Plant lateral branch number correlated transcription factor AtDOF 4.2, its encoding gene and application
A plant and gene technology, applied in the direction of plant gene improvement, application, plant peptides, etc., can solve problems such as loss and reduction of side branch development
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Embodiment 1
[0068] Embodiment 1, the cloning of Arabidopsis AtDOF4.2 gene
[0069] 1. Cloning of AtDOF4.2 gene
[0070]To clone the AtDOF4.2 gene, Arabidopsis thaliana cv Columbia-0, Col-0, hereinafter referred to as wild-type Arabidopsis thaliana, was purchased from Arabidopsis Center Arabidopsis Biological Resource Center) total RNA and mRNA were purified using a Promega kit (purchased from Promega) PolyAT tract mRNA isolation system IV to separate, and the isolated mRNA was quantified with an ultraviolet spectrophotometer. The cDNA obtained by reverse transcription was used as a template for PCR amplification, and the primer sequences were 5'-ATGAATAATTTGAATGTTTT-3' (sequence 3) and 5'-TTATGATTCATATTCAAAT-3' (sequence 4).
[0071] The PCR product was detected by 0.8% agarose gel electrophoresis, and the result showed that the size of the PCR amplification product was about 0.5 kb, which was consistent with the expected result. This fragment was recovered using an agarose gel recovery...
Embodiment 2
[0085] Example 2, AtDOF4.2 transgenic Arabidopsis and its functional research
[0086] 1. Identification of T-DNA mutants of AtDOF4.2 gene
[0087] T-DNA insertion mutant dof4.2 (CS813276) was purchased from ABRC (Arabidopsis Biological Resource Center).
[0088] Seeds of wild-type Arabidopsis and dof4.2 mutants were grown in petri dishes for 2 weeks and then transplanted (grown at 22°C with a daily light of 16h / 8h (light / dark)) into pots. After flowering, the whole plants were taken Extract RNA for RT-PCR identification.
[0089] Primer is AtDof4.2 5'-ATGAATAATTTGAATGTTTT-3' (SEQ ID NO: 3)
[0090] AtDof4.2 5'-TTATGATTCATATTCAAAT-3' (SEQ ID NO: 4)
[0091] Actin was used as an internal reference, and the primers were
[0092] Actin F 5'-GAAGATTAAGGTCGTTGCACCACCTG3'
[0093] Actin R 5'-ATTAACATTGCAAAAGAGTTTCAAGGT3'
[0094] The result is as figure 2 As shown, Col-0 is wild-type Arabidopsis, and dof4.2 is a mutant. It can be seen from the figure that A...
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