Abstract
Tomato zonate spot virus (TZSV), a member of the genus orthotospovirus, causes severe damage to vegetables and ornamental crops in southwest China. The NSs protein is an RNA silencing suppressor in various orthotospovirus like TZSV, but its mechanism and role in virus infection are poorly understood. Here, we observed that an NSs-GFP fusion protein was transiently expressed on the plasma membrane and Golgi bodies in Nicotiana benthamiana plants. The TZSV NSs gene was silenced and infiltrated into N. benthamiana and N. tabacum cv. K326. RT-qPCR and Indirect enzyme-linked immunosorbent assay (ID-ELISA) showed that the transcription and the protein expression of the NSs gene were inhibited by more than 90.00%, and the symptoms on silenced plants were alleviated. We also found that the expression of the Zingipain-2-like gene significantly decreased when the NSs gene was silenced, resulting in co-localization of the NSs-GFP and the Zingipain-2-like-mCherry fusion protein. The findings of this study provide new insights into the mechanism of silencing suppression by NSs, as well as its effect on systemic virus infection, and also support the theory of disease resistance breeding and control and prevention of TZSV in the field.
Highlights
Tomato zonate spot virus (TZSV), the dominant species, which belongs to the orthotospovirus genus of the Bunyaviridae family in Yunnan Province, is transmitted by thrips [1]
Previous studies have shown that confocal laser scanning microscopy (CLSM) was used to analyze the protein localized in living cells [22]
N. benthamiana is a model plant species to assess the subcellular localization of viral proteins
Summary
Tomato zonate spot virus (TZSV), the dominant species, which belongs to the orthotospovirus genus of the Bunyaviridae family in Yunnan Province, is transmitted by thrips [1]. TZSV has been prevalent in Yunnan, Guizhou, Guangxi, and other regions of China and Southeast Asia. TZSV has a very broad host range, infecting more than 20 types of economically important crops and weed species, which belong to 7 families [2, 3]. This prevalence has led to production losses and quality problems for important vegetable and ornamental plants in Southwest China and seriously threatened economically important crops for local farmers [4].
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