Anti-stress Related Expression Analysis of TaSKP2A Gene in Wheat (Triticum aestivum) and Its Interaction Protein Screening
MENG Yu-Yu1, LI Hu-Ying1, XU Yuan1, WEI Chun-Ru1, FAN Run-Qiao1, YU Xiu-Mei1,2*, ZHAO Wei-Quan2, KANG Zhen-Sheng3, LIU Da-Qun2,4
1 College of Life Sciences, Hebei Agricultural University / Key Laboratory of Hebei Province of Plant Physiology and Molecular Pathology, Baoding 071001, China; 2 Biological Control Centre of Plant Disease and Plant Pests of Hebei Province, Baoding 071001, China; 3 College of Plant Protection, Northwest A&F University / State Key Laboratory of Crop Stress Biology for Arid Areas, Yangling 712100, China; 4 Graduate School of Chinese Academy of Agricultural Sciences, Beijing 100081, China
Abstract：S-phase kinase-associated protein 2A (SKP2A) is a F-box protein, which is involved in regulating cell division and improving tolerance to osmotic stress in Arabidopsis thaliana. In order to study the role of SKP2A in the process of wheat (Triticum aestivum) response to abiotic stresses and fungi pathogen infection, the present study further analyzed TaSKP2A gene and its encoded protein based on the full length ORF of wheat TaSKP2A. The results showed that TaSKP2A encoded a polypeptide of 381 amino acids, predicted isoelectric point of TaSKP2A was 6.83 and the molecular weight was 40.87 kD. TaSKP2A was non-secreted protein according to the signal peptide prediction, and its subcellular localization was nuclear. TaSKP2A belonged to F-box/LRR protein based on the C-terminal domain. qRT-PCR results showed that TaSKP2A was significantly higher expressed in pistil and flag leave, while lower expressed in young stem and stamen of wheat. After treating with salicylic acid (SA) and NaCl, the expression of TaSPK2A was up-regulated at 24 and 48 h, respectively. When treating with H2O2, TaSPK2A showed gradually down-regulation. The relative expression of TaSPK2A in resistance combination at all inoculation timepoints were higher than that in susceptible one, and the highest expression (8.1 times higher when comparing to sample at 0 h) occurred in resistance combination at 12 h after inoculation with leaf rust pathogen (Puccinia triticina). Yeast (Saccharomyces cerevisiae) two hybrid was used to screen the target protein interacting with TaSPK2A, the results showed that chitinase (CHI) and S-phase kinase-associated protein 1 (SKP1), a scaffolded protein of Skp1-Cullins-F-box (SCF) complex, could interact with TaSPK2A. These results suggested that TaSKP2A might play an important role in response to salt stress and leaf rust pathogen infection, the TaSPK2A may be a member of SCF complex. The results lay basic foundation for analyzing the functional knowledge of SKP2A and further exploration of the regulatory network and mechanism of SKP2A.
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