Application of soybean transcription factor GmZF351 in vegetable oil and fat metabolism regulation
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A technology of plant tissue and oil content, applied in the biological field, can solve problems such as little known
Active Publication Date: 2014-10-29
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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However, in plants, the regulatory mechanism of lipid sy
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Embodiment 1
[0051] Example 1, cDNA cloning of the transcription factor GmZF351 encoding gene related to regulation of soybean and lipid metabolism and construction of plant expression vector
[0052] 1. Acquisition of transcription factor GmZF351
[0053] Soybean material: Soybean Heinong 44;
[0054] Total RNA was extracted from Heinong 44 seedlings, and the RNA was reverse-transcribed with reverse transcriptase to synthesize cDNA.
[0055] According to the information of GmZF351 full-length cDNA sequence in the soybean genome sequence of PlantGDB, design primers, and the primer sequences are as follows:
[0056] GmZF351-up: 5'-ATGAGTAGTGTTTTTTTCAG (SEQ ID NO: 3);
[0057] GmZF351-dp: 5'-CTACATCAGCAATTCATT (SEQ ID NO: 4).
[0058] Using the cDNA of HN44 as a template and using GmZF351-up and GmZF351-dp as primers, PCR amplification was performed to obtain a PCR product of about 1Kb. After sequencing, the PCR product is 1056bp, which has the nucleotide shown in sequence 1 in the seque...
Embodiment 2
[0069] Embodiment 2, the application of GmZF351 in the regulation and control of plant tissue total oil or fatty acid content
[0070] 1. Obtaining recombinant Agrobacterium
[0071] The recombinant vector pGWB412-GmZF351 obtained in 2 of Example 1 was introduced into Agrobacterium GV3101 by electric shock method to obtain recombinant bacteria.
[0072] The plasmid of the recombinant bacterium was extracted, and after sequencing, the plasmid was pGWB412-GmZF351, and the recombinant bacterium containing the plasmid was named GV3101 / GmZF351, namely the recombinant Agrobacterium.
[0073] 2. Acquisition and identification of transgenic GmZF351 Arabidopsis
[0074] The recombinant Agrobacterium GV3101 / GmZF351 was cultivated to the logarithmic phase, and then transformed into flowers of Arabidopsis thaliana (col-0) (seeds were purchased from Arabidopsis Biological Resource Center (ABRC)) by vacuum method. Harvest the seeds, sow the seeds on the MS screening medium containing kana...
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Abstract
The invention discloses oil and fat metabolism regulation related soybean transcription factor GmZF351 and a coding gene and application thereof. The protein is named as GmZF351, is derived from papilionaceae glycine soybean (Glycine max (L.) Merrill), is a protein as follows (a) or (b): (a) protein is composed of an amino acid sequence shown as the sequence 2 in the sequence table; and (b) protein is a protein obtained by substitution and / or deletion and / or addition of one or a plurality of amino acid residues of the amino acid sequence shown as the sequence 2 and is in relevant to plant tissue total oil and fat content and derived from the sequence 2. Experiments show that, the transcription factor GmZF351 and the coding gene can regulate the oil and fat content in plant seeds, and after over expression, can improve the oil and fat content in the plant seeds. The coding gene has important theoretical and practical significances in the aspects of increasing and improving crop oil and fat ingredients, especially improving oil and fat ingredients in soybean and other oil plant seeds, and cultivating high oil and fat containing varieties.
Description
technical field [0001] The invention relates to the field of biotechnology, in particular to the application of a soybean transcription factor GmZF351 in the regulation and control of vegetable oil metabolism. Background technique [0002] 71% of the fat in the human diet comes from plants. Among several major oil-producing crops in the world, the total oil production of soybean accounts for about 30%, ranking first in the world's vegetable oil production (Table 1). [0003] Table 1 shows the main oil-producing crops in the world [0004] [0005] The synthesis of fatty acids is one of the most important metabolic pathways in plants. It exists in any cell of plants and is necessary for growth and development. Blocking it leads to cell death, so a plant mutant that blocks fatty acid synthesis has not been found so far. [0006] Plants differ greatly from other eukaryotes in the enzymes involved in the fatty acid synthesis pathway. Synthesis of fatty acids with 16 or 18...
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