A proteomics sample preparation method for mature, recalcitrant leaves of perennial plants

Deng Gang; Zhong Xinyue; Zhang Na; Lao Chengying; Wang Bo; Peng Dingxiang; Liu Lijun

doi:10.1371/journal.pone.0102175

A proteomics sample preparation method for mature, recalcitrant leaves of perennial plants

PLoS One. 2014 Jul 16;9(7):e102175. doi: 10.1371/journal.pone.0102175. eCollection 2014.

Authors

Deng Gang¹, Zhong Xinyue², Zhang Na³, Lao Chengying², Wang Bo², Peng Dingxiang², Liu Lijun²

Affiliations

¹ MOA Key Laboratory of Crop Ecophysiology and Farming System in the Middle Reaches of the Yangtze River, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, Hubei, China; School of Agricultural Science, Yunnan University, Kunming, Yunnan, China.
² MOA Key Laboratory of Crop Ecophysiology and Farming System in the Middle Reaches of the Yangtze River, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, Hubei, China.
³ Wuhan Agriculture and Science Institute, Huangpi District, Wuhan, Hubei, China.

Abstract

Sample preparation is key to the success of proteomics studies. In the present study, two sample preparation methods were tested for their suitability on the mature, recalcitrant leaves of six representative perennial plants (grape, plum, pear, peach, orange, and ramie). An improved sample preparation method was obtained: Tris and Triton X-100 were added together instead of CHAPS to the lysis buffer, and a 20% TCA-water solution and 100% precooled acetone were added after the protein extraction for the further purification of protein. This method effectively eliminates nonprotein impurities and obtains a clear two-dimensional gel electrophoresis array. The method facilitates the separation of high-molecular-weight proteins and increases the resolution of low-abundance proteins. This method provides a widely applicable and economically feasible technology for the proteomic study of the mature, recalcitrant leaves of perennial plants.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Analytic Sample Preparation Methods / methods*
Plant Leaves / chemistry*
Plant Leaves / growth & development
Plant Proteins / analysis
Plants / chemistry*
Proteomics / methods*

Substances

Plant Proteins

Grants and funding

This research was supported by a grant from National science and technology support (2010BAD02B01), the National Natural Science Foundation of China (31000731) and China Agriculture Research System (CARS-19-E12). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.