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      Draft Sequencing of the Heterozygous Diploid Genome of Satsuma ( Citrus unshiu Marc.) Using a Hybrid Assembly Approach

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          Abstract

          Satsuma ( Citrus unshiu Marc.) is one of the most abundantly produced mandarin varieties of citrus, known for its seedless fruit production and as a breeding parent of citrus. De novo assembly of the heterozygous diploid genome of Satsuma (“Miyagawa Wase”) was conducted by a hybrid assembly approach using short-read sequences, three mate-pair libraries, and a long-read sequence of PacBio by the PLATANUS assembler. The assembled sequence, with a total size of 359.7 Mb at the N 50 length of 386,404 bp, consisted of 20,876 scaffolds. Pseudomolecules of Satsuma constructed by aligning the scaffolds to three genetic maps showed genome-wide synteny to the genomes of Clementine, pummelo, and sweet orange. Gene prediction by modeling with MAKER-P proposed 29,024 genes and 37,970 mRNA; additionally, gene prediction analysis found candidates for novel genes in several biosynthesis pathways for gibberellin and violaxanthin catabolism. BUSCO scores for the assembled scaffold and predicted transcripts, and another analysis by BAC end sequence mapping indicated the assembled genome consistency was close to those of the haploid Clementine, pummel, and sweet orange genomes. The number of repeat elements and long terminal repeat retrotransposon were comparable to those of the seven citrus genomes; this suggested no significant failure in the assembly at the repeat region. A resequencing application using the assembled sequence confirmed that both kunenbo-A and Satsuma are offsprings of Kishu, and Satsuma is a back-crossed offspring of Kishu. These results illustrated the performance of the hybrid assembly approach and its ability to construct an accurate heterozygous diploid genome.

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          Most cited references68

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          MCScanX: a toolkit for detection and evolutionary analysis of gene synteny and collinearity

          MCScan is an algorithm able to scan multiple genomes or subgenomes in order to identify putative homologous chromosomal regions, and align these regions using genes as anchors. The MCScanX toolkit implements an adjusted MCScan algorithm for detection of synteny and collinearity that extends the original software by incorporating 14 utility programs for visualization of results and additional downstream analyses. Applications of MCScanX to several sequenced plant genomes and gene families are shown as examples. MCScanX can be used to effectively analyze chromosome structural changes, and reveal the history of gene family expansions that might contribute to the adaptation of lineages and taxa. An integrated view of various modes of gene duplication can supplement the traditional gene tree analysis in specific families. The source code and documentation of MCScanX are freely available at http://chibba.pgml.uga.edu/mcscan2/.
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            LTRharvest, an efficient and flexible software for de novo detection of LTR retrotransposons

            Background Transposable elements are abundant in eukaryotic genomes and it is believed that they have a significant impact on the evolution of gene and chromosome structure. While there are several completed eukaryotic genome projects, there are only few high quality genome wide annotations of transposable elements. Therefore, there is a considerable demand for computational identification of transposable elements. LTR retrotransposons, an important subclass of transposable elements, are well suited for computational identification, as they contain long terminal repeats (LTRs). Results We have developed a software tool LTRharvest for the de novo detection of full length LTR retrotransposons in large sequence sets. LTRharvest efficiently delivers high quality annotations based on known LTR transposon features like length, distance, and sequence motifs. A quality validation of LTRharvest against a gold standard annotation for Saccharomyces cerevisae and Drosophila melanogaster shows a sensitivity of up to 90% and 97% and specificity of 100% and 72%, respectively. This is comparable or slightly better than annotations for previous software tools. The main advantage of LTRharvest over previous tools is (a) its ability to efficiently handle large datasets from finished or unfinished genome projects, (b) its flexibility in incorporating known sequence features into the prediction, and (c) its availability as an open source software. Conclusion LTRharvest is an efficient software tool delivering high quality annotation of LTR retrotransposons. It can, for example, process the largest human chromosome in approx. 8 minutes on a Linux PC with 4 GB of memory. Its flexibility and small space and run-time requirements makes LTRharvest a very competitive candidate for future LTR retrotransposon annotation projects. Moreover, the structured design and implementation and the availability as open source provides an excellent base for incorporating novel concepts to further improve prediction of LTR retrotransposons.
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              GenomeScope: fast reference-free genome profiling from short reads.

              GenomeScope is an open-source web tool to rapidly estimate the overall characteristics of a genome, including genome size, heterozygosity rate and repeat content from unprocessed short reads. These features are essential for studying genome evolution, and help to choose parameters for downstream analysis. We demonstrate its accuracy on 324 simulated and 16 real datasets with a wide range in genome sizes, heterozygosity levels and error rates.
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                Author and article information

                Contributors
                Journal
                Front Genet
                Front Genet
                Front. Genet.
                Frontiers in Genetics
                Frontiers Media S.A.
                1664-8021
                05 December 2017
                2017
                : 8
                : 180
                Affiliations
                [1] 1Division of Citrus Research, Institute of Fruit Tree and Tea Science, National Agriculture and Food Research Organization , Shimizu, Japan
                [2] 2Genome Informatics Laboratory, Center for Information Biology, National Institute of Genetics , Mishima, Japan
                [3] 3Comparative Genomics Laboratory, Center for Information Biology, National Institute of Genetics , Mishima, Japan
                Author notes

                Edited by: Xiaowu Wang, Biotechnology Research Institute (CAAS), China

                Reviewed by: Qiang Xu, Huazhong Agricultural University, China; Liu Min, Biomarker Technologies Corporation (China), China

                *Correspondence: Tokurou Shimizu tshimizu@ 123456affrc.go.jp

                This article was submitted to Plant Genetics and Genomics, a section of the journal Frontiers in Genetics

                †Present Address: Hideki Nagasaki, Department of Frontier Research, Kazusa DNA Research Institute, Kisarazu, Japan

                Article
                10.3389/fgene.2017.00180
                5723288
                29259619
                860413f0-53b0-45ac-b10c-16a71fae23c7
                Copyright © 2017 Shimizu, Tanizawa, Mochizuki, Nagasaki, Yoshioka, Toyoda, Fujiyama, Kaminuma and Nakamura.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 07 August 2017
                : 06 November 2017
                Page count
                Figures: 7, Tables: 8, Equations: 0, References: 102, Pages: 19, Words: 13044
                Funding
                Funded by: Ministry of Agriculture, Forestry and Fisheries 10.13039/501100003993
                Award ID: NGB-1006
                Award ID: NGB-2009
                Funded by: Japan Society for the Promotion of Science 10.13039/501100001691
                Award ID: 24580061
                Categories
                Genetics
                Original Research

                Genetics
                citrus,satsuma,draft genome assembly,gene prediction,genome synteny,gibberellic acid biosynthesis,carotenoid biosynthesis,parentage analysis

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