ERF transcription factor related to plant stress resistance and encoding gene and application thereof
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A transcription factor, coding gene technology, applied in the direction of plant genetic improvement, botanical equipment and methods, applications, etc.
Inactive Publication Date: 2016-01-20
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI +1
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Embodiment 1
[0063] Example 1 Isolation, Identification and Cloning of the Gene Encoding the Transcription Factor of Lotus japonicus LcERF056
[0064] 1 Materials and methods
[0065] 1.1 Materials
[0066] 1.1.1 Plant material cultivation and stress treatment
[0067] Using lotus root "Leo" as the material, select mature and plump seeds that are free from diseases and insect pests, soak them in 2% NaClO for 15-20 minutes, wash them with sterile water for 3 times, dry them, and put the sterilized seeds on the B5 medium , cultured in a light incubator for 30 days, treated with 150mM sodium chloride and 20% PEG for 0h, 3h, 12h, and 24h, collected leaves were immediately frozen in liquid nitrogen, and stored in a -80°C refrigerator for later use.
[0068] Analysis of tissue expression specificity: The roots, stems, leaves, flowers, and seeds of J. japonicus were taken, quick-frozen in liquid nitrogen, and stored at -80°C until use.
[0069] Hormone-treated japonicus japonicus, the seedling...
Embodiment 2
[0113] Example 2 Construction of plant expression vector containing LcERF056 gene and its salt tolerance function verification in Arabidopsis
[0114] 1 Materials and methods
[0115] 1.1 Materials
[0116] 1.1.1 Plant material
[0117] Arabidopsis thaliana Columbia (Col-0) ecotype: preserved in the inventor's laboratory.
[0118] 1.1.2 Strain and carrier
[0119] Agrobacterium tumefaciens strain GV3101 was preserved in the inventor's laboratory; gateway entry vector pDNOR201 and expression vector pH7WG2D were purchased from Invitrogen.
[0120] 1.1.3 Medium
[0121] 1.1.3.1YEB medium (1L)
[0122] Peptone 5g, yeast extract 1g, sucrose 5g, MgSO 4 ·7H 2 O0.5g, dilute to 1L with deionized water, if it is a solid medium, add 15g agar powder, and sterilize under high temperature and high pressure at 121°C for 15min.
[0123] 1.1.3.2LB medium (1L)
[0124] Tryptone 10g, sodium chloride 10g, yeast extract 5g, deionized water to 1L, if it is a solid medium, add 15g agar powd...
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Abstract
The invention discloses an ERF transcription factor related to the plant stress resistance and an encoding gene and application thereof. The invention firstly provides an ERF transcription factor gene LcERF056 separated from lotus japonicas, the amino acid sequence of the gene LcERF056 is shown as SEQ ID No.1, and the nucleotide sequence of the encoding transcription factor is shown as SEQ ID No.2. The invention further provides an ERF transcription factor structure domain which can be combined with a cis-acting element to start stress resistance response gene expression and is shown as SEQ ID No.3. Function analysis experiments prove that the resistance of a plant to high salt stress can be effectively enhanced or improved by overexpressing the LcERF056 gene in the plant, and therefore the protein encoded by the LcERF056 gene has the functions of the EFR transcription factor. The invention further provides application of the ERF transcription factor and the encoding gene on the aspects of enhancing the adversity stress resistance of the plant, breeding new transgenic plant varieties with the adversity stress resistance and the like.
Description
technical field [0001] The present invention relates to transcription factors, in particular to ERF transcription factors and their encoding genes isolated from Lotus corniculatus L., which are related to plant resistance to adversity stress, and the present invention further relates to their application in improving the ability of plants to resist adversity stress, The invention belongs to the field of ERF transcription factor and its application. Background technique [0002] Salinity is the primary factor that reduces crop yields. Soil salinization and secondary salinization problems have been widespread in the world, especially in arid and semi-arid areas, where the problem is more serious. According to incomplete statistics from UNESCO and FAO, the global saline-alkali land area has reached 950 million hm 2 , accounting for about 10% of the total land area of the earth, of which China is 99.13 million hm 2 , accounting for one-ninth of the world's saline soil, equiv...
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