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Colletotrichum transcription factor CsATF1 and application thereof

A transcription factor, anthracnose technology, applied in applications, fungicides, fungi, etc., to achieve the effect of improving sterilization efficiency

Active Publication Date: 2020-08-18
HAINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are still few studies on the related ATF transcription factors in Hevea anthracnose

Method used

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  • Colletotrichum transcription factor CsATF1 and application thereof
  • Colletotrichum transcription factor CsATF1 and application thereof
  • Colletotrichum transcription factor CsATF1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Cloning of embodiment 1 rubber tree anthracnose bacteria CsATF1 gene

[0026] According to the bZIP gene sequence of C. graminicola transcription factor (accession number XM008094996.1) obtained in NCBI, the homologous sequence was obtained by using the local BLAST method in the transcriptome database of Hevea anthracis HN08, and the primer pair CsATF1- F(5'-ATGGGAACTTCGCCGACCGAC-3') / CsATF1-R(5'-TCATGAGAAACGTCGCTGGA-3'), using the cDNA and DNA of C. siamense HN08 as templates, respectively amplified to obtain the target bands. Sequence analysis showed that the obtained sequence contained a complete coding open reading frame. The size of the DNA sequence is 1758bp, and the size of the cDNA sequence is 1611bp. The gene contains 2 introns, encodes 536 amino acids, and contains three Aft1 domains and a BRLZ (basic leucine zipper) domain, indicating that CsATF1 is bZIP ATF transcription factor in the transcription factor family. The gene was named CsATF1.

Embodiment 2

[0027] Example 2 Construction of CsATF1 gene knockout vector and acquisition of CsATF1 gene knockout mutant

[0028] Before and after the coding reading frame of the CsATF1 gene, design primer pairs CsATF1-UP-F / CsATF1-UP-R and CsATF1-D-F / CsATF1-D-R, use PCR amplification to obtain the upper arm sequence of the CsATF1 gene and the lower arm sequence after the C-terminal, using The method of homologous recombination, the upper arm and the lower arm sequence are joined into the vector pCX62-S, and the knockout vector pCX62-S-CsATF1 is obtained. See the diagram for details figure 1 .

[0029] The knockout vector pCX62-S-CsATF1 obtained from the above construction was introduced into protoplasts of Hevea anthracis HN08 by PEG-mediated protoplast transformation method, and screened by DCM medium containing chlorimuron-methyl (100 μg / mL) . A total of 3 batches were transformed and 159 transformants were obtained.

[0030] Genomic DNA sequences of 159 transformants were extracted ...

Embodiment 3

[0032] Example 3 CsATF1 gene deletion affects the sensitivity of fungi to the fungicide fludioxonil

[0033]In the medium containing different concentrations of fludioxonil, with the increase of fludioxonil concentration, the growth rate of the wild-type strain containing CsATF1 gene decreased gradually. However, in the CsATF1 gene deletion mutant, on the medium with the same concentration of fludioxonil, the growth rate of the mutant is greater than that of the wild type; see image 3 . The results indicated that the transcription factor CsATF1 can regulate the sensitivity of fungi to pyrrole-like agents such as fludioxonil, and the expression of this gene can increase the sensitivity of fungi such as anthracnose to fludioxonil and other pyrrole-like agents.

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Abstract

The invention provides a colletotrichum transcription factor CsATF1 and an application, the gene contains two introns, codes 536 amino acids, contains three Aft1 structural domains and a BRLZ (basic leucine zipper) structural domain, and is an ATF transcription factor in a bZIP transcription factor family. Experiments prove that the transcription factor participates in regulating the sensitivity of fungi to pyrrole medicaments such as fludioxonil and the like, which indicates that CsATF1 can be applied to preparation of a sterilization enhancer.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an anthrax transcription factor CsATF1 and its application. Background technique [0002] (1) Transcription factors can inhibit or activate the transcription of certain genes, regulate the physiological metabolism and growth and development of organisms [0003] Transcription factors (TFs) play crucial roles in almost all biological processes. Transcription factors, also known as trans-acting factors, refer to DNA-binding proteins that can specifically interact with cis-acting elements in the gene promoter region, and activate or inhibit certain genes through the interaction between them and other related proteins. Gene transcription, thereby regulating the expression of the target gene at a specific intensity, at a specific time and space, and regulating the physiological metabolism and growth and development of organisms (Latchman, 1997). There are many types of transcription fac...

Claims

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Application Information

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IPC IPC(8): C07K14/37C12N15/31C12N1/15A01N47/44A01P3/00C12R1/645
CPCC07K14/37A01N47/44
Inventor 林春花方思齐宋苗廖卫国刘文波李潇
Owner HAINAN UNIVERSITY
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