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A Comparative Proteomic Analysis of Pinellia ternata Leaves Exposed to Heat Stress
Institute of Chinese Medicinal Materials, Nanjing Agricultural University, Nanjing 210095, China
Institute of Morden Chinese Medical Materials, Guizhou Academy of Agricaltural Sciences, Guiyang 550006, China
Key Laboratory of Agricultural Biotechnology, Guizhou Academy of Agricultural Sciences, Guiyang 550006, China
* Authors to whom correspondence should be addressed.
Received: 20 July 2013; in revised form: 22 September 2013 / Accepted: 29 September 2013 / Published: 15 October 2013
Abstract: Pinellia ternata is an important traditional Chinese medicinal plant. The growth of P. ternata is sensitive to high temperatures. To gain a better understanding of heat stress responses in P. ternata, we performed a comparative proteomic analysis. P. ternata seedlings were subjected to a temperature of 38 °C and samples were collected 24 h after treatment. Increased relative ion leakage and lipid peroxidation suggested that oxidative stress was frequently generated in rice leaves exposed to high temperature. Two-dimensional electrophoresis (2-DE) was used to analyze heat-responsive proteins. More than 600 protein spots were reproducibly detected on each gel; of these spots, 20 were up-regulated, and 7 were down-regulated. A total of 24 proteins and protein species were successfully identified by MALDI-TOF/TOF MS. These proteins and protein species were found to be primarily small heat shock proteins (58%) as well as proteins involved in RNA processing (17%), photosynthesis (13%), chlorophyll biosynthetic processes (4%), protein degradation (4%) and defense (4%). Using 2-DE Western blot analysis, we confirmed the identities of the cytosolic class II small heat shock protein (sHSPs-CII) identified by MS. The expression levels of four different proteins [cytosolic class I small heat shock protein (sHSPs-CI), sHSPs-CII, mitochondrial small heat shock protein (sHSPs-MIT), glycine-rich RNA-binding protein (GRP)] were analyzed at the transcriptional level by quantitative real-time PCR. The mRNA levels of three sHSPs correlated with the corresponding protein levels. However, GRP was down-regulated at the beginning of heat stress but then increased substantially to reach a peak after 24 h of heat stress. Our study provides valuable new insight into the responses of P. ternata to heat stress.
Keywords: proteomics; quantitative real-time PCR; heat stress; Pinellia ternata
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Zhu, Y.; Zhu, G.; Guo, Q.; Zhu, Z.; Wang, C.; Liu, Z. A Comparative Proteomic Analysis of Pinellia ternata Leaves Exposed to Heat Stress. Int. J. Mol. Sci. 2013, 14, 20614-20634.
Zhu Y, Zhu G, Guo Q, Zhu Z, Wang C, Liu Z. A Comparative Proteomic Analysis of Pinellia ternata Leaves Exposed to Heat Stress. International Journal of Molecular Sciences. 2013; 14(10):20614-20634.
Zhu, Yunhao; Zhu, Guosheng; Guo, Qiaosheng; Zhu, Zaibiao; Wang, Changlin; Liu, Zuoyi. 2013. "A Comparative Proteomic Analysis of Pinellia ternata Leaves Exposed to Heat Stress." Int. J. Mol. Sci. 14, no. 10: 20614-20634.