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Acta Agronomica Sinica ›› 2021, Vol. 47 ›› Issue (8): 1522-1530.doi: 10.3724/SP.J.1006.2021.04194


Sugarcane PsbR subunit response to SCMV infection and its interaction with SCMV-6K2

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

  1. National Engineering Research Center for Sugarcane/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/Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian, China
  • Received:2020-08-25 Accepted:2020-11-13 Online:2021-08-12 Published:2020-12-22
  • Contact: XU Jing-Sheng E-mail:zhanghai940410@163.com;xujingsheng@126.com
  • Supported by:
    National Natural Science Foundation of China(31971991);Science and Technology Innovation Project of Fujian Agriculture and Forestry University(CXZX2018026);Guiding Project of Science and Technology Department of Fujian Province(2017N0003)


The PsbR subunit of photosystem II (PSII) plays a vital role in the assembly and stability of the oxygen-evolving complex. In the previous study, we cloned the coding sequence of the PsbR subunit from sugarcane (Saccharum spp. hybrid) and designated it as ScPsbR. The interaction between ScPsbR and 6K2 protein encoded by Sugarcane mosaic virus (SCMV) was verified by yeast two-hybrid technology. In this study, bioinformatics analysis indicated ScPsbR protein was found to possess a canonical subunit domain of PsbR and a transmembrane domain without signal peptide, and be a stable hydrophobic protein. Phylogenetic tree analysis indicated obvious divergence between C3 and C4 plants for the PsbRs. Subcellular localization experiments suggested that ScPsbR was localized and co-localized with SCMV-6K2 to the chloroplast. The interaction of ScPsbR with the SCMV-6K2 was further verified by bimolecular fluorescence complementation assays. Real-time quantitative PCR results indicated that ScPsbR gene was tissue-specific in sugarcane plants. There was almost no expression of ScPsbR gene in the roots or stems, with increased expression level in the senescing leaves and immature leaves, and the highest expression level in mature leaves. However, the expression level of ScPsbR was significantly changed under the challenged of SCMV. During the infection of SCMV, ScPsbR was significantly up-regulated at 0-12 hour(s) and reduced to a level slightly lower than that of the control at 1-5 days with no significant differences, then significantly down-regulated in 7-15 days.

Key words: PsbR subunit, Sugarcane mosaic virus, 6K2, protein interaction

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

Nucleotide sequence of the ScPsbR gene and its encoded amino acid sequence *: stop codon. The sequence underlined in black indicates the transit peptide and the sequence underlined in red indicates the transmembrane domains. ↓ indicates the digestion site. The border between transit sequence and mature protein is marked with an arrowhead."

Fig. 2

Amino acid sequence alignment of ScPsbR and PsbRs from other monocotyledon species SbPsbR (XP_002444002.1): Sorghum bicolor; ZmPsbR (XP_020408082.1): Zea mays; PhPsbR (XP_025821160.1): Panicum hallii; SiPsbR (XP_004972630.1): Setaria italic; BdPsbR (XP_010234446.1): Brachypodium distachyon; OsPsbR (AAB46718.1): Oryza sativa."

Fig. 3

Phylogenetic tree of ScPsbR protein and PsbR proteins from other plant species"

Fig. 4

Subcellular localization of ScPsbR fused with YFP in the epidermal cells of N. benthamiana A: subcellular localization of ScPsbR-YFP; B: subcellular colocalization of ScPsbR-YFP with 6K2-CFP. White arrows indicate the chloroplasts, Bar: 25 μm."

Fig. 5

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

Fig. 6

Relative expression level of ScPsbR in different sugarcane tissues The error bars represent the standard error of each treating group (n = 3). LR: leaf roll; +1 L: 1st leaf; +7 L: 7th leaf; +3 I: 3rd internode; +8 I: 8th internode; R: root. Different lowercase letters above the bars are significantly different at the 5% probability level."

Fig. 7

Expression profile of ScPsbR under the infection of SCMV The error bars represent the standard error of each treating group (n = 3). Different lowercase letters above the bars are significantly different at the 5% probability level."

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