Journal of Integrative Agriculture ›› 2022, Vol. 21 ›› Issue (3): 751-761.DOI: 10.1016/S2095-3119(21)63681-X

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  • 收稿日期:2021-01-11 接受日期:2021-03-08 出版日期:2022-03-01 发布日期:2021-03-08

PnSCR82, a small cysteine-rich secretory protein of Phytophthora nicotianae, can enhance defense responses in plants

WANG Ya-di1*, LI Fei1*, ZHANG Xin2, LIU Ting-li3, LIANG Wen-xing1, LI De-long1   

  1. 1 College of Plant Health and Medicine, the Key Laboratory of Integrated Crop Pest Management of Shandong Province, Qingdao Agricultural University, Qingdao 266109, P.R.China
    2 Hunan Academy of Agricultural Sciences, Changsha 410125, P.R.China
    3 Provincial key laboratory of agrobiology, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, P.R.China
  • Received:2021-01-11 Accepted:2021-03-08 Online:2022-03-01 Published:2021-03-08
  • About author:WANG Ya-di, E-mail: 314785383@qq.com; Correspondence LI De-long, Tel: +86-532-58957776, E-mail: ldlong@qau.edu.cn * These authors contributed equally to this study.
  • Supported by:
    This work was supported by the National Natural Science Foundation of China (31972218, 31501590, and 31601615), the Program for Talents in Qingdao Agricultural University, China (6631114307), and the Independent Innovation of Agricultural Sciences in Jiangsu Province, China (CX(18)3012).  

摘要:

植物病原菌侵染植物过程中分泌大量的外泌蛋白调控植物免疫反应,这些外泌蛋白中包含一类富含半胱氨酸(small cysteine-rich, SCR)的小分泌蛋白,它能帮助病原菌在寄主植物中定殖。烟草疫霉中同样存在富含半胱氨酸外泌蛋白,但其功能尚不清楚。本研究通过生物信息学分析,发现烟草疫霉PnSCR82同源于恶疫霉的植物毒性蛋白PcF (Phytophthora cactorum Fragaria),其具有外泌信号肽,同时富含半胱氨酸。为研究烟草疫霉PnSCR82功能,将PnSCR82构建于马铃薯X病毒(PVX)表达载体(pGR106),利用农杆菌介导的转化方法注射本氏烟草叶片和番叶片茄,发现在烟草叶片瞬时表达PnSCR82,能够诱导烟草叶片和番茄叶片出现过敏性坏死,并且能增加胼胝质和活性氧在叶片的积累,接种辣椒疫霉后发现其增强对辣椒疫霉的抗性。通过构建其亚细胞定位载体PnSCR82-GFP,利用农杆菌介导的转化注射烟草叶片发现其定位于烟草细胞膜。同时RT-PCR分析发现在烟草叶片中瞬时表达PnSCR82能够激活烟草水杨酸,茉莉酸和乙烯途径的部分相关基因上调表达。烟草疫霉PnSCR82作为激发子一员,能够诱导烟草细胞过敏性坏死同时也能激活不同免疫信号通路相关基因上调表达,能够增强植物对辣椒疫霉的抗性,具有潜在的提高植物免疫应用价值。本文首次验证了烟草疫霉富含半胱氨酸蛋白PnSCR82功能,作为激发子,PnSCR82能够诱导植物产生过敏性坏死反应,能够激活部分免疫相关基因的上调表达,具有潜在的应用价值。


Abstract: A number of plant pathogenic species of Phytophthora are known to produce different classes of secretory proteins during interactions with their hosts.  Although several small cysteine-rich (SCR) secretory proteins, conserved in oomycete pathogens, have been identified in Phytophthora, their specific involvement in these interactions remains unknown.  In this study, an SCR effector encoded by Pnscr82 in P. nicotianae was identified and shown to have similarities to P. cactorum phytotoxic protein, PcF (Phytophthora cactorum Fragaria).  Agroinfection with potato virus X vector, PnSCR82, was capable of inducing plant hypersensitive cell death in Nicotiana benthamiana and Solanum lycopersicum.  Real-time PCR results indicated that transiently expressed PnSCR82 in N. benthamiana leaves activated the jasmonate, salicylic acid and ethylene signaling pathways.  Transient expression of PnSCR82 enhanced plant resisitance to P. capsici.  In summary, our results demonstrated that P. nicotianae PnSCR82 elicits defensive responses in N. benthamiana and may potentially play a significant role in future crop protection programs.

Key words: small cysteine-rich secretory protein ,  defense responses ,  resistance