Application of protein TaNADH-GoGAT for controlling plant root development
A plant root system, 1. tanadh-gogat technology, applied in the biological field, can solve the problems of not knowing much about the mechanism of food crops
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Embodiment 1
[0055] Embodiment 1, the construction of transgenic TaNADH-GoGAT gene wheat
[0056] 1. Preparation of transgenic TaNADH-GoGAT plants
[0057] (1) Acquisition of TaNADH-GoGAT gene
[0058] 1. The total RNA of wheat variety KN199 was extracted, and its genomic cDNA was obtained by reverse transcription.
[0059] 2. Use the cDNA obtained in step 1 as a template, and use the following primers as primers to perform PCR amplification to construct the sequence required for overexpressing the TaNADH-GoGAT transgenic line wheat:
[0060] TaNADH-GoGAT-OE-F: 5'- AAGCTT ATGCCGACGGCGCAGGGGATTGG-3' (the underlined sequence is the recognition site for Hind III digestion);
[0061] TaNADH-GoGAT-OE-R: 5'- GAATTC CTATGTAAGGCCTGCGACATTTTG-3' (the underlined sequence is the recognition site for EcoR I digestion).
[0062] Use the following primers as primers to perform PCR amplification to construct the required sequence for reducing the expression of TaNADH-GoGAT transgenic line wheat: ...
Embodiment 2
[0105] Embodiment 2, root phenotype identification of TaNADH-GoGAT transgenic wheat
[0106] Tested wheat: three T2 generation TaNADH-GoGAT overexpressed wheat GoOE1, GoOE17 and GoOE18, three T2 generation TaNADH-GoGAT downexpressed wheat GoR1, GoR2 and GoR3, wild-type wheat KN199, and empty control plants (NC ).
[0107] Root phenotype identification was carried out under hydroponic conditions, and the specific steps were as follows:
[0108] The seeds of each tested wheat were treated overnight with 1% hydrogen peroxide to break dormancy, and then transplanted to seedling trays and cultivated for 7 days. When they grew to two leaves and one heart, the wheat seedlings with the same growth were selected and transplanted to high-nitrogen nutrient solution. (2mM N) and low-nitrogen nutrient solution (0.2mM N), and the formula of the culture solution is shown in Table 1 below. After 14 days of cultivation, the aboveground part and the root system were separated, and the dry wei...
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