An Optimized Protein Extraction Method for Gel-Free Proteomic Analysis of Opuntia Ficus-Indica
1
Faculty of Medicine, University of Tsukuba, Tsukuba 305-8575, Japan
2
School of Nursing and Medical Care, Yokkaichi Nursing and Medical Care University, Yokkaichi 512-8045, Japan
3
Institute for Comprehensive Medical Science, Fujita Health University, Toyoake 470-1192, Japan
4
Tsukuba-Plant Innovation Research Center, University of Tsukuba, Tsukuba 305-8577, Japan
5
Faculty of Life and Environmental Sciences, University of Tsukuba, Tsukuba 305-8577, Japan
*
Author to whom correspondence should be addressed.
Plants 2021, 10(1), 115; https://doi.org/10.3390/plants10010115
Received: 10 December 2020
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Revised: 4 January 2021
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Accepted: 4 January 2021
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Published: 8 January 2021
Opuntia spp. is an economically important vegetable crop with high stress-tolerance and health benefits. However, proteomic analysis of the plant has been difficult due to the composition of its succulent cladodes; the abundant polysaccharides interfere with protein extraction. To facilitate proteomic analysis of this plant, we present a rapid and simple protein extraction method for Opuntia ficus-indica (L.) Miller. The optimized method produced highly reproducible protein patterns and was compatible with a gel-free quantitative workflow without the need for additional purification. We successfully analyzed the cladode mesocarp and exocarp tissues, resulting in the identification of 319 proteins. In addition, we used this method to examine the relative changes in the Opuntia proteome in response to salt stress to determine whether physiological changes could be captured. Qualified observations were obtained, revealing that salt stress increased phosphoenolpyruvate carboxylase abundance and decreased ribulose-bisphosphate carboxylase in young O. ficus-indica plants. These findings suggest that Crassulacean acid metabolism is promoted under salinity. This study highlights the efficacy of our optimized protein extraction method for elucidating the metabolic adaptations of Opuntia using gel-free proteomic analysis.
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Keywords:
Opuntia; succulent; protein extraction; gel-free proteomic analysis; abiotic stress; environmental stress
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MDPI and ACS Style
Hashiguchi, A.; Yamaguchi, H.; Hitachi, K.; Watanabe, K. An Optimized Protein Extraction Method for Gel-Free Proteomic Analysis of Opuntia Ficus-Indica. Plants 2021, 10, 115. https://doi.org/10.3390/plants10010115
AMA Style
Hashiguchi A, Yamaguchi H, Hitachi K, Watanabe K. An Optimized Protein Extraction Method for Gel-Free Proteomic Analysis of Opuntia Ficus-Indica. Plants. 2021; 10(1):115. https://doi.org/10.3390/plants10010115
Chicago/Turabian StyleHashiguchi, Akiko, Hisateru Yamaguchi, Keisuke Hitachi, and Kazuo Watanabe. 2021. "An Optimized Protein Extraction Method for Gel-Free Proteomic Analysis of Opuntia Ficus-Indica" Plants 10, no. 1: 115. https://doi.org/10.3390/plants10010115
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