Identification of the Cysteine Protease Family and Corresponding miRNAs in Nicotiana tabacum L. and Their Responses to PVY

doi:10.13560/j.cnki.biotech.bull.1985.2023-0297

Abstract

Abstract:

Plant cysteine proteases（CPs）widely affect plant physiological processes and disease resistance regulation. To study the functional mechanism of tobacco cysteine proteases in regulating potato virus Y（PVY）, the CPs family genes of Nicotiana tabacum L. at the genome-wide level were identifed. The evolutionary relationships, motifs and promoter cis-acting elements of CPs family genes were analyzed by bioinformatics. Then the miRNAs corresponding to CPs were screend, the expression characteristics of CPs genes and their corresponding miRNAs after PVY infection were analyzed. The results showed that a total of 70 CPs genes were identified in Nicotiana tabacum L. genome, which were divided into five subfamilies, of which C1A contained 39 CPs, C2A contained 2 CPs, C12 contained 5 CPs, C13 contained 8 CPs and C14A contained 16 CPs. Motif analyses showed that the CPs of subfamily had similar motif distribution. Promoter cis-acting elements analysis showed that several CPs family gene promoters contained light response elements, hormone response elements and low temperature, drought, high temperature, salt stress response elements. Based on the sequencing results of transcriptome and small RNA, it was found that 38 CPs were up-regulated while 18 CPs were down-regulated after PVY infection.Total 51 miRNAs were found to be targeted to 28 members of cysteine protease gene family, 38 miRNAs were up-regulated while 13 were down-regulated after PVY infection,7 of them were negatively correlated with miRNAs.The results of RT-qPCR showed that 15 CPs genes were significantly up-regulated after PVY infection. This study is conducive to better understanding the functions of CPs genes and their corresponding miRNAs in regulating tobacco PVY responses, which may provide evidence for the molecular mechanism of anti-PVY in Solanaceae crops.

Key words: tobacco, cysteine protease, gene family, miRNA, interaction

YIN Guo-ying, LIU Chang, CHANG Yong-chun, YU Wang-jie, WANG Bing, ZHANG Pan, GUO Yu-shuang. Identification of the Cysteine Protease Family and Corresponding miRNAs in Nicotiana tabacum L. and Their Responses to PVY[J]. Biotechnology Bulletin, 2023, 39(10): 184-196.

Figures/Tables 7

Fig. 1 Phylogenetic tree of cysteine protease family in tobacco

Fig. 2 Conserved domain pattern of cysteine protein in tobacco

Fig. 3 Cis-acting element mode diagram of cysteine protein in tobacco

Fig. 4 Networks of corresponding cysteine protease family genes and their relatived miRNAs in tobacco Yellow represents target gene and blue represents miRNAs

Fig. 5 Expressions of cysteine protease family genes and miRNAs infected PVY in tobacco A, B indicate the expression heatmap of cysteine protease family genes and miRNAs; C, D indicate the scatter plot of cysteine protease family genes and miRNAs, respectively; PVY refers to infection with PVY, while CK refers to control

Fig. 6 Negative correlation network of expressions of cysteine protease family genes and corresponding miRNAs after PVY infection in tobacco

Fig. 7 RT-qPCR analysis of the cysteine protease genes in PVY-infected tobacco “CK” and “PVY” indicate the control and infected PVY, respectively; * and * * respectively indicate significant differences in gene expression levels at the P<0.05 and P<0.01 levels before and after infection with PVY

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