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Application of rice OsWRKY21 transcription factor gene to plant insect resistance improvement

A technology of transcription factor and insect resistance, applied in the field of agricultural biology, can solve the problems of rising insect resistance, unsatisfactory control effect, and environmental pollution.

Active Publication Date: 2018-10-09
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among the main cultivated rice varieties promoted on a large scale in my country, there are few varieties resistant to rice planthoppers, which mainly rely on chemical insecticides for control. However, rice planthoppers are characterized by migration, suddenness and rampantness. , which makes the prevention and control difficult and costly, and the resulting problems such as the increase of pest resistance, harmful residues, and environmental pollution also make the control effect less and less ideal

Method used

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  • Application of rice OsWRKY21 transcription factor gene to plant insect resistance improvement
  • Application of rice OsWRKY21 transcription factor gene to plant insect resistance improvement
  • Application of rice OsWRKY21 transcription factor gene to plant insect resistance improvement

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1 Contains the construction of the expression vector of OsWRKY21 gene

[0032] The construction steps of the overexpression vector pDT-W21 containing the OsWRKY21 gene are as follows: The T vector pGEM-OsWRKY21 containing the full-length coding sequence of the OsWRKY21 gene constructed in the laboratory earlier (has been patented as "201210183789.X", and the name is "rice WRKY transcription In the application of factor gene OsWRKY21 in cultivating plant disease-resistant varieties", the OsWRKY21 DNA fragment was obtained by double digestion with XbaI and KpnI, and inserted into the pCDTNGUS vector (already in the document "Chen SB et al. Green fluorescent protein as a vital elimination marker to easily screenmarker-free transgenic progeny derived from plants co-transformed with double T-DNA binary vector system.Plant Cell Rep.2005,23(9):625-31." Disclosed in XbaI and KpnI restriction sites For specific steps, please refer to the instructions of restriction en...

Embodiment 2

[0034] Embodiment 2 Transformation of OsWRKY21 gene

[0035] (1) Preparation and transformation of Agrobacterium competent

[0036] 1) Agrobacterium freezing transformation method

[0037] A. Take 50 μL of Agrobacterium EHA105 bacterial liquid and add it to 5 mL of YEB liquid medium containing 50 μg / mL rifampicin, and culture at 26°C and 250 r / min for 12 hours to activate the strain;

[0038] B. Take 250uL of the activated bacterial strain to be transformed and add it to 25mL of YEB liquid medium containing 50μg / mL rifampicin, and culture at 26°C and 250r / min for 12h for expansion;

[0039] C. Take 20mL of the cultured bacterial solution and divide it into two 25mL refrigerated centrifuge tubes, centrifuge at 4000r / min for 10min at 4°C; discard the supernatant, add 10mL of pre-cooled 1×TE to each tube to resuspend the pellet, and put it on ice 15min;

[0040] D. Centrifuge at 4000r / min for 10min at 4°C;

[0041] E. Add 1mL YEB liquid medium to each tube to resuspend the pe...

Embodiment 3

[0096] Embodiment 3 rice cultivation and identification method for resisting rice planthopper

[0097] 1. Rice culture and identification method for rice planthopper resistance

[0098] (1) Rice seedling raising and breeding

[0099] 1) After rinsing the wild-type material (indica rice 9311) and the transgenic material seeds several times with tap water, place them in a culture bottle filled with 200 mL of pure water, soak for 36 hours at 33°C in total darkness to accelerate germination;

[0100] 2) Put the germinated seeds in a petri dish filled with 10mL of purified water, about 50 seeds per dish, gently shake the dish to make the seeds evenly distributed, and cultivate for 4-5 days at 28°C, 12h light, 12h dark;

[0101] 3) Transplant the seedlings into seedling pots, one seedling per hole, and match the corresponding number of wild-type plants in each pot at a ratio of at least 1 / 7, and move the pots to the plant cultivation room. The daytime temperature of the cultivation...

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Abstract

The invention belongs to the field of agricultural biotechnologies, and discloses application of a rice OsWRKY21 transcription factor gene in plant insect resistance improvement. An inventor discoversthat the OsWRKY21 gene has a positive regulating effect in a defense reaction of plant diseases and insect pests, namely the OsWRKY21 gene can improve the capacity of rice for resisting the insect pests of rice planthopper. The OsWRKY21 gene is a transcription factor gene. While the OsWRKY21 gene is over-expressed in a plant, the expression of a series of downstream plant disease and insect pestresistant functional genes can be transcriptionally-activated, and a transgenic plant is endowed with a good disease-resistant effect, so a defect that an individual functional gene is transcribed isovercome. The application is capable of providing a foundation for a follow-up WRKY transcription factor insect-resistant mechanism research, and providing a theoretical foundation for enriching brown-planthopper-resistant gene resources, and cultivating high-yield high-resistance rice varieties.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology, and particularly relates to the application of rice OsWRKY21 transcription factor gene in improving plant insect resistance. Background technique [0002] Pests and diseases are one of the main factors that cause rice yield reduction. The more serious pests and diseases in rice production mainly include rice planthopper (including brown planthopper, white-backed planthopper, etc.), rice leaf roller, rice stem borer, rice blast and grain There are five kinds of blight. Among them, the etiology of rice planthopper, leaf roller and rice blast has the characteristics of large annual fluctuations, rapid onset, and easy outbreaks, and poses the greatest threat to rice production. [0003] Rice planthopper belongs to Homoptera planthopper family, commonly known as fire midge, rattle insect, is a monophagous rice pest. There are mainly three kinds of rice planthoppers that harm rice in my coun...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8286
Inventor 许新萍龙永全温博冠
Owner SUN YAT SEN UNIV
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