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The Storm by Kate Chopin

Hideki Hirakawa and Katsuhiko Sumitomo authors contributed equally to this work. Cultivated chrysanthemum Chrysanthemum morifolium Ramat. The diploid Chrysanthemum seticuspe is often used as a model of cultivated chrysanthemum, since the two species are closely related.

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To expand our knowledge of the cultivated chrysanthemum, we here performed de novo whole-genome assembly in C. XMRS10, a C. The 2. The N50 length of scaffolds was 44, bp. Next, based on the assembled genome sequences, we performed linkage map construction, gene discovery and comparative analyses for C. The generated C. Moreover, single nucleotide polymorphism identification and annotation on the C. The genome sequences assembled herein are expected to contribute to future chrysanthemum studies. In addition, our approach demonstrated the Chryswnthemums of short-read genome assembly and the importance of choosing an appropriate next genome sequencing technology based on the purpose of the post-genome analysis.

Chrysanthemum Chrsyanthemums first cultivated in China and developed for horticultural purposes in East Asia. Soon after the discovery of the response of plants to day length, i. Although C. The recent advances in next genome sequencing NGS technology have brought de novo whole-genome sequencing to various organisms. The sequencing cost has been dramatically decreasing while the quality of the assembled Analysis Of The Chrysanthemums And Chopin s has been increasing along with the growth of long-read sequencing technologies. However, de novo whole-genome sequencing is still costly in species with large genomes, and thus many of these species have not benefitted from the new NGS technologies.

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In this study, we performed de novo whole-genome assembly in C. Based on the assembled genome, gene discovery analysis was conducted for genes related to flowering, which is the most important trait in chrysanthemum.

Genetic analysis was also performed such as linkage map construction, comparative phylogenetic investigation between C. Our approach suggests a potential strategy for advancing genetic and genomic studies in species that are not candidates for high quality de novo whole-genome assembly due to biological or other difficulties.

1. Introduction

Three C. AEV2 is a natural self-compatible mutant that was selfed for five generations to reduce genomic heterogeneity. NIFS-3 was used for genome size estimation by flow cytometry and for transcript sequencing for simple sequence repeat SSR marker development for linkage map construction. Four MP libraries were constructed with expected insert sizes of 2, 5, 10 and 15 Kb and sequenced by HiSeq with a Analysis Of The Chrysanthemums And Chopin s length of nt.

Potential contaminated sequences on the assembled scaffolds were identified and removed using BLASTN searches against the chloroplast and mitochondrial genome sequence of A. Known repetitive sequences registered in Repbase and de novo repetitive sequences defined by RepeatModeler 1. The predicted genes were used as a training set for ab initio gene prediction by Augustus in the MAKER-P pipeline, along with the additional training sets continue reading from the following amino acid sequences: A. In parallel, evidence-based gene prediction was performed by mapping the C.

The A. The genes related to flowering of C. Bootstrap values for 1, resampling are shown on each branch.

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After growing the plants under the respective conditions Supplementary Fig. S3shoot tips from each group were sampled for RNA extraction. The expression levels of the three genes were investigated using quantitative real-time PCR. The primers used were as follows. Segregation analysis was also performed using the SSR markers developed in this study.

Primers were designed by Primer3 for the SSRs that had 8, 5, 4, 3 and 3 repeats in the di- tri- tetra- penta- and hexa-motifs, respectively.]