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Salt-induced functional element of high-efficiency soybean promoter and application of salt-induced functional element

A functional element, high-efficiency start-up technology, applied in applications, angiosperms/flowering plants, introduction of foreign genetic material using vectors, etc.

Active Publication Date: 2017-06-09
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

After searching, there is no relevant report about the functional element of the salt-inducible promoter

Method used

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  • Salt-induced functional element of high-efficiency soybean promoter and application of salt-induced functional element
  • Salt-induced functional element of high-efficiency soybean promoter and application of salt-induced functional element
  • Salt-induced functional element of high-efficiency soybean promoter and application of salt-induced functional element

Examples

Experimental program
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Effect test

Embodiment 1

[0021] Example 1 Obtaining of Salt-Inducible Functional Elements of High-efficiency Promoter of Soybean Shengdou No. 9 Gene

[0022] According to the reported RNAseq results of GmNAC4-transformed GmNAC4 soybean plants under salt / non-salt conditions, the genes up-regulated by the transcription factor GmNAC4 under high-salt stress were found, using the http: / / meme-suite.org / website The conserved sequence—CCTCCACCC—commonly contained in the promoter segments of these genes was compared, and specific primers were designed to fuse this sequence with the Arabidopsis TUB2 promoter, and named it pGmBM1.

[0023] 1.1 Cloning of pGmBM1

[0024] The reaction system for high-fidelity enzyme HIFI amplification is as follows (20 μl system):

[0025]

[0026] The amplification conditions are as follows:

[0027]

[0028] After the reaction, the reaction solution was detected by electrophoresis on 0.8% TAE agarose gel.

[0029] 1.2 Purification and recovery of cloned gene fragments ...

Embodiment 2

[0067] Example 2. Using the Arabidopsis protoplast transient transformation technology to verify that the key element pGmBM1 is induced by salt.

[0068] 2.1 Construction of pGmBM1-pGreen II-0800LUC vector

[0069] 2.1.1 Cloning of the pGmBM1 promoter fragment containing restriction sites

[0070] The dual fluorescent reporter vector pGmBM1-pGreen II-0800LUC of pGmBM1 was constructed by double enzyme digestion-ligation method. Using the restriction enzyme site analysis on Primer5pGmBM1, and the multiple cloning site on pGreen II-0800LUC (such as figure 2 shown) were compared, and the available enzyme cutting sites were screened out as Sal I and Pst I.

[0071] Use pGmBM1 primers containing restriction sites (F: 5'- ACG CGT CGA C AC CGT CGT GTT CAC AAATC-3', R: 5'- AAC TGC AGT T TT TCA AAA ACA CAA TTT CG-3') PCR amplification of pGmBM1.

[0072] The reaction system for high-fidelity enzyme HIFI amplification is as follows (20 μl system):

[0073]

[0074] The amplif...

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Abstract

The invention discloses a salt-induced functional element of a high-efficiency soybean promoter. The element is named as the salt-induced functional element GmBM1 of the high-efficiency soybean promoter, and a nucleotide sequence of the element is as shown as SEQ ID No. 1 (sequence identifier number 1). The invention further discloses a plant bifluorescence report carrier pGmBM1-pGreen II-0800 LUC comprising the salt-induced functional element and an application of the salt-induced functional element in improving salt stress tolerance of a plant. Experiments prove that the salt-induced functional element can quickly and efficiently respond to high salt induction, which indicates that the salt-induced functional element can be widely used for relevant functional studies of cultivating salt-resistant plant species and genetically modified salt-tolerant crops.

Description

technical field [0001] The invention relates to a salt-inducible functional element and its application, in particular to a new high-efficiency promoter salt-inducible functional element of soybean and its application. Background technique [0002] Soybean (Glycine max (L.) Merill) is a traditional edible leguminous crop and is also recognized as an important dicotyledonous plant. The seeds provide human beings with rich protein, high-quality vegetable oil, minerals and vitamins and other resources. In addition, soybean isoflavones, saponins, phosphatidylcholine, etc. also exist in soybean seeds as important functional compounds. [0003] According to the normal growth rate of plants under high salt stress, the plants were graded for salt stress tolerance. The results of the classification showed that cultivated soybean is a moderately salt-tolerant crop, and the high salinity content of the land limits the development of soybean planting. [0004] The growth and developmen...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/82A01H5/00
CPCC12N15/113C12N15/8222C12N15/8273
Inventor 向凤宁王楠李朔刘振华
Owner SHANDONG UNIV
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