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Thinopyrum intermedium ERF-transcription factor and its coding gene and use

A technology for transcription factors and coding genes, which can be used in applications, genetic engineering, plant genetic improvement, etc.

Inactive Publication Date: 2007-03-28
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the currently available genetic resources related to disease resistance and stress resistance are limited, and there are problems such as short lifespan of disease-resistant varieties and unsatisfactory stress-resistant varieties, which are difficult to meet the needs of current production and ecological improvement.

Method used

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  • Thinopyrum intermedium ERF-transcription factor and its coding gene and use
  • Thinopyrum intermedium ERF-transcription factor and its coding gene and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1, Cloning and Sequence Analysis of ERF Transcription Factor Gene of Echinopsis intermedia

[0034] Three weeks of growth at room temperature (26 degrees) for Thiopyrum intermedia, some plants were inoculated with wheat sheath blight bacteria on the leaf sheaths and leaves of Thiopyrum intermedia seedlings by toothpick and friction method, and the roots of some plants were soaked in 250mm NaCl aqueous solution deal with. Take Trizol (Invitrogen) method to extract total RNA from the leaves of Thiopyrum intermedium inoculated with sheath blight for 24 hours or treated with NaCl for 5 hours, dissolved in DEPC water, and detected by agarose gel electrophoresis. Take 5ug of total RNA with good quality and no DNA pollution, and use the Superscript First-Strand Synthesis System for RT-PCR kit (Invitrogen) from Invitrogen Company, and use Oligo d(T) 12-18 As primers, the first strand of cDNA was synthesized according to the kit instructions. According to the wheat EST...

Embodiment 2

[0040] Embodiment 2, the induced expression analysis of TiERF1 gene

[0041] The following stress treatments were carried out on Thiopyrum intermedia: pathogenic bacteria, 250mM NaCl, drought, low temperature, 2mM ethephon solution (ET), and the specific treatment methods were as follows:

[0042] Pathogen treatment: Inoculate wheat sheath blight mycelium on three-week-old Thinopyrum intermedium leaves by rubbing and dental inspection, and take leaves at 0h, 2h, 5h, 12h, and 24h after inoculation, freeze in liquid nitrogen, and store at -70 degree of preservation.

[0043] Salt treatment: the roots of Thiopyrum intermedia plants growing around four weeks were placed in 250mM NaCl aqueous solution, and the leaves were taken after 0h, 2h, 5h, 10h, and 24h, quickly frozen in liquid nitrogen, and stored at -70°C for later use.

[0044] Drought treatment: wash the three-week-old Thiopyrum intermedia seedlings with water, put them on filter paper enough to absorb the water, dehydra...

Embodiment 3

[0048] Embodiment 3, in vitro expression of Tierf1 gene in Escherichia coli

[0049] Design a pair of specific primers according to sequence 1 in the sequence table, and add enzyme recognition sites EcoRI and XhoI at both ends of the upstream and downstream primers. The specific sequences are as follows AP1 5'-CCGAATTCCCGACGACTCCGACGACA-3', AP25'-TTCTCGAGCTAGCTGACCAGCTGCTGC-3 '; Amplify TiERF1 by PCR comprising the AP2 DNA domain and the sequences on both sides to obtain a cDNA fragment of about 700bp, and use EcoRI and XhoI (Takara) to digest the target gene amplification fragment and the fusion protein expression vector PGEX-4T-1 ( purchased from Amersham Company), purify and recover the target gene fragment and vector fragment. by T 4 DNA ligase inserts TiERF1 into the GST site of PGEX-4T-1, then transforms it into E. coli BL21 cells, picks a single colony, cultures, extracts the plasmid, and uses AP1 and AP2 primers for PCR identification and sequencing. The identified ...

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Abstract

The invention discloses a middle couch grass ERF transcription factor and its coding gene. The middle couch grass ERF transcription factor can be the protein with one of the following amino acid residues sequences that SEQ ID No: 2 in sequence list; or its amino acid residues sequence which is replaced and / or missed and / or added with one or many amino acid residues, and relevant to plant against disease against reverse performance. The transcription expression of the coding gene is greatly induced by rhizoctonia cerealis, salt, cold, ethylene etc. The TiERF1 of the invention supplies the base for the relevant gene expression, gene resources for gene engineering breeding, and will play an importance role in increasing plant against disease against reverse performance for gene engineering breeding.

Description

technical field [0001] The present invention relates to a plant ERF transcription factor and its encoding gene and application, in particular to an ERF transcription factor of Echinopsis intermedia and its encoding gene, and the application of the gene in cultivating disease-resistant plants and stress-resistant plants. Background technique [0002] At present, biotic stress such as various diseases and insect pests, and abiotic adversity stress such as drought, salinity, and low temperature are important factors restricting the production and development of crops, forest grass, and vegetables. For example, with changes in farming systems, fertilizer and water conditions, and climate conditions, the damage caused by soil-borne fungal diseases such as wheat sheath blight and scab and air-borne fungal diseases such as wheat powdery mildew and stripe rust in my country's main wheat producing areas It is becoming more and more serious, ranging from causing a 20-30% reduction in w...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63A01H1/00
Inventor 张增艳姚乌兰辛志勇
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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