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Chinese Journal of Biological Control ›› 2021, Vol. 37 ›› Issue (2): 323-330.DOI: 10.16409/j.cnki.2095-039x.2021.02.006

• RESEARCH REPORTS • Previous Articles    

Resistance to TMV in Tobacco Induced by Eukaryotic Expression Production of Y3

ZHANG Zhiyun, BIAN Yeyu, HUANG Hang, HAN Xiaoxiao, WU Lan, WU Liping   

  1. School of Life Sciences, Nanchang University/Key Laboratory of Poyang Lake Environment and Resource, Ministry of Education, Nanchang University, Nanchang 330031, China
  • Received:2020-07-29 Published:2021-04-14

Abstract: Protein Y3, an alkaline glycoprotein extracted from Coprinus comatus, could reduce TMV infection to tobacco. Vector pPIC-9k-Y3 was constructed based on the cDNA of Y3 and the expression plasmid pPIC-9k, and then transformed into Pichia pastoris GS115 by eletroporation successfully. The positive transformants was induced for 6 d by 1.0% methanol at the condition of 28℃ and 250 r/min. After sprayed with 300 μg/mL Y3 protein, tobacco plants were inoculated by TMV, then suffered with the activity detection of the disease-resistant defense enzymes such as polyphenol oxidase (PPO), phenylalanine ammonialyase (PAL) and β-1,3 glucanase. The results showed that they were enhanced, and the maximum activity of PPO reached to 2 times compared with control after 24 h of protein treatment. Analysis of resistance-related genes expression by Quantitative real-time PCR (qRT-PCR) showed that basic pathogenesis-related gene 1 (PR1-b), nonexpressor of pathogenesis-related gene 1 (NPR1) and PAL gene were significantly up regulated after Y3 treatment (P<0.05). Thus, we can infer that the recombinant Y3 induce systemic disease resistance of tobacco by increasing the activity of defense enzyme and transcription of resistance genes related to salicylic acid signal transduction pathway.

Key words: protein Y3, eukaryotic expression, tobacco mosaic virus, induced resistance

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