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A carrier for increasing plant methionine content and its construction method and application

A technology of methionine and carrier, applied in the field of plant molecular breeding, can solve the problems such as failure of corn methionine level to achieve the effect of reducing cost, high protein quality and reducing demand

Active Publication Date: 2019-06-14
SAAS BIOTECH & NUCLEAR TECH RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional Breeding Fails to Increase Methionine Levels in Maize

Method used

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  • A carrier for increasing plant methionine content and its construction method and application
  • A carrier for increasing plant methionine content and its construction method and application
  • A carrier for increasing plant methionine content and its construction method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Embodiment 1 Objective gene cloning and the construction of vector pTF102 / RbcS1P:AtSAT1

[0074] 1. Construction of vector pMD19-T: RbcS1

[0075] The promoter used is Bundle sheath-specific Rubisco small subunit 1 (RBCS1gene) referred to as RbcS1 (GenBank accession number: AH005359.3, see http: / / www.ncbi.nlm.nih.gov / nuccore / 339635306 for sequence), the promoter comes from Maize, 1030bp long, (Sattarzadeh A, Fuller J, Moguel S, et al. Transgenic maize lines with cell-type specific expression of fluorescent proteins in plastids[J]. Plant biotechnology journal, 2010,8(2):112-125.) . There is a signal peptide of about 20bp behind the promoter, which enables the expressed protein to target the bundle sheath cells, realizes tissue cell-specific expression, and avoids the toxicity of exogenous proteins accumulated in non-functional tissues to the plant. Use primers (primer nucleotide sequences are shown in the nucleotide sequences of SEQ ID NO.3 and SEQ ID NO.4 in the seque...

Embodiment 2

[0105] Preparation of embodiment two transgenic strains

[0106] 1. Obtaining of transgenic strains

[0107] The expression vector pTF102 / RbcS1P:AtSAT1 prepared in Example 1 was introduced into commonly used Agrobacterium tumefaciens strains; the immature embryos (1.5-2.0 mm) of the corn HiII hybrids about 13 days after pollination were taken, and the young embryos were infected with pre-activated Agrobacterium. Embryo. Aspirate the bacterial solution and transfer the immature embryos to the co-culture medium. After three days of co-cultivation at 20°C, transfer to recovery medium. After 7 days of dark culture at 28 degrees, the immature embryos with growth ability were transferred to the first selection medium, and cultured at 28 degrees dark for 2 weeks. Transfer to the second selection medium again, and culture in the dark for 2 weeks. The well-grown callus was transferred to the first differentiation medium for differentiation, and differentiated in the dark at 28 degr...

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Abstract

The invention provides an expression carrier for improving methionine and / or cysteine content in corns and like plant. The carrier uses the overexpression of Arabidopsis serine acetyl transferase gene mediated by Rbcs promoter in the crops to finish the vascular bundle sheath cell specific expression through the serine acetyl transferase, thereby improving the content of total methionine and cysteine in the corn grains, and improving the content of 10kDa-zein enriched in methionine and glutathione content in the plant. The invention further provides a construction method and application of the carrier, and a method for cultivating the transgenic corn with high methionine and cysteine content using the carrier.

Description

[0001] Technical field: The present invention belongs to the field of plant molecular breeding, and specifically provides an expression vector, its use and construction method, and the vector can increase the content of methionine and glutamine in plants. Background technique: [0002] Protein shortage, including the shortage of protein content and low nutritional quality, is a serious global problem in the 21st century. It is of great significance to increase the protein content of crops as soon as possible, and actively improve the amino acid composition of protein and improve nutritional value. [0003] Sulfur is the fourth most essential nutrient element for plants after nitrogen, phosphorus, and potassium. When sulfur is deficient, protein synthesis is blocked, resulting in the accumulation of non-protein nitrogen, which not only affects crop growth, but also reduces agricultural products; quality; plant sulfur content is 0.1 %-0.5%, and its variation is obviously affected...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82C12N15/66C12N15/84A01H5/00A01H4/00A01H6/46
CPCA01H4/008C12N15/66C12N15/8205C12N15/8253
Inventor 向小利巫永睿潘光堂路史迪克·托马斯梅辛·约阿希姆
Owner SAAS BIOTECH & NUCLEAR TECH RES INST
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