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Acta Agronomica Sinica ›› 2022, Vol. 48 ›› Issue (2): 332-341.doi: 10.3724/SP.J.1006.2022.14001


Sugarcane ubiquitin-like protein UBL5 responses to SCMV infection and interacts with SCMV-6K2

YANG Zong-Tao(), LIU Shu-Xian, CHENG Guang-Yuan, ZHANG Hai, ZHOU Ying-Shuan, SHANG He-Yang, HUANG Guo-Qiang, XU Jing-Sheng*()   

  1. Sugarcane Research & Development Center, China Agricultural Technology System, Fujian Agriculture and Forestry University / Key Laboratory of Sugarcane Biology and Genetic Breeding, Ministry of Agriculture and Rural Affairs / Key Laboratory of Ministry of Education for Genetics, Breeding and Multiple Utilization of Crops, Fuzhou 350002, Fujian, China
  • Received:2021-01-05 Accepted:2021-04-14 Online:2022-02-12 Published:2021-06-16
  • Contact: XU Jing-Sheng E-mail:fafuyangzongtao@163.com;xujingsheng@126.com
  • Supported by:
    This study was supported by the National Natural Science Foundation of China(31971991);the Science and Technology Innovation Project of Fujian Agriculture and Forestry University(CXZX2018026);Guiding Project of Science and Technology Department of Fujian Province(2017N0003)


Ubiquitylation plays key roles in the regulation of protein function, growth and development, and response to stress. Ubiquitin-like proteins (UBLs) are the main components of the ubiquitin-proteasome system (UPS). In our previous study, the UBL5 homologue was isolated from sugarcane (Saccharum spp. hybrid) by yeast two-hybrid (Y2H) with the 6K2 of Sugarcane mosaic virus (SCMV) as bait, and then designated as ScUBL5 with 73 aa in length. In the present study, the interaction of ScUBL5 with the SCMV-6K2 was further confirmed by bimolecular fluorescence complementation assays (BiFC). Bioinformatics analysis showed that ScUBL5 is a stable hydrophilic non-secretory protein without signal peptide or transmembrane domain. Phylogenetic tree analysis showed that ScUBL5 is species specific. Subcellular localization analysis showed that ScUBL5 is localized in cytoplasm and nucleus. ScUBL5 gene shows obvious tissue specificity in sugarcane by real-time quantitative PCR analysis. The expression levels of ScUBL5 gene in the established morphogenesis tissues such as the 1st leaf, the 7th leaf, the 8th internode and the root were significantly higher than those in the immature tissues such as leaf roll and the 3rd internode. The expression of ScUBL5 gene is significantly affected by SCMV infection. ScUBL5 was significantly upregulated in the early stage of SCMV infection, then downregulated but significantly higher than the control at the late stage of SCMV infection.

Key words: sugarcane, ubiquitin-like protein, Sugarcane mosaic virus, 6K2

Table 1

Primers used in this study"

Primer name
Primer sequence (5′-3′)
Vector generation for BiFC
Reference gene
Reference gene
Detection of SCMV

Fig. 1

Three-dimensional structure of ScUBL5 protein based on SWISS-MODEL ScUBL5 (MN167908) is from Saccharum spp. hybrid; SbUBL5 (XP_002448973.1) is from Sorghum bicolor; OsUBL5 (XP_015619383.1) is from Oryza sativa; AtUBL5 (NP_176030.1) is from Arabidopsis thaliana."

Fig. 2

The deduced amino acid sequence of ScUBL5 Oryza sativa: OsUBL5 (XP_015619383.1); Saccharum: ScUBL5 (QHD26889.1); Brachypodium distachyon: BdUBL5 (XP_003578938.1); Sorghum bicolor: SbUBL5 (XP_002448973.1); Arabidopsis thaliana: AtUBL5 (NP_199045.1); Nicotiana tabacum: NtUBL5 (XP_ 016510251.1); Triticum aestivum: TaUBL5 (AJA91631.1)."

Fig. 3

Phylogenetic analysis UBL5 proteins from different plant species"

Fig. 4

Subcellular localization of ScUBL5 fused with YFP in the epidermal cells of N. benthamiana Bar: 25 μm."

Fig. 5

BiFC assays for protein-protein interaction between ScUBL5 and SCMV-6K2 A: the C-terminal half of YFP was fused to the C-terminus of ScUBL5 to generate ScUBL5-YC, while the N-terminus half of YFP was fused to the N-terminus of SCMV-6K2 to generate 6K2-YN; B: the N-terminus half of YFP was fused to the N-terminus of ScUBL5 to generate ScUBL5-YN, while the C-terminal half of YFP was fused to the C-terminal of SCMV-6K2 to generate 6K2-YC. Plasmids combination of 6K2-YN plus ScUBL5-YC (A), ScUBL5-YN plus 6K2-YC (B) were individually co-injected into N. benthamiana leaves for transient expression. The fluorescent signal was monitored by confocal microscopy at 48 h post infiltration. Bar: 25 μm."

Fig. 6

Expression profile of ScUBL5 in different sugarcane tissues LR: leaf roll; +1 L: +1 leaf; +7 leaf; +3 I: +3 internode; +8 I: +8 internode; R: root. The error bars represent the standard error of each treating group (n = 3). Bars superscripted by different lowercase letters are significantly different at P < 0.05."

Fig. 7

Expression profile of ScUBL5 under the infection of SCMV The error bars represent the standard error of each treating group (n = 3). Bars superscripted by different lowercase letters are significantly different at P < 0.05."

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