HD-Zip I type transcription factor GmHDL 57 genes and application

A technology of transcription factors and genes, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problems of lack of practical application value, stable transformation of japonicus japonicus seeds, etc., and achieve the effect of improving salt resistance

Active Publication Date: 2018-08-31
XINYANG NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the stable transformation of japonicus japonicus seeds has not been obtained

Method used

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  • HD-Zip I type transcription factor GmHDL 57 genes and application
  • HD-Zip I type transcription factor GmHDL 57 genes and application
  • HD-Zip I type transcription factor GmHDL 57 genes and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] A preparation method of the HD-Zip class I transcription factor gene GmHDL57 that improves the salt resistance of legumes, the steps of which are:

[0035] The seeds of cultivated soybean Williams82 were sterilized and placed in 1 / 2 Hoagland medium for germination and growth. The parameters of the light incubator were set at a temperature of 25 °C, a relative humidity of 60%, 18 hours of light and 6 hours of darkness. The root tissue of the seedlings was taken, and after quick freezing in liquid nitrogen, the total RNA of the root tissue of the seedlings was extracted according to the operation instructions of the RNA extraction kit of TaKaRa Company. The first-strand cDNA was prepared using the reverse transcription kit from TIANGEN. According to the GmHDL57 gene sequence published on the NCBI website (GenBank: XP_006574472.1), primers F-GmHDL57 (5'-ATGGCGAGTGGCAAGCTT TATGC-3') and R-GmHDL57 (5'-TCAATAGGGCCAGAAACAG-3') were designed to target the first strand of cDNA ...

Embodiment 2

[0037] The stress expression detection of GmHDL57 gene, its step is:

[0038] The seeds of cultivated soybean Williams82 were sterilized and placed in 1 / 2 Hoagland medium for germination and growth. The parameters of the light incubator were set at a temperature of 25 °C, a relative humidity of 60%, 18 hours of light and 6 hours of darkness. When the seedlings grow to the V1 stage (the first compound leaf stage), the abiotic stress treatment is carried out. The seedlings in the three experimental groups were treated with 1 / 2 Hoagland culture solution containing 100 μmol / L ABA, 100 mmol / L NaCl, and 30% PEG-6000, respectively. Another group of seedlings were transferred to a 4 °C incubator for cold treatment. There were 5 seedlings in each group, and 3 replicates were set up. The root tissues of the seedlings were collected at 0 h before the stress and 1, 6, 12, 24 and 48 h after the stress, and were quickly frozen in liquid nitrogen and stored in a -80 °C refrigerator for late...

Embodiment 3

[0042] A kind of HD-Zip class I transcription factor gene GmHDL57 that improves the salt resistance ability of legumes is applied in japonicus japonicus, and the specific implementation steps are as follows:

[0043] japonicus stable transformation:

[0044] (1) Construction of overexpression fusion vector

[0045] Insert the correctly sequenced target gene into the plant overexpression vector p1301U, the upstream primer is F-GmHDL57-OX (5'-CGggatccATGGCGAGTGGCAAG-3'), the downstream primer is R-GmHDL57-OX (5'-GGggtaccTCAATAGGGCCAGAAAC-3'), The lowercase letter area is the restriction site sequence BamH I and Kpn I, and the overexpression vector p1301U-GmHDL57 was constructed ( figure 1 shown in B). The extracted plasmid was electroporated to transform Agrobacterium tumefaciens EHA105.

[0046] (2) Preparation of Agrobacterium liquid

[0047] A single colony of Agrobacterium EHA105 was cultured in YEB liquid medium supplemented with 40 mg / L rifampicin and 50 mg / L kanamycin...

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Abstract

The invention discloses a HD-Zip I type transcription factor GmHDL 57 gene with ability of improving salt resistance of leguminous plants. The sequence of the gene is a nucleotide sequence shown as SEQ ID NO:1, and a coded protein sequence is an amino acid sequence shown as SEQ ID NO: 2. The invention further discloses application of the HD-Zip I type transcription factor GmHDL 57 gene with ability of improving salt resistance of leguminous plants in lotus corniculatus. By utilizing an overexpression technique, the salt resistance of the gene in the leguminous plant lotus corniculatus is researched. Results show that compared with control plants, the salt resistance of the plants is obviously enhanced after overexpression, and the gene has great application prospects for widening a planting range of the leguminous plants in saline-alkali soil.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, in particular to the HD-Zip class I transcription factor GmHDL57 gene and its application in enhancing the salt-resistant ability of japonicus japonicus. Background technique [0002] Homeodomain leucine zipper protein HD-Zip (homeodomain leucine-zipper) is a plant-specific transcription factor, including a homeodomain HD-Zip composed of 60 or 61 amino acids and a leucine zipper structure Domain LZ. HD-Zip class I proteins are mainly involved in the response to abiotic stress. There are 17 HD-Zip class I proteins in Arabidopsis, among them, ATHB5-7 and ATHB12 are induced by drought and exogenous ABA, and the expression of ATHB7 can be induced by salt stress and osmotic stress. Tobacco HD-Zip class I transcription factor NaHD20 is induced by dehydration stress and positively regulates ABA accumulation in leaves. MtHB1 of Medicago truncatula was induced by NaCl stress, and its ...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/06A01H6/54
CPCC07K14/415C12N15/8273
Inventor 柯丹霞舒勇彭昆鹏
Owner XINYANG NORMAL UNIVERSITY
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