Application of protein TaNADH-GoGAT for controlling plant root development
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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
Active Publication Date: 2018-11-20
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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Therefore, although great progress has been made in the study of Arabidopsis root development in recent years, the relevant mechanisms of food crops such as wheat are not well understood.
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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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Abstract
The invention discloses application of a protein TaNADH-GoGAT for controlling plant root development. The invention provides application of the protein TaNADH-GoGAT or related biological materials thereof in control of plant root development. The related biological materials are nucleic acid molecules capable of expressing the protein TaNADH-GoGAT or an expression cassette, a recombinant vector, arecombinant bacterium or a transgenic cell line containing the nucleic acid molecules. Compared with a wild plant, the transgenic plant over-expressed by the TaNADH-GoGAT has the advantage that the dry weight of the root system and/or the lateral root length are/is obviously increased; the transgenic plant under-expressed by the TaNADH-GoGAT has the advantage that the dry weight of the root system and/or the lateral root length are/is obviously decreased. Therefore, the plant root development can be controlled by utilizing the protein TaNADH-GoGAT. The protein TaNADH-GoGAT disclosed by the invention has important application value for breeding novel high-yield plant materials.
Description
technical field [0001] The invention relates to the field of biotechnology, in particular to the application of the protein TaNADH-GoGAT to regulate the development of plant roots. Background technique [0002] The root is an important organ for plant growth and development, and its main functions are absorption, transportation, support, synthesis and storage. Plants absorb water, minerals and other nutrients from the soil through the root system, so the root system is extremely important to the growth and development of plants. There are significant differences between the root systems of monocotyledonous and dicotyledonous plants. Therefore, although great progress has been made in the study of Arabidopsis root development in recent years, the relevant mechanisms of food crops such as wheat are not well understood. [0003] Wheat is the main food crop, but in semi-arid dry farming areas, the lack of water resources has become the main factor limiting the high yield of wh...
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