cluster analysis based on phylogenetic tree showed that the Egyptian isolates branched as a separate divergent group from the others retrieved from GenBank, reflecting the presence of sequence polymorphism between the current isolates of B. cinerea and those previously identified.

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Light Microscopic Observation of Nuclei and Mitotic Chromosomes ofBotrytisSpecies

Cited by 47 publications

References 0 publications

A Real-Time, Quantitative PCR Seed Assay for Botrytis spp. that Cause Neck Rot of Onion

A Real-Time, Quantitative PCR Seed Assay for Botrytis spp. that Cause Neck Rot of Onion

Cytological and electrophoretic karyotyping of the chestnut blight fungus Cryphonectria parasitica

Molecular and Pathological Variability Associated with Transposable Elements of Botrytis cinerea Isolates Infecting Grape and Strawberry in Egypt

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