Grape endophytic fungus and application thereof in preventing and controlling grape gray mold rot

A technology for botrytis cinerea and endophytic fungi, applied in the field of plant disease prevention and control, can solve problems such as botrytis cinerea that has not yet appeared, achieve good inhibition, improve commerciality, and save costs

Inactive Publication Date: 2017-06-27
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no research report on the biological cont

Method used

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  • Grape endophytic fungus and application thereof in preventing and controlling grape gray mold rot
  • Grape endophytic fungus and application thereof in preventing and controlling grape gray mold rot
  • Grape endophytic fungus and application thereof in preventing and controlling grape gray mold rot

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Isolation and identification of the endophytic fungus Myromyces verrucosus

[0031] The inventor found that the Chinese wild grape variety 'Shuangyou' has strong resistance to grape gray mold. Therefore, the leaves of the Chinese wild grape variety 'Shuangyou' in the field were rinsed with sterile water for 3 times, and the concentration Disinfect the surface with 70% alcohol for 30 seconds, disinfect with 2.5% sodium hypochlorite for 3 minutes, and then rinse with sterile water for 3 times. The leaves were cut into 3mm×5mm tissue, placed on a PDA plate and cultured at 25°C. After the endophytic hyphae grew around the leaf tissue, the hyphae at the edge of the colony were picked and transferred to a new PDA medium. After sporulation, the endophytic fungi were isolated and purified by single spores.

[0032]The DNA of the endophytic fungal strain was extracted, amplified by ITS primers (forward primer: 5'-TCCGTAGGTGAACCTGCGGA-3', reverse primer: 5'-TCCTACCTGA...

Embodiment 2

[0036] Embodiment 2: the preparation of verrucous verrucous spot bacteria biological control bacterium agent

[0037] 1. Inoculate Verrucous verrucosa on PDA medium and cultivate for 15 days. Irregular black spots will appear on the surface of the colony to be cultured. Gently scrape the paint-like black spots with a sterile scalpel, dissolve them in sterile water, and use Calculate the number of spores with a hemocytometer and adjust to 1×10 6 individual / mL, to obtain the biological control fungicide for use ( figure 2 A).

[0038] 2. Inoculate the Mycobacterium verrucosum on the PDA medium and cultivate it for 5 days, punch holes at the edge of the colony with a hole puncher with a diameter of 4mm, inoculate 8 bacterium cakes per 150mL Chapei liquid medium, and shake in the dark at 25°C ( 100rpm) for 15 days ( figure 2 B). The culture solution of Chapei's liquid medium was filtered through 4 layers of gauze, centrifuged at 12000rpm for 10min, and then sterilized by 0.2...

Embodiment 3

[0040] Embodiment 3: Determination of antagonistic activity of Mycocystis verrucosus to Botrytis cinerea

[0041] Due to the slow growth of Mycobacterium verrucosus, inoculate Mycocystis verrucosus on PDA medium in advance, about 2-3cm away from the center, after growing at 25°C for 5 days, when the colony directly reaches 2cm, inoculate Botrytis cinerea on On the other side of the PDA culture dish, take the PDA not inoculated with Myromycosporum verrucosus as a control, take pictures 4 days after inoculation and calculate the colony area, and observe continuously for 1 month. The results showed that Mycocystis verrucosus had a strong inhibitory effect on the growth of Botrytis cinerea, and the colony area of ​​Botrytis cinerea after 4 days of culture was significantly lower than that of the control, and after one month of culture, it was found that Mycosporum verrucosae could produce a significant zone of inhibition ( image 3 ).

[0042] In order to further confirm whether...

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Abstract

The invention discloses a grape endophytic fungus and an application thereof in preventing and controlling grape gray mold rot. The grape endophytic fungus is named as Myrothecium verrucaria. The biological control strain Myrothecium verrucaria is the endophytic fungus carried by the grape, can completely overcome various problems caused by the usage of the chemical pesticide for preventing and controlling, is beneficial to harmless and green production of the grape, is beneficial to the promotion of the commodity of the grape fruit and the reduction of the cost and has a potential application prospect. Confrontation culture and bacteriostatic experiment both prove that the biological control strain has a better restraining effect to the growth development of the grape botrytis cinerea. In vivo experiment also proves that the preventing and controlling effect is obvious after the fungicide is applied to the grape variety; the grape endophytic fungus has the advantages of safety, no environmental pollution, and the like; and the grape endophytic fungus can be used for preventing and controlling grape gray mold rot.

Description

technical field [0001] The invention relates to the technical field of plant disease prevention and control, in particular to a grape endophytic fungus and its application in preventing and controlling grape cinerea. Background technique [0002] Grapes occupy an important position in the world's fruit production. The annual output of table grapes in my country has been ranking first in the world for many years. Botrytis cinerea is not only a common disease in production fields, but also a devastating disease in post-harvest storage. Therefore, botrytis cinerea has become a major obstacle restricting the production of grapes in my country, and it is one of the most important diseases in the production and storage of grapes in protected areas. The disease has caused huge economic losses every year. [0003] Botrytis cinerea, with the form of Botryotinia fuckeliana (de Bary) Whetzel.], belonging to the genus Botryotinia fuckeliana (de Bary) Whetzel. The anamorphic form is Bot...

Claims

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Application Information

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IPC IPC(8): C12N1/14A01N63/04A01P3/00C12R1/645
CPCA01N63/30C12N1/14
Inventor 李智王西平
Owner NORTHWEST A & F UNIV
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