Biotechnology Bulletin ›› 2023, Vol. 39 ›› Issue (10): 184-196.doi: 10.13560/j.cnki.biotech.bull.1985.2023-0297

Previous Articles     Next Articles

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

YIN Guo-ying1(), LIU Chang2, CHANG Yong-chun1,3, YU Wang-jie1,4, WANG Bing1, ZHANG Pan1, GUO Yu-shuang1()   

  1. 1. Guizhou Academy of Tobacco Science, Guiyang 550081
    2. College of Agronomy and Biotechnology, Southwest University, Chongqing 400715
    3. College of Agriculture, Shanxi Agricultural University, Jinzhong 030800
    4. College of Advanced Agriculture and Ecological Environment of Heilongjiang University, Harbin 150006
  • Received:2023-04-04 Online:2023-10-26 Published:2023-11-28
  • Contact: GUO Yu-shuang E-mail:yinyinghygy6239@126.com;yshguo@126.com

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