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Construction and characterization of a bacterial artificial chromosome library for Camellia sinensis

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Abstract

Tea is a popular and natural non-alcoholic beverage, and is produced from fresh leaves of Camellia sinensis. Tea leaves contain many bioactive compounds that have significant health benefits. We constructed a high quality bacterial artificial chromosome (BAC) library by using the fresh petals of C. sinensis “Shuchazao” for genome sequencing and improvement of genomic assembly. BAC library is still a significant tool for studies of functional genomes and preservation of precious genetic resources. The BAC library contains 161,280 clones with an average insert size of 113 kb, which represents approximately 6.2-fold coverage of haploid genome equivalents of C. sinensis. We characterized 20 complete BAC clones and 738 BAC end sequences (BESs) ranging from 105 to 917 bp. In addition, we predicted cis-regulatory elements of LAR (leucoanthocyanidin reductase), TCS (caffeine synthase), and TS (theanine synthetase) involved in tea characteristic metabolite synthesis and identified a larger number of light-responsive cis-acting elements in these three genes. Meanwhile, we analyzed alternative splicing of these three genes. Furthermore, 12 pairs of SSR primers were successfully amplified in tea plant DNA. The tea BAC library was a critical resource to accomplish de novo whole-genome sequencing, accelerate gene discovery and enhance molecular breeding of C. sinensis.

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Acknowledgments

This work received financial support from the Science and Technology Project of AnHui Province, China (Project 13Z03012), Tea Genome Project of AnHui Province, China, the Special Innovative Province Construction in Anhui province in 2015 (15czs08032), the Central Guiding the Science and Technology Development of the Local (2016080503B024), the Major Project of Chinese National Programs for Fundamental Research and Development (2012CB722903), the Natural Science Foundation of Anhui Province (No.1608085QC60), and the Youth Foundation of Anhui Agricultural University (2016ZR012). We appreciate Chun Liu (Beijing Genome Institute at Shenzhen, China) for technical support and analysis. We are grateful to the unknown editor at the elixigen editing service (ID151023-6181) for the English polishing.

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Author notes

  1. Yuling Tai and Hao Wang contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, Hefei, China

    Yuling Tai, Chaoling Wei, Min Li, Liqun Wang, Jiayue Jiang, Hua Yang, Weiwei Deng, Bei Huang, Xiaochun Wan & Jun Sun

  2. School of Life Science, Anhui Agricultural University, Hefei, China

    Yuling Tai & Ling Su

  3. College of Life Sciences and Technology, Huazhong Agricultural University, Wuhan, China

    Hao Wang, Zhaozhao Dai, Lingyun Shao & Meizhong Luo

  4. College of Horticulture, Anhui Agricultural University, Hefei, China

    Jun Sun

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  1. Yuling Tai
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Contributions

Conceived and designed the experiment: WC and SJ; analyzing data: WL, WH, TL, JY, LZ, YH, and DW; experiment: SL, DZ, SY, HB, WQ, and LM. TL and WH contributed to writing the text.

Corresponding authors

Correspondence to Xiaochun Wan or Jun Sun.

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The authors declare that they have no competing interests.

Additional information

Communicated by W.-W. Guo

Data achieving statement

The raw data of tea plant BAC are now are available in the NCBI SRA under project accession number PRJNA385558. The experiment accession numbers were SRX2982355. The BAC sequences of tea plant will be submitted to the European Nucleotide Archive database (ENA; http://www.ebi.ac.uk/ena) under project number accession PRJEB21650 if the manuscript is accepted for publication in the tree genetics and genomes prior to publication.

The Illumina RNA-seq data used in this study has been deposited in the NCBI SRA (http://trace.ncbi.nlm.nih.gov/Traces/sra) with accessions SRR1928149.

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Tai, Y., Wang, H., Wei, C. et al. Construction and characterization of a bacterial artificial chromosome library for Camellia sinensis . Tree Genetics & Genomes 13, 89 (2017). https://doi.org/10.1007/s11295-017-1173-5

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