Whole genome amplification of the rust Puccinia striiformis f. sp. tritici from single spores

Wang, Yanchun; Zhu, Ming-Qiang; Zhang, Rong; Yang, Hanli; Wang, Yan; Sun, Guangyu; She-lin, Jin; Hsiang, Tom

doi:10.1016/j.mimet.2009.02.007

Cited by 11 publications

(5 citation statements)

References 32 publications

(40 reference statements)

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“…It has been observed that a quota of GBM CD31+ endothelial cells shares with tumors cells chromosomal aberrations [140] and that a quota of GBM CD105+ endothelial cells harbours the same somatic mutations identified within tumor cells, such as amplification of EGFR and chromosome 7 [141]. In a GBM model, it was demonstrated that the tumor-derived endothelial cells originated from TICs [138].…”

Section: Perivascular Niches (Pvn) Relationship Between Npcs/gscs Anmentioning

confidence: 99%

Tumor Microenvironment — Perivascular and Perinecrotic Niches

Schiffer¹,

Mellai²,

Annovazzi³

et al. 2015

Molecular Considerations and Evolving Surgical Management Issues in the Treatment of Patients With a Brain Tumor

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Section: Perivascular Niches (Pvn) Relationship Between Npcs/gscs Anmentioning

confidence: 99%

Tumor Microenvironment — Perivascular and Perinecrotic Niches

Schiffer¹,

Mellai²,

Annovazzi³

et al. 2015

Molecular Considerations and Evolving Surgical Management Issues in the Treatment of Patients With a Brain Tumor

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“…MDA was evaluated for amplification of genomic DNA from single urediospores in a previous study ). Wang et al (2009) tested the fidelity of the MDA process by sequencing of two genetic markers: portions of the multicopy nuclear ribosomal DNA internal transcribed spacer (rDNA-ITS) region and the single-copy β-tubulin gene and found that sequences of these two markers were highly consistent between DNA synthesized via MDA and DNA extracted via CTAB method. However, whether this method can amplify other regions of the pathogen genome equally well remains undetermined.…”

Section: Introductionmentioning

confidence: 99%

Multiple displacement amplification of whole genomic DNA from urediospores of Puccinia striiformis f. sp. tritici

Zhang

2015

Curr Genet

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Biotrophic fungi, such as Puccinia striiformis f. sp. tritici, because they cannot be cultured on nutrient media, to obtain adequate quantity of DNA for molecular genetic analysis, are usually propagated on living hosts, wheat plants in case of P. striiformis f. sp. tritici. The propagation process is time-, space- and labor-consuming and has been a bottleneck to molecular genetic analysis of this pathogen. In this study we evaluated multiple displacement amplification (MDA) of pathogen genomic DNA from urediospores as an alternative approach to traditional propagation of urediospores followed by DNA extraction. The quantities of pathogen genomic DNA in the products were further determined via real-time PCR with a pair of primers specific for the β-tubulin gene of P. striiformis f. sp. tritici. The amplified fragment length polymorphism (AFLP) fingerprints were also compared between the DNA products. The results demonstrated that adequate genomic DNA at fragment size larger than 23 Kb could be amplified from 20 to 30 urediospores via MDA method. The real-time PCR results suggested that although fresh urediospores collected from diseased leaves were the best, spores picked from diseased leaves stored for a prolonged period could also be used for amplification. AFLP fingerprints exhibited no significant differences between amplified DNA and DNA extracted with CTAB method, suggesting amplified DNA can represent the pathogen's genomic DNA very well. Therefore, MDA could be used to obtain genomic DNA from small precious samples (dozens of spores) for molecular genetic analysis of wheat stripe rust pathogen, and other fungi that are difficult to propagate.

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“…The MDA of single cells for preimplantation genetic diagnosis was successfully used for analyzing Duchenne muscular dystrophy (Ren et al, 2009). MDA of gDNA from single spores of a rust fungus Puccinia striiformis can be used in molecular genetic analysis of the wheat yellow rust fungus (Wang et al, 2009). Moreover, MDA can convert complex mixtures of DNA fragments to long linear and circular concatamers, *Corresponding author.…”

Section: Introductionmentioning

confidence: 99%

Application of multiple displacement amplification in detection of five events of genetically modified organisms

Zhou¹,

Gao²,

Zhao³

et al. 2013

Int. J. Genet. Mol. Biol.

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In this study, multiple displacement amplification (MDA) was used for transgene detection, and proved to effectively improve the sensitivity of most fluorescence polymerase chain (PCR). Five events of genetically modified plants were chosen for detection in different plant species by MDA and real-time PCRs. Individual primer/probe systems based on each transgene target were used. The extracted DNA of each sample was diluted and used as a template for MDA. All extracted DNA and diluted mdaDNA were detected by real-time PCR. The results were evaluated with respect to DNA quantity, DNA purity and difference of Ct values of the isogenetic DNA extract before and after MDA (D-values). The MDA amplifying effects were largely different among genes and plants. Organelle DNA has more copies than genomic DNA by MDA.

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Whole genome amplification of the rust Puccinia striiformis f. sp. tritici from single spores

Cited by 11 publications

References 32 publications

Tumor Microenvironment — Perivascular and Perinecrotic Niches

Tumor Microenvironment — Perivascular and Perinecrotic Niches

Multiple displacement amplification of whole genomic DNA from urediospores of Puccinia striiformis f. sp. tritici

Application of multiple displacement amplification in detection of five events of genetically modified organisms

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