High‐resolution melting of the <i>cytochrome B</i> gene in fecal DNA: A powerful approach for fox species identification of the <i>Lycalopex</i> genus in Chile

Anabalón, Leonardo; Encina‐Montoya, Francisco; Sánchez, Pedro Mascaro; Solano, Jaime; Benavente, Felipe; Guiñez, Basilio; Olivares, Flavio; Oberti, Carlos; Vega, Rolando

doi:10.1002/ece3.5230

Cited by 3 publications

(1 citation statement)

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“…DNA sequence differences, such as single nucleotide polymorphisms and small insertions and deletions (Indels), can be detected based on the location of a differential peak and the shape of the melting transition curves. 15,24 The ITS2 region barcode marker was chosen because it is widely used for phylogenetic reconstructions at both genus and species levels. 25,26 In addition, the chloroplast ribulose bisphosphate carboxylase large chain gene (rbcL) is another barcode region used in most land plants due to the ease of amplification, the availability of databases 21,[27][28][29] and its potential application for authenticity testing.…”

Section: Introductionmentioning

confidence: 99%

Cannabis Seeds Authentication by Chloroplast and Nuclear DNA Analysis Coupled with High-Resolution Melting Method for Quality Control Purposes

Anabalón

Solano

Encina‐Montoya

et al. 2022

Cannabis and Cannabinoid Research

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Background: Cannabis plants and their seed have been used in many cultures as a source of medicine and feeding during history. Today, there is an increasing demand for cannabis seeds for medical use. Moreover, a seed sales market with no legal regulations has also grown. This may pose some issues if a quality control is not set in place. Identification of cannabis strains is important for quality control purposes in a nonregulated growing market and in cases of illegal traffic and medical use. Owing to the high price as a pharmacological drug, commercial products of cannabis plants and seeds for medical users are often subjected to adulterations, either when packing or distributing certified seeds in the market. Materials and Methods: Cannabis commercial seeds and cannabis seeds for medical use were analyzed with high-resolution melting (HRM) analysis using barcoding markers. Humulus lupulus L. plants from a local market were used as outgroup control. DNA barcoding uses specific regions of the genome to identify differences in the genetic sequence of conserved regions such as internal transcribed spacer (ITS) and rbcL. DNA barcoding data can be generated with real-time polymerase chain reaction combined with HRM analysis to distinguish specific conserved DNA regions of closely related species. HRM analysis is the method of choice for rapid analysis of sequence variation. Results: The melting temperature (T m ) of homogeneous packages was consistent with single genotypes. However, packages containing contaminating seeds showed T m differences of 0.2°C on average. Conclusions: An effective, rapid, and low-cost method based on ITS nuclear DNA and on chloroplast rbcL regions for screening and detection of contamination in commercial cannabis seeds was developed and applied for the analysis of different samples. This approach can be used as a quality control tool for cannabis seeds or other plant material.

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Section: Introductionmentioning

confidence: 99%

Cannabis Seeds Authentication by Chloroplast and Nuclear DNA Analysis Coupled with High-Resolution Melting Method for Quality Control Purposes

Anabalón

Solano

Encina‐Montoya

et al. 2022

Cannabis and Cannabinoid Research

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A forest‐specialist carnivore in the middle of the desert?Comments on Anabalon et al. 2019

Moreira‐Arce

Silva‐Rodríguez

Napolitano

et al. 2020

Ecology and Evolution

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We present comments on an article recently published in Ecology and Evolution ("High-resolution melting of the cytochrome B gene in fecal DNA: A powerful approach for fox species identification of the Lycalopex genus in Chile") by Anabalon et al. that reported the presence of Darwin's fox (Lycalopex fulvipes), a temperate forest specialist, in the hyperarid Atacama Desert of northern Chile. We argue that this putative record lacks ecological support in light of ongoing research on this endangered species, and contains numerous methodological flaws and omissions related to the molecular identification of the species. Based on these issues, we suggest the scientific community and conservation decision-makers disregard the alleged presence of the Darwin's fox in the Atacama Desert. The ongoing vertebrate population declines reported in the Living Planet Report (WWF, 2018) challenges conservationists and environmental agencies to provide reliable information, particularly on endangered species, to inform decisions and help to achieve conservation goals. The discovery of new populations is particularly important for conservation planning, especially when laying outside the known geographic distributions (Guisan et al., 2013; Margules & Pressey, 2000). In a recent issue of Ecology and Evolution, Anabalón et al. (2019) used high-resolution melting (HRM), based on thermal denaturation of DNA amplicons, as a method to identify different Chilean fox species (Lycalopex spp.) from scat samples collected in the Atacama Desert, northern Chile (ca. 26°31′S, 70°30′W; Figure 1), one of the most arid environments in the world (McKay et al., 2003). The study reports a remarkable discovery: the presence of the endangered Darwin's fox (Lycalopex fulvipes), in the hyperarid Atacama Desert, more than 1,200 km further north than the limit of its currently known distribution (Figure 1). We argue this record is South American gray fox (L. griseus) is present on Chiloe Island (a stronghold for the Darwin's fox); this is incorrect (see González delThis is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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Comparative Analyses of the Gut Microbiome of Two Fox Species, the Red Fox (Vulpes Vulpes) and Corsac Fox (Vulpes Corsac), that Occupy Different Ecological Niches

et al. 2021

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High‐resolution melting of the cytochrome B gene in fecal DNA: A powerful approach for fox species identification of the Lycalopex genus in Chile

Cited by 3 publications

References 31 publications

Cannabis Seeds Authentication by Chloroplast and Nuclear DNA Analysis Coupled with High-Resolution Melting Method for Quality Control Purposes

Cannabis Seeds Authentication by Chloroplast and Nuclear DNA Analysis Coupled with High-Resolution Melting Method for Quality Control Purposes

A forest‐specialist carnivore in the middle of the desert?Comments on Anabalon et al. 2019

Comparative Analyses of the Gut Microbiome of Two Fox Species, the Red Fox (Vulpes Vulpes) and Corsac Fox (Vulpes Corsac), that Occupy Different Ecological Niches

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