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      Chromosome‐scale assembly of the genome of Salix dunnii reveals a male‐heterogametic sex determination system on chromosome 7

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          Abstract

          Sex determination systems in plants can involve either female or male heterogamety (ZW or XY, respectively). Here we used Illumina short reads, Oxford Nanopore Technologies (ONT) long reads and Hi‐C reads to assemble the first chromosome‐scale genome of a female willow tree ( Salix dunnii), and to predict genes using transcriptome sequences and available databases. The final genome sequence of 328 Mb in total was assembled in 29 scaffolds, and includes 31,501 predicted genes. Analyses of short‐read sequence data that included female and male plants suggested a male heterogametic sex‐determining factor on chromosome 7, implying that, unlike the female heterogamety of most species in the genus Salix, male heterogamety evolved in the subgenus Salix. The S. dunnii sex‐linked region occupies about 3.21 Mb of chromosome 7 in females (representing its position in the X chromosome), probably within a pericentromeric region. Our data suggest that this region is enriched for transposable element insertions, and about one‐third of its 124 protein‐coding genes were gained via duplications from other genome regions. We detect purifying selection on the genes that were ancestrally present in the region, though some have been lost. Transcriptome data from female and male individuals show more male‐ than female‐biased genes in catkin and leaf tissues, and indicate enrichment for male‐biased genes in the pseudo‐autosomal regions. Our study provides valuable genomic resources for further studies of sex‐determining regions in the family Salicaceae, and sex chromosome evolution.

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          Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2

          In comparative high-throughput sequencing assays, a fundamental task is the analysis of count data, such as read counts per gene in RNA-seq, for evidence of systematic changes across experimental conditions. Small replicate numbers, discreteness, large dynamic range and the presence of outliers require a suitable statistical approach. We present DESeq2, a method for differential analysis of count data, using shrinkage estimation for dispersions and fold changes to improve stability and interpretability of estimates. This enables a more quantitative analysis focused on the strength rather than the mere presence of differential expression. The DESeq2 package is available at http://www.bioconductor.org/packages/release/bioc/html/DESeq2.html. Electronic supplementary material The online version of this article (doi:10.1186/s13059-014-0550-8) contains supplementary material, which is available to authorized users.
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            The Sequence Alignment/Map format and SAMtools

            Summary: The Sequence Alignment/Map (SAM) format is a generic alignment format for storing read alignments against reference sequences, supporting short and long reads (up to 128 Mbp) produced by different sequencing platforms. It is flexible in style, compact in size, efficient in random access and is the format in which alignments from the 1000 Genomes Project are released. SAMtools implements various utilities for post-processing alignments in the SAM format, such as indexing, variant caller and alignment viewer, and thus provides universal tools for processing read alignments. Availability: http://samtools.sourceforge.net Contact: rd@sanger.ac.uk
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              STAR: ultrafast universal RNA-seq aligner.

              Accurate alignment of high-throughput RNA-seq data is a challenging and yet unsolved problem because of the non-contiguous transcript structure, relatively short read lengths and constantly increasing throughput of the sequencing technologies. Currently available RNA-seq aligners suffer from high mapping error rates, low mapping speed, read length limitation and mapping biases. To align our large (>80 billon reads) ENCODE Transcriptome RNA-seq dataset, we developed the Spliced Transcripts Alignment to a Reference (STAR) software based on a previously undescribed RNA-seq alignment algorithm that uses sequential maximum mappable seed search in uncompressed suffix arrays followed by seed clustering and stitching procedure. STAR outperforms other aligners by a factor of >50 in mapping speed, aligning to the human genome 550 million 2 × 76 bp paired-end reads per hour on a modest 12-core server, while at the same time improving alignment sensitivity and precision. In addition to unbiased de novo detection of canonical junctions, STAR can discover non-canonical splices and chimeric (fusion) transcripts, and is also capable of mapping full-length RNA sequences. Using Roche 454 sequencing of reverse transcription polymerase chain reaction amplicons, we experimentally validated 1960 novel intergenic splice junctions with an 80-90% success rate, corroborating the high precision of the STAR mapping strategy. STAR is implemented as a standalone C++ code. STAR is free open source software distributed under GPLv3 license and can be downloaded from http://code.google.com/p/rna-star/.
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                Author and article information

                Contributors
                heli198724@163.com , jianfeng.mao@bjfu.edu.cn
                jianfeng.mao@bjfu.edu.cn
                Journal
                Mol Ecol Resour
                Mol Ecol Resour
                10.1111/(ISSN)1755-0998
                MEN
                Molecular Ecology Resources
                John Wiley and Sons Inc. (Hoboken )
                1755-098X
                1755-0998
                16 March 2021
                August 2021
                : 21
                : 6 ( doiID: 10.1111/men.v21.6 )
                : 1966-1982
                Affiliations
                [ 1 ] Beijing Advanced Innovation Center for Tree Breeding by Molecular Design National Engineering Laboratory for Tree Breeding Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants Ministry of Education College of Biological Sciences and Technology Beijing Forestry University Beijing China
                [ 2 ] College of Forestry Fujian Agriculture and Forestry University Fuzhou China
                [ 3 ] Ori (Shandong) Gene Science and Technology Co., Ltd Weifang China
                [ 4 ] Department of Systematics, Biodiversity and Evolution of Plants (with Herbarium) University of Goettingen Göttingen Germany
                [ 5 ] Department of Ecology and Evolutionary Biology University of California Irvine Irvine CA USA
                [ 6 ] Institute of Evolutionary Biology School of Biological Sciences University of Edinburgh Edinburgh UK
                Author notes
                [*] [* ] Correspondence

                Li He and Jian‐Feng Mao, Beijing Advanced Innovation Center for Tree Breeding by Molecular Design, National Engineering Laboratory for Tree Breeding, Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants, Ministry of Education, College of Biological Sciences and Technology, Beijing Forestry University, Beijing, China.

                Emails: heli198724@ 123456163.com (LH); jianfeng.mao@ 123456bjfu.edu.cn (JFM)

                Author information
                https://orcid.org/0000-0002-4591-8056
                https://orcid.org/0000-0001-6623-7623
                https://orcid.org/0000-0002-7600-1128
                https://orcid.org/0000-0001-9735-8516
                Article
                MEN13362
                10.1111/1755-0998.13362
                8359994
                33609314
                1e4af143-cebd-4d10-9557-2e6370161a50
                © 2021 The Authors. Molecular Ecology Resources published by John Wiley & Sons Ltd

                This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 10 February 2021
                : 01 November 2020
                : 16 February 2021
                Page count
                Figures: 4, Tables: 3, Pages: 0, Words: 13640
                Funding
                Funded by: National Natural Science Foundation of China , doi 10.13039/501100001809;
                Award ID: 31800466
                Funded by: Natural Science Foundation of Fujian Province of China
                Award ID: 2018J01613
                Categories
                Resource Article
                RESOURCE ARTICLES
                Permanent Genetic Resources
                Custom metadata
                2.0
                August 2021
                Converter:WILEY_ML3GV2_TO_JATSPMC version:6.0.5 mode:remove_FC converted:12.08.2021

                Ecology
                gene expression,genome‐wide association,long terminal repeat‐retrotransposons,sex‐linked region,xx/xy

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