A simplified method to isolate rice mitochondria
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(2020) 16:149 Xu et al. Plant Methods https://doi.org/10.1186/s13007-020-00690-6
Open Access
RESEARCH
A simplified method to isolate rice mitochondria Yanghong Xu1 , Xiaoyi Li1, Jishuai Huang1, Leilei Peng1, Dinghui Luo1, Qiannan Zhang1, Zhiwu Dan1, Haijun Xiao2, Fang Yang1* and Jun Hu1*
Abstract Background: Mitochondria play critical roles in plant growth, development and stress tolerance. Numerous researchers have carried out studies on the plant mitochondrial genome structure, mitochondrial metabolism and nuclearcytoplasmic interactions. However, classical plant mitochondria extraction methods are time-consuming and consist of a complicated ultracentrifugation procedure with expensive reagents. To develop a more rapid and convenient method for the isolation of plant mitochondria, in this study, we established a simplified method to isolate rice mitochondria efficiently for subsequent studies. Results: To isolate rice mitochondria, the cell wall was first disrupted by enzymolysis to obtain the protoplast, which is similar to animal mitochondria. Rice mitochondria were then isolated with a modified method based on the animal mitochondria isolation protocol. The extracted mitochondria were next assessed according to DNA and protein levels to rule out contamination by the nucleus and chloroplasts. Furthermore, we examined the physiological status and characteristics of the isolated mitochondria, including the integrity of mitochondria, the mitochondrial membrane potential, and the activity of inner membrane complexes. Our results demonstrated that the extracted mitochondria remained intact for use in subsequent studies. Conclusion: The combination of plant protoplast isolation and animal mitochondria extraction methods facilitates the extraction of plant mitochondria without ultracentrifugation. Consequently, this improved method is cheap and time-saving with good operability and can be broadly applied in studies on plant mitochondria. Keywords: Mitochondria isolation, Protoplast, Rice Background Mitochondria were first discovered in 1850 and are considered to have evolved from the engulfment of an α-proteobacterium by a precursor of the modern eukaryotic cell [1]. The differences between plant mitochondria and animal mitochondria are small regarding their size and function since both of them evolved from the
*Correspondence: [email protected]; [email protected] 1 State Key Laboratory of Hybrid Rice, Engineering Research Center for Plant Biotechnology and Germplasm Utilization of Ministry of Education, College of Life Sciences, Wuhan University, Wuhan 430072, Hubei, China Full list of author information is available at the end of the article
same microorganism [2, 3], but the plant mitochondrial genome can be 100 times larger than those of animals [4, 5]. Mitochondria produce more than 90% of the cellular energy (ATP) required for an organism’s growth, reproduction, and maintenance [6]. In plants, dysfunction of the mitochondria always leads to retardation of plant growth, hypersensitivity to disease, em
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