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A plant expression vector and its application in improving cotton yield

A plant expression vector, cotton technology, applied in the field of genetic engineering, can solve problems such as simultaneous improvement of cotton, achieve the effect of achieving yield, and promoting targeted transportation and distribution

Inactive Publication Date: 2019-07-12
SOUTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is a genetic negative correlation between the yield and quality of cotton. Relying on traditional breeding methods, it is impossible to greatly increase the yield and quality of cotton in a short period of time. Therefore, the use of transgenic technology can break the genetic negative correlation of cotton. Enhance the disease resistance, herbicide resistance, and insect resistance of cotton and simultaneously improve the yield and quality of cotton

Method used

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  • A plant expression vector and its application in improving cotton yield
  • A plant expression vector and its application in improving cotton yield
  • A plant expression vector and its application in improving cotton yield

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] [Example 1] Cloning of Ustilago maize sucrose transporter (UmSRT1) gene and construction of its expression vector

[0048] 1. Acquisition of a specific promoter

[0049] According to the Arabidopsis BANYULS gene (GenBank accession number: AF092912) and its genome sequence, BAN promoter primers (SEQ ID No.1, SEQ ID No.2) were designed. The uppercase letters are the added restriction sites, and a fragment of about 300bp was obtained by PCR amplification from the Arabidopsis genome. After cloning the amplified DNA fragment, sequencing analysis showed that it was the BAN-specific promoter of Arabidopsis thaliana, and its nucleotide sequence was shown in SEQ ID No.3.

[0050] 2. Acquisition of Ustilago maize sucrose transporter (UmSRT1) gene

[0051] Primers were designed according to the Ustilago maydis sucrose transporter (UmSRT1) gene sequence (GenBank accession number: XM_011390372.1), and the bacterium solution of Ustilago maydis (purchased from China Microorganism Re...

Embodiment 2

[0054] [Example 2] Preparation of transformants and transgenic plants

[0055] 1. Introduce the constructed plant expression vector plasmid into Agrobacterium LBA4404 by electric shock method.

[0056] Referring to the Bio-RAD MicroPulser user manual, the above-mentioned vectors were introduced into Agrobacterium LBA4404 by electric shock transformation.

[0057] 2. The gene vector specifically expressing Ustilago maize sucrose transporter (UmSRT1) was integrated into the genome of upland cotton Jimian 14 (given to Professor Ma Zhiying of Hebei Agricultural University) through the method mediated by Agrobacterium tumefaciens.

[0058] Medium for genetic transformation of cotton mediated by Agrobacterium tumefaciens

[0059]

[0060]

[0061] MS: Murashige & Skoog, 1962

[0062] B5: Gamborg, 1986

[0063] Gelrite: Sigma, Item No.: G1910

[0064] SH: Schenk & Hildebrandt, 1972

[0065] The above-mentioned expression vectors were mediated by Agrobacterium and used cott...

Embodiment 3

[0069] [Example 3] Using real-time PCR to detect the expression of the imported UmSRT1 gene in cotton ovules

[0070] Refer to the instructions of the EASYspin Plant RNA Rapid Extraction Kit (Beijing Aidelai Biotechnology Co., Ltd.) to extract and rapidly extract the total RNA of ovules on the day of cotton flowering. After extraction, take 1 μL RNA agar gel electrophoresis, estimate the concentration and store it in a -80°C refrigerator.

[0071] use The RT reagent Kit with gDNA Eraser Kit (TaKaRa) was used to synthesize the first strand of cDNA in strict accordance with the reagent instructions, and the RNA concentration was estimated by agarose gel electrophoresis, and finally the reaction product was stored in a -20°C refrigerator.

[0072] Quantification of cDNA template: add 80 μL deionized water to cDNA, dilute it to 100 μL, perform quantitative PCR amplification on a real-time quantitative PCR instrument (Bio-Rad, CFX96), check the expression of cotton GhHIS (interna...

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Abstract

The invention relates to the field of genetic engineering and in particular relates to an ustilago maydis sucrose transportation protein UmSRT1 gene, an expression vector and an application of the expression vector in increase of cotton yield. According to the invention, a specific promoter and the ustilago maydis sucrose transportation protein UmSRT1 gene are fused, a plant expression vector specifically expressing a sucrose transportation protein gene is constructed, the plant expression vector is transformed into a host, so that a transformant is obtained, then the host is used for transforming a plant, and obtained transgenic cotton specifically expressing the ustilago maydis sucrose transportation protein UmSRT1 gene can be improved in yields of unginned cotton and ginned cotton of cotton.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a plant expression vector for specifically expressing the sucrose transporter UmSRT1 gene of Ustilago maize and a method for improving cotton yield through genetic engineering technology. Background technique [0002] Plants fix CO through photosynthesis 2 The synthetic carbohydrates are partitioned as assimilates mainly in the form of sucrose. Sucrose not only provides a carbon source for plant growth and development, but also provides transport signals for other components in plants, such as phytohormones (Turgeon R et al, 2008). Sucrose can be transported passively between cells through plasmodesmata, and can also be released into the apoplast for active transport through proteins dependent on ATP and protons (Norbert Sauer, 2007). Sucrose is loaded on the sieve tube molecules of the "source" organ, transported through the phloem for a long distance, and then unloaded to t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82C12N15/31A01H5/00A01H6/60
CPCC07K14/37C12N15/8205C12N15/8261
Inventor 裴炎丁晓艳钱山山侯磊李先碧
Owner SOUTHWEST UNIV
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