A method for rapid identification of disease resistance by using powdery mildew of grapes to inoculate detached leaves of grapes

A technology of detached leaves and powdery mildew, applied in the field of bioengineering, to achieve the effect of fresh spores, good effect, and rich interaction theory

Inactive Publication Date: 2017-01-18
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the shortcomings of the current field natural disease identification method and indoor artificial spray inoculation identification method to identify grape powdery mildew resistance, and disclose a method for quickly identifying the disease resistance of isolated grape leaves by inoculating grape powdery mildew

Method used

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  • A method for rapid identification of disease resistance by using powdery mildew of grapes to inoculate detached leaves of grapes
  • A method for rapid identification of disease resistance by using powdery mildew of grapes to inoculate detached leaves of grapes
  • A method for rapid identification of disease resistance by using powdery mildew of grapes to inoculate detached leaves of grapes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Identification of disease resistance of isolated leaves of grape germplasm resources planted in the field with the powdery mildew strain NAFU1 inoculated

[0034] a. Isolation and purification of Erysiphe spp. strain NAFU1

[0035] During the onset period of grape powdery mildew, the leaves infected with grape powdery mildew were collected from serious vineyards, placed in a curler and brought back to the laboratory, and fresh diseased leaves with obvious symptoms of pathogenic bacteria were selected and placed in sealed bags until the disease occurred After it becomes particularly obvious, sweep the powdery mildew from the diseased leaves infected with powdery mildew to fresh grape leaves, and then place the inoculated grape leaves in a light incubator at a temperature of 22±1°C, light intensity of 10000LX, and relative humidity 50-75%, after 10-15 days, a monoclonal colony is obtained. When the monoclonal colony of powdery mildew grows to a diameter of 1 cm...

Embodiment 2

[0049] Example 2: Identification of disease resistance of isolated leaves of grape hybrid progeny plants inoculated with grape powdery mildew strain NAFU1

[0050] a. Isolation and purification of Erysiphe spp. strain NAFU1

[0051] During the peak period of grape powdery mildew, the leaves seriously infected with grape powdery mildew were collected in the vineyard, and the single spore isolation method was used to isolate and purify grape powdery mildew from the picked diseased leaves, and the isolated grape powdery mildew strains were pathogenic Sexual identification, one of the strains was rapidly susceptible and highly pathogenic, meeting the identification requirements, and named the strain NAFU1;

[0052] b. Infection with Erysiphe spp. strain NAFU1

[0053] The healthy leaves of grapes were inoculated with preserved fresh powdery mildew NAFU1 by tablet inoculation method, and the infection process of NAFU1 on grapes was detected;

[0054] c. Sequence amplification of ...

Embodiment 3

[0069] Example 3: Identification of disease resistance of isolated leaves of transgenic grape plants inoculated with Vitis vinifera strain NAFU1

[0070] a. Isolation and purification of Erysiphe spp. strain NAFU1

[0071] During the peak period of grape powdery mildew, the leaves seriously infected with grape powdery mildew were collected in the vineyard, and the single spore isolation method was used to isolate and purify grape powdery mildew from the picked diseased leaves, and the isolated grape powdery mildew strains were pathogenic Sexual identification, one of the strains was rapidly susceptible and highly pathogenic, meeting the identification requirements, and named the strain NAFU1;

[0072] b. Infection with Erysiphe spp. strain NAFU1

[0073] The healthy leaves of grapes were inoculated with preserved fresh powdery mildew NAFU1 by tablet inoculation method, and the infection process of NAFU1 on grapes was detected;

[0074] c. Sequence amplification of NAFU1 rDNA...

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Abstract

The invention discloses a method for rapidly identifying disease resistance by inoculating grape powdery mildew to detached grape leaves. According to the technical scheme, the method comprises the following steps: separating grape powdery mildew strain microspecies, purifying and infecting; performing rDNA sequence amplification and evolution analysis so as to obtain a grape powdery midew NAFU1 with high pathogenicity; analyzing the preservation and the expanding propagation of the strain; inoculating the strain with the detached grape leaves, dyeing by using trypan blue, inoculating to sick leaves in different periods, and observing indexes such as the time of spore germination, growth velocity, hypha length and necrocytosis, thereby rapidly and accurately detecting the disease resistance of grape. The grape powdery mildew preserved and amplified by using the method is high in sporulation quantity, the spore is fresh and high in pathogenicity, the defects that the conventional field identification period is long, the repeatability is poor and the weather influence can be caused are overcome, not only is the efficiency in detecting the disease resistance of a grape germplasm resource in large scale improved, but also direct reference values in rapidly identifying the disease resistance of the grape germplasm resource as well as hybrid progeny and transgenosis plants of the grape germplasm resource in large scale are achieved.

Description

technical field [0001] The invention relates to a method for identifying plant disease resistance, in particular to a method for identifying disease resistance of isolated grape leaves inoculated with powdery mildew of grapes, and belongs to the field of bioengineering. Background technique [0002] Grape powdery mildew is one of the important fungal diseases that seriously endanger grape production. The widely cultivated European grapes are very susceptible to diseases, which seriously affects the yield and quality of grapes. Although the traditional chemical control method is effective, its application is limited due to its high residue and heavy pollution, which seriously affects the food safety of grape products, threatens human health, and pollutes the ecological environment. Production practice shows that the most economical, effective, and environmentally friendly measure to prevent and control grape powdery mildew is to breed disease-resistant varieties, so resistan...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/18C12Q1/68C12Q1/04C12R1/645
Inventor 文颖强韩永涛高玉荣
Owner NORTHWEST A & F UNIV
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