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A Comparative Proteomic Analysis of Pinellia ternata Leaves Exposed to Heat Stress

1
Institute of Chinese Medicinal Materials, Nanjing Agricultural University, Nanjing 210095, China
2
Institute of Morden Chinese Medical Materials, Guizhou Academy of Agricaltural Sciences, Guiyang 550006, China
3
Key Laboratory of Agricultural Biotechnology, Guizhou Academy of Agricultural Sciences, Guiyang 550006, China
*
Authors to whom correspondence should be addressed.
Int. J. Mol. Sci. 2013, 14(10), 20614-20634; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms141020614
Received: 20 July 2013 / Revised: 22 September 2013 / Accepted: 29 September 2013 / Published: 15 October 2013
(This article belongs to the Section Biochemistry)
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. View Full-Text
Keywords: proteomics; quantitative real-time PCR; heat stress; Pinellia ternata proteomics; quantitative real-time PCR; heat stress; Pinellia ternata
MDPI and ACS Style

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.

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