Isolation of Dna From Fungal Mycelia and Single Spores

Lee, Steven B.; Taylor, John W.

doi:10.1016/b978-0-12-372180-8.50038-x

Cited by 650 publications

(387 citation statements)

References 14 publications

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“…Phylogenetic analysis.-DNA was extracted from the fungal cultures according to Lee and Taylor (1990), and the ITS and ␤-tubulin regions were amplified (Kang et al 2001). The ITS1 region, 5.8S rRNA gene and the ITS2 region of the nuclear-encoded ribosomal RNA gene were amplified with primers ITS1 and ITS4 (White et al 1990), and part of the ␤-tubulin gene was amplified with primers T1 (O'Donnell and Cigelnik 1997) and ␤t-2b (Glass and Donaldson 1995).…”

Section: Methodsmentioning

confidence: 99%

Characterization of Colletotrichum Species Associated with Diseases of Proteaceae

et al. 2004

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Colletotrichum spp. are known to occur on and cause diseases of Proteaceae, but their identities are confused and poorly understood. The aim of the present study thus was to identify accurately the Colletotrichum spp. associated with diseases of cultivated Proteaceae. Colletotrichum spp. associated with proteaceous hosts growing in various parts of the world were identified based on morphology, sequence data of the internal transcribed spacer region (ITS-1, ITS-2), the 5.8S gene, and partial sequences of the ␤-tubulin gene. Four species of Colletotrichum were found to be associated with Proteaceae. Colletotrichum gloeosporioides, a cosmopolitan species known to occur on numerous hosts, was isolated from Protea cynaroides cultivated in South Africa and Zimbabwe, and from a Leucospermum sp. in Portugal. A recently described species, C. boninense was associated with Zimbabwean and Australian Proteaceae but also occurred on a Eucalyptus sp. in South Africa. This represents a major geographical and host extension for the species and a description of the African strains is

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Section: Methodsmentioning

confidence: 99%

Characterization of Colletotrichum Species Associated with Diseases of Proteaceae

et al. 2004

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“…The standardized amplification assay was as follows: template DNA, 25 ng; Taq DNA polymerase (Genei, Bangalore, India), 0.5 units; MgCl 2 , 5 mm; dNTP (Genei), 100 μm each of dATP, dGTP, dCTP, dTTP; primer (Operon Bio-technologies, Cologne, Germany), 1 μm; buffer (Genei), 1¥ in a reaction volume of 25 μl. Different PCR protocols given by Pascual et al (2000) and Lee and Taylor (1990) were tested for obtaining the best amplification of nucleic acids of the isolates under investigation. The PCR was performed using a palmcycler (Carbett Research, Mortalake, Australia) with the following temperature profile: initial denaturation at 94 ∞C for 2 min, followed by 45 cycles of denaturation at 92 ∞C for 1 min; annealing at 37 ∞C for 1 min; extension at 72 ∞C for 2 min with final elongation at 72 ∞C for 5 min.…”

Section: Optimization Of Polymerase Chain Reaction (Pcr)mentioning

confidence: 99%

Genetic Variability and Relationships among Seventeen Trichoderma Isolates to Control Dry Root Rot Disease Using RAPD Markers

Gopal

Sreenivasulu

Gopi

et al. 2008

Zeitschrift Für Naturforschung C

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Trichoderma spp. has been identified as potential antagonist of Fusarium solani, which is causing dry root rot of Citrus. A random amplified polymorphic DNA (RAPD) marker was used to estimate the genetic variation among 17 isolates of Trichoderma. These isolates were characterized using 20 random primers of the OPM series, out of which 16 primers gave a total of 145 DNA fragments, showing 91.8% polymorphism. The genetic distance between each isolate was calculated, and cluster analysis was used to generate a dendrogram showing the relationship among them. The isolates grouped into two major clusters, the first major cluster consisted of TCT 14 , TCT 17 , TCT 13 , TCT 12 and TCT 16 . The remaining isolates in the second major cluster separated in two sub-clusters; the first cluster consisted of TCT 4 , TCT 10 , TCT 2 , TCT 3 , TCT 8 , TCT 6 , TCT 9 , and the second sub-cluster consisted of TCT 1 TCT 15 TCT 5 , TCT 11 , and TCT 7 . The similarity matrix indicated that TCT 6 and TCT 13 were genetically distinct as they showed only 22.6% similarity followed by TCT 5 and TCT 16 ; TCT 6 and TCT 16 (25%), while the isolates TCT 4 and TCT 10 were found to be genetically similar, as 66.7% similarity was observed between the isolates followed by 61.3% similarity between the TCT 2 and TCT 4 isolates.

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“…The mycelium was then scraped off the cellophane membrane and ground to a fine powder in liquid nitrogen with a mortar and pestle. The DNA was purified as described by Lee & Taylor (1990), except that a further phenol-chloroform extraction step and RNAase treatment were included (Karjalainen & Kammiovirta, 1994). The genomic DNA pellet was dissolved in TE buffer (10 mM Tris-HCl, 1 mM EDTA, pH 8 .…”

Section: Dna Extractionmentioning

confidence: 99%

Geographical cline of DNA variation within the F intersterility group of Heterobasidion annosum in Italy

et al. 1997

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Eighty‐six Heterobasidion annosum isolates, mainly belonging to the F intersterility group and obtained from 32 different geographical localities in Italy, were subjected to genetic analysis by the Random Amplified Polymorphic DNA (RAPD) markers. The similarity between F and S groups was higher than that between F and P. In UPGMA Cluster Analysis, the F isolates originating from the same locality usually grouped in the same cluster. The isolates also showed a tendency to group at the level of larger geographical areas. Within the F group, isolates from the south of the Italian peninsula showed the highest genetic variation and northern isolates from the Alpine regions showed the lowest. This indicates a gradual cline along the peninsula. The genetic variability in the Italian F group is discussed in relation to the past and present distribution of the host species in Italy and Europe.

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Isolation of Dna From Fungal Mycelia and Single Spores

Cited by 650 publications

References 14 publications

Characterization of Colletotrichum Species Associated with Diseases of Proteaceae

Characterization of Colletotrichum Species Associated with Diseases of Proteaceae

Genetic Variability and Relationships among Seventeen Trichoderma Isolates to Control Dry Root Rot Disease Using RAPD Markers

Geographical cline of DNA variation within the F intersterility group of Heterobasidion annosum in Italy

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