化学诱导后白木香转录组文库的构建与测序

生物技术通报 ›› 2013, Vol. 0 ›› Issue (8): 63-67.

化学诱导后白木香转录组文库的构建与测序

吴宏清^{1, 2}, 王磊¹, 陶美华¹, 高晓霞³, 白玲², 章卫民¹

（1. 广东省微生物研究所广东省菌种保藏与应用重点实验室广东省微生物应用新技术公共实验室广东省华南应用微生物重点实验室 -省部共建国家重点实验室培育基地，广州 510070；2.江西农业大学理学院，南昌 330045；3.广东药学院，广州 510006）

收稿日期:2013-08-11 修回日期:2013-08-11 出版日期:2013-08-11 发布日期:2013-09-02
作者简介:吴宏清，硕士研究生，研究方向：植物分子生物学；E-mail：whq0401@163.com
基金资助:
国家自然科学基金项目（31100496，81102418，81203006），广东省中国科学院全面战略合作项目（2011B090300078），广东省科技计划项目（2012A030100014）

Transcriptome Library Construction and Sequencing from Chemically Induced Aquilaria sinensis

Wu Hongqing^{1, 2}, Wang Lei¹, Tao Meihua¹, Gao Xiaoxia³, Bai Ling², Zhang Weimin¹

（1. State Key Laboratory of Applied Microbiology Ministry-Guangdong Province Jointly Breeding Base，South China，Guangdong Open Laboratory of Applied Microbiology，Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application，Guangdong Institute of Microbiology，Guangzhou 510070；2. College of Science，Jiangxi Agricultural University，Nanchang 330045；3. Guangdong Pharmaceutical University，Guangzhou 510006）

Received:2013-08-11 Revised:2013-08-11 Published:2013-08-11 Online:2013-09-02

摘要/Abstract

摘要： 采用改良异硫氰酸胍-CTAB法对5年生白木香树干经化学诱导后1年的各部分组织进行总RNA提取，提取到的总RNA经富集mRNA、打断、构建测序用cDNA文库后用于转录组测序，测序质量较高，Q20高达97.45%，共获得54 685 634条Clean reads，总测序长度达4 921 707 060 nt，经初步组装，获得190 109条Contigs序列，进一步组装，获得83 467条Unigenes序列，总长度为58 569 625 nt，平均长度为702 nt，N50值高达1 120，大于等于3 000 nt的Unigenes有1 691条，占总Unigenes的2.03%，组装质量较高，使白木香的转录组信息得到较好的保存，为进行白木香结香相关的表达谱分析奠定基础。

关键词: 白木香, 化学诱导, 转录组文库, 测序, 组装, Unigenes

Abstract: Modified guanidinium isothiocyanate-CTAB method was used to isolate the total RNA from five-year-old Aquilaria sinensis treated by chemical induction one year ago. The mRNA of A. sinensis was enriched from the total RNA and broken into short fragments, and then the cDNA library was established for RNA-Seq. As a result, 54 685 634 clean reads was obtained after sequencing with a total length of 4 921 707 060 nt. The value of Q20 was up to 97.45%, exhibiting good sequencing quality. After the initial assembly of sequence data, these clean reads were assembled to 190 109 contigs, which were then assembled to 83 467 unigenes with a total length of 58 569 625 nt and an average length of 702 nt after the further assembly. The value of N50 was up to 1 120. There were 1 691 unigenes longer than 3 000 nt, accounted for 2.03% of all unigenes. The good quality of assembly showed the information on the transcriptome of A. sinensis was well preserved, which laid the foundation for digital gene expression analysis associated with agarwood formation of A. sinensis.

Key words: Aquilaria sinensis, Chemical induction, Transcriptome library, Sequencing, Assembly, Unigenes

吴宏清, 王磊, 陶美华, 高晓霞, 白玲, 章卫民,. 化学诱导后白木香转录组文库的构建与测序[J]. 生物技术通报, 2013, 0(8): 63-67.

Wu Hongqing, Wang Lei, Tao Meihua, Gao Xiaoxia, Bai Ling, Zhang Weimin. Transcriptome Library Construction and Sequencing from Chemically Induced Aquilaria sinensis[J]. Biotechnology Bulletin, 2013, 0(8): 63-67.

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