Method for increasing nitrogen-use efficiency in plants
A plant-efficient technology, applied in botany equipment and methods, biochemical equipment and methods, plant peptides, etc., can solve problems such as agricultural losses
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[0215] 1.1. Materials and methods
[0216] Vector construction and rice transformation
[0217] We use Figure 10 The primers listed in amplified the OsNRT2.1 / OsNRT2.2 open reading frame (ORF) sequence from cDNA isolated from japonica rice Nipponbare (Oryza sativa L. ssp. Japonicacv. Nipponbare), in which the two genes were identical. We use Figure 11 The primers listed in amplify the OsNAR2.1 and ubiquitin promoters from the pOsNAR2.1(1698bp):GUS (Feng et al., 2011) and pUbi:OsPIN2 (Chen et al., 2012) constructs, respectively. The PCR product was cloned into pMD19-T vector (TaKaRa) and confirmed by restriction enzyme digestion and DNA sequencing. Such as Figure 15 The pUbi:OsNRT2.1 and pOsNAR2.1:OsNRT2.1 vectors were constructed as indicated. These constructs were introduced into Agrobacterium tumefaciens strain EHA105 by electroporation, which was then transformed into rice as previously described (Tang et al., 2012).
[0218] 1.1.1. Southern blot analysis
[0219] ...
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