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Post, R. von; Post, Lars von; Dayteg, Christophe; Nilsson, Marie; Forster, B. P.; Tuvesson, Stine

doi:10.1023/a:1022863006134

Cited by 45 publications

(10 citation statements)

References 12 publications

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“…It has been calculated that 60% of the total time required from leaf collection to PCR reaction is used for DNA extraction (Dilworth and Frey 2000). Thus, simple, rapid, robust and inexpensive DNA isolation method is needed for high-throughput MAS (von Post et al 2003; Karakousis and Langridge 2003). Unlike other plant DNA extraction protocols, the method implemented in this study does not include the use of liquid nitrogen or freeze-drying for initial grinding of the tissue.…”

Section: Discussionmentioning

confidence: 99%

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Molecular and genetic characterization of the Ryadg locus on chromosome XI from Andigena potatoes conferring extreme resistance to potato virus Y

et al. 2018

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Key messageWe have elucidated the Andigena origin of the potato Ryadg gene on chromosome XI of CIP breeding lines and developed two marker assays to facilitate its introgression in potato by marker-assisted selection.AbstractPotato virus Y (PVY) is causing yield and quality losses forcing farmers to renew periodically their seeds from clean stocks. Two loci for extreme resistance to PVY, one on chromosome XI and the other on XII, have been identified and used in breeding. The latter corresponds to a well-known source of resistance (Solanum stoloniferum), whereas the one on chromosome XI was reported from S. stoloniferum and S. tuberosum group Andigena as well. To elucidate its taxonomic origin in our breeding lines, we analyzed the nucleotide sequences of tightly linked markers (M45, M6) and screened 251 landraces of S. tuberosum group Andigena for the presence of this gene. Our results indicate that the PVY resistance allele on chromosome XI in our breeding lines originated from S. tuberosum group Andigena. We have developed two marker assays to accelerate the introgression of Ryadg gene into breeding lines by marker-assisted selection (MAS). First, we have multiplexed RYSC3, M6 and M45 DNA markers flanking the Ryadg gene and validated it on potato varieties with known presence/absence of the Ryadg gene and a progeny of 6,521 individuals. Secondly, we developed an allele-dosage assay particularly useful to identify multiplex Ryadg progenitors. The assay based on high-resolution melting analysis at the M6 marker confirmed Ryadg plex level as nulliplex, simplex and duplex progenitors and few triplex progenies. These marker assays have been validated and can be used to facilitate MAS in potato breeding.

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Section: Discussionmentioning

confidence: 99%

“…Plates were put on ice and stored at − 70 °C. DNA extraction was carried out based on the method of Dayteg et al (1998) described by von Post et al (2003) and adapted to the 96-well plate for medium-throughput extraction. Forty microliters of 0.25 M NaOH was added to each well plate directly on the tissue.…”

Section: Methodsmentioning

confidence: 99%

Molecular and genetic characterization of the Ryadg locus on chromosome XI from Andigena potatoes conferring extreme resistance to potato virus Y

et al. 2018

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“…The extraction of high-quality DNA from plant tissue is time consuming, arduous, and costly due to multiple steps and the high cost of liquid nitrogen. In addition, the problems associated with the available commercial kits are their high cost and low yield of DNA [5, 7]. Several methods to isolate DNA from plant tissues are available; however, these methods produce either small amounts or DNA of inconsistent quality.…”

Section: Introductionmentioning

confidence: 99%

Comparison of three genomic DNA extraction methods to obtain high DNA quality from maize

2017

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BackgroundThe world’s top three cereals, based on their monetary value, are rice, wheat, and corn. In cereal crops, DNA extraction is difficult owing to rigid non-cellulose components in the cell wall of leaves and high starch and protein content in grains. The advanced techniques in molecular biology require pure and quick extraction of DNA. The majority of existing DNA extraction methods rely on long incubation and multiple precipitations or commercially available kits to produce contaminant-free high molecular weight DNA.ResultsIn this study, we compared three different methods used for the isolation of high-quality genomic DNA from the grains of cereal crop, Zea mays, with minor modifications. The DNA from the grains of two maize hybrids, M10 and M321, was extracted using extraction methods DNeasy Qiagen Plant Mini Kit, CTAB-method (with/without 1% PVP) and modified Mericon extraction. Genes coding for 45S ribosomal RNA are organized in tandem arrays of up to several thousand copies and contain codes for 18S, 5.8S and 26S rRNA units separated by internal transcribed spacers ITS1 and ITS2. While the rRNA units are evolutionary conserved, ITS regions show high level of interspecific divergence and have been used frequently in genetic diversity and phylogenetic studies. In this study, the genomic DNA was then amplified with PCR using primers specific for ITS gene. PCR products were then visualized on agarose gel.ConclusionThe modified Mericon extraction method was found to be the most efficient DNA extraction method, capable to provide high DNA yields with better quality, affordable cost and less time.

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“…Extraction of the genetic material was performed by alkaline lysis (von Post et al, 2003). A colony of the axenic culture was taken and placed in an Eppendorf tube with 40 μl of NaOH at 0.20 M. It was then heated in a microwave at 10% power or 700 W for 1 min.…”

Section: Dna Extractionmentioning

confidence: 99%

Rhizospheric rhizobia identification in maize (Zea mays L.) plants

et al. 2019

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Rhizobia have been studied for the symbiosis that they establish with the roots of legumes. However, the colonization and promotion of growth in non-leguminous plants has also been demonstrated. The aim of this work was the biochemical and molecular identification of rhizosphere rhizobia present inthe rhizosphere of two commercial maize cultivars. Cultivableisolates were obtained in yeast-mannitol-agar (YMA) mediumfrom rhizospheric soil and the rhizoplane. The cultural (size,color, mucus, etc.), morphological, and staining (cell shape,response to staining and sporulation) characteristics weredetermined as well as isolate responses to eight biochemicaltests (acid-base production, citrate, oxidase, catalase, H2Sproduction, urease, gelatinase and the oxidative-fermentativeassay) that are valuable for rhizobia identification. The genuswas determined by 16S rDNA gene sequencing. We obtained 81total isolates of which 30.86% showed the cultural, morphological and staining characteristics expected for rhizobia and only 20% of these corresponded to the genus Rhizobium.

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Cited by 45 publications

References 12 publications

Molecular and genetic characterization of the Ryadg locus on chromosome XI from Andigena potatoes conferring extreme resistance to potato virus Y

Molecular and genetic characterization of the Ryadg locus on chromosome XI from Andigena potatoes conferring extreme resistance to potato virus Y

Comparison of three genomic DNA extraction methods to obtain high DNA quality from maize

Rhizospheric rhizobia identification in maize (Zea mays L.) plants

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