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      Genome sequence of the agarwood tree Aquilaria sinensis (Lour.) Spreng: the first chromosome-level draft genome in the Thymelaeceae family

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          Abstract

          Backgroud

          Aquilaria sinensis (Lour.) Spreng is one of the important plant resources involved in the production of agarwood in China. The agarwood resin collected from wounded Aquilaria trees has been used in Asia for aromatic or medicinal purposes from ancient times, although the mechanism underlying the formation of agarwood still remains poorly understood owing to a lack of accurate and high-quality genetic information.

          Findings

          We report the genomic architecture of A. sinensis by using an integrated strategy combining Nanopore, Illumina, and Hi-C sequencing. The final genome was ∼726.5 Mb in size, which reached a high level of continuity and a contig N50 of 1.1 Mb. We combined Hi-C data with the genome assembly to generate chromosome-level scaffolds. Eight super-scaffolds corresponding to the 8 chromosomes were assembled to a final size of 716.6 Mb, with a scaffold N50 of 88.78 Mb using 1,862 contigs. BUSCO evaluation reveals that the genome completeness reached 95.27%. The repeat sequences accounted for 59.13%, and 29,203 protein-coding genes were annotated in the genome. According to phylogenetic analysis using single-copy orthologous genes, we found that A. sinensis is closely related to Gossypium hirsutum and Theobroma cacao from the Malvales order, and A. sinensis diverged from their common ancestor ∼53.18–84.37 million years ago.

          Conclusions

          Here, we present the first chromosome-level genome assembly and gene annotation of A. sinensis. This study should contribute to valuable genetic resources for further research on the agarwood formation mechanism, genome-assisted improvement, and conservation biology of Aquilaria species.

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          Most cited references32

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          Exploiting EST databases for the development and characterization of gene-derived SSR-markers in barley (Hordeum vulgare L.).

          A software tool was developed for the identification of simple sequence repeats (SSRs) in a barley ( Hordeum vulgare L.) EST (expressed sequence tag) database comprising 24,595 sequences. In total, 1,856 SSR-containing sequences were identified. Trimeric SSR repeat motifs appeared to be the most abundant type. A subset of 311 primer pairs flanking SSR loci have been used for screening polymorphisms among six barley cultivars, being parents of three mapping populations. As a result, 76 EST-derived SSR-markers were integrated into a barley genetic consensus map. A correlation between polymorphism and the number of repeats was observed for SSRs built of dimeric up to tetrameric units. 3'-ESTs yielded a higher portion of polymorphic SSRs (64%) than 5'-ESTs did. The estimated PIC (polymorphic information content) value was 0.45 +/- 0.03. Approximately 80% of the SSR-markers amplified DNA fragments in Hordeum bulbosum, followed by rye, wheat (both about 60%) and rice (40%). A subset of 38 EST-derived SSR-markers comprising 114 alleles were used to investigate genetic diversity among 54 barley cultivars. In accordance with a previous, RFLP-based, study, spring and winter cultivars, as well as two- and six-rowed barleys, formed separate clades upon PCoA analysis. The results show that: (1) with the software tool developed, EST databases can be efficiently exploited for the development of cDNA-SSRs, (2) EST-derived SSRs are significantly less polymorphic than those derived from genomic regions, (3) a considerable portion of the developed SSRs can be transferred to related species, and (4) compared to RFLP-markers, cDNA-SSRs yield similar patterns of genetic diversity.
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            Fast and accurate long-read assembly with wtdbg2

            Existing long-read assemblers require thousands of CPU hours to assemble a human genome and are being outpaced by sequencing technologies in terms of both throughput and cost. We developed a long-read assembler wtdbg2 (https://github.com/ruanjue/wtdbg2) that is 2–17 times as fast as published tools while achieving comparable contiguity and accuracy. It paves the way for population-scale long-read assembly in future.
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              The sunflower genome provides insights into oil metabolism, flowering and Asterid evolution

              The domesticated sunflower, Helianthus annuus L., is a global oil crop that has promise for climate change adaptation, because it can maintain stable yields across a wide variety of environmental conditions, including drought. Even greater resilience is achievable through the mining of resistance alleles from compatible wild sunflower relatives, including numerous extremophile species. Here we report a high-quality reference for the sunflower genome (3.6 gigabases), together with extensive transcriptomic data from vegetative and floral organs. The genome mostly consists of highly similar, related sequences and required single-molecule real-time sequencing technologies for successful assembly. Genome analyses enabled the reconstruction of the evolutionary history of the Asterids, further establishing the existence of a whole-genome triplication at the base of the Asterids II clade and a sunflower-specific whole-genome duplication around 29 million years ago. An integrative approach combining quantitative genetics, expression and diversity data permitted development of comprehensive gene networks for two major breeding traits, flowering time and oil metabolism, and revealed new candidate genes in these networks. We found that the genomic architecture of flowering time has been shaped by the most recent whole-genome duplication, which suggests that ancient paralogues can remain in the same regulatory networks for dozens of millions of years. This genome represents a cornerstone for future research programs aiming to exploit genetic diversity to improve biotic and abiotic stress resistance and oil production, while also considering agricultural constraints and human nutritional needs.
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                Author and article information

                Journal
                Gigascience
                Gigascience
                gigascience
                GigaScience
                Oxford University Press
                2047-217X
                March 2020
                02 March 2020
                02 March 2020
                : 9
                : 3
                : giaa013
                Affiliations
                [1 ] Hainan Engineering Research Center of Agarwood, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences , Rd. Xueyuan No. 4, Haikou 571101, China
                [2 ] Guangdong Laboratory of Lingnan Modern Agriculture, Shenzhen; Genome Analysis Laboratory of the Ministry of Agriculture; Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences , Rd. Pengfei No. 7, Shenzhen 518120, China
                [3 ] Key Laboratory of Biology and Genetic Resources of Tropical Crops of Ministry of Agriculture and Rural Affairs, Institute of Tropical Bioscience and Biotechnology; Chinese Academy of Tropical Agriculture Sciences , Rd. Xueyuan No. 4, Haikou 571101, China
                Author notes
                Correspondence address. Peng Cui, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Rd. Pengfei No. 7, Shenzhen 518120, China. Tel: +86-13828743816; E-mail: cuipeng@ 123456caas.cn
                Correspondence address. Haofu Dai, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Rd. Xueyuan No. 4, Haikou 571101, China. Tel: +86-89866961869; E-mail: daihaofu@ 123456itbb.org.cn

                Contributed equally to this work.

                Author information
                http://orcid.org/0000-0001-9195-7112
                http://orcid.org/0000-0002-4076-3497
                http://orcid.org/0000-0003-2882-7758
                http://orcid.org/0000-0001-6838-6550
                http://orcid.org/0000-0003-2699-5037
                http://orcid.org/0000-0002-7279-516X
                http://orcid.org/0000-0001-9531-5504
                http://orcid.org/0000-0002-2201-9250
                http://orcid.org/0000-0001-5650-1304
                http://orcid.org/0000-0002-0221-7858
                http://orcid.org/0000-0002-7028-7858
                http://orcid.org/0000-0002-3583-3884
                http://orcid.org/0000-0001-6132-6758
                http://orcid.org/0000-0001-5249-8945
                http://orcid.org/0000-0002-1559-9824
                http://orcid.org/0000-0003-3076-0070
                http://orcid.org/0000-0002-5422-8137
                Article
                giaa013
                10.1093/gigascience/giaa013
                7050300
                32118265
                d795ef4a-0f6d-4de3-a620-ef93fa2b8e2c
                © The Author(s) 2020. Published by Oxford University Press.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 08 August 2019
                : 01 December 2019
                : 03 February 2020
                Page count
                Pages: 10
                Funding
                Funded by: Chinese Academy of Tropical Agricultural Sciences 10.13039/501100005206
                Award ID: 17CXTD-15
                Award ID: 1630052020003
                Funded by: National Natural Science Foundation of China 10.13039/501100001809
                Award ID: 31870668
                Categories
                Data Note

                aquilaria sinensis,agarwood,chromosome-level genome assembly,hi-c sequencing,annotation

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