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Molecular & Cellular Proteomics 8:2443-2460, 2009.
© 2009 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

Label-free Quantitative Proteomics Analysis of Etiolated Maize Seedling Leaves during Greening^*,

Zhuo Shen^,, Ping Li^,¶, Rui-Juan Ni^,, Mark Ritchie^||, Chuan-Ping Yang, Gui-Feng Liu, Wei Ma^,**, Guan-Jun Liu, Ling Ma, Shu-Juan Li, Zhi-Gang Wei, Hong-Xia Wang^¶, and Bai-Chen Wang^,¶¶

From the Key Laboratory of Forest Tree Genetic Improvement and Biotechnology, Ministry of Education and
School of Forestry, Northeast Forestry University, 26 Hexing Road, Harbin 150040, China,
¶National Center of Biomedical Analysis, 27 Taiping Road, Beijing 100850, China,
||Waters Asia Limited, 1 Science Park Road, Singapore Science Park II, Singapore 117528, Singapore and
**Heilongjiang University of Chinese Medicine, 24 Heping Road, Harbin 150040, China

To better understand light regulation of C₄ plant maize development, we investigated dynamic proteomic differences between green seedlings (control), etiolated seedlings, and etiolated seedlings illuminated for 6 or 12 h using a label-free quantitative proteomics approach based on nanoscale ultraperformance liquid chromatography-ESI-MS^E. Among more than 400 proteins identified, 73 were significantly altered during etiolated maize seedling greening. Of these 73 proteins, 25 were identified as membrane proteins that seldom had been identified with two-dimensional electrophoresis methods, indicating the power of our label-free method for membrane protein identification; 31 were related to light reactions of chlorophyll biosynthesis, photosynthesis, and photosynthetic carbon assimilation. The expression of photosystem II subunits was highly sensitive to light; most of them were not identified in etiolated maize seedlings but drastically increased upon light exposure, indicating that the complex process of biogenesis of the photosynthetic apparatus correlates with the transition from a dark-grown to a light-grown morphology. However, transcriptional analysis indicated that most transcripts encoding these proteins were not regulated by light. In contrast, the levels of mRNAs and proteins for enzymes involved in carbon assimilation were tightly regulated by light. Additionally phosphoenolpyruvate carboxykinase, the key enzyme of the phosphoenolpyruvate carboxykinase C₄ pathway, was more tightly regulated by light than the key enzymes of the NADP-malic enzyme C₄ pathway. Furthermore phosphoenolpyruvate carboxylase 1C, which was originally reported to be specifically expressed in roots, was also identified in this study; expression of this enzyme was more sensitive to light than its isoforms. Taken together, these results represent a comprehensive dynamic protein profile and light-regulated network of C₄ plants for etiolated seedling greening and provide a basis for further study of the mechanism of gene function and regulation in light-induced development of C₄ plants.

¶¶ To whom correspondence may be addressed. Tel.: 86-451-82190607-12; Fax: 86-451-82190607-11; E-mail: wangbc{at}nefu.edu.cn.

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