Abstract
Alternaria alternata (tobacco pathotype) is a necrotrophic fungus causing severe losses in Nicotiana species by infection of mature leaves. Similar to what has been observed in cultivated tobacco, N. tabacum, young leaves of wild tobacco, N. attenuata, were more resistant to A. alternata than mature leaves, and this was correlated with stronger blue fluorescence induced after infection. However, the nature of the fluorescence-emitting compound, its role in defence, and its regulation were not clear. Silencing feruloyl-CoA 6'-hydroxylase 1 (F6'H1), the gene encoding the key enzyme for scopoletin biosynthesis, by virus-induced gene silencing (VIGS) revealed that the blue fluorescence was mainly emitted by scopoletin and its β-glycoside form, scopolin. Further analysis showed that scopoletin exhibited strong antifungal activity against A. alternata in vitro and in vivo. Importantly, jasmonic acid (JA) levels were highly elicited in young leaves but much less in mature leaves after infection; and fungus-elicited scopoletin was absent in JA-deficient plants, but was largely restored with methyl jasmonate treatments. Consistent with this, plants strongly impaired in JA biosynthesis and perception were highly susceptible to A. alternata in the same way scopoletin/scopolin-depleted VIGS F6'H1 plants. Furthermore, silencing MYC2, a master regulator of most JA responses, reduced A. alternata-induced NaF6'H1 transcripts and scopoletin. Thus, it is concluded that JA signalling is activated in N. attenuata leaves after infection, which subsequently regulates scopoletin biosynthesis for the defence against A. alternata partly through MYC2, and higher levels of scopoletin accumulated in young leaves account for their strong resistance.
Highlights
In many plant–pathogen systems, the resistance of plants usually depends on the developmental stage at which the plant is infected
Similar to what has been observed in cultivated tobacco, N. tabacum, young leaves of wild tobacco, N. attenuata, were more resistant to A. alternata than mature leaves, and this was correlated with stronger blue fluorescence induced after infection
Silencing feruloyl-CoA 6ʹ-hydroxylase 1 (F6ʹH1), the gene encoding the key enzyme for scopoletin biosynthesis, by virus-induced gene silencing (VIGS) revealed that the blue fluorescence was mainly emitted by scopoletin and its β-glycoside form, scopolin
Summary
In many plant–pathogen systems, the resistance of plants usually depends on the developmental stage at which the plant is infected. Plants are generally more susceptible to disease in early than in late phases, including rice against Xanthomonas oryzae, tobacco to Phytophthora parasitica, and Arabidopsis to Pseudomonas syringae (Century et al, 1999; Hugot et al, 1999; Kus et al, 2002; Develey-Riviere and Galiana, 2007). This is not true in the Nicotiana tabacum–Alternaria alternata interaction (Cheng and Sun, 2001; Zhang et al, 1998).
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