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MYB transcription factor for improving drought resistance of populus sutchuenensis and application of MYB transcription factor

A technology of transcription factor and Populus japonicus, applied in application, genetic engineering, plant genetic improvement, etc., can solve problems that restrict forestry output and development, and achieve the effect of improving drought resistance

Active Publication Date: 2022-03-25
SHENYANG AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moreover, since the arid and semi-arid area accounts for more than half of the country's land area, it seriously restricts the output and development of forestry. Therefore, cultivating new drought-resistant varieties is a key problem to be solved in forestry production.

Method used

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  • MYB transcription factor for improving drought resistance of populus sutchuenensis and application of MYB transcription factor
  • MYB transcription factor for improving drought resistance of populus sutchuenensis and application of MYB transcription factor
  • MYB transcription factor for improving drought resistance of populus sutchuenensis and application of MYB transcription factor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: Expression analysis of MYB6 transcription factor of Populus montana

[0025] The two-month-old poplar seedlings were subjected to drought stress treatment at 7 time points (water treatment was used as the control), and then the RNA of roots, stems, and leaves were extracted, reverse-transcribed into cDNA, and MYB6 transcription factor real-time fluorescent quantitative PCR (qRT-PCR) analysis.

[0026] The result is as figure 1 As shown, MYB6 was up-regulated in the roots, stems and leaves of Populus japonicus, and the expression level in the leaves treated for 12h was 3.6 times that of the control group, and the expression level in the roots and stems treated for 6h was 3.6 times that of the control group. group 5.625 times and 4.48 times, the results show that MYB6 is related to the drought resistance of Populus montana.

Embodiment 2

[0027] Example 2: Transient infection of MYB6 transcription factor of Populus montana and detection of physiological indicators

[0028] Using Populus japonicus cDNA as a template, primers MYB6-F (5'-CTCTAGAGGATCCCCGGGATGGATGTTAAAG-3') and MYB6-R (5'-TCGAGCTCGGTACCCGGGTCACAAGTTGTT-3') were used to amplify the target gene sequence shown in SEQ ID NO.1. The target gene sequence was recombined into the pROK II vector to obtain a recombinant overexpression vector, which was transformed into Agrobacterium competent cell EHA105 by electric shock transformation method, and then transformed into Populus montana by high-efficiency transient transformation technology for drought stress After processing, the physiological indicators were detected, and the specific experimental steps were as follows:

[0029] 1) High-efficiency transient transformation of Populus montana with MYB6 overexpression vector:

[0030] Use 50ml of 1 / 2MS liquid medium (adding 150 μM acetosyringone) to resuspend ...

Embodiment 3

[0036] Example 3: Stable genetic transformation and histochemical staining analysis of the MYB6 transcription factor of Populus montana

[0037] 1) Transform the recombinant overexpression vector of Example 2 into the Agrobacterium competent cell EHA105 by electric shock transformation, and then use the Agrobacterium-mediated method to stabilize the EHA105 Agrobacterium containing the pROK II-MYB6 overexpression vector genetically transformed into Populus montana, and selected by kanamycin to obtain kanamycin-resistant seedlings ( image 3 ), and extract the DNA of the resistant seedlings, and use the carrier primers pROK II-F (5'-TCGAGCTCGGTACCCGGGTCACAAGTTGTT-3') and pROK II-R (5'-GATGTGCTGCAAGGCGATT-3') for PCR detection ( Figure 4 ), the vector primer amplified empty load was 447bp, and the gene band was 975bp, so the length of the target band was 1422bp. It can be seen from the figure that the position is correct, indicating that 4 transgenic lines of Populus montana wer...

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Abstract

The invention relates to the technical field of biology, in particular to an MYB transcription factor capable of improving drought resistance of Populus sutchuenensis and application of the MYB transcription factor. The MYB transcription factor is MYB6, and the nucleotide sequence is shown as SEQ ID NO.1. The transcription factor MYB6 positively regulates the drought resistance of Populus sutchuenensis, the drought-resistant Populus sutchuenensis can be obtained by overexpressing the transcription factor MYB6 through a transgenic technology, and the application of the MYB transcription factor to the drought resistance of Populus sutchuenensis is achieved. A foundation can be provided for cultivating a new drought-resistant variety of populus sutchuenensis

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a MYB transcription factor for improving the drought resistance of Populus japonicus and its application. Background technique [0002] At present, desertification, aridity, semi-aridity, and salinization of my country's land are relatively serious. Among the proposed new afforestation forest land, difficult sites account for more than 60%, which makes afforestation more difficult. Therefore, breeding high-quality, high-resistant new varieties of forest trees is an urgent problem to be solved, which plays an important supporting role in the restoration of the ecological environment, the improvement of forest functions, and the construction of ecological security. Moreover, since the arid and semi-arid areas account for more than half of the country's land area, which seriously restricts the output and development of forestry, the cultivation of new drought-resistant varieties is a ke...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/84C12N1/21A01H5/00A01H6/00
CPCC07K14/415C12N15/8205C12N15/8273
Inventor 国会艳李卓燃及晓宇王玉成李丹丹
Owner SHENYANG AGRI UNIV
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