Identification and Expression Analysis of Wheat (Triticum aestivum) SKP1 Family Members and Their Interaction with F-box Proteins
FAN Run-Qiao1, WEI Chun-Ru1, YANG Yi-Ming1, WANG Xiao-Dong2, WEI Xin-Yan2, LIU Da-Qun2, ZHAO Wei-Quan2,*, YU Xiu-Mei1,2,*
1 College of Life Sciences/Hebei Key Laboratory of Plant Physiology and Molecular Pathology, Hebei Agricultural University, Baoding 071001, China; 2 Technological Innovation Center for Biological Control of Crop Diseases and Insect Pests of Hebei Province, Baoding 071001, China
Abstract：As an adaptor protein of the SCF (Skp1-Cul1-F-box) complex, SKP1 (S-phase kinase-related protein 1) connects with Cullin-1 (Cul1) and F-box proteins, respectively. SKP1 family proteins play their roles in many cytological processes, such as growth and development of plants, transduction of hormone signal and response to biotic/abiotic stresses etc. To uncover the basic biological characteristics and functional mechanism of SKP1 in wheat (Triticum aestivum), a systemic identification and analysis on wheat SKP1 genes in the present study was conducted. 115 sequences were identified in the wheat proteome, which were encoded by 101 SKP1 genes, among them, 9 genes (TSK37, TSK41, TSK44, TSK50, TSK52, TSK59, TSK60, TSK85 and TSK99) were first identified in the present study. The length of 115 deduced SKP1 proteins ranged from 104 to 530 aa, and the theoretical pI showed that all SKP1 members were acidic proteins. The wheat SKP1 proteins were present in the cytoplasm, nucleus and chloroplast by subcellular localization prediction, most of them were localized in nucleus. All members of wheat SKP1 family contained at least one SKP1 and/or SKP1_POZ domain, and the SKP1 domain shared higher conservation when comparing to the SKP1_POZ domain. Phylogenetic analysis of the SKP1 protein from wheat, Arabidopsis thaliana, and rice (Oryza sativa) revealed that the SKP1 sequences from the same plant species were grouped in one branch, which indicated that these sequences shared closer phylogenetic relationship. Location of 101 SKP1 genes was identified on all 21 wheat chromosomes but showed an uneven distribution. The density on chromosome of No. 3 and No. 5 was relatively higher than that of the others. Gene expression patterns indicated that a few SKP1 genes (TSK37, TSK41, TSK54.1, TSK75, TSK97) had higher expression levels in different developmental stages of 5 wheat tissues based on heat map analysis, some of them, such as TSK62, TSK71, TSK56, TSK91.1, and TSK101.1, showed relative higher expression levels in a certain tissue or a specific stage of development, but most of them were not expressed or expressed at lower level in 5 tissues at all developmental stages. The expression patterns of SKP1 family genes in wheat varied greatly under drought and/or heat stresses, and heat stress brought stronger effect on the expression of SKP1 genes in wheat. Three genes TSK41, TSK43.2, and TSK62, which belonged to different type of SKP1 proteins, were obtained from wheat isogenic line TcLr15 based on the strategy of homologous cloning. qPCR showed that these three genes were significantly up-regulated in wheat response to salt stress, and down-regulated with different degrees after treatment by H2O2, whereas 3 stress-related hormones ABA, SA and MeJA had little effect on the expression of the three TSK genes (no more than 2.5 times), and inoculation with leaf rust pathogen (Puccinia triticina) caused less effects on their expressions, too, moreover their expression showed no obvious changes to the different virulent strains. Yeast two hybrid (Y2H) was used to screen the target proteins interacting with TSK41, TSK43.2 and TSK62. Results presented that 3 wheat SKP1 proteins and 8 F-box proteins had multiple interaction modes: TSK62 interacted with TaFBK19, TaFBK34, TaFBK71, TaSKP2A, and TaSKIP27, these 5 F-box proteins belong to 3 types: Kelch (FBK), LRR (FBL) and unknown functional domains (FBU), and TSK41 only interacted with 2 Kelch-type proteins TaFBK19 and TaFBK34, but TSK43.2 did not interact with any of these 8 F-box proteins. These findings supplied some basic and important knowledge for further dissecting the detailed mechanism of wheat SKP1 family members in response to biotic and abiotic stresses.
范润侨, 魏春茹, 杨一鸣, 王逍冬, 魏新燕, 刘大群, 赵伟全, 于秀梅. 小麦SKP1家族成员鉴定、基因表达及其与F-box蛋白的互作分析[J]. 农业生物技术学报, 2022, 30(2): 207-221.
FAN Run-Qiao, WEI Chun-Ru, YANG Yi-Ming, WANG Xiao-Dong, WEI Xin-Yan, LIU Da-Qun, ZHAO Wei-Quan, YU Xiu-Mei. Identification and Expression Analysis of Wheat (Triticum aestivum) SKP1 Family Members and Their Interaction with F-box Proteins. 农业生物技术学报, 2022, 30(2): 207-221.
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