Method for identifying phenotype of indoor wheat dwarf virus resistance of triticeae crops and application

A technology of wheat dwarf virus and crops, applied in the field of agricultural biology, can solve the problems of large-scale identification in the field, poor control effect of wheat dwarf virus disease, difficulty in identification of disease resistance and research on pathogenic mechanism , to achieve the effect of low cost and high efficiency

Inactive Publication Date: 2016-12-07
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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AI Technical Summary

Problems solved by technology

Conventional pest control and disease control measures are very ineffective against wheat dwarf virus disease once the vector has completed the virus transmission process once the pest is seen, so cultivating disease-resistant varieties is the most effective control strategy
[0004] Due to the specialized transmission of wheat dwarf virus by the leafhopper, the disease occurs sporadically in the field, and large-scale identification in the field is difficult, which makes the identification of disease resistance and the mechanism of pathogenicity very difficult. Therefore, it is necessary to establish a simple and feasible method for phenotypic identification of dwarf virus resistance in wheat crops, which can be used for indoor inoculation with leafhoppers carrying WDV virus to achieve rapid and effective identification of disease resistance phenotypes and the purpose of multiplying the source of the virus

Method used

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  • Method for identifying phenotype of indoor wheat dwarf virus resistance of triticeae crops and application
  • Method for identifying phenotype of indoor wheat dwarf virus resistance of triticeae crops and application
  • Method for identifying phenotype of indoor wheat dwarf virus resistance of triticeae crops and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1. Preparation of toxin source and mediator

[0035] The source of the wheat dwarf virus (WDV) virus is the isolate of wheat dwarf virus in Hancheng, Shaanxi. After the plants with obvious symptoms are identified as positive by PCR and serology, the living bodies are preserved on the host wheat. The virus-transmitting agent-Psammotettix striatus (Linnaeus) was collected from Hancheng, Shaanxi Province, and was bred and bred on healthy wheat all year round after detoxification and purification. The medium breeding and the wheat variety Yangmai No. 12 used as feed and virus transmission host were all raised in a light incubator at 22°C, with 16 hours of light and 8 hours of darkness, and the light intensity was 20000Lx.

Embodiment 2

[0036] Embodiment 2. Feeding poison and inoculation of mediator

[0037] Draw 60-80 headline sand leafhoppers with an insect sucker, place it on the wheat dwarf virus source plant and feed it with a cover for 3 days ( figure 1 ), on the other hand, the work is to plant the wheat material to be identified 3 days in advance, that is, sow 25 seeds of the material to be identified in each glass cup filled with sterilized soil of 1 / 4 volume, and sow the 7th seed of the identified material to grow. The day is the 4th day of vector feed poisoning, after all the leafhoppers fed poison are sucked out with an insect sucker, they are placed in the glass cups of the seedlings to be identified, the mouth of the bottle is immediately covered with gauze cloth, and the rubber band is tightened ( figure 2 ). Then place it in a light incubator for 3 days and inoculate it for 3 days. On the 4th day, suck out all the leafhoppers, carefully scoop out the material to be identified in the glass cu...

Embodiment 3

[0038] Example 3. The rate of poisoning in the population of leafhoppers detected by PCR after being fed with poison

[0039] The poisoned leafhoppers were taken out with a suction device, and the total DNA of the leafhoppers was extracted from each head. The Wizard Genomic DNA Extraction Kit from Promega Company was used, and the reference manual of the extraction method was slightly changed. The extracted DNA samples were stored at -20°C for future use. PCR amplification primers were CP / F:5'-ATGGTGACCAACAAGGACTCC-3'; CP / R:5'-TTACTGAATGCCGATGGCTTTG-3', which were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd. Taking the total system 50 μL as an example, the PCR amplification system is as follows: rTaq 0.25 μL; 10×PCR Buffer 5 μL; dNTP Mixture (2.5 mmol / L each) 4 μL; upstream primer and downstream primer (10 μmol / L) 2 μL each; / μL) 2 μL, ddH 2 O 34.75 μL. PCR amplification program: 94°C for 3min; (94°C for 45sec, annealing temperature for 50sec, 72°C for 30sec) ...

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Abstract

The invention relates to a method for identifying the phenotype of indoor wheat dwarf virus resistance of triticeae crops and application, and belongs to the field of agricultural biological variety resources resistance identification. The identifying method disclosed by the invention comprises the following steps: transmitting the virus out from a living body virus source plant by utilizing a virus transmitting amboceptor, namely, Psammotettix striatus L, using the obtained amboceptor which carries the WDV (Wheat Dwarf Virus) as to-be-identified material for group inoculating, then separating seedlings, and transplanting the separated seedlings which are used for the identification of the indoor disease resistance of the WDV; setting the disease grading standards to be level 0, level 1, level 2, level 3, level 4 and level 5 according to the resistance identified plant phenotypes, wherein the level 0 is immune, namely, disease-free, the level 1 is high-resistant, the level 2 is disease-resistant, the level 3 is medium susceptible, the level 4 is susceptible to disease and the level 5 is high-susceptible. The resistance resource obtained through the identification can be further used for variety hybridizing and breeding, and can be used for preventing and controlling the wheat dwarf virus through field demonstration and popularization; the disease susceptible plants can also be used for researching the mechanisms, such as pathogenicity and the interaction of the amboceptor and a host.

Description

technical field [0001] The invention belongs to the technical field of agricultural biology, and in particular relates to a phenotypic identification technology and application for identifying indoor resistance to wheat dwarf virus of wheat crops with relatively simple operations. Background technique [0002] Wheat dwarf disease caused by wheat dwarf virus (WDV) was first discovered in the former Czechoslovakia in 1961. In recent years, the damage caused by the virus has increased year by year. At present, it has become one of the most threatening diseases in wheat production in Europe, Africa and Asia. Wheat dwarf virus is a member of the genus Mastrevirus in the family Geminiviridae, and its virions are in the form of twin particles. The virus has a wide range of hosts, including wheat (Triticum aestivum), barley (Hordeum vulgare), oats (Avena sativa) and other grass crops, as well as some weeds, causing severe dwarfing, yellowing, streaks and increased tillers of plants...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01G7/06
CPCA01G7/06
Inventor 刘艳吴慧娟王锡锋
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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