Construction and characterization of a bacterial artificial chromosome library for Camellia sinensis
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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.
KeywordsTea plant BAC library BAC end sequencing SSR Alternative splicing
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.
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.
Compliance with ethical standards
The authors declare that they have no competing interests.
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