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Application of OsWRKY28 transcription factor gene of rice in improvement of plant disease resistance

A transcription factor and disease resistance technology, applied in the application field of rice OsWRKY28 transcription factor gene in improving plant disease resistance, can solve problems such as difficult research, slow process, and lack of resistance sources

Active Publication Date: 2012-12-12
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the strong saprophytic and wide host range of Rhizoctonia solani, genetic resources for resistance to rice sheath blight with a high level of resistance have not been found, and it is difficult and slow to carry out research on conventional disease resistance genetic breeding.
The lack of sources of resistance has also led to the lack of substantial progress in the research on the R gene cloning of sheath blight resistance, and the research on the molecular mechanism of the defense response against sheath blight is still in its infancy

Method used

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  • Application of OsWRKY28 transcription factor gene of rice in improvement of plant disease resistance
  • Application of OsWRKY28 transcription factor gene of rice in improvement of plant disease resistance
  • Application of OsWRKY28 transcription factor gene of rice in improvement of plant disease resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Isolation and Cloning of OsWRKY28 Gene

[0025] Using rice variety 9311 (He Guangming et al. Studies on the aggregation of rice anti-aging IPT gene and bacterial blight resistance gene Xa23. Acta Genetica Sinica, 2004, 31(8)) as material, inoculated D. Strong pathogenic strain RH-9 of Rhizoctonia solani (Li Aihong et al. Chitinase activity and sheath blight resistance of transgenic lines and different rice varieties. Acta Crops Sinica, 2003, 29(4)) . Inoculate the mycelial block of Rhizoctonia solani on the PDA medium, culture in the dark at 25°C for 3 days, use the tip of a Gilson 10mL pipette (D10mL, Gilson Company) as a hole puncher, and place on the outer edge of the mycelial culture Take mycelium agar strips (diameter 1.8mm, length 3mm), and use tweezers to insert the mycelium agar strips into the inner side of the third leaf sheath from top to bottom of the stem, so that the leaf sheaths maintain the original phimosis state. After 24 hours, the rice le...

Embodiment 2

[0027] Example 2 Expression profile analysis of OsWRKY28 gene

[0028]Using the rice variety 9311 as the material, use the same method as in Example 1 to inoculate Rhizoctonia solani strain RH-9 on tillering stage (10-week-old) rice, and insert blank agar strips into the mock control group (mock), respectively before inoculation. (0h) and 6h, 12h, 24h, 48h, 72h after inoculation, the rice leaf sheaths were cut and stored in liquid nitrogen. 3-week-old rice seedlings were sprayed with 2 mM salicylic acid (SA, prepared with pH 6.5 double-distilled water) and 100 μM jasmonic acid (JA, prepared with 0.1% (v / v) ethanol solution), and the simulated control group were respectively sprayed. The solvent components in these two solutions were sprayed, and all solutions were added with 0.05% (v / v) Tween20 to increase the adhesion of the leaf surface, respectively before spraying (0h) and after spraying 0.5h, 1h, 3h, Cut off rice leaves at 6h, 9h, 12h, and 24h, and store them in liquid n...

Embodiment 3

[0029] Example 3 Production and disease resistance identification of transgenic rice overexpressed with OsWRKY28 gene

[0030] The plasmid pGEM-OsWRKY28 obtained in Example 1 was double digested with AscI and BamHI, the full-length coding frame sequence of the OsWRKY28 gene was isolated and purified, and the sequence was connected to Figure 4 The shown plant expression vector pFGC1008 (Kerschen et al. Effectiveness of RNA interference in transgenic plants. FEBS letters. 2004, 566 (1)) between the AscI and BamHI restriction sites, the detailed steps of recombination are restricted by TaKaRa company Endonuclease and T4 ligase manual standard operation, transfer the recombinant plasmid into Escherichia coli DH5α, screen positive clones by enzyme digestion and confirm by sequencing, and name the correct recombinant plasmid pRSE-OsWRKY28, which is the overexpression of the OsWRKY28 gene carrier. The plasmid was transformed into Agrobacterium strain LBA4404 (Hiei et al. Efficient ...

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Abstract

The invention belongs to the field of plant genetic engineering and discloses application of OsWRKY28 transcription factor gene of rice in improvement of plant disease resistance. In research of functions of the WRKY transcription factor gene OsWRKY28 of rice as shown inthe SEQ ID NO.1 by means of a reverse genetics method, the OsWRKY28 transcription factor gene is found to have positive regulation function for resistance to rice banded sclerotial blight. The OsWRKY28 transcription factor gene is planted on a proper plant expressing carrier to convert cells or rice, and resistance of obtained rice with OsWRKY28 excessive-expression transcription factor to the banded sclerotial blight is evidently improved. Accordingly, disease resistance of rice and other crops can be improved by the OsWRKY28 gene, and new varieties of transgenosis plants resistant to the banded sclerotial blight can be cultured.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, in particular to the application of rice OsWRKY28 transcription factor gene in improving plant disease resistance. Background technique [0002] Plant diseases have serious negative effects on plant growth, development and reproduction, which can lead to reduced yield and quality of plants. However, plants have developed a series of active defense mechanisms to protect themselves during the long-term evolution of interactions with pathogenic microorganisms. Plant defense system mainly consists of local defense (Local defense) and systemic acquired resistance (Systemic acquired resistance, SAR). The recognition of pathogen-associated molecular patterns (PAMPs) by pattern recognition receptors (PRRs) on the surface of plant cells directly leads to PAMP-triggered immune responses (PAMP-triggered immunity, PTI). However, during the long evolutionary process, pathogens have evolved...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29A01H5/00
Inventor 许新萍陈凯张岭
Owner SUN YAT SEN UNIV
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