Virus-induced gene silencing in soybean seeds and the emergence stage of soybean plants with Apple latent spherical virus vectors

Abstract

Virus-induced gene silencing (VIGS) has great potential as a reverse-genetics tool in plant genomics. In this study, we examined the potential of VIGS in soybean seeds and the emergence stage of soybean plants using Apple latent spherical virus (ALSV) vectors. Inoculation of an ALSV vector (soyPDS-ALSV) carrying a fragment of the soybean phytoene desaturase (soyPDS) gene into soybean seedlings resulted in a highly uniform photo-bleached phenotype, typical of PDS inhibition, on the upper leaves throughout plant growth. The photo-bleached phenotype was also found on all immature pods, all seed coats, and about 50% embryos of seeds on soybean plants infected with soyPDS-ALSV. Infection with an ALSV vector (soyIFS2-ALSV) having a fragment of soybean isoflavone synthase 2 (soyIFS2) gene also led to a reduction of the levels of both soyIFS2- and soyIFS1- mRNAs and an isoflavone content in the cotyledons of about 36% mature seeds of infected soybean plants. Furthermore, VIGS of soyPDS was induced in the next generation plants by the seed transmission of soyPDS-ALSV. Thus ALSV vectors will be useful for studying gene functions in the reproductive stages and early growth stages, such as emergence and cotyledon stages, in addition to the vegetative stages of soybean plants.

Application of somatic embryos to rapid and reliable analysis of soybean seed components by RNA interference-mediated gene silencing

“…Bean pod mottle virus-based vectors are an efficient and well-established tool for analysis of gene function in soybean, but such vectors were found not to induce gene silencing in seeds (Kachroo et al 2008;Zhang and Ghabrial 2006). In contrast, cucumber mosaic virus-and apple latent spherical virus-based vectors induced gene silencing in both the vegetative and reproductive stages in soybean and were shown to affect metabolite concentrations in soybean seeds (Liu et al 2010;Nagamatsu et al 2007;Yamagishi and Yoshikawa 2009). However, the efficacy of induced gene silencing and the extent of induced symptoms varied with the specific combinations of each of these three types of viral vector and soybean cultivars.…”

Virus-induced Gene Silencing in Apricot (Prunus armeniaca L.) and Japanese Apricot (P. mume Siebold ^|^amp; Zucc.) with the Apple Latent Spherical Virus Vector System

“…), pear (Pyrus communis L.), and Japanese pear (P. pyrifolia Nakai) (Igarashi et al, 2009;Sasaki et al, 2011;Yamagishi and Yoshikawa, 2009). ALSV, which is classified into the genus Cheravirus, consists of isometric virus particles ca.…”

Functional genomics to study stress responses in crop legumes: progress and prospects

“…In soybean, a bean pod mottle virus (BPMV)-based system was used to identify genes participating in basal, resistance gene-mediated, and systemic immunity (Kachroo and Ghabrial 2012). In another study, the apple latent spherical virus (ALSV) vector was used to study gene function in the reproductive and early growth stages (emergence and cotyledon) in addition to the vegetative stages (Yamagishi and Yoshikawa 2009). The pea early-browning virus (PEBV) has been developed as a VIGS vector and used for functional analysis of several genes involved in the symbiosis.…”