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A Simple Method for Isolation of Soybean Protoplasts and Application to Transient Gene Expression Analyses
Published on: January 25, 2018
Development of resistant transgenic soybeans with inverted repeat-coat protein genes of soybean dwarf virus
Makoto Tougou1, Noriyuki Furutani, Noriko Yamagishi
1National Agricultural Research Center for Tohoku Region, Morioka, Iwate, 020-0198, Japan.
Abstract:
In an attempt to generate soybean plants resistant to soybean dwarf virus (SbDV), we transformed a construct containing inverted repeat-SbDV coat protein (CP) genes spaced by beta-glucuronidase (GUS) sequences into soybean somatic embryos via microprojectile bombardment. Three T(0) plants with an introduced CP gene were obtained, and one generated T(1) seeds. The presence of the transgene in T(1) plants was confirmed by PCR and Southern blot hybridization analysis, but expression of CP was not detected by northern blot hybridization analysis. Two months after inoculation of SbDV by aphid, T(2) plants contained little SbDV-specific RNA and remained symptomless. These plants contained SbDV-CP-specific siRNA. These results suggest that the T(2) plants achieved resistance to SbDV by an RNA-silencing-mediated process.
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