title: A proteomics sample preparation method for mature, recalcitrant leaves of perennial plants.
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.
authors.affiliation: 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.
authors.forename: Deng
authors.initials: D
authors.lastname: Gang
authors.affiliation: 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.
authors.forename: Zhong
authors.initials: Z
authors.lastname: Xinyue
authors.affiliation: Wuhan Agriculture and Science Institute, Huangpi District, Wuhan, Hubei, China.
authors.forename: Zhang
authors.initials: Z
authors.lastname: Na
authors.affiliation: 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.
authors.forename: Lao
authors.initials: L
authors.lastname: Chengying
authors.affiliation: 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.
authors.forename: Wang
authors.initials: W
authors.lastname: Bo
authors.affiliation: 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.
authors.forename: Peng
authors.initials: P
authors.lastname: Dingxiang
authors.affiliation: 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.
authors.forename: Liu
authors.initials: L
authors.lastname: Lijun
mesh_terms: D053000:Analytic Sample Preparation Methods / Q000379:methods*; D018515:Plant Leaves / Q000737:chemistry* / Q000254:growth & development; D010940:Plant Proteins / Q000032:analysis; D010944:Plants / Q000737:chemistry*; D040901:Proteomics / Q000379:methods*
chemical_list: D010940:Plant Proteins