Application of Arabidopsis gene MYB73 on the aspect of plant sclerotinia sclerotiorum proof disease

A technology of transgenic plants, Arabidopsis thaliana, applied in the field of application of the gene MYB73 in cultivating transgenic plant varieties resistant to Sclerotinia sclerotiorum, can solve problems such as unreported

Inactive Publication Date: 2013-01-09
HEBEI AGRICULTURAL UNIV.
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Problems solved by technology

[0004]Existing studies have shown that inducing callose deposition in plants can stimulate the basic disease resistance response in plants (Clay et al, 2009), MYB73 gene is the MYB gene in plants One of the members of the transcription factor family, when the myb73 mutant is infected with Pst DC3000, the production of callose is sig

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  • Application of Arabidopsis gene MYB73 on the aspect of plant sclerotinia sclerotiorum proof disease
  • Application of Arabidopsis gene MYB73 on the aspect of plant sclerotinia sclerotiorum proof disease
  • Application of Arabidopsis gene MYB73 on the aspect of plant sclerotinia sclerotiorum proof disease

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[0030] Example one

[0031] Screening of Arabidopsis disease-resistant mutants and analysis of disease resistance

[0032] In order to obtain disease-resistant mutants, 4-week-old Arabidopsis was inoculated with Pseudomonas syringae (Pst DC3000) for 1.5 hours, and then the genes whose expression changes were analyzed by gene chip technology, and the corresponding genes were obtained from the Arabidopsis ABRC library. Homozygous mutants of the gene. After the inventors connected these homozygous mutants to Pst DC3000, they found that the mutant myb73( figure 1 ), 4 days after inoculation, the number of pathogens in wild-type Arabidopsis is 100 times that of the mutant ( figure 2 ), using the Trypan Blue staining method, it was found that a large area of ​​necrotic lesions appeared on the wild type 2 days after inoculation, and the necrotic area of ​​the mutant leaf was significantly less than that of the wild type ( image 3 ), using DAB staining method, it was found that a large am...

Example Embodiment

[0033] Example two

[0034] Expression analysis of disease resistance related genes

[0035] After Pst DC3000 was inoculated with wild-type Arabidopsis for 0, 0.5, 1.5, 4, and 12 hours, Real time-PCR analysis revealed that the expression of MYB73 gene was significantly down-regulated after inoculation ( Image 6 ), and dropped to a minimum at 1.5h, which indicates that the MYB73 gene plays a negative regulatory role in the process of Arabidopsis resistance to Pst DC3000. At the same time, the inventor analyzed the expression changes of the resistance-related genes PR1 and PDF1.2, and found that PDF1 .2 The expression of the gene reached the maximum 1.5h after inoculation ( Figure 7 ), the expression of PR1 gene increased significantly at 0.5h of inoculation, and maintained at a fairly high level at 1.5h ( Figure 8 ), which shows that MYB73 gene is negatively correlated with the expression of resistance-related genes PR1 and PDF1.2.

Example Embodiment

[0036] Example three

[0037] Phenotype analysis of MYB73 gene overexpression mutant

[0038] The CDS region of MYB73 gene was first cloned into the pMD-19 simple vector, and then the CDS region of MYB73 gene was cut out with Xba I and BamH I, and the CDS region of MYB73 gene was connected to the vector pCAMBIA1300 by Xba I and BamH I. Between Xba I and BamH I restriction sites. The vector contains the CaMV 35S promoter, which can drive the overexpression of the target gene. The pCAMBIA1300 vector containing the CDS region of the MYB73 gene was transferred to Arabidopsis thaliana by using Agrobacterium to dip into the flowers. The over-expressed myb73 strain, mutant and wild-type Arabidopsis thaliana were cultured under long-day light for 4 weeks, and the Pst DC3000 strain was connected with a needle-free syringe for 3 days, and the disease was observed. The results are as follows Picture 9 As shown, the leaves inoculated with wild-type Arabidopsis and the overexpression mutant ...

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Abstract

The sclerotinia sclerotiorum is an important disease of commercial crops, such as soybean and rape to cause the production reduction even total crop failure of the crop so as to cause severe economic loss. A sclerotinia sclerotiorum sensitive mutant myb73 for a crucifer plant disease is obtained by screening, the over-expression vector of a MYB73 gene (of which the gene number is AT4G37260) is built, and the over-expression mutant of the gene is obtained by agrobacterium conversion. An experiment shows that the MYB73 gene is induced and expressed by pathogenic bacteria to negatively regulate the expression of the in vivo anti-disease gene of the plant, the generation of the in vivo callose of the plant is regulated, and a theory and production practice foundation is laid for obtaining the new species of the sclerotinia sclerotiorum proof transgene plant by the analysis and the identification of the MYB73 gene function.

Description

technical field [0001] The present invention relates to the application of a kind of Arabidopsis gene MYB73 in anti-sclerotinia sclerotiorum, in particular to the application of the gene MYB73 in anti-sclerotinia sclerotiorum (Sclerotinia sclerotiorum), and also relates to the application of the gene MYB73 in cultivating anti-sclerotinia transgene The application in plant varieties belongs to the field of genetic engineering. Background technique [0002] For a long time, the infection of pathogenic bacteria has been the main reason for the huge loss of crop yield. Instead of passively waiting for the infection of pathogenic bacteria, plants will produce a series of disease-resistant defense responses, such as synthesizing phytodefensins, producing hydrolytic enzymes and some antibacterial proteins to resist the invasion of pathogenic bacteria. Using molecular biology theory and technology to study the interaction mechanism between plants and pathogenic bacteria at the mole...

Claims

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Application Information

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IPC IPC(8): C12N15/84C12N15/09A01H5/00
Inventor 贾娇董金皋邢继红闫淑娟
Owner HEBEI AGRICULTURAL UNIV.
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