<i>Fusarium solani </i>and use of same in prevention and treatment of southern blight of <i>Dendrobium officinale </i>Kimura et Migo

Abstract

A Fusarium solani strain and a use thereof in the control of southern blight disease of Dendrobium officinale Kimura et Migo, and the strain JSNL007-2 has been identified as Fusarium solani and has been deposited in China Center for Type Culture Collection on Mar. 10, 2017 with a deposit number of CGMCC NO. 13687. The Fusarium solani JSNL007-2 of the present invention has a significant effect on the disease resistance of Dendrobium officinale Kimura et Migo, especially for improving the ability of Dendrobium officinale Kimura et Migo to resist southern blight disease, and has a certain effect on reducing the use of chemical pesticides, which is an important means to ensure food safety.

Claims

1. A method for improving the resistance of Dendrobium officinale Kimura et Migo plants to southern blight disease comprising a step of administrating an effective amount of a microbial inoculant comprising Fusarium solani to a Dendrobium officinale Kimura et Migo plant subject in need of improving resistance to southern blight disease.

2. The method according to claim 1, wherein the plant subject is a surface of a Dendrobium officinale Kimura et Migo seedling, and the effective amount of the microbial inoculant is 5-8 grams per the Dendrobium officinale Kimura et Migo seedling.

3. The method according to claim 2, wherein relative water content of the plant is maintained above 60% for 6-10 days after the microbial inoculant is administrated on the surface of the Dendrobium officinale Kimura et Migo seedling.

4. The method according to claim 1, wherein the Fusarium solani is Fusarium solani strain JSNL007-2, which has been deposited in China Center for Type Culture Collection on Mar. 10, 2017 with a deposit number of CGMCC NO. 13687.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 is a colony photograph of the strain JSNL007-2 of the present invention;

(2) FIG. 2 is an analysis diagram of the ability of mycorrhizal Dendrobium officinale Kimura et Migo to resist southern blight disease with the strain JSNL007-2 of the present invention.

DETAILED DESCRIPTION

(3) The present invention is further illustrated below with reference to the drawings and examples.

Example 1

(4) A southern blight disease resistant strain JSNL007-2 of Dendrobium officinale Kimura et Migo is isolated and screened from plants with good growth conditions in the Dendrobium officinale Kimura et Migo planting area in Jurong City, Jiangsu Province, and cultured on a PDA plate, of which the front mycelia are off-white, fleece, and are attached to the surface of the substrate; the reverse mycelia are white to pale yellow with slight pigmentation, as shown in FIG. 1.

(5) The full sequence of the ITS region of the strain JSNL007-2 is amplified and sequenced, and the full sequence of the rDNA ITS sequence obtained by PCR amplification is shown in SEQ ID No 1. By alignment on genebank, the results show that the strain JSNL007-2 has the closest homology with Fusarium solani strain, and has 99% homology with the strains Fusarium solani LYF019 and Fusarium solani CDR3P2F2, combining the morphological and physiological and biochemical characteristics, the strain JSNL007-2 is initially identified as Fusarium solani and named as Fusarium solani JSNL007-2, which belongs to Fusarium genus of Tuberculariaceae of Fungi Imperfecti subgenus. The strain JSNL007-2 has been deposited in China Center for Type Culture Collection of Institute of Microbiology, Chinese Academy of Sciences located in Beijing, China, on Mar. 10, 2017 with a deposit number of CGMCC NO. 13687.

Example 2

(6) The microbial inoculant produced by the Fusarium solani JSNL007-2:

(7) (1) preparing a solid medium using cottonseed husk dregs as culture substrate, ammonium nitrate as exogenous nitrogen source, and sucrose as exogenous carbon source, which was put into a autoclaving bag to sterilize for 20-25 minutes under a condition of 121° C. and a high pressure of 101.3 kPa, after the medium is cooled, inoculating the Fusarium solani strain JSNL007-2 under sterile conditions, and then performing artificial culture;

(8) (2) specific culture conditions for artificial culture are culturing for 5-8 days under a condition of 25-30° C., 8 hours of light and 16 hours of darkness per day, and pH of 6.0-6.5;

(9) (3) cutting the solid microbial inoculant cultured in step (2) into 1-1.5 cm3 blocks and setting aside.

Example 3

(10) Effect analysis of the microbial inoculant produced by the Fusarium solani JSNL007-2 on the ability of Dendrobium officinale Kimura et Migo to resist southern blight disease.

(11) (1) Artificial culture on PDA medium to obtain the sclerotia of southern blight disease pathogen;

(12) (2) inoculating the solid microbial inoculant cultured in Example 2 and the sclerotia of southern blight disease pathogen cultured in step (1) simultaneously on the cultivation substrate surface of the Dendrobium officinale Kimura et Migo seedling, in which the inoculation amount of the solid inoculant per Dendrobium officinale Kimura et Migo seedling is 5-8 g, and the number of the sclerotia of southern blight disease pathogen is 10;

(13) (3) after applying the inoculant, managing the Dendrobium officinale Kimura et Migo plants tested in step (2) under the same conditions, wherein the relative water content of the substrate is maintained above 60% for 6-10 days to ensure that the mycelia survive in the substrate;

(14) the temperature in the greenhouse is 28-30° C., the humidity is 40%-50%, each treatment is 100 strains, and three groups are repeated; at the same time, plants inoculated with only the sclerotia of southern blight disease pathogen and without Fusarium solani JSNL007-2 solid inoculant are used as controls (CK).

(15) (4) Monitoring the incidence degree, statistical incidence, and incidence index of southern blight disease every day after inoculation.

(16) Diseased plants grading criteria: disease grading is mainly based on the number of rotten roots after inoculation, and the initial grading criteria are:

(17) 0: normal;

(18) I: there are a few rotten roots;

(19) II: the number of rotten roots accounts for less than ¼ of the total number of roots;

(20) III: the number of rotten roots accounts for ¼-½ of the total number of roots;

(21) IV: the number of rotten roots accounts for ½-¾ of the total number of roots;

(22) IV: the number of rotten roots accounts for more than ¾ of the total number of roots.
Diseased index=Σ(number of diseased plants at each grading×representative value of the grading)×100/(total number of plants×representative value of the highest grading);
Diseased rate=(number of diseased plants/number of inoculated plants)×100%.

(23) The diseased rate and diseased index of the test plants not inoculated with JSNL007-2 and the plants inoculated with JSNL007-2 inoculant are shown in FIG. 2.

(24) It can be seen from FIG. 2 that within 30 days after artificial inoculation, the diseased rate and diseased index of Dendrobium officinale Kimura et Migo inoculated with Fusarium solani JSNL007-2 inoculant are significantly lower than those inoculated without Fusarium solani JSNL007-2 inoculant.