title: Expression of the moss PpLEA4-20 gene in rice enhances membrane protection and client proteins stability.
abstract: Green vegetative tissues of the moss Physcomitrella patens possess a powerful ability to tolerate severe drought stress. Proteomics analysis have revealed that a large number of late embryogenesis abundant (LEA) proteins were key players in the drought tolerance of the photosynthetic tissues. PpLEA4-20, a member of the moss LEA protein family, was selected for further function study using an ectopic expression method in rice. Through molecular identification via PCR, southern blotting and TAIL-PCR, we demonstrated that the PpLEA4-20 gene was transformed and inserted into a non-encoded region in chromosome 4 of rice and expressed stably in transgenic rice. Unexpectedly, PpLEA4-20 protein emerged as two high-expressed spots on 2-D gels generated from transgenic rice, suggesting that PpLEA4-20 proteins are complete compatible and might be modified in rice. Both growth and physiological analysis showed that seedlings of transgenic PpLEA4-20 rice displayed altered phenotypes and tolerance to salt. In addition, electrolyte leakage was reduced in transgenic PpLEA4-20 compared to wild type under stress conditions. Anti-aggregation analysis found that the PpLEA4-20 protein expressed in rice remained soluble at high temperature and in addition to some native proteins from transgenic PpLEA4-20 rice. Based on Nano LC MS/MS analysis, we identified several proteins from transgenic PpLEA4-20 rice of increased heat-stability. Our results provide evidence for a role of PpLEA4-20 in salt tolerance and stabilization of client proteins.
authors: Key Laboratory of Photobiology, Institute of Botany, The Chinese Academy of Sciences, Beijing 100093, China; University of the Chinese Academy of Sciences, Beijing 100049, China; College of Life Sciences, Capital Normal University, Beijing 100048, China.
authors: Li
authors: L
authors: Li
authors: College of Life Sciences, Capital Normal University, Beijing 100048, China.
authors: Dandan
authors: D
authors: Deng
authors: College of Life Sciences, Capital Normal University, Beijing 100048, China.
authors: Xi
authors: X
authors: Chen
authors: College of Life Sciences, Capital Normal University, Beijing 100048, China.
authors: Baomei
authors: B
authors: Wu
authors: College of Life Sciences, Capital Normal University, Beijing 100048, China.
authors: Ke
authors: K
authors: Hu
authors: College of Life Sciences, Capital Normal University, Beijing 100048, China.
authors: Tianhang
authors: T
authors: Qiu
authors: College of Life Sciences, Capital Normal University, Beijing 100048, China. Electronic address: sxcui@cnu.edu.cn.
authors: Suxia
authors: S
authors: Cui
authors: Key Laboratory of Photobiology, Institute of Botany, The Chinese Academy of Sciences, Beijing 100093, China. Electronic address: fhuang@ibcas.ac.cn.
authors: Fang
authors: F
authors: Huang
mesh_terms: D001483:Base Sequence; D019068:Bryopsida / Q000235:genetics*; D002853:Chromatography, Liquid; D017931:DNA Primers; D017343:Genes, Plant; D012275:Oryza / Q000235:genetics*; D010940:Plant Proteins / Q000502:physiology*; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D055550:Protein Stability; D053719:Tandem Mass Spectrometry; D019076:Transgenes
chemical_list: D017931:DNA Primers; D010940:Plant Proteins
keywords: 2-DE; Membrane protection; PpLEA4-20; Protein targets; Rice; Transformation