Rhee Sun-Ju, Kwon Taehyung, Seo Minseok, Jang Yoon Jeong, Sim Tae Yong, Cho Seoae, Han Sang-Wook, Lee Gung Pyo
Department of Integrative Plant Science, Chung-Ang University, Ansung, Republic of Korea.
Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, Republic of Korea.
PLoS One. 2017 Nov 2;12(11):e0187147. doi: 10.1371/journal.pone.0187147. eCollection 2017.
The whole-genome sequence of watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai), a valuable horticultural crop worldwide, was released in 2013. Here, we compared a de novo-based approach (DBA) to a reference-based approach (RBA) using RNA-seq data, to aid in efforts to improve the annotation of the watermelon reference genome and to obtain biological insight into male-sterility in watermelon. We applied these techniques to available data from two watermelon lines: the male-sterile line DAH3615-MS and the male-fertile line DAH3615. Using DBA, we newly annotated 855 watermelon transcripts, and found gene functional clusters predicted to be related to stimulus responses, nucleic acid binding, transmembrane transport, homeostasis, and Golgi/vesicles. Among the DBA-annotated transcripts, 138 de novo-exclusive differentially-expressed genes (DEDEGs) related to male sterility were detected. Out of 33 randomly selected newly annotated transcripts and DEDEGs, 32 were validated by RT-qPCR. This study demonstrates the usefulness and reliability of the de novo transcriptome assembly in watermelon, and provides new insights for researchers exploring transcriptional blueprints with regard to the male sterility.
西瓜(Citrullus lanatus (Thunb.) Matsum. & Nakai)是一种在全球范围内具有重要价值的园艺作物。其全基因组序列于2013年发布。在此,我们使用RNA测序数据将基于从头测序的方法(DBA)与基于参考基因组的方法(RBA)进行比较,以助力改进西瓜参考基因组的注释,并深入了解西瓜雄性不育的生物学机制。我们将这些技术应用于两个西瓜品系的现有数据:雄性不育系DAH3615-MS和雄性可育系DAH3615。使用DBA,我们新注释了855个西瓜转录本,并发现了预测与刺激反应、核酸结合、跨膜运输、稳态以及高尔基体/囊泡相关的基因功能簇。在DBA注释的转录本中,检测到138个与雄性不育相关的从头测序独有的差异表达基因(DEDEG)。在随机选择的33个新注释转录本和DEDEG中,32个通过RT-qPCR得到验证。本研究证明了西瓜从头转录组组装的实用性和可靠性,并为研究人员探索雄性不育的转录蓝图提供了新的见解。