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Highly effective sequencing whole chloroplast genomes of angiosperms by nine novel universal primer pairs

文献类型:期刊论文

作者Yang, Jun-Bo; Li, De-Zhu; Li, Hong-Tao
刊名MOLECULAR ECOLOGY RESOURCES
出版日期2014-09-01
卷号14期号:5页码:1024-1031
关键词angiosperm chloroplast genomes next-generation sequencing novel universal primer pairs phylogenomics species identification
ISSN号1755-098X
产权排序第一
通讯作者Li, DZ (reprint author), Chinese Acad Sci, Kunming Inst Bot, Germplasm Bank Wild Species, Kunming 650201, Peoples R China. ; dzl@mail.kib.ac.cn ; lihongtao@mail.kib.ac.cn
英文摘要Chloroplast genomes supply indispensable information that helps improve the phylogenetic resolution and even as organelle-scale barcodes. Next-generation sequencing technologies have helped promote sequencing of complete chloroplast genomes, but compared with the number of angiosperms, relatively few chloroplast genomes have been sequenced. There are two major reasons for the paucity of completely sequenced chloroplast genomes: (i) massive amounts of fresh leaves are needed for chloroplast sequencing and (ii) there are considerable gaps in the sequenced chloroplast genomes of many plants because of the difficulty of isolating high-quality chloroplast DNA, preventing complete chloroplast genomes from being assembled. To overcome these obstacles, all known angiosperm chloroplast genomes available to date were analysed, and then we designed nine universal primer pairs corresponding to the highly conserved regions. Using these primers, angiosperm whole chloroplast genomes can be amplified using long-range PCR and sequenced using next-generation sequencing methods. The primers showed high universality, which was tested using 24 species representing major clades of angiosperms. To validate the functionality of the primers, eight species representing major groups of angiosperms, that is, early-diverging angiosperms, magnoliids, monocots, Saxifragales, fabids, malvids and asterids, were sequenced and assembled their complete chloroplast genomes. In our trials, only 100 mg of fresh leaves was used. The results show that the universal primer set provided an easy, effective and feasible approach for sequencing whole chloroplast genomes in angiosperms. The designed universal primer pairs provide a possibility to accelerate genome-scale data acquisition and will therefore magnify the phylogenetic resolution and species identification in angiosperms.
学科主题Biochemistry & Molecular Biology ; Ecology ; Evolutionary Biology
类目[WOS]Biochemistry & Molecular Biology ; Ecology ; Evolutionary Biology
研究领域[WOS]Biochemistry & Molecular Biology ; Environmental Sciences & Ecology ; Evolutionary Biology
关键词[WOS]PLASTID GENOMES ; PHYLOGENETIC IMPLICATIONS ; CONSENSUS PRIMERS ; FLOWERING PLANTS ; GENUS CUSCUTA ; DNA ; GENES ; AMPLIFICATION ; IDENTIFICATION ; PHYLOGENOMICS
收录类别SCI
资助信息National Natural Science Foundation of China [31170182, 30870169]; National 863 Project of China [2012AA021801]; Instruments Development and Technology Innovation Project of the Chinese Academy of Sciences; Yunnan Provincial Natural Science Foundation [2010CD108]
语种英语
WOS记录号WOS:000340425100013
公开日期2014-10-11
源URL[http://ir.kib.ac.cn/handle/151853/18317]  
专题昆明植物研究所_中国西南野生生物种质资源库
作者单位Chinese Acad Sci, Kunming Inst Bot, Germplasm Bank Wild Species, Kunming 650201, Peoples R China
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GB/T 7714
Yang, Jun-Bo,Li, De-Zhu,Li, Hong-Tao. Highly effective sequencing whole chloroplast genomes of angiosperms by nine novel universal primer pairs[J]. MOLECULAR ECOLOGY RESOURCES,2014,14(5):1024-1031.
APA Yang, Jun-Bo,Li, De-Zhu,&Li, Hong-Tao.(2014).Highly effective sequencing whole chloroplast genomes of angiosperms by nine novel universal primer pairs.MOLECULAR ECOLOGY RESOURCES,14(5),1024-1031.
MLA Yang, Jun-Bo,et al."Highly effective sequencing whole chloroplast genomes of angiosperms by nine novel universal primer pairs".MOLECULAR ECOLOGY RESOURCES 14.5(2014):1024-1031.

入库方式: OAI收割

来源:昆明植物研究所

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