Article

Soybean dwarf virus-resistant transgenic soybeans with the sense coat protein gene.

National Agricultural Research Center for Tohoku Region, Morioka, Iwate 020-0198, Japan.
Plant Cell Reports (Impact Factor: 2.94). 12/2007; 26(11):1967-75. DOI: 10.1007/s00299-007-0404-x
Source: PubMed

ABSTRACT We transformed a construct containing the sense coat protein (CP) gene of Soybean dwarf virus (SbDV) into soybean somatic embryos via microprojectile bombardment to acquire SbDV-resistant soybean plants. Six independent T(0) plants were obtained. One of these transgenic lines was subjected to further extensive analysis. Three different insertion patterns of Southern blot hybridization analysis in T(1) plants suggested that these insertions introduced in T(0) plants were segregated from each other or co-inherited in T(1) progenies. These insertions were classified into two types, which overexpressed SbDV-CP mRNA and accumulated SbDV-CP-specific short interfering RNA (siRNA), or repressed accumulation of SbDV-CP mRNA and siRNA by RNA analysis prior to SbDV inoculation. After inoculation of SbDV by the aphids, most T(2) plants of this transgenic line remained symptomless, contained little SbDV-specific RNA by RNA dot-blot hybridization analysis and exhibited SbDV-CP-specific siRNA. We discuss here the possible mechanisms of the achieved resistance, including the RNA silencing.

0 Bookmarks
 · 
63 Views
  • [Show abstract] [Hide abstract]
    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.
    Plant Molecular Biology 07/2009; 71(1-2):15-24. · 3.52 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Faba bean is an excellent candidate crop to provide nitrogen input into temperate agricultural systems. However, its growth is hampered by several factors including environmental stresses and the presence of anti-nutritional factors. To solve these limitations, breeding programs have been initiated that were successful for monogenic traits but not so for multigenic traits. The large genome size of faba bean has slowed down breeding processes. Several other legumes have emerged as model legumes including Medicago truncatula, Lotus japonicus, Glycine max and Pisum sativum. The establishment of these models has already boosted our understanding of important processes such as the nitrogen-fixing symbiotic interaction. The high level of synteny and collinearity existing between legumes makes possible the transfer of key knowledge from model legumes to faba bean. Here we review the most recent knowledge gained from model legumes on grain quality, resistance to biotic and abiotic stresses, nitrogen-fixing symbiosis and how this knowledge can be employed for faba bean breeding.
    Field Crops Research 02/2010; 115(3):253-269. · 2.47 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Transgenic plants expressing double-stranded RNA (dsRNA) of virus origin have been previously shown to confer resistance to virus infections through the highly conserved RNA-targeting process termed RNA silencing or RNA interference (RNAi). In this study we applied this strategy to soybean plants and achieved robust resistance to multiple viruses with a single dsRNA-expressing transgene. Unlike previous reports that relied on the expression of one long inverted repeat (IR) combining sequences of several viruses, our improved strategy utilized a transgene designed to express several shorter IRs. Each of these short IRs contains highly conserved sequences of one virus, forming dsRNA of less than 150 bp. These short dsRNA stems were interspersed with single-stranded sequences to prevent homologous recombination during the transgene assembly process. Three such short IRs with sequences of unrelated soybean-infecting viruses (Alfalfa mosaic virus, Bean pod mottle virus, and Soybean mosaic virus) were assembled into a single transgene under control of the 35S promoter and terminator of Cauliflower mosaic virus. Three independent transgenic lines were obtained and all of them exhibited strong systemic resistance to the simultaneous infection of the three viruses. These results demonstrate the effectiveness of this very straight forward strategy for engineering RNAi-based virus resistance in a major crop plant. More importantly, our strategy of construct assembly makes it easy to incorporate additional short IRs in the transgene, thus expanding the spectrum of virus resistance. Finally, this strategy could be easily adapted to control virus problems of other crop plants.
    Phytopathology 11/2011; 101(11):1264-9. · 2.97 Impact Factor