A New Chloroplast DNA Extraction Protocol Significantly Improves the Chloroplast Genome Sequence Quality of Foxtail Millet (Setaria italica (L.) P. Beauv.)
文献类型： 外文期刊
作者： Liu, Dan 2 ; Cui, Yanjiao 1 ; Li, Suying 1 ; Bai, Guihua 3 ; Li, Qiang 1 ; Zhao, Zilong 1 ; Liang, Dan 2 ; Wang, Conglei; 1 ;
作者机构： 1.Tangshan Normal Univ, Dept Life Sci, Tangshan 063000, Peoples R China
2.Tianjin Acad Agr Sci, Tianjin Crop Res Inst, Tianjin 300384, Peoples R China
3.ARS, USDA, Hard Winter Wheat Genet Res Unit, 4008 Throckmorton Hall, Manhattan, KS 66506 USA
期刊名称：SCIENTIFIC REPORTS （ 影响因子：4.379； 五年影响因子：5.133 ）
ISSN： 2045-2322
年卷期： 2019 年 9 卷
页码：
收录情况： SCI
摘要： The complexity of the leaf constitution of foxtail millet (Setaria italica (L.) P. Beauv.) makes it difficult to obtain high-purity cpDNA. Here, we developed a protocol to isolate high-quality cpDNA from foxtail millet and other crops. The new protocol replaces previous tissue grinding and homogenization by enzyme digestion of tiny leaf strips to separate protoplasts from leaf tissue and protects chloroplasts from damage by undue grinding and homogenization and from contamination of cell debris and nuclear DNA. Using the new protocol, we successfully isolated high-quality cpDNAs for whole-genome sequencing from four foxtail millet cultivars, and comparative analysis revealed that they were approximately 27 parts per thousand longer than their reference genome. In addition, six cpDNAs of four other species with narrow and thin leaf blades, including wheat (Triticum aestivum L.), maize (Zea may L.), rice (Oryza sativa L.) and sorghum (Sorghum bicolor (L.) Moench), were also isolated by our new protocol, and they all exhibited high sequence identities to their corresponding reference genomes. A maximum-likelihood tree based on the chloroplast genomes we sequenced here was constructed, and the result was in agreement with previous reports, confirming that these cpDNA sequences were available for well-supported phylogenetic analysis and could provide valuable resources for future research.
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