CAMELLIA SINENSIS ENDOPHYTE PENICILLIUM EHRLICHII AND APPLICATION THEREOF

Abstract

The present disclosure belongs to the technical field of plant pest control, and particularly relates to a Camellia sinensis endophyte Penicillium ehrlichii and an application thereof. The present disclosure relates to a novel endophyte separated from a Camellia sinensis body and identified as Penicillium ehrlichii by means of morphology and molecular biology. Biological activity evaluation indicates that Penicillium ehrlichii shows good antagonistic antifungal activity to common pathogenic fungi on several plants. It means that the strain used as a biocontrol fungus for phytopathogens can be applied to environmental-friendly and safe prevention and control of plant diseases.

Claims

1. A Camellia sinensis endophyte Penicillium ehrlichii, with a preservation number: CCTCC NO: M 2021635.

2. An application of the Camellia sinensis endophyte Penicillium ehrlichii according to claim 1 in preparation of a reagent in preventing and treating plant anthracnose wherein the plant anthracnose comprises at least one of tea anthracnose, melon anthracnose, pear anthracnose and apple anthracnose.

3. (canceled)

4. An application of the Camellia sinensis endophyte Penicillium ehrlichii according to claim 1 in preparation of a reagent in preventing and treating leaf spot disease of plants, wherein the leaf spot disease of plant comprises at least one of tea leaf spot disease and Curvularia oryzae leaf spot disease.

5. (canceled)

6. An application of the Camellia sinensis endophyte Penicillium ehrlichii according to claim 1 in preparation of a reagent in preventing and treating plant wilt disease wherein the plant wilt disease comprises at least one of cotton wilt disease and watermelon wilt disease.

7. (canceled)

8. An application of the Camellia sinensis endophyte Penicillium ehrlichii according to claim 1 in preparation of a reagent in preventing and treating grey seedling disease or Sclerotinia of plants or fruit disease of plants, wherein the grey seedling disease of plants comprises grey seedling disease of grapes, the Sclerotinia includes Sclerotinia rot of colza, and the fruit disease of the plants includes at least one of melon anthracnose, pear anthracnose and apple anthracnose.

9. (canceled)

10. A biological control inoculant, comprising the Camellia sinensis endophyte Penicillium ehrlichii according to claim 1.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0026] FIG. 1 shows inhibitory rate of an endophyte P. ehrlichii to 10 phytopathogens.

[0027] FIG. 2 shows an antifungal effect of an endophyte P. ehrlichii antagonistic to mycelial growth of an original pathogenic fungus of melon anthracnose for 7d.

[0028] FIG. 3 shows an antifungal effect of an endophyte P. ehrlichii antagonistic to mycelial growth of an original pathogenic fungus of tea zonate spot for 7d.

[0029] FIG. 4 shows an antifungal effect of an endophyte P. ehrlichii antagonistic to mycelial growth of an original pathogenic fungus of pear anthracnose for 7d.

[0030] FIG. 5 shows an antifungal effect of an endophyte P. ehrlichii antagonistic to mycelial growth of an original fungus of cotton wilt disease for 7d.

[0031] FIG. 6 shows an antifungal effect of an endophyte P. ehrlichii antagonistic to mycelial growth of an original pathogenic fungus of apple anthracnose for 7d.

[0032] FIG. 7 shows an antifungal effect of an endophyte P. ehrlichii antagonistic to mycelial growth of an original pathogenic fungus of grape grey mould for 7d.

[0033] FIG. 8 shows an antifungal effect of an endophyte P. ehrlichii antagonistic to mycelial growth of curvularia oryzae for 7 days.

[0034] FIG. 9 shows an antifungal effect of an endophyte P. ehrlichii antagonistic to mycelial growth of a pathogenic fungus of watermelon wilt disease for 7d.

[0035] FIG. 10 shows an antifungal effect of an endophyte P. ehrlichii antagonistic to mycelial growth of a pathogenic fungus of sclerotinia rot of colza for 7d.

[0036] FIG. 11 shows an antifungal effect of an endophyte P. ehrlichii antagonistic to mycelial growth of a pathogenic fungus of tea anthracnose for 7d.

DETAILED DESCRIPTION OF THE EMBODIMENTS

[0037] In order to better understand the present disclosure rather than to limit the scope of the present disclosure, all numbers and other numerical values representing dosage and percent used in the present disclosure shall be understood as being modified by a word “approximate” under all circumstances. Therefore, unless otherwise specified, numerical parameters listed in the description and attached claims are approximate values which may be changed according to different ideal properties attempted to be obtained. All numerical parameters shall be regarded as being obtained by a conventional rounding method according to reported effective numbers. In the present disclosure, “approximate” refers to a set value or a value within 10% of a scope, preferably within 5% of a scope.

[0038] Unless otherwise defined, temperatures in the embodiments of the present disclosure are normal temperatures. Normal temperature refers to a natural room temperature condition in the four seasons without additional cooling or heating treatment. Normal temperature is generally controlled at 10-30° C., 15-25° C. the best.

[0039] The present disclosure discloses a Camellia sinensis endophyte Penicillium ehrlichii and an application thereof, specifically shown in the following embodiments.

Example 1 Separation and Identification of an Endophyte Strain

[0040] Healthy Camellia sinensis root, stem and leaf tissues were taken, surface soil was cleaned with tap water, the healthy Camellia sinensis root, stem and leaf tissues were first immersed in 70% alcohol for 1 min, then placed in a 3% sodium hypochlorite solution to be immersed for 4 min, then placed in 70% alcohol to be rinsed for 1 s and finally washed with sterile water for 5 times, and 100 uL of the sterile water for last washing was absorbed and coated to a PDA culture medium to prepare a control culture medium.

[0041] A tissue separation method: a surface-disinfected Camellia sinensis tissue was sliced into pieces of 5 mm*5 mm, sections were attached to the PDA culture, the pieces were cultured at a constant temperature of 25° C. for 7d, observation was made every day, and new mycelia were picked out with a fungus moving ring and cultured independently.

[0042] ITS and 18S genetic locus sequencing was performed on the separated endophytic fungus, and BLAST comparison was made on the sequencing results on NCBI to obtain a similar strain named Penicillium ehrlichii (table 1).

TABLE-US-00001 TABLE 1 ITS sequencing identification result Reference Accession Domain Phylum Class Order Family Genus species NR_121229.1 Fungi Ascomycota Eurotiomycetes Eurotiales Aspergillaceae Penicillium Penicillium ehrlichii

Example 2 an Antagonistic Experiment of Endophyte

[0043] In the example, plate antagonistic experiments were performed on the endophytic fungus P. ehrlichii against nine plant disease pathogenic fungi (melon anthracnose (Colletorichum lagenerium (Pass.) Ell. et Halst), tea zonate spot (Pseudopestalotiopsis theae), pear anthracnose (Colletrichum gloeosporioides Penz.), cotton wilt disease (Fusarium oxysporum), apple anthracnose (Colletotrichum gloeosporioides (Penz.) Penz. et Sacc.), grape grey mould (Botrytis cinerea Pers), curvularia oryzae leaf spot disease (Culvularia lunata), watermelon wilt disease (Fusarium oxysporum), and sclerotinia rot of colza (Sclerotinia sclerotiorum (Lib.) de Bary)), diameters of growing mycelia were counted at different culture times, and the inhibitory rates were calculated. The specific contents are as follows:

[0044] 1. Plate Antagonistic Method:

[0045] A fungal cake was cut from the edge of a fungal colony of a pathogenic fungus and inoculated to a center point of a plate of the PDA culture medium with mycelia facing downwards, four endophyte P. ehrlichii fungal cakes were cut by the same method and inoculated to four directions that were 2.5 cm from the fungal cake of the pathogenic fungus respectively, a plate without P. ehrlichii fungal cakes was used as control, the fungal cake was cultured at a constant temperature of 25° C. for 7d, the diameter of the fungal colony of the pathogenic fungus was measured every day, and the inhibitory rate was calculated. A calculating method for inhibitory rate:

[0046] The inhibitory rate of an antagonistic fungus (%)=((the diameter of the fungal colony in the control group-5)−(the diameter of the fungal colony in the treatment group-5))/(the diameter of the fungal colony in the control group-5)

[0047] 2. An antagonistic experiment result: researches showed that on the seventh day, the inhibitory rates of P. ehrlichii to the selected 10 pathogenic fungi were 50% or above (FIG. 1), nearly 80% or above for prevention and treatment of anthracnose. Effect diagrams of the endophytic fungus P. ehrlichii to growth inhibition effect of mycelia of 10 phytopathogens were shown in FIG. 2-11. The effect diagrams of the control group were on the upper side and the effect diagrams of the treatment group were on the lower side.

[0048] The above mentioned is merely the preferred embodiment of the present disclosure and is not used to limit the present disclosure. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present disclosure shall be regarded as within the protection scope of the present disclosure.