A Fast, Simple, and Reliable High-Yielding Method for DNA Extraction From Different Plant Species

Tapia-Tussell, Raúl; Quijano-Ramayo, Andrés; Rojas-Herrera, Rafael; Larqué-Saavedra, Alfonso; Pérez‐Brito, Daisy

doi:10.1385/mb:31:2:137

Cited by 24 publications

(18 citation statements)

References 2 publications

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“…About 50 mg of fresh tissue from infected lesion was processed according to the method described previously [28]. In order to detect C. capsici, PCR reactions were carried out with the C. capsici-specific primers developed in this study and under the reaction conditions described for this primer set.…”

Section: Detection Of C Capsici From Naturally Infected Papayamentioning

confidence: 99%

A Species-Specific Polymerase Chain Reaction Assay for Rapid and Sensitive Detection of Colletotrichum capsici

et al. 2011

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Colletotrichum capsici is an important fungal species that causes anthracnose in many genera of plants causing severe economic losses worldwide. A primer set was designed based on the sequences of the ribosomal internal transcribed spacer (ITS1 and ITS2) regions for use in a conventional PCR assay. The primer set (CcapF/CcapR) amplified a single product of 394 bp with DNA extracted from 20 Mexican isolates of C. capsici. The specificity of primers was confirmed by the absence of amplified product with DNA of four other Colletotrichum species and eleven different fungal genera. This primer set is capable of amplifying only C. capsici from different contaminated tissues or fungal structures, thereby facilitating rapid diagnoses as there is no need to isolate and cultivate the fungus in order to identify it. The sensitivity of detection with this PCR method was 10 pg of genomic DNA from the pathogen. This is the first report of a C. capsici-specific primer set. It allows rapid pathogen detection and provides growers with a powerful tool for a rational selection of fungicides to control anthracnose in different crops and in the post-harvest stage.

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Section: Detection Of C Capsici From Naturally Infected Papayamentioning

confidence: 99%

A Species-Specific Polymerase Chain Reaction Assay for Rapid and Sensitive Detection of Colletotrichum capsici

et al. 2011

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“…Although the quantity and quality of DNAs obtained by these methods are generally satisfactory, these protocols are not universally applicable to all plants (e.g., Prosopis spp). Additionally, although some reported methods have minimized the DNA extraction steps, they still need a large amount of plant tissue, lysis steps, and liquid nitrogen (Tussell et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

Fast protocol for extraction of DNA from Prosopis spp leaves (plant adapted to arid environment) without liquid nitrogen

2013

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ABSTRACT. The extraction of high-quality genomic DNA from Prosopis spp for polymerase chain reaction (PCR) amplification is complicated, owing to the presence of a high percentage of secondary metabolites that bind to or co-precipitate with nucleic acids. In the present study, we report a modified sodium dodecyl sulfate/phenol protocol that eliminates the use of liquid nitrogen in the maceration process, β-mercaptoethanol in the buffer extraction, and the ethanol precipitation step. The A 260 /A 280 absorbance ratios of the isolated DNA were approximately 2.0 to 1.9, suggesting that the DNA fraction was pure and can be used for further PCR analysis. The DNA isolated by this protocol is of sufficient quality for molecular applications; this technique could be applied to other organisms that have similar substances that hinder DNA extraction. Finally, this proposal represents an alternative fast, cheap, and effective method for the isolation of genomic DNA from fresh leaves of Prosopis spp, even in low-technology laboratories.

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“…Total genomic DNA used as templates in PCR reaction was extracted from approximately 1.5 g of cut midrib tissues of tomato plants with disease symptoms, according to a method developed in the GeMBio laboratory and this was used in all the tests performed (Tapia-Tussell et al, 2005).…”

Section: Nucleic Acid Extractionmentioning

confidence: 99%

Molecular characterization of Yucatan tomato phytoplasma (Group 16Sr III)s

Tapia-Tussell¹

2012

Afr. J. Biotechnol.

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Tomato (Lycopersicon esculentum) is an important vegetable crop in Mexico. Recently, a phytoplasma associated with leaf yellowing and curling, severe stunting and little leaf in tomato plant was identified as Yucatan tomato phytoplasma (16SrIII group). DNAs extracted from tomato leaves with symptoms were examined for the presence of this phytoplasma by nested polymerase chain reaction (PCR). Positive results were obtained in 44% of samples, yielding an rDNA product of 1.25 kb. In vitro and in silico restriction fragment length polymorphism (RFLP) patterns obtained with endonucleases HpaII, MseI, RsaI and TaqI were characteristics of group 16SrIII, according to the classification scheme of phytoplasmas. The pattern with AluI and HaeIII discriminated between these phytoplasmas and the members of 16SrIII group. Molecular characterization of the causal agent of Yucatan tomato phytoplasma will facilitate the study of this disease's epidemic aspects and its phytosanitary management. In addition, it will contribute to a greater knowledge of the genetic diversity of phytoplasmas present in Mexico.

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A Fast, Simple, and Reliable High-Yielding Method for DNA Extraction From Different Plant Species

Cited by 24 publications

References 2 publications

A Species-Specific Polymerase Chain Reaction Assay for Rapid and Sensitive Detection of Colletotrichum capsici

A Species-Specific Polymerase Chain Reaction Assay for Rapid and Sensitive Detection of Colletotrichum capsici

Fast protocol for extraction of DNA from Prosopis spp leaves (plant adapted to arid environment) without liquid nitrogen

Molecular characterization of Yucatan tomato phytoplasma (Group 16Sr III)s

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