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Acta Agronomica Sinica ›› 2023, Vol. 49 ›› Issue (1): 97-104.doi: 10.3724/SP.J.1006.2023.14239

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Gene cloning and expression analysis of ScPR10 in sugarcane under Acidovorax avenae subsp. avenae infection

LI Juan, ZHOU Jing-Ru, CHU Na, SUN Hui-Dong, HUANG Mei-Ting, FU Hua-Ying, GAO San-Ji()   

  1. National Engineering Research Center for Sugarcane, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian, China
  • Received:2021-12-17 Accepted:2022-03-25 Online:2023-01-12 Published:2022-04-20
  • Contact: GAO San-Ji E-mail:gaosanji@fafu.edu.cn
  • Supported by:
    Earmarked Fund of the China Agriculture Research System of MOF and MARA(CARS-170302);Director’s Innovation Fund in National Engineering Research Center for Sugarcane, FAFU, China(KJG16005M)

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Abstract:

PR10 is a pathogenesis-related protein (PR), which plays critical roles in the growth and development and various environmental stress responses in plants. This study demonstrated that two ScPR10 genes were cloned from the leaf samples of ROC22 (resistant to red stripe) and Mintang 11-610 (susceptible to red stripe) by RT-PCR, and then RT-qPCR, transcriptome, and proteomics assays were carried out to explore the expression patterns of two genes in varieties ROC22 and Mintang 11-610 post inoculation with red stripe pathogen Acidovorax avenae subsp. avenae (Aaa). Sequence analysis showed that the two ScPR10 genes encoding 187 amino acids had more 27 amino acids than PR10 proteins from other plants, which contained the conserved domains P-loop and Bet v 1. ScPR10 obtained in this study shared high homology with these published PR10 proteins encoded by Saccharum spp., Sorghum bicolor, and Erianthus arundinaceus. The RT-qPCR analysis showed that the relative expression levels of ScPR10 genes were significantly increased by 27.2 times and 39.7 times in the resistant and susceptible varieties under Aaa infection, especially at 24 hours post inoculation (24 hpi), compared with the controls (0 hpi), respectively. Meanwhile, the transcriptome data revealed that the relative expression levels of ScPR10 genes were significantly increased by 5.3-5.4 folds [log2(Fold Change)] in the two varieties under Aaa infection, particular at 72 hours post inoculation (72 hpi). Prediction of protein interaction revealed that ScPR10 would interact with the pleiotropic drug resistance (PDR) protein of ABCG transporter subfamily (Cluster-13677.282407) and protein kinase (Cluster-13677.166559). Proteomics analysis referred that the relative expression of ScPR10 proteins were increased by 1.65-1.69 folds (log2FC) evaluated by 24 hpi versus 0 hpi in resistant and susceptible varieties. Similarly, the relative expression levels of the ABC transporter (Cluster-13677.282407) were upregulated to a certain extent in two varieties, but the relative expression levels of protein kinase (Cluster-13677.166559) were only enhanced in Mintang 11-610, not in ROC22. In conclusion, the ScPR10 may synergize with the ABC transporter (Cluster-13677.282407) involving in the defense response in sugarcane in response to Aaa infection.

Key words: PR10 protein, Acidovorax avenae subsp. avenae, biotic stress, gene expression, Saccharum spp.

Table 1

Physical and chemical properties of ScPR10 proteins"

NCBI登录号
NCBI
accession ID.		 基因长度
Gene length (bp)		 CDS长度
CDS length
(bp)		 氨基酸
No. of amino acid (aa)		 分子式
Molecular
formula		 分子量
Molecular weight (kD)		 等电点
pI		 不稳定系数
Instability index		 亲水性
Hydrophilicity
OK290572 697 564 187 C883H1401N231O274S5 19,797.52 4.98 32.02 0.042
OK290571 695 564 187 C885H1406N230O275S5 19,828.58 4.98 31.93 0.067

Fig. 1

Structure prediction for ScPR10 proteins A and B present the secondary structures of ScPR10 proteins from ROC22 and MT11-610 varieties, respectively, and C shows the tertiary structures of ScPR10 proteins. α-helix, irregular curl, extended chain, and β-sheet are shown in blue, pink, red, and green colors, respectively. α (1-3): alpha-helix; β (1-7): beta-sheet; L (1-9): loop."

Fig. 2

Multiple sequence alignment of PR10 proteins from different plants The conserved domains of P-loop and Bet v 1 are indicated by a red box and underline, respectively. The hydrophobic ligand binding site is marked with yellow triangles."

Fig. 3

Phylogenetic tree (A) and sequence identity (B) based on PR10 proteins from various plants"

Fig. 4

Relative expression profiling of ScPR10 genes of sugarcane under Aaa attack A: qRT-PCR assay; B: the transcriptome data (log2FC). ROC22 and MT11-610 are resistant and susceptible to red stripe disease, respectively. Different lowercase letters above the bars indicate significant differences at P < 0.05."

Fig. 5

Post-transcriptional expression levels of ScPR10 and their interaction proteins The values of log2 FC in blue and red boxes are the ratios of protein expression level of resistant (R24) or susceptible (S24) variety at 24 hours post inoculation (hpi) versus at 0 hpi (CK0), respectively."

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