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Nobody likes a bearer of bad news, a gloomy Gus or a glass-half-empty attitude.
Various other reporter constructs with promoters of other terpene synthases- SlTPS3, SlTPS7 and SlTPS8- driving expression of GUS or a GUSsYFP1 fusion (SlTPS9) were included in the analyses.
A number of transgenic lines were created in which the DIR1 promoter region was placed upstream of GUS or a DIR1-GUS fusion in Ws and dir1-1, respectively.
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GUS staining of transgenic rice plants harboring an OsNCED3::GUS or OsNCED4::GUS construct was carried out to determine the expression patterns of these two genes in developing seeds and seedlings under salt stress.
GUS staining revealed that PtoWUSa and PtoPWOX4a were both strongly expressed in the CAM zone of stems; they were also weakly expressed in roots as GUS staining of roots of transgenic lines expressing either P PtoWUSa ::GUS or P PtoWOX4a ::GUS occurred after a prolonged period.
However, analysis of transformants containing either β-glucuronidase (GUS) or luciferase (LUC) genes, with and without an intron revealed no detectable marker gene expression.
a PCR analysis of OsNCED3::GUS or OsNCED4::GUS constructs in N3P or N4P transgenic rice plants, respectively.
Cells infected with either A. tumefaciens containing pCaMV35S: GUS or A. tumefaciens only was used as positive and negative control, respectively.
The presence of OsNCED3::GUS or OsNCED4::GUS constructs in the transgenic rice plants were verified by PCR.
The binary vector was then used to introduce the OsNCED3::GUS or OsNCED4::GUS constructs into Japonica rice variety TNG67.
Rice transformation involved introducing p OsADF1i ::GUS or p OsADF3i ::GUS by electroporation with Agrobacterium tumefaciens strain EHA101.
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