Tephrosia purpurea endophytic fungus TPL25 and application thereof in plant disease prevention and treatment

A technology of endophytic fungus and edamame, applied in the field of microorganisms, can solve the problems of agricultural product safety threats, chemical pesticide pollution, residual toxicity, etc.

Active Publication Date: 2015-04-22
HUNAN AGRICULTURAL UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Plant fungal diseases not only directly cause the decline in crop yield and quality, but also some pathogenic fungi can secrete and produce a variety of toxins and metabolites harmful to humans and animals during the process of infecting crops, posing a great threat to the safety of agricultural products
[0004]Chemical control

Method used

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  • Tephrosia purpurea endophytic fungus TPL25 and application thereof in plant disease prevention and treatment
  • Tephrosia purpurea endophytic fungus TPL25 and application thereof in plant disease prevention and treatment
  • Tephrosia purpurea endophytic fungus TPL25 and application thereof in plant disease prevention and treatment

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1: Penicillium grisea ( Penicillium griseofulvum ) Separation and purification of TPL25

[0036] Penicillium grisea of ​​the present invention ( Penicillium griseofulvum ) TPL25 was obtained from gray edamame leaves on the campus of Hunan Agricultural University in Hunan Province, China. Isolation method: Disinfection of the tissue surface: Rinse the gray edamame leaves with sterile water, and cut them into small pieces of 0.5cm×0.5cm. Disinfect the surface of the above materials in an ultra-clean bench, the procedure is: rinsing with 75% alcohol by volume fraction for 3-5min→1g·L -1Rinse with mercury chloride for 30s-1min→5 times with sterile water. Separation and purification: Inoculate the sterilized tissue material on the poured PDA plate medium, 3-4 pieces per plate, and place it at 28±1°C for 3-7 days in the dark. When mycelia (colonies) grow out of the cuts along the outer edge of the material, the mycelium is picked by tip mycelium, and the myce...

Embodiment 2

[0037] Embodiment 2: Penicillium grisea ( Penicillium griseofulvum ) Identification of TPL25 strain

[0038] The strains were identified by combining morphological and molecular biological methods. Penicillium grisea TPL25 can grow on PDA medium, cultivated at a constant temperature of 28±1°C, the colony is light green at first, the edge is white, and later turns dark green, the texture is fluffy and granular, and conidia are produced in large quantities. Pale orange, soluble pigment similar to the lighter color on the reverse side (eg figure 1 ). Microscopic morphological features: conidia strung together, colorless, spherical (eg figure 2 ). The strain TPL25 was amplified by PCR using ITS sequence general primers ITS4 and ITS5 (ITS4: TCCTCCGCTTATTGATATGC; ITS5: GGAAGTAAAAGTCGTAACAAGG). Using TPL25 total DNA as a template, the PCR reaction conditions were: pre-denaturation at 95°C for 5 min; denaturation at 95°C for 30 s, annealing at 56°C for 30 s, extension at 72°C...

Embodiment 3

[0039] Embodiment 3: Penicillium grisea ( Penicillium griseofulvum ) Preliminary screening of TPL25 anti-pathogenic fungal activity.

[0040] Under sterile conditions, use the plate confrontation method for primary screening (such as Figure 4 ), the endophytic fungus strain TPL25 and the pathogenic fungus were respectively made into cakes with a 6mm hole punch, placed on 1 / 3 of the PDA plate, cultured at 28°C, and observed whether the endophytic strains and the tested pathogenic fungi had Antagonism, and measure the width of the inhibition zone, and compare the activity of each endophytic fungal strain.

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Abstract

The invention discloses tephrosia purpurea endophytic fungus penicillium griseofulvum TPL25 and application of a fermentation product of the tephrosia purpurea endophytic fungus penicillium griseofulvum TPL25 in plant disease prevention fungal pathogens. The tephrosia purpurea endophytic fungus penicillium griseofulvum TPL25 is preserved in China Center for Type Culture Collection, and the preservation number is CCTCC NO: M2014607. Fungus fermentation liquor, fermentation liquor extractives and mycelium extractives have the strong mycelium grow inhibition function on sclerotinia rot of colza, tobacco blackleg and other phytopathogen, excellent strains are provided for development of microbial pesticide, and in addition, the tephrosia purpurea endophytic fungus is studied for the first time.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and relates to Penicillium grisea ( Penicillium griseofulvum ) TPL25 and its application in the control of plant diseases. Background technique [0002] gray edamame ( Tephrosia purpurea ) is a genus of edamame (Leguminosae) ( Tephrosia Pers .) Perennial woody plants, mainly distributed in tropical and subtropical regions, such as India, Pakistan, Vietnam, Cambodia, Laos, Myanmar, and Guangdong, Guangxi, Yunnan, Fujian and Hunan in China. The branches and leaves of gray edamame can be used as green manure, smashed and thrown into the water to poison fish, and it is also a good sand-fixing and embankment soil-conserving plant. At present, the research on edamame mainly focuses on phytochemistry and pharmacology, and there are few reports on endophytic fungi. [0003] Plant diseases have always been one of the important factors restricting high-quality and high-yield crops, and among p...

Claims

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Application Information

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IPC IPC(8): C12N1/14A01N63/04A01P3/00C12R1/80
CPCA01N63/30C12N1/145C12R2001/80
Inventor 李有志罗仄平丁文兵邹凯
Owner HUNAN AGRICULTURAL UNIV
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