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.
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Acknowledgments
We are grateful to Dr. Soh Hidaka, Dr. Seiji Kanematsu, and Dr. Makoto Tougou (National Agricultural Research Center Tohoku region) for providing the Helios Gene Gun System and supplying Jack, Enrei, Suzukari, and Hatayutaka seeds; and National Agricultural Research Center Tohoku region for supplying the Dewamusume seeds. We gratefully acknowledge Associate Professor Chigen Tsukamoto, Iwate University, for detailed advice in HPLC analysis. We thank to Miss Aki Igarashi, Miss Yukari Takahashi, and Mr Daichi Yamane for their technical support. This work was supported by Grant-in-Aids for KAKENHI (no. 20380025).
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Yamagishi, N., Yoshikawa, N. Virus-induced gene silencing in soybean seeds and the emergence stage of soybean plants with Apple latent spherical virus vectors. Plant Mol Biol 71, 15–24 (2009). https://doi.org/10.1007/s11103-009-9505-y
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DOI: https://doi.org/10.1007/s11103-009-9505-y