Apple latent spherical virus vectors for reliable and effective virus-induced gene silencing among a broad range of plants including tobacco, tomato, Arabidopsis thaliana, cucurbits, and legumes. Virology

Plant Pathology Laboratory, Faculty of Agriculture, Iwate University, Morioka 020-8550, Japan.
Virology (Impact Factor: 3.32). 03/2009; 386(2):407-16. DOI: 10.1016/j.virol.2009.01.039
Source: PubMed


Apple latent spherical virus (ALSV) vectors were evaluated for virus-induced gene silencing (VIGS) of endogenous genes among a broad range of plant species. ALSV vectors carrying partial sequences of a subunit of magnesium chelatase (SU) and phytoene desaturase (PDS) genes induced highly uniform knockout phenotypes typical of SU and PDS inhibition on model plants such as tobacco and Arabidopsis thaliana, and economically important crops such as tomato, legume, and cucurbit species. The silencing phenotypes persisted throughout plant growth in these plants. In addition, ALSV vectors could be successfully used to silence a meristem gene, proliferating cell nuclear antigen and disease resistant N gene in tobacco and RCY1 gene in A. thaliana. As ALSV infects most host plants symptomlessly and effectively induces stable VIGS for long periods, the ALSV vector is a valuable tool to determine the functions of interested genes among a broad range of plant species.

Download full-text


Available from: Hajime Yaegashi
  • Source
    • "Some changes in protein levels would precede the transcriptomic changes, for example, the physical interaction of some host proteins with Tav, the degradation of Tav or some host proteins or both, and post-translational modifications such as phosphorylation, ubiquitination, or nitrosylation of Tav or some host proteins or both. In addition to such analytical studies, the present system is also useful for testing gene function using virus-induced gene silencing (VIGS), because VIGS in tobacco plants is possible using the Apple latent spherical virus vector [38]. Furthermore, the experimental system would also be useful for analyzing the process of chloroplast damage, because the chlorophyll decrease was not rapid and therefore, chloroplasts at different stages of damage would be available using the system. "
    [Show abstract] [Hide abstract]
    ABSTRACT: The underlying molecular mechanism of chlorosis, a typical symptom of plant viral diseases, remains poorly understood. To establish an experimental system to determine the molecular changes during chlorosis, especially in the early phase, we generated transgenic tobacco plants expressing Cauliflower mosaic virus Transactivator/viroplasmin (Tav) under the control of a chemically inducible promoter. Induction of Tav resulted in visible chlorosis in ten days, a statistically significant decrease in chlorophyll content in two days, decreased expression of chloroplast protein genes, and abnormal thylakoid stacks, indicating that this system reproduces the common features of chlorosis in virus-infected plants.
    Full-text · Article · Oct 2014 · Physiological and Molecular Plant Pathology
  • Source
    • "Because most vectors have limited host ranges, vector-mediated HGT from Brassicaceae (or from Fabaceae, in the case of albumin 1[35]) to Orobanchaceae and Convolvulaceae appears to be unlikely. Only vectors with a wide host range, such as apple latent spherical virus [57], may be able to mediate these transfer events. However, no plant viruses carrying the foreign genes have been discovered so far. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Besides gene duplication and de novo gene generation, horizontal gene transfer (HGT) is another important way of acquiring new genes. HGT may endow the recipients with novel phenotypic traits that are important for species evolution and adaption to new ecological niches. Parasitic systems expectedly allow the occurrence of HGT at relatively high frequencies due to their long-term physical contact. In plants, a number of HGT events have been reported between the organelles of parasites and the hosts, but HGT between host and parasite nuclear genomes has rarely been found. A thorough transcriptome screening revealed that a strictosidine synthase-like (SSL) gene in the root parasitic plant Orobanche aegyptiaca and the shoot parasitic plant Cuscuta australis showed much higher sequence similarities with those in Brassicaceae than with those in their close relatives, suggesting independent gene horizontal transfer events from Brassicaceae to these parasites. These findings were strongly supported by phylogenetic analysis and their identical unique amino acid residues and deletions. Intriguingly, the nucleus-located SSL genes in Brassicaceae belonged to a new member of SSL gene family, which were originated from gene duplication. The presence of introns indicated that the transfer occurred directly by DNA integration in both parasites. Furthermore, positive selection was detected in the foreign SSL gene in O. aegyptiaca but not in C. australis. The expression of the foreign SSL genes in these two parasitic plants was detected in multiple development stages and tissues, and the foreign SSL gene was induced after wounding treatment in C. australis stems. These data imply that the foreign genes may still retain certain functions in the recipient species. Our study strongly supports that parasitic plants can gain novel nuclear genes from distantly related host species by HGT and the foreign genes may execute certain functions in the new hosts.
    Full-text · Article · Jan 2014 · BMC Plant Biology
  • Source
    • "Our finding also implies that many other available viral vectors, such as Apple latent spherical virus (Igarashi et al., 2009; Yamagishi et al., 2011), Brome mosaic virus (Ding et al., 2006), and Barley stripe mosaic virus (Holzberg et al., 2002), could be used in a similar strategy to the TRV VbMS vector for functional analysis of miRNAs in a diverse range of eudicot and monocot crops. In addition, besides IPS1-based miRNA target mimicry and STTM methods, several other techniques, such as transcriptional gene silencing of miRNA gene promoters (Vaistij et al., 2010), artificial miRNA-directed silencing of miRNA precursors (Eamens et al., 2011), and miRNA decoy (Ivashuta et al., 2011), have successfully been used to investigate miRNA/target interactions in transgenic plants. "
    [Show abstract] [Hide abstract]
    ABSTRACT: MicroRNAs (miRNAs) play pivotal roles in various biological processes across kingdoms. Many plant miRNAs have been experimentally identified or predicted by bioinformatics mining of small RNA databases. However, functions of these miRNAs remain largely unknown due to the lack of effective genetic tools. Here, we report a virus-based miRNA silencing (VbMS) system that can be used for functional analysis of plant miRNAs. VbMS is performed through Tobacco rattle virus (TRV)-based expression of miRNA target mimics to silence endogenous miRNAs. VbMS of either miR172 or miR165/166 caused developmental defects in Nicotiana benthamiana. VbMS of miR319 reduced the complexity of tomato compound leaves. These results demonstrate that TRV-based VbMS is a powerful tool to silence endogenous miRNAs and to dissect their functions in different plant species.
    Full-text · Article · Dec 2013 · Plant physiology
Show more

We use cookies to give you the best possible experience on ResearchGate. Read our cookies policy to learn more.