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| A Simple Method for Isolating Chloroplast DNA and Mitochondria DNA from the Same Rapeseed Green Leaf Tissue |
| HU Zhi-yong, ZHAN Gao-miao, WANG Han-zhong, HUA Wei |
| Key Laboratory for Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture/Oil Crops Research Institute, Chinese Academy of Agriculture Sciences, Wuhan 430062, P.R.China |
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摘要 In the study, we present a fast, simple and inexpensive protocol for isolating chloroplast and mitochondrial DNA from one rapeseed leaf tissue sample. The chloroplast and mitochondria were separated from the same green leaf tissue by differential centrifugations. The protocol is the first report that isolates plant chloroplast DNA (cpDNA) and mitochondrial DNA (mtDNA) from the same sample homogenate. The organelle DNA yield is 2-10 micrograms per gram of tissue; the DNA was fully restrictable and was successfully used for sequencing.
Abstract In the study, we present a fast, simple and inexpensive protocol for isolating chloroplast and mitochondrial DNA from one rapeseed leaf tissue sample. The chloroplast and mitochondria were separated from the same green leaf tissue by differential centrifugations. The protocol is the first report that isolates plant chloroplast DNA (cpDNA) and mitochondrial DNA (mtDNA) from the same sample homogenate. The organelle DNA yield is 2-10 micrograms per gram of tissue; the DNA was fully restrictable and was successfully used for sequencing.
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Received: 09 January 2011
Accepted:
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| Fund: This work was supported by the Ministry of Agriculture of China Genetically Modified Organisms Breeding Major Projects (2009ZX08009-018B). |
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Corresponding Authors:
HUA Wei, Tel: +86-27-86711806, E-mail: huawei@oilcrops.cn
E-mail: huawei@oilcrops.cn
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| About author: HU Zhi-yong, Tel: +86-27-86712076; E-mail: oilhzy@yahoo.com.cn; |
Cite this article:
HU Zhi-yong, ZHAN Gao-miao, WANG Han-zhong, HUA Wei.
2012.
A Simple Method for Isolating Chloroplast DNA and Mitochondria DNA from the Same Rapeseed Green Leaf Tissue. Journal of Integrative Agriculture, 12(7): 1212-1215.
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| [1]Itani K, Handa H. 1998. Rapeseed mitochondrial ccb206, a gene involved in cytochrome c biogenesis, is cotranscribed with the nad3 and rps12 genes: organization, transcription, and RNA editing of the nad3/rps12/ccb206 locus. Current Genetics, 34, 318-325. [2]Kemble R J. 1987. A rapid single leaf, nucleic acid assay for determining the cytoplasmic organelle complement of rapeseed and related Brassica species. Theoretical and Applied Genetics, 73, 364-370. [3]Pay A, Smith M A. 1988. A rapid method for purification of organeiles for DNA isolation: self-generated percoll gradients. Plant Cell Reports, 7, 96-99. [4]Scotti N, Cardi T, Marechaldrouard L. 2001. Mitochondrial DNA and RNA isolation from small amounts of potato tissue. Plant Molecular Biology Reporter, 19, 67a-67h. [5]Triboush S O, Danilenko N G, Davydenko O G. 1998. A method for isolation of chloroplast DNA and mitochondrial DNA from sunflower. Plant Molecular Biology Reporter, 16, 183-189. [6]Virupakshi S, Naik G R. 2007. Purification of DNA from chloroplast and mitochondria of sugarcane. Current Science, 92, 1613-1619. [7]Wang X, Liu G, Yang Q, Hua W, Liu J, Wang H. 2010. Genetic analysis on oil content in rapeseed (Brassica napus L.). Euphytica, 173, 17-24. |
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