Zhang Ying-Li, Jia Qing-Li, Li Da-Wei, Wang Jun-E, Yin Yan-Xu, Gong Zhen-Hui
College of Horticulture, Northwest A&F University, Yangling 712100, Shaanxi, China.
Int J Mol Sci. 2013 Apr 25;14(5):8985-9004. doi: 10.3390/ijms14058985.
The most significant threat to pepper production worldwide is the Phytophthora blight, which is caused by the oomycete pathogen, Phytophthora capsici Leonian. In an effort to help control this disease, we isolated and characterized a P. capsici resistance gene, CaRGA2, from a high resistant pepper (C. annuum CM334) and analyzed its function by the method of real-time PCR and virus-induced gene silencing (VIGS). The CaRGA2 has a full-length cDNA of 3,018 bp with 2,874 bp open reading frame (ORF) and encodes a 957-aa protein. The protein has a predicted molecular weight of 108.6 kDa, and the isoelectric point is 8.106. Quantitative real-time PCR indicated that CaRGA2 expression was rapidly induced by P. capsici. The gene expression pattern was different between the resistant and susceptible cultivars. CaRGA2 was quickly expressed in the resistant cultivar, CM334, and reached to a peak at 24 h after inoculation with P. capsici, five-fold higher than that of susceptible cultivar. Our results suggest that CaRGA2 has a distinct pattern of expression and plays a critical role in P. capsici stress tolerance. When the CaRGA2 gene was silenced via VIGS, the resistance level was clearly suppressed, an observation that was supported by semi-quantitative RT-PCR and detached leave inoculation. VIGS analysis revealed their importance in the surveillance to P. capsici in pepper. Our results support the idea that the CaRGA2 gene may show their response in resistance against P. capsici. These analyses will aid in an effort towards breeding for broad and durable resistance in economically important pepper cultivars.
全球辣椒生产面临的最重大威胁是由卵菌病原体辣椒疫霉(Phytophthora capsici Leonian)引起的疫病。为了帮助控制这种病害,我们从高抗辣椒(C. annuum CM334)中分离并鉴定了一个辣椒疫霉抗性基因CaRGA2,并通过实时荧光定量PCR和病毒诱导基因沉默(VIGS)方法分析了其功能。CaRGA2的全长cDNA为3018 bp,开放阅读框(ORF)为2874 bp,编码一个957个氨基酸的蛋白质。该蛋白质预测分子量为108.6 kDa,等电点为8.106。实时荧光定量PCR表明,辣椒疫霉能快速诱导CaRGA2表达。抗性和感病品种之间的基因表达模式不同。CaRGA2在抗性品种CM334中快速表达,接种辣椒疫霉后24 h达到峰值,比感病品种高5倍。我们的结果表明,CaRGA2具有独特的表达模式,在辣椒对辣椒疫霉的胁迫耐受性中起关键作用。当通过VIGS沉默CaRGA2基因时,抗性水平明显受到抑制,半定量RT-PCR和离体叶片接种实验也证实了这一观察结果。VIGS分析揭示了它们在辣椒对辣椒疫霉监测中的重要性。我们的结果支持CaRGA2基因可能在抗辣椒疫霉反应中发挥作用的观点。这些分析将有助于培育经济价值高的辣椒品种的广谱持久抗性。