A Simple and Efficient Method for Isolation of DNA in High Mucilaginous Plant Tissues

Echevarría‐Machado, Ileana; Sánchez-Cach, Lucila A.; Hernández‐Zepeda, Cecilia; Rivera‐Madrid, Renata; Moreno‐Valenzuela, Oscar A.

doi:10.1385/mb:31:2:129

Cited by 46 publications

(24 citation statements)

References 5 publications

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“…Another possibility is that the techniques (DNA extraction, PCR primers) employed were ineffective for many of the samples. It is well-known that many eudicot species contain abundant secondary metabolites [12,25], that are known to interfere with nucleic acid isolation or PCR amplification.…”

Section: Discussionmentioning

confidence: 99%

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Preliminary identification and coat protein gene phylogenetic relationships of begomoviruses associated with native flora and cultivated plants from the Yucatan Peninsula of Mexico

et al. 2007

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A number of native and cultivated eudicots in the Yucatan Peninsula of Mexico (YPM) exhibit symptoms associated with virus infection. Symptomatic leaves were collected and assessed for begomoviral detection using polymerase chain reaction (PCR), and universal primers that amplify a fragment of the coat protein gene (core Cp). Begomovirus were detected in nine native and seven cultivated species, representing seven eudicot families. DNA extracts from the 16 hosts were used for PCR amplification and sequencing of a fragment containing the coat protein (Cp) gene. The complete Cp sequence was used to establish provisional species identification. Results indicated that 13 distinct begomovirus species were represented. Among these, five potentially new begomovirus species were identified, for which we propose the names Anoda golden mosaic virus (AnGMV), Boerhavia yellow spot virus (BoYSV), Papaya golden mosaic virus (PaGMV), Desmodium leaf distortion virus (DeLDV), and Hibiscus variegation virus (HiVV). Five previously described begomoviral species were provisionally identified for the first time in the YPM; these include Euphorbia mosaic virus (EuMV), Melon chlorotic leaf curl virus (MCLCuV), Okra yellow mosaic Mexico virus (OkYMMV), Sida golden mosaic virus (SiGMV), and Tobacco apical stunt virus (TbASV). Additionally, viruses previously reported from this region, Bean golden yellow mosaic virus (BGYMV), Pepper golden mosaic virus (PepGMV), and Tomato mottle virus (ToMoV) were provisionally identified in cultivated hosts. Phylogenetic analysis provisionally placed all isolates from the YPM in a Western Hemisphere begomovirus clade.

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Section: Discussionmentioning

confidence: 99%

“…Total plant nucleic acids were extracted from 0.10 g of leaf tissue according to Echevarria-Machado et al [12]. The resulting nucleic acid pellets were resuspended in sterile water and stored at -20°C.…”

Section: Virus Collectionsmentioning

confidence: 99%

Preliminary identification and coat protein gene phylogenetic relationships of begomoviruses associated with native flora and cultivated plants from the Yucatan Peninsula of Mexico

et al. 2007

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“…The method reported by Echevarría-Machado et al (2005) [37] was used for genomic DNA extraction. The concentration and purity of DNA were determined by spectrophotometry (Thermo Scientific NanoDrop™ 1000), and its integrity was verified by electrophoresis on a 1% agarose gel.…”

Section: Dna Extractionmentioning

confidence: 99%

Analysis of Two Clonal Lines (Embryogenic and Non-Embryogenic) of <i>Agave fourcroydes</i> Using AFLP and MSAP

Monja-Mio¹,

Quiroz-Moreno²,

Herrera-Herrera³

et al. 2018

AJPS

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Somatic embryogenesis is a very efficient way to propagate economically important plants; however, not all genotypes within a species can be propagated using this method, as a combined effect of both genetic and epigenetic mechanisms may be involved in the response. The aim of the present study was to perform a comparative analysis of the genetic differences through amplified fragment length polymorphism (AFLP) and the epigenetic differences through methylation-sensitive amplified polymorphism (MSAP) of two Agave fourcroydes clonal lines, one highly embryogenic (K33) and the other non-embryogenic (K7). Genetic and epigenetic variabilities existed within each clonal line; however, the polymorphic profiles from the two marker systems allowed us to clearly distinguish the two clonal lines before somatic embryogenesis induction. During the induction, the changes detected were mainly 1) unmethylated fragments in the initial explants that were methylated during induction (methylation events) and 2) fragments with different methylation states in the initial explant that were unmethylated in some stages of the process (demethylation events). K33 showed greater dynamism in relation to methylation/demethylation events, while K7 presented the methylation events in a more constant range and at higher levels during all process.

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“…Nuclei integrity was confirmed by optical microscopy, and their quantification was carried out using a hemocytometer. Genomic DNA was obtained by following the silica method described by Echeverria-Machado et al (2005). Quantity, quality, and purity of the DNA were evaluated using NANODROP equipment (NANODROP, Promega, USA).…”

Section: Ribosomal Gene Copy Numbermentioning

confidence: 99%

Comparative Characterization of Ribosomal DNA Regions in Different Agave Accessions with Economical Importance

2015

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Two ribosomal DNA regions (5S and 18S) were characterized in three economically important species of Agave Linnaeus, 1753 namely Agave tequilana Weber, 1902; Agave angustifolia Haworth, 1915; and Agave fourcroydes Lemaire, 1864 which are used to produce several products such as tequila, mezcal, and hard fibers. Characterization included Agave L. accessions with different ploidy levels (2n=2x=60 to 2n=6×=180) in order to relate this factor with copy number, haplotype number, expression profile, and predictable functionality of ribosomal DNA (rDNA) sequences. Only total rDNA copy number (5S and 18S) was related with ploidy level. Main differences were found in the 5S rDNA gene since it exhibited different genetic traits of Agave L. accession. In this gene, four different allelic groups (I, 105; II, 107; III, 110; and IV, 111 bp) were detected, which have probably evolved separately, thus exhibiting different expression profiles and different haplotype occurrence. Allelic groups III and IV exhibit the highest number of total and expressed copies in all Agave L. accessions. Nonredundant haplotypes were probably more functional in these allelic groups. Differences between the Agave L. accessions were more clearly observed in the most cultivated accession, A. tequilana (2n=2×=60), where the allelic group III shows non-redundant haplotypes and is transcriptionally upregulated suggesting a different evolutionary pressure on this Agave L. accession.

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A Simple and Efficient Method for Isolation of DNA in High Mucilaginous Plant Tissues

Cited by 46 publications

References 5 publications

Preliminary identification and coat protein gene phylogenetic relationships of begomoviruses associated with native flora and cultivated plants from the Yucatan Peninsula of Mexico

Preliminary identification and coat protein gene phylogenetic relationships of begomoviruses associated with native flora and cultivated plants from the Yucatan Peninsula of Mexico

Analysis of Two Clonal Lines (Embryogenic and Non-Embryogenic) of <i>Agave fourcroydes</i> Using AFLP and MSAP

Comparative Characterization of Ribosomal DNA Regions in Different Agave Accessions with Economical Importance

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A Simple and Efficient Method for Isolation of DNA in High Mucilaginous Plant Tissues

Cited by 46 publications

References 5 publications

Preliminary identification and coat protein gene phylogenetic relationships of begomoviruses associated with native flora and cultivated plants from the Yucatan Peninsula of Mexico

Preliminary identification and coat protein gene phylogenetic relationships of begomoviruses associated with native flora and cultivated plants from the Yucatan Peninsula of Mexico

Analysis of Two Clonal Lines (Embryogenic and Non-Embryogenic) of &lt;i&gt;Agave fourcroydes&lt;/i&gt; Using AFLP and MSAP

Comparative Characterization of Ribosomal DNA Regions in Different Agave Accessions with Economical Importance

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Analysis of Two Clonal Lines (Embryogenic and Non-Embryogenic) of <i>Agave fourcroydes</i> Using AFLP and MSAP