%0 Journal Article
%T Efficient virus-induced gene silencing in apple, pear and Japanese pear using Apple latent spherical virus vectors
%A Shintarou Sasaki
%A Noriko Yamagishi
%A Nobuyuki Yoshikawa
%J Plant Methods
%D 2011
%I BioMed Central
%R 10.1186/1746-4811-7-15
%X Inoculation of ALSV vectors carrying a partial sequence of endogenous genes from apple [ribulose-1, 5-bisphosphate carboxylase small subunit (rbcS), alpha subunit of chloroplast chaperonin (CPN60a), elongation factor 1 alpha (EF-1a), or actin] to the cotyledons of seeds by a particle bombardment induced highly uniform knock-down phenotypes of each gene on the true leaves of seedlings from 2~3 weeks after inoculation. These silencing phenotypes continued for several months. Northern blot and RT-PCR analyses of leaves infected with ALSV containing a fragment of rbcS gene showed that the levels of rbcS-mRNA drastically decreased in the infected apple and pear leaves, and, in reverse, rbcS-siRNAs were generated in the infected leaves. In addition, some of apple seedlings inoculated with ALSV vector carrying a partial sequence of a TERMINAL FLOWER 1 gene of apple (MdTFL1) showed precocious flowering which is expected as a knock-down phenotype of the silencing of MdTFL1 gene.The ALSV-based VIGS system developed have provides a valuable new addition to the tool box for functional genomics in apple, pear, and Japanese pear.The infection of virus vector carrying sequences of plant genes triggers virus-induced gene silencing (VIGS) that results in the degradation of endogenous mRNA homologous to the plant genes through a homology-dependent RNA degradation mechanism [1,2]. Because VIGS offers an easy way to determine the functions of the genes in a short time, and it can also be applied to high throughput functional genomics in plants [1,3,4], the technology is an important tool for functional genomics in plants and used routinely for the analysis of gene function in many laboratories around the world. Though there are many reports on virus vectors for VIGS in plants, most are useful for the analysis of gene function in a limited range of dicot plants, e.g., Arabidopsis thaliana, Nicotiana benthamiana, N. tabacum, tomato, potato, legume species, cucurbits, and cassava etc, and
%U http://www.plantmethods.com/content/7/1/15