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      Evidence of facultative parthenogenesis in three Neotropical pitviper species of the Bothrops atrox group

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          Abstract

          We examined four suspected cases of facultative parthenogenesis in three species of a neotropical lineage of pitvipers of the Bothrops atrox group. Reproduction without mating was observed in captive females of B. atrox, B. moojeni and B. leucurus housed alone for seven years (the two former species) and nine years (the latter one). In addition to the observation of captivity data, we investigated molecularly this phenomenon using heterologous microsatellites. DNA was extracted from the mothers’ scales or liver, from embryo and newborn fragments, and yolked ova. Four of the microsatellites showed good amplification using Polymerase Chain Reaction and informative band segregation patterns among each mother and respective offspring. Captivity information, litter characteristics (comparison of the number of newborns, embryos and yolked ova) and molecular data altogether agreed with facultative parthenogenesis predictions in at least three out of the four mothers studied: B. atrox (ID#933) was heterozygous for three out of the four markers, and the sons S1 and S2 were homozygous; B. moojeni (BUT86) was heterozygous for two out of four markers, offspring S1, S3, E2, and E4, and O1 to O6 were homozygous; and B. leucurus (MJJS503) was heterozygous for three out of four markers, and son E1 and O1 were homozygous. B. moojeni (BUT44) was homozygous for all loci analyzed in the mother and offspring, which although not informative is also consistent with parthenogenesis. This study represents the first molecular confirmation of different pitviper species undergoing facultative parthenogenesis among Neotropical endemic snakes.

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          Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material.

          Procedures utilizing Chelex 100 chelating resin have been developed for extracting DNA from forensic-type samples for use with the PCR. The procedures are simple, rapid, involve no organic solvents and do not require multiple tube transfers for most types of samples. The extraction of DNA from semen and very small bloodstains using Chelex 100 is as efficient or more efficient than using proteinase K and phenol-chloroform extraction. DNA extracted from bloodstains seems less prone to contain PCR inhibitors when prepared by this method. The Chelex method has been used with amplification and typing at the HLA DQ alpha locus to obtain the DQ alpha genotypes of many different types of samples, including whole blood, bloodstains, seminal stains, buccal swabs, hair and post-coital samples. The results of a concordance study are presented in which the DQ alpha genotypes of 84 samples prepared using Chelex or using conventional phenol-chloroform extraction are compared. The genotypes obtained using the two different extraction methods were identical for all samples tested.
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            Purging inbreeding depression and the probability of extinction: full-sib mating.

            Inbreeding depression has been a topic of interest in recent years from a number of perspectives, particularly in the captive breeding of endangered species. Generally, the goal of captive breeding is to avoid the detrimental effects of inbreeding depression and to retain genetic variation for future adaptation. However, an important component of another suggested approach to captive breeding is to purge rapidly the population of its genetic load so that its long-term fitness is not compromised. I have examined the effectiveness of purging the genetic load by documenting both the reduction in inbreeding depression and the increase of the probability of extinction when there is continuous full-sib mating. When the genetic load is the result of lethals, the inbreeding depression is quickly purged without a high probability of extinction, except when the total genetic load is high. On the other hand, if the load is due to detrimentals of relatively small effect, the genetic load becomes fixed, the mean fitness is reduced, and the probability of extinction may be greatly increased. In other words, the success of such a programme to purge genetic load without an increase in the probability of extinction is highly dependent upon the genetic basis of inbreeding depression, information that is not readily available for most species.
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              Unisexual reproduction among vertebrates.

              The past decade has seen a remarkable revision of perspectives on unisexual reproduction in vertebrates. One can no longer view it as a rare curiosity far outside the mainstream of evolution. More than 80 taxa of fish, amphibians, and reptiles are now known to reproduce by parthenogenesis (Greek for 'virgin birth') or its variants, and they persist in nature as all-female lineages. Other lower vertebrates that ordinarily rely on sexual reproduction can resort to facultative parthenogenesis under extenuating circumstances that isolate females from males. Molecular tools have now been applied to the study of unisexual organisms, and fascinating insights have emerged regarding the molecular mechanisms that preserve heterozygosity and increase genetic diversity in all-female populations. A deeper understanding of the underlying genetics increasingly calls into question the assumption that unisexuality in vertebrates is an evolutionary dead-end. Copyright © 2010 Elsevier Ltd. All rights reserved.
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                Author and article information

                Contributors
                Journal
                PeerJ
                PeerJ
                peerj
                peerj
                PeerJ
                PeerJ Inc. (San Diego, USA )
                2167-8359
                18 November 2020
                2020
                : 8
                : e10097
                Affiliations
                [1 ]Laboratório de Ecologia e Evolução, Instituto Butantan , São Paulo, Brazil
                [2 ]Laboratório Especial de Ciclo Celular, Instituto Butantan , São Paulo, Brazil
                [3 ]Aquário Municipal de Campinas , Campinas, São Paulo, Brazil
                [4 ]Parque Ecológico Municipal de Americana , Americana, São Paulo, Brazil
                [5 ]Museu Biológico, Instituto Butantan , São Paulo, Brazil
                Article
                10097
                10.7717/peerj.10097
                7680053
                ed6c68c7-6650-457a-8290-8b26328beedc
                ©2020 Cubides-Cubillos et al.

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ) and either DOI or URL of the article must be cited.

                History
                : 15 October 2019
                : 14 September 2020
                Funding
                Funded by: FAPESP (Jovem Pesquisador 2005/04557-3 and Auxílio Regular 2014/02885-2 for Maria José de J. Silva)
                Funded by: INCTTOX (Proc. 573790/2008-6 and a Post-Doctoral CNPq Fellowship for José S.L. Patané #151069/2009-3)
                Funded by: CNPq Grant #306490/2012-9
                Funded by: CAPES (Finance Code 001) fellowship for Sergio D. Cubides-Cubillos
                Funded by: Fundacão Butantan
                This work was supported by FAPESP (Jovem Pesquisador 2005/04557-3 and Auxílio Regular 2014/02885-2 for Maria José de J. Silva), INCTTOX (Proc. 573790/2008-6 and a Post-Doctoral CNPq Fellowship for José Salvatore Leister Patané #151069/2009-3), CNPq Grant #306490/2012-9, and CAPES (Finance Code 001) fellowship for Sergio D. Cubides-Cubillos, and Fundacão Butantan (payment of publication fee). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Biodiversity
                Developmental Biology
                Genetics
                Molecular Biology
                Zoology

                serpentes,squamata,neotropical snakes,b. moojeni,b. leucurus,automixis,genetic markers,microsatellites

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