Vegetable oil and fat metabolism related regulatory protein GmMYB73 as well as coding gene and application thereof
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A technology for vegetable oils and related proteins, which is applied in the fields of plant genetic improvement, botanical equipment and methods, and applications, and can solve problems such as poorly understood lipid synthesis.
Active Publication Date: 2011-07-20
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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However, the regulatory mechanism of lipid synthesis and its associated genes are still poorly understood
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Embodiment 1
[0066] Example 1. Screening and cDNA Cloning of Transcription Factor GmMYB73 Encoding Gene Related to Regulation of Soybean and Lipid Metabolism
[0067] 1. Fatty acid accumulation and metabolism analysis during soybean seed development
[0068] Two soybean varieties or germplasms with different fatty acid contents: Heinong 44 (HN44) and ZYD7. Among them, the oil content of HN44 grains was 23%, while that of ZYD7 grains was 12%. Such as figure 1 As shown, the seven different stages of the seed development process of the two varieties were collected and preserved separately.
[0069] The oil content of HN44 and ZYD7 seeds in each of the seven developmental stages was measured by gas chromatography, and the oil accumulation curve during seed development was analyzed ( figure 2 ). Such as figure 2 The two inflection points of the oil content growth curve indicated by the arrows were taken respectively: HN44-5 and HN44-6 in HN44; ZYD7-4 and ZYD7-5 in ZYD7. The total RNA of t...
Embodiment 2
[0077] Embodiment 2, the acquisition of GmMYB73 transgenic Arabidopsis
[0078] 1. Construction of plant expression vector pPROKII-GmMYB73
[0079] 1. Primer design
[0080] Primers were designed according to the cDNA sequence (DQ822927) of GmMYB73, and the primer sequences were as follows:
[0081] GmMYB73 up (upstream primer with BamHI restriction site):
[0082] 5'-ggatccAATGGCTGACATAGATC-3';
[0083] GmMYB73dp (downstream primer with KpnI restriction site):
[0084] 5'-ggtACCGATCATTGGCTAGTCG-3'.
[0085] 2. Construction of plant expression vector pPROKII-GmMYB73
[0086] ①Develop Heinong 44 to figure 1 The grains of the 6th stage shown (that is, the 37th day after flowering) were ground in liquid nitrogen, suspended in 4mol / L guanidine thiohydrogen, extracted with acidic phenol and chloroform, and absolute ethanol was added to the supernatant to precipitate the total RNA. cDNA was synthesized from RNA using reverse transcriptase.
[0087] ② Using cDNA as a templat...
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Abstract
The invention discloses a vegetable oil and fat metabolism related regulatory protein GmMYB73 (transcription factor) as well as a coding gene and application thereof. The protein provided by the invention is protein (a) or (b): the protein (a) is formed by amino acid sequences shown in a sequence 2 in a sequence table; and the protein (b) is related to vegetable oil and fat metabolism, derived from (a) and formed by enabling the amino acid sequences in the sequence 2 to be replaced and/or deleted and/or added through one or more amino acid residues. The invention also provides the coding geneof the protein and a recombinant expression vector containing the coding gene. The activity of the coding gene is related to the oil content of soybeans and the oil and fat accumulation velocity, andthe coding gene has a function of increasing the oil and fat content. The oil and fat content in a genetically modified seed is remarkably higher than that of a non-generically modified plant, and for example, in genetically modified arabidopsis, the oil and fat content in the seed is increased by about 14 percent through comparison. The coding gene has important theoretic and realistic significance on improving the oil and fat ingredients in oil plant kernels, such as soybean, and the like and cultivating varieties with high oil and fat content.
Description
technical field [0001] The invention relates to a regulatory protein GmMYB73 related to plant oil metabolism and its coding gene and application. 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, palm oil and rapeseed oil ranking second and third respectively (the world's main Oil-producing crops are shown in Table 1). [0003] Table 1 Major oil-producing crops in the world [0004] type Production volume (million tons) % of total production relative order Soybean 15.50 29.1 1 Palm 8.52 16.0 2 Rapeseed 7.03 13.2 3 Sunflower 7.00 13.1 4 Cottonseed 3.31 6.2 5 Coconut 2.71 5.1 6 Peanuts...
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