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Gene for controlling content of soybean vitamin B1 and application of gene

A gene and soybean technology, applied in the field of plant molecular biology and genetic engineering, can solve the problems affecting the normal growth and development of plants, affecting the plant glycolysis pathway, activity and content, and achieve the regulation of chloroplast development and the improvement of soybean amino acid content , the effect of good application prospects

Active Publication Date: 2019-09-27
NORTHEAST INST OF GEOGRAPHY & AGRIECOLOGY C A S
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The activity and content of pyruvate, oxaloacetate and α-ketoglutarate dehydrogenase are affected, which will affect the plant's glycolysis pathway, tricarboxylic acid cycle, and synthesis of branched amino acids, thereby affecting the normal growth and development of plants

Method used

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  • Gene for controlling content of soybean vitamin B1 and application of gene
  • Gene for controlling content of soybean vitamin B1 and application of gene
  • Gene for controlling content of soybean vitamin B1 and application of gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Isolation and structural analysis of GmPGL1 gene in soybean

[0041] (1) Isolation of genes

[0042]The mRNA was extracted from young leaves of soybean variety Williams 82, and the first strand of cDNA was synthesized using the mRNA as a template and Oligo(T)17 as a primer. Then, using the first strand of the cDNA as a template, primers OL8172 (5'-TCCCCCCGGGATGGCTGCCATGGCAACCACA-3') and primers OL8173 (5'-CGCGGATCCTTAAGCATCAACAATTTCCTCAGTG-3') were used for PCR amplification to obtain a GmPGL1 gene with a length of 1056 bp The cDNA fragment was cloned into the pEASY-Blunt vector (TransGen, Beijing, China) and named pEASY-Blunt-GmPGL1.

[0043] The GmPGL1 CDS sequence contained in pEASY-Blunt-GmPGL1 is shown in SEQ ID NO: 1, with a total of 1056 bp, which encodes a protein of 351 amino acids (SEQ ID NO: 2).

[0044] (2) Gene structure analysis

[0045] Using the young leaves of Williams 82 as materials, extract the DNA from the young leaves, use the genomic ...

Embodiment 2

[0046] Example 2 Soybean GmPGL1 gene regulates the content of vitamin B1

[0047] In order to study the influence of vitamin B1 on the growth and development of plants, the present invention obtained a mutant with lighter leaf color through EMS mutagenesis, which was named Glycine max pale green leaf 1 (Gmpgl1), sequenced by BulkedSegregant Analysis (BSA) Methods A mutant of the target gene GmPGL1 was obtained, which was missing a base T in the 292nd position of the CDS region, which resulted in premature termination of the gene coding.

[0048] The content of vitamin B1 in wild type and Gmpgl1 mutant leaves and seeds was determined by HPLC (15 leaf repeats from 3 wild type Williams82, 30 seed repeats, 15 leaf repeats from Gmpgl1 mutant, 30 seed repetitions), the average vitamin B1 content in the wild type Williams82 and Gmpgl1 mutant leaves were 43.87μg / g, 37.04μg / g, respectively, in the wild type 15 leaves, 30 seed repetitions, the vitamin B1 content of each leaf There was ...

Embodiment 3

[0049] Example 3 Functional Verification of Soybean GmPGL1 Gene

[0050] (A) Soybean GmPGL1 gene regulates plant chloroplast development

[0051] Take the leaves of wild type Williams82 and Gmpg1 mutants at the same growth period and at the same growth position in the growth chamber (20 wild-type and mutant leaves were collected), and cut the leaves into 1cm×1cm long strips with a scalpel , put it into 4% glutaraldehyde, press the sample on the bottom of the fixative with cotton, put it in 4°C overnight; fix the sample with osmic acid, 2h, 4°C; rinse the sample 3 times with distilled water, 15min each time; %, 70%, 80%, and 95% ethanol for gradient dehydration of the sample, 10 min each time, and then mixed with 95% ethanol and 95% acetone 1:1 for 10 min. Dehydrate with 100% acetone twice, 20min each time. Finally, replace acetone with propylene oxide and dehydrate once for 20 minutes; mix 1:1 with propylene oxide and embedding agent (SPI-PON 812), and pre-soak the sample fo...

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Abstract

The invention provides a gene for controlling content of soybean vitamin B1 and an application of the gene and belongs to the technical field of gene engineering. The gene GmPGL1 for controlling the content of soybean vitamin B1 has a nucleotide sequence shown in SEQ ID No.1 or a sequence coded with the same functional protein by replacing, deleting or adding one or more nucleotides in the sequence. It is found that the GmPGL1 gene participates in regulation of the content of soybean vitamin B1, development of chloroplast, amino acid metabolism, glycolysis and pentose phosphoric acid ways. The GmPGL1 gene has good application prospects in plant breeding and germplasm resource improvement.

Description

technical field [0001] The invention belongs to the technical fields of plant molecular biology and genetic engineering, and specifically relates to a soybean GmPGL1 gene and the application of the gene in regulating soybean vitamin B1 content regulation, chloroplast development, amino acid metabolism, glycolysis and pentose phosphate pathways. Background technique [0002] Vitamin B1 is very important to the growth and development of humans, animals and plants. Plants can synthesize vitamin B1 by themselves, but humans and animals must obtain vitamin B1 from food. Vitamin B1 is widely found in fruits, vegetables, nuts, seafood, grains and beans. The lack of vitamin B1 in the human body will produce various diseases, such as: nerve inflammation, edema, beriberi and so on. At present, in developing and underdeveloped countries, vitamin B1 deficiency in the human body is very common. It is imminent to effectively improve the content of vitamin B1 in food to solve the symptom...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82C07K14/415A01H5/00A01H6/54
CPCC07K14/415C12N15/8251C12N15/8243
Inventor 杨素欣冯星星
Owner NORTHEAST INST OF GEOGRAPHY & AGRIECOLOGY C A S
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