甘蔗PsbR亚基应答SCMV侵染及其与SCMV-6K2的互作

doi:10.3724/SP.J.1006.2021.04194

Abstract

Abstract:

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

ZHANG Hai, CHENG Guang-Yuan, YANG Zong-Tao, LIU Shu-Xian, SHANG He-Yang, HUANG Guo-Qiang, XU Jing-Sheng. Sugarcane PsbR subunit response to SCMV infection and its interaction with SCMV-6K2[J].Acta Agronomica Sinica, 2021, 47(8): 1522-1530.

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引物名称 Primer name	引物序列 Primer sequence (5′-3′)	策略 Strategy
221-ScPsbR-F 221-ScPsbR-R	GGGGACAAGTTTGTACAAAAAAGCAGGCTTCATGGCGGCCTCCATGATCTC GGGGACCACTTTGTACAAGAAAGCTGGGTCGCCGGAGAGGGCGCTGGTGT	双分子荧光互补载体构建 Vector generation for BiFC
ScPsbR-qF ScPsbR-qR	TCCATGATCTCGTCGTCAGCTCTG ACGGTCAGGCCACCTCCAATC	定量PCR Real-time-qPCR
GAPDH-F GAPDH-R	CACGGCCACTGGAAGCA TCCTCAG GGTTCCTGATGCC	内参基因 Reference gene
eEF-1α-F eEF-1α-R	TTTCACACTTGGAGTGAAGCAGAT GACTTCCTTCACAATCTCATCATAA	内参基因 Reference gene
SCMV-CP-F SCMV-CP-R	TACAGAGAGACACACAGCTG ACGCTACACCAGAAGACACT	SCMV检测 Detection of SCMV

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

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