Applications of TaNAC2 protein and encoding gene thereof
A technology that encodes genes and proteins, and is used in applications, genetic engineering, plant genetic improvement, etc.
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Embodiment 1
[0066] Embodiment 1, the construction of transgenic TaNAC2 wheat
[0067] 1. Preparation of transgenic TaNAC2 plants
[0068] (1) Acquisition of TaNAC2 gene
[0069] 1. The total RNA of Chinese spring wheat was extracted, and its genomic cDNA was obtained by reverse transcription.
[0070] 2. Use the cDNA obtained in step 1 as a template, and perform PCR amplification with the following primers as primers.
[0071] Upstream primer: 5'- GGATCC ATGGGGATGCCGGCCGTG-3' (SEQ ID No. 1)
[0072] (The underlined sequence is the recognition site for BamHI digestion)
[0073] Downstream primer: 5'- GGTACC GAACGGGGCCGGCATGC-3' (SEQ ID No. 2)
[0074] (The underlined sequence is the recognition site of KpnI enzyme digestion)
[0075] PCR system (40μl): template cDNA 4μl, KOD plus DNA polymerase 1μl, 10×PCR buffer for KOD plus 4μl, dNTPs (2mM each) 4μl, 25mM MgSO 4 2μl, 20mM each of the upstream and downstream primers, and then make up the reaction system to 40μl with double disti...
Embodiment 2
[0115] Example 2, Phenotype detection of transgenic TaNAC2 wheat
[0116] 1. Root phenotype identification is carried out under hydroponic conditions, and the specific steps are as follows:
[0117] The seeds of T3 transgenic TaNAC2-transferred wheat (OE1 and OE2) and wild-type wheat Longchun 23 were germinated in a 23°C incubator (tap water culture) for 7 days, and then transferred to high-nitrogen nutrient solution or low-nitrogen nutrient solution after removing the embryo. Cultured in nutrient solution.
[0118] Two, after the plant of step 1 is cultivated for 14 days, measure the main root length (longest root length) with a ruler, and analyze the number of lateral roots (number of lateral root branches) with the WinRHIZO root system scanning system.
[0119] The result is as Figure 4 shown. Figure 4 Among them, LC represents the wild type wheat Longchun 23, LN represents the result of culture in low-nitrogen nutrient solution, and HN represents the result of culture...
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