Regulation and control system for enhancing disease resistance of tomato plants and regulation and control method and application thereof

A control system and disease resistance technology, applied in the field of plant genetic engineering, can solve problems affecting tomato yield and quality, leaf wilting, environmental and human health risks and impacts, and achieve enhanced regulation, enhanced disease resistance, and improved resistance. disease effects

Pending Publication Date: 2022-03-01
山东狮克现代农业投资有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In tomato, Pseudomonas syringae frequently infects the leaves and veins, usually starting from the lower mature leaves, causing the leaves to wither, seriously affecting the yield and quality of tomatoes
[0003] At

Method used

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  • Regulation and control system for enhancing disease resistance of tomato plants and regulation and control method and application thereof
  • Regulation and control system for enhancing disease resistance of tomato plants and regulation and control method and application thereof
  • Regulation and control system for enhancing disease resistance of tomato plants and regulation and control method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0049] Example 1: Getting and Phenotypic Analysis of Each Transgenic Line

[0050] Experimental method:

[0051] 1.1, WRKY75 turn base

[0052]Get the special sequence of the WRKY75 coding area and the WRKY75 full CDS sequence (https: / / phytozome.jgi.doe.gov / pz / portal.jgi.doe.gov / pz / portal.jgi.doe.gov / pzportal.jgi.doe.gov / pz / portal.jgi.doe.gov / pz / portal.jgi.doe.gov / pz / portal.jgi.doe.gov / pzportal.html database, the number of WRKY75 is Solyc05G015850), and the sequence is amplified separately Plant RNAI interference vector and PSUPER1300 over the expression vector. The above carrier is converted to Agrobacterium strain LBA4404 by freezing, and then transferred into tomato by Agrobacterium, and the RNAi silence strain of WRKY75 and its overexpression strain were obtained. The obtained strain was detected by QRT-PCR.

[0053] 1.2, SLWRKY75 Phenotypic observation and determination of bacterial biomass

[0054] Use 10mm MGCL 2 The hemacterium bacterium bacteria to the bacteria is OD600 =...

Example Embodiment

[0060] Example 2:

[0061] experimental method:

[0062] 1.1, yeast single hybridization experiment

[0063] The full length sequence of WRKY75 coding regions was respectively obtained into a GAD vector; the 4-stage start subsequence (P1-P4) of GH3.3 was brought into a PlacZi2u yeast expression vector. The WRKY75-GAD + P1-LACZ, WRKY75-GAD + P2-LACZ, WRKY75-GAD + P3-LACZ and WRKY75-GAD + P3-LACZ and WRKY75-GAD + P4-Lacz were then transferred to yeast EGY48 strains, respectively.

[0064] 1.2, luciferase (LUC) activation experiment

[0065] The WRKY75 coding region full-length sequence was constructed into a PZP211-Flag expression vector; a promoter sequence (P2 region) of GH3.3 was brought into a PZP211-LUC plant expression vector. The constructing WRKY75-FLAG / -FLAG + P2PRO-LUC was transferred to the Agrobacterium GV3101 and instantaneously transformed tobacco experiments.

[0066] 1.3, gel migration block experiment (EMSA)

[0067] The experiment was mainly carried out by the EMS...

Example Embodiment

[0074] Example 3:

[0075] experimental method:

[0076] 1.1, yeast double hybrid experiment

[0077] The full length sequence of the WRKY75 coding region was respectively formed into the AD vector; the full length sequence of the coding region of the VQ10 was bd into a BD yeast expression vector. The constructing WRKY75-AD and VQ10-BD were then transferred to yeast Y2H strain.

[0078] 1.2, luciferase complementary experiment

[0079] The WRKY75 coding region full-length sequence is constructed into a PCambia-NLUC plant expression vector; a full length sequence of the coding region of the VQ10 is brought into a pCambia-Cluc plant expression vector. The constructing WRKY75-NLUC and VQ10-NLUC were transferred to the Agrobacterium GV3101 for instantaneous transformation tobacco experiments.

[0080] Planting: Tobacco (Island Smoke) planting in the greenhouse, the sunshine is 16 hours, the dark culture is 8 hours, the culture temperature is 25 ° C at normal temperature, the air humid...

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Abstract

The invention discloses a regulation and control system for enhancing disease resistance of tomato plants and a regulation and control method and application thereof, and relates to the technical field of plant genetic engineering. The invention provides a regulation and control system for enhancing disease resistance of tomato plants. The regulation and control system comprises a transcription factor WRKY75 and auxin amino synthase GH3.3, the amino acid sequence of the transcription factor WRKY75 is SEQ ID NO. 1; the amino acid sequence of the auxin amino synthase GH3.3 is SEQ ID NO. 2. In the prevention and treatment of tomato diseases caused by pseudomonas syringae, a WRKY75-mediated auxin-related disease-resistant regulation and control path is found and confirmed in tomatoes for the first time by utilizing reverse genetics; the transcription factor WRKY75 can activate the expression of the encoding auxin amino synthase gene GH3.3 and reduce the level of active auxin (free state), thereby improving the disease resistance of tomato plants. In the process, the transcription cofactor VQ10 interacts with the transcription factor WRKY75, so that the regulation of the WRKY75 on GH3.3 is enhanced, and the disease resistance of tomato plants is enhanced.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, in particular to a control system for enhancing the disease resistance of tomato plants, a control method and an application thereof. Background technique [0002] Pseudomonas syringae (Pseudomonas syringae) is an aerobic, saprophytic Gram-negative bacteria, and the incidence of plant diseases caused by it ranks first among the top ten bacterial plant diseases. In tomato, Pseudomonas syringae often occurs on the leaves and veins, usually from the lower mature leaves, causing the leaves to wither and seriously affecting the yield and quality of tomatoes. [0003] At present, the control of tomato diseases caused by Pseudomonas syringae is mostly chemical control, but chemical control of diseases will also bring potential risks and impacts on the environment and human health. Therefore, how to adopt environmentally friendly control measures to improve tomato disease resistance is...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N9/00C12N15/29C12N15/52C12N15/84A01H5/00A01H6/82
CPCC07K14/415C12N9/93C12N15/8205C12N15/8218C12N15/8281C12N15/8294C12Y603/02
Inventor 孟伦庄焜扬陈飞勇史庆华许思思李彦刘汉敬
Owner 山东狮克现代农业投资有限公司
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