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      Recombinants from the crosses between amphidiploid and cultivated peanut ( Arachis hypogaea) for pest-resistance breeding programs

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          Abstract

          Peanut is a major oilseed crop worldwide. In the Brazilian peanut production, silvering thrips and red necked peanut worm are the most threatening pests. Resistant varieties are considered an alternative to pest control. Many wild diploid Arachis species have shown resistance to these pests, and these can be used in peanut breeding by obtaining hybrid of A and B genomes and subsequent polyploidization with colchicine, resulting in an AABB amphidiploid. This amphidiploid can be crossed with cultivated peanut (AABB) to provide genes of interest to the cultivar. In this study, the sterile diploid hybrids from A. magna V 13751 and A. kempff-mercadoi V 13250 were treated with colchicine for polyploidization, and the amphidiploids were crossed with A. hypogaea cv. IAC OL 4 to initiate the introgression of the wild genes into the cultivated peanut. The confirmation of the hybridity of the progenies was obtained by: (1) reproductive characterization through viability of pollen, (2) molecular characterization using microsatellite markers and (3) morphological characterization using 61 morphological traits with principal component analysis. The diploid hybrid individual was polyploidized, generating the amphidiploid An 13 ( A. magna V 13751 x A. kempff-mercadoi V 13250) 4x. Four F 1 hybrid plants were obtained from IAC OL 4 × An 13, and 51 F 2 seeds were obtained from these F 1 plants. Using reproductive, molecular and morphological characterizations, it was possible to distinguish hybrid plants from selfed plants. In the cross between A. hypogaea and the amphidiploid, as the two parents are polyploid, the hybrid progeny and selves had the viability of the pollen grains as high as the parents. This fact turns the use of reproductive characteristics impossible for discriminating, in this case, the hybrid individuals from selfing. The hybrids between A. hypogaea and An 13 will be used in breeding programs seeking pest resistance, being subjected to successive backcrosses until recovering all traits of interest of A. hypogaea, keeping the pest resistance.

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          Genetic linkage maps of Eucalyptus grandis and Eucalyptus urophylla using a pseudo-testcross: mapping strategy and RAPD markers.

          We have used a "two-way pseudo-testcross" mapping strategy in combination with the random amplified polymorphic DNA (RAPD) assay to construct two moderate density genetic linkage maps for species of Eucalyptus. In the cross between two heterozygous individuals many single-dose RAPD markers will be heterozygous in one parent, null in the other and therefore segregate 1:1 in their F1 progeny following a testcross configuration. Meiosis and gametic segregation in each individual can be directly and efficiently analyzed using RAPD markers. We screened 305 primers of arbitrary sequence, and selected 151 to amplify a total of 558 markers. These markers were grouped at LOD 5.0, theta = 0.25, resulting in the maternal Eucalyptus grandis map having a total of 240 markers into 14 linkage groups (1552 cM) and the paternal Eucalyptus urophylla map with 251 markers in 11 linkage groups (1101 cM) (n = 11 in Eucalyptus). Framework maps ordered with a likelihood support > or = 1000:1 were assembled covering 95% of the estimated genome size in both individuals. Characterization of genome complexity of a sample of 48 mapped random amplified polymorphic DNA (RAPD) markers indicate that 53% amplify from low copy regions. These are the first reported high coverage linkage maps for any species of Eucalyptus and among the first for any hardwood tree species. We propose the combined use of RAPD markers and the pseudo-testcross configuration as a general strategy for the construction of single individual genetic linkage maps in outbred forest trees as well as in any highly heterozygous sexually reproducing living organisms. A survey of the occurrence of RAPD markers in different individuals suggests that the pseudo-testcross/RAPD mapping strategy should also be efficient at the intraspecific level and increasingly so with crosses of genetically divergent individuals. The ability to quickly construct single-tree genetic linkage maps in any forest species opens the way for a shift from the paradigm of a species index map to the heterodox proposal of constructing several maps for individual trees of a population, therefore mitigating the problem of linkage equilibrium between marker and trait loci for the application of marker assisted strategies in tree breeding.
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            Princípios de resistência de plantas a insetos

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              A study of the relationships of cultivated peanut (Arachis hypogaea) and its most closely related wild species using intron sequences and microsatellite markers.

              The genus Arachis contains 80 described species. Section Arachis is of particular interest because it includes cultivated peanut, an allotetraploid, and closely related wild species, most of which are diploids. This study aimed to analyse the genetic relationships of multiple accessions of section Arachis species using two complementary methods. Microsatellites allowed the analysis of inter- and intraspecific variability. Intron sequences from single-copy genes allowed phylogenetic analysis including the separation of the allotetraploid genome components. Intron sequences and microsatellite markers were used to reconstruct phylogenetic relationships in section Arachis through maximum parsimony and genetic distance analyses. Although high intraspecific variability was evident, there was good support for most species. However, some problems were revealed, notably a probable polyphyletic origin for A. kuhlmannii. The validity of the genome groups was well supported. The F, K and D genomes grouped close to the A genome group. The 2n = 18 species grouped closer to the B genome group. The phylogenetic tree based on the intron data strongly indicated that A. duranensis and A. ipaënsis are the ancestors of A. hypogaea and A. monticola. Intron nucleotide substitutions allowed the ages of divergences of the main genome groups to be estimated at a relatively recent 2·3-2·9 million years ago. This age and the number of species described indicate a much higher speciation rate for section Arachis than for legumes in general. The analyses revealed relationships between the species and genome groups and showed a generally high level of intraspecific genetic diversity. The improved knowledge of species relationships should facilitate the utilization of wild species for peanut improvement. The estimates of speciation rates in section Arachis are high, but not unprecedented. We suggest these high rates may be linked to the peculiar reproductive biology of Arachis.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                19 April 2017
                2017
                : 12
                : 4
                : e0175940
                Affiliations
                [1 ]Departamendo de Morfologia e Patologia, Universidade Federal de São Carlos, São Carlos, São Paulo, Brasil
                [2 ]Embrapa Pecuária Sudeste, São Carlos, São Paulo, Brasil
                Bhabha Atomic Research Centre, INDIA
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                • Conceptualization: AFP BBZV APF.

                • Data curation: AFP.

                • Formal analysis: AFP APF.

                • Funding acquisition: APF.

                • Investigation: AFP NBD BBZV APF.

                • Methodology: AFP BBZV APF.

                • Project administration: APF.

                • Resources: BBZV APF.

                • Supervision: BBZV APF.

                • Validation: AFP NBD BBZV APF.

                • Writing – original draft: AFP.

                • Writing – review & editing: AFP NBD BBZV APF.

                [¤a]

                Current address: Departamento de Morfologia e Patologia, Universidade Federal de São Carlos, Centro de Ciências Biológicas e da Saúde, São Carlos, São Paulo, Brasil

                [¤b]

                Current address: Embrapa Pecuária Sudeste, Fazenda Canchim, São Carlos, São Paulo, Brasil

                Article
                PONE-D-16-32179
                10.1371/journal.pone.0175940
                5396913
                28423007
                a08a9aa6-68fb-4cae-86a0-13908bf7d5ed
                © 2017 de Paula et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 11 August 2016
                : 3 April 2017
                Page count
                Figures: 6, Tables: 4, Pages: 15
                Funding
                Funded by: Embrapa
                Award ID: 0211080060008
                Award Recipient :
                Funded by: CNPq
                Award ID: 471657/2011-5
                Award Recipient :
                Funded by: CAPES
                Award Recipient :
                APF received funds by Embrapa (no. 0211080060008) and National Council for Scientific and Technological Development (CNPq, grant number 471657/2011-5). AFP received a grant by Coordination for the Improvement of Higher Education Personnel (CAPES). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Plant Science
                Plant Anatomy
                Pollen
                Biology and Life Sciences
                Organisms
                Plants
                Legumes
                Peanut
                Biology and Life Sciences
                Plant Science
                Plant Anatomy
                Seeds
                Medicine and Health Sciences
                Pharmacology
                Drugs
                Colchicine
                Biology and Life Sciences
                Plant Science
                Plant Anatomy
                Trichomes
                Biology and Life Sciences
                Organisms
                Animals
                Invertebrates
                Arthropoda
                Mites
                Biology and Life Sciences
                Agriculture
                Agronomy
                Plant Breeding
                Biology and Life Sciences
                Genetics
                Plant Genetics
                Biology and Life Sciences
                Plant Science
                Plant Genetics
                Custom metadata
                All relevant data are within the paper and its Supporting Information files.

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                Uncategorized

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