Applications of TaNAC2 protein and encoding gene thereof
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A technology that encodes genes and proteins, and is used in applications, genetic engineering, plant genetic improvement, etc.
Active Publication Date: 2014-08-27
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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In summary, NAC proteins, as a class of important transcription factors, play an important role in regulating plant growth and development and various plant stress defense responses. There are few studies reported on
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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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Abstract
The invention discloses applications of a TaNAC2 protein and an encoding gene thereof. The TaNAC2 protein can be used for promoting the absorption of a plant to nitrogen. The invention further discloses a method for improving the absorption transshipment and / or assimilation efficiency of the nitrogen element in the plant. The method comprises the following step: feeding the encoding gene of the protein shown as SEQ ID No.4 into a primary plant so as to obtain a transgenic plant. Compared with the primary plant, the absorption transshipment and / or assimilation efficiency of the nitrogen element in the transgenic plant are improved. The method has the advantages that 1, the TaNAC2 gene taken as a regulatory factor for response of nitrate nitrogen is used for regulating and controlling a series of gene expressions on absorption and utilization ways of the nitrogen element in wheat, thereby greatly promoting the mechanism study of illuminating the metabolic utilization of the nitrogen element in the plant; 2, the expression quantity of the TaNAC2 is improved by adopting a genetic engineering technology, so that the assimilation efficiency of the nitrogen element is improved, thereby realizing the purposes of reduction in fertilizer application, yield increasing and the like.
Description
technical field [0001] The invention relates to a transcription regulation factor and the application of its coding gene; in particular, it relates to the application of TaNAC2 protein and its coding gene in regulating plant nitrogen uptake and utilization, and belongs to the field of biotechnology. Background technique [0002] Wheat is an important food crop in my country. my country is the largest wheat producer in the world, both in terms of cultivated area and total output. In the early days of the founding of new China, China's wheat yield was very low, only 0.73 tons / ha (48.8 kg / mu) in 1952, and reached 4.55 tons / ha (303.3 kg / mu) in 2006, an increase of 5.2 times, one of the important The reason is the increase in the use of chemical fertilizers. According to statistics, chemical fertilizers account for 32% of agricultural production, 25% of agricultural production costs, and 50% of all agricultural production material costs. At present, the annual nitrogen fertili...
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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82C07K14/415A01H5/00
CPCC07K14/415Y02A40/146C12N15/8261
Inventor 童依平何雪赵学强曲宝原马文英李文静洪霞史计李滨李振声
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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