KR102090758B1 - Composition for promoting growth of plant and Method for promoting growth of plant - Google Patents

Abstract

The present invention relates to a composition for promoting plant growth, comprising at least one selected from the group consisting of a culture medium, culture filtrate and spores of Isaria javanica pf185 KACC93241, and to a method for promoting plant growth using the composition, wherein the composition is eco-friendly, is very useful when being easily obtainable, and can contribute to the development of plant related businesses.

Description

Composition for promoting growth of plant and method for promoting growth of plant

The present invention relates to a composition for promoting plant growth and a method for promoting plant growth.

Insect pathogenic fungi have the potential to control pests. These fungi are commercially available as biopesticides for controlling arthropod pests in various crop systems. Recent studies have shown that insect-pathogenic fungi have a positive effect on plant health through induction of systemic resistance to plant diseases and abiotic stress, promotion of crop growth, and direct inhibition of plant pathogens. Consequently, these entomopathogenic fungi represent an attractive topic in accumulated pest management studies. Their cross-kingdom effect indicates that in protecting crops, insect pathogenic fungi can have better effects than simply pest control.

Several fungal insect pathogens, including Beauveria spp., Metarhizium spp., And Lecanicillum spp., Promote plant growth after colonization of the host plant. . Proteomic analysis of palm trees colonized by the insect-pathogenic fungi, Beauveria bassiana and Lecanicillium spp., Shows that photosynthesis, energy metabolism, induction of plant defense and stress protection proteins are improved. Show that. Mechanisms of plant growth promotion may include changes in plant growth hormones or improved nutrition. For example, colonization of M. anisopliae on soybeans grown in saline conditions reduces the production of abscisic acid and enhances the production of jasmonic acid. Insectogenic M. robertsii secretes a siderophore that can promote iron nutrients in the host plant in culture conditions free of iron. Changed plant nutrients can be observed in colonized plants of B. bassiana . Microbial volatile compounds also enhance plant growth. The isolate of Fusarium oxysporum produces volatile organic compounds that affect plant growth. Rhizobacteria such as Bacillus subtilis and Pseudomonas chlororaphis produce 2R, 3R-butanediol, which promotes plant growth and induces systemic resistance to biotic or abiotic stresses.

In the present invention, the growth of plants by the insect pathogenic fungus Isaria javanica pf185 was investigated. The fungus has a biocontrol effect against insects and plant diseases. In addition, we tested whether this fungal metabolite affects plant growth. I. It was confirmed that the culture medium, culture filtrate, and spores of javanica pf185 promote growth.

Korean Patent Publication No. 1997-0058555

An object of the present invention is to provide a composition for promoting plant growth comprising at least one selected from the group consisting of a culture medium, a culture filtrate, and spores of Isaria javanica pf185 KACC93241.

In addition, an object of the present invention is to provide a method for promoting plant growth by spraying at least one selected from the group consisting of a culture medium, a culture filtrate, and spores of Isaria javanica pf185 on a plant or a planted soil.

1. Isaria javanica pf185 ( Isaria javanica pf185) A composition for promoting plant growth, comprising at least one selected from the group consisting of culture medium, culture filtrate, and spores of KACC93241.

2. In the above 1, wherein the plant is at least one selected from the group consisting of pepper, tomato potato, bell pepper, goji, tomato, tobacco, a composition for promoting plant growth.

3. In the above 1, wherein the plant is tobacco ( Nicotiana ), a composition for promoting plant growth.

4. The composition of 1 above, wherein the growth is growth of at least one site selected from the group consisting of shoots, roots, and stems.

5. A method for promoting growth of plants, including; spraying the composition of the above 1 onto plants or soil planted in plants.

6. The method of 5 above, wherein the plant is at least one selected from the group consisting of pepper, tomato potato, bell pepper, goji berry, tomato, and tobacco, and the method for promoting plant growth.

7. The method of 5 above, wherein the plant is tobacco ( Nicotiana ).

8. The method of 5 above, wherein the growth is growth of at least one site selected from the group consisting of shoots, roots, and stems.

The composition or method of the present invention is that the composition comprising at least one selected from the group consisting of a culture medium, a culture filtrate, and spores of the Isaria javanica pf185 strain has an effect of promoting plant growth, so that an environment-friendly composition is used. Therefore, it has an advantage that can be used in various industries.

1 is a view confirming the effect of the culture medium of Isaria javanica pf185 on the growth of tobacco ( Nicotiana tabaccum cv. Xanthi) plant in the pot.
2 is a view confirming the effect of the culture filtrate of Isaria javanica pf185 on the growth of tobacco ( Nicotiana tabaccum cv. Xanthi) plants in the pot.
3 is a view confirming the effect of direct treatment of spores of Isaria javanica pf185 on the growth of tobacco ( Nicotiana tabaccum cv. Xanthi) plants in pots.
4 is a view confirming the effect of indirect treatment of spores of Isaria javanica pf185 on the growth of tobacco ( Nicotiana tabaccum cv. Xanthi) plants in pots.

Hereinafter, the present invention will be described in detail.

The present invention relates to a composition for promoting plant growth, comprising at least one selected from the group consisting of a culture medium, a culture filtrate, and spores of Isaria javanica pf185 KACC93241.

The culture medium is a mixture of nutrients required by the culture medium to cultivate the tissues of microorganisms or animals and plants as a main component, and mixed with substances for special purposes, and contains microorganisms and metabolites of microorganisms. Therefore, the culture solution of Isaaria Javanica pf185 is a liquid or solid substance that contains all of the nutrients for the culture of Isaaria Javanica pf185, the corresponding strain, and the metabolites of Isaaria Javanica pf185.

The culture filtrate is a filtrate removed by filtering the cells in a liquid medium in which microorganisms are cultured. The culture filtrate of Isaria japonica pf185 means that only the isaria japonica pf185 cells are removed from the culture medium in which Isaria japonica pf185 is cultured. do.

The spore refers to germ cells formed by fungi or plants as a means of asexual reproduction, and the spores of Isaria japonica pf185 are germ cells of isaria japonica pf185.

The plant may be a family of branches. Examples of the branch plant may be, for example, red pepper, tomato potato, green pepper, goji, tomato, tobacco, etc., but is not limited thereto. Further, the plant may preferably be tobacco ( Nicotiana ), and plants belonging to the tobacco ( Nicotiana ) include, for example, Nicotiana acuminate, Nicotiana alata, Nicotiana attenuata , Nicotiana benthamiana, Nicotiana clevelandii, Nicotiana excelsior, Nicotiana forgetiana, and the like Nicotiana glauca, Nicotiana glutinosa, Nicotiana langsdorffii , Nicotiana longiflora, Nicotiana obtusifolia, Nicotiana paniculata, Nicotiana plumbagifolia, Nicotiana quadrivalvis, Nicotiana repanda, Nicotiana rustica, Nicotiana suaveolens, Nicotiana sylvestris, cultivated tobacco (Nicotiana tabacum), Nicotiana tomentosa, more Preferably, it may be cultivated tobacco ( Nicotiana tabacum ), but is not limited thereto.

The growth means that the number of cells constituting the organism increases, and the size of the organism increases or the weight increases. For example, the weight or length of the plant cell shoots, roots, stems, etc. becomes heavier or longer. , May be an increase in root hair, and the like, preferably, may be growth of at least one site selected from the group consisting of shoots or roots and stems, but is not limited thereto.

The composition for promoting plant growth of the present invention can directly promote the growth of a plant by directly treating the plant, and indirectly, the volatile material may be contacted with the plant to promote plant growth, but is not limited thereto. .

In addition, the present invention relates to a method for promoting growth of a plant comprising; spraying the composition on a plant or soil planted therein.

When the composition is sprayed on a plant, the growth of the plant may be promoted according to the direct absorption of the plant, but is not limited thereto.

When the composition is sprayed on the plant-planted soil to promote the growth of the plant, the composition in the plant-planted soil may be absorbed by the plant through the soil, and the material volatilized in the air is absorbed by contact with the plant. It may be, and may be absorbed by being directly in contact with the plant in the air when spraying, but is not limited thereto.

The plant may be a plant belonging to the tobacco genus, as described above, more preferably, it may be a cultivation tobacco ( Nicotiana tabacum ), but is not limited thereto.

The growth may be growth, such as weight or size (length) of each part of the plant, such as a shoot, root, or stem, as described above, and preferably growth of at least one site selected from the group consisting of roots and stems. However, it is not limited thereto.

Hereinafter, examples will be described in detail to specifically describe the present invention.

Example

1. for tobacco plants Isaria javanica  Preparation of experiments of pf185 culture, culture filtrate, and spore growth promoting effect

(One) Isaria javanica  Cultivation of pf185

Isaria javanica pf185 (KACC93241P) was purchased from the National Agricultural Promotion Agency (Wonju, Korea). The pf185 strain was cultured on potato agar (Patoto dextrose agar, PDA; Difco Inc. Detroit, MI, USA).

(2) Preparation of tobacco plants

Nicotiana tabacum L. cv. Xanthi seeds were surface sterilized using ethanol and sodium hypochlorite solution (Han, Anderson, et al., 2006). Murashig and Skoog (MS, Sigma-Aldrich Inc., St. Louis, MO, added 0.5% (w / v) agar and 3% (w / v) sucrose (Murashige and Skoog, 1962) to sterilized tobacco seeds USA) Petri dishes (SPL Life Science Co., Pecheon, Korea). Tobacco plants were grown for 2 weeks under a 40-W fluorescent lamp (2,000 lux, 80 μmol photons m ^-2 s ^-1 ) with a period of 16 hours of light and 8 hours of darkness. The temperature was maintained at 25 ± 3 ° C and the humidity was 54 to 60%.

(3) Isaria javanica  Analysis of pf185 culture medium and culture filtrate to promote tobacco plant growth

Two weeks after planting tobacco plants, tobacco seedlings were planted on the side of I plates (100 x 15 mm ² , SPL Life Sci.) Containing MS solid medium. Three seedlings were planted per plate. PDA medium was spread with 0.1 ml of conidia (1Х10 ⁷ spores / ml) suspension and inoculated in distilled water 24 hours later. As a control, a PDA medium treated with only 0.1 ml distilled water was used. The inoculated I-plate was sealed with Parafilm, placed in a growth chamber under the above-mentioned fluorescent lamp for 2 weeks, and tobacco growth parameters were measured.

Root growth of the treated tobacco plants was confirmed by M165 FC (Leica Microsystems, Tokyo, Japan) microscope.

Other tobacco seedlings were planted in pots containing 500 cm ³ of sterile, soil-free medium (chocoal: vermiculite: pearlite, 7: 3: 3, vol / vol) with one seedling transplanted per pot. These pots were given 10 ml of distilled water every 2 days. After 2 weeks of growth, the seedlings had mold.

For the treatment of the intact culture, I. javanica pf185 contained in PDB was shake cultured at 150 rpm in a shake incubator at 25 ° C. The culture solution was diluted with distilled water 1: 1 by volume. For preparation of cell-free supernatant (culture filtrate), fungal debris was removed by centrifuging the culture for 7 days and filtered through a 0.2 μm filter (Millipore Filter Corp., Bedford, MA, USA). The culture was incubated with distilled water on a 7-day-old plate. The suspension was filtered through two layers of sterile cheese cloth to remove mycelial debris.

Tobacco plants were treated with supernatant (culture filtrate) 4 times each 15ml per week. Intact culture medium (20 ml) was applied once. Distilled water or PDB was used for control treatment. The experiment was repeated twice with 6 plants per treatment. Plant growth factors during treatment were measured one week after the final treatment.

(4) Isaria javanica  Analysis of pf185 spores promoting tobacco plant growth

In the same manner as the effect analysis of the culture medium and the culture filtrate, two weeks after planting tobacco plants, tobacco seedlings were planted on the side of I plates (100 x 15 mm ² , SPL Life Sci.) Containing MS solid medium. Three seedlings were planted per plate. PDA medium was spread with 0.1 ml of a conidia (1Х10 ⁷ spores / ml) suspension and inoculated in distilled water 24 hours later. As a control, a PDA medium treated with only 0.1 ml distilled water was used. The inoculated I-plate was sealed with Parafilm, placed in a growth chamber under the above-mentioned fluorescent lamp for 2 weeks, and tobacco growth parameters were measured. Root growth of the treated tobacco plants was confirmed by M165 FC (Leica Microsystems, Tokyo, Japan) microscope.

Other tobacco seedlings were planted in pots containing 500 cm ³ of sterile, soil-free medium (chocoal: vermiculite: pearlite, 7: 3: 3, vol / vol) with one seedling transplanted per pot. These pots were given 10 ml of distilled water every 2 days. After 2 weeks of growth, the seedlings had mold.

To obtain spores to be treated on plants, fungal agar plugs were cultured in PDA plates at 25 ° C., and the culture was incubated with distilled water on a 7-day-old plate. The suspension was filtered through two layers of sterile cheese cloth to remove mycelial debris. Spore concentrations were determined using a hemocytometer (Marienfeld Superior, Lauda-Kφnigshofen, Germany) under a microscope (Olympus BX41, Japan) and adjusted to 1 x 10 ⁸ spores / ml.

Tobacco plants were treated with spore suspensions 4 times every 15 ml each week, spore suspensions were directly treated to plants to confirm the effect of spores promoting tobacco growth, and spore suspensions were used to confirm the tobacco growth promoting effect of volatile substances contained in the spore suspensions. It was treated on the side of the medium so as not to touch the plants. In addition, distilled water or PDB was used for the control treatment. The experiment was repeated twice with 6 plants per treatment. Plant growth factors during treatment were measured one week after the final treatment.

(5) Growth analysis of tobacco plants

To determine the effectiveness of treatment on tobacco plants, the chlorophyll content of the shoots was measured using a Chlorophyll meter (SPAD-502Plus, Minolta Camera Co., Osaka, Japan) at least 10 times of 3-4 fully developed tobacco shoots. Measured at different locations. After that, each plant was cultivated. The weight of plants was measured using analytic balance (A & D Korea Limited, Seoul, Korea). The diameter and length of the plant stem were measured using a digital ruler (Mitutoyo Corp., Kanagawa, Japan). The density of the portal root was measured using image analysis software ASSESS 2.0 (APS Press, St. Paul, M, USA). Six plants per treatment were repeated twice.

(6) Statistical analysis

Isaria javanica pf185 data for analyzing plant growth effects by culture and filtrate were analyzed by using SPSS (Statistical Package for Social Sciences, SPSS Inc., Chicago, IL, USA) of Student Software Package Program Version 23. Significance test was done at p <0.05 level. Average values were made within the standard deviation.

2. For tobacco plants Isaria javanica  Experimental results of pf185 culture, culture filtrate, and spores promoting plant growth

(One) Isaria javanica  Confirmation of the effect of promoting culture of tobacco plants on the culture medium and filtrate of pf185

In pot experiments, tobacco was promoted by fungi and its metabolites. Seedlings treated with the complete fungal culture of pf185 were more grown than control plants treated with 1/2 PDB without culture (FIG. 1). I. The root length (p = 0.004), shoot weight (p = 0.02), and shoot height (p = 0.001) increased significantly to 28 days after soaking the roots in javanica pf185 complete culture. There was no effect. Even when the culture filtrate was treated, it was confirmed that growth of shoots was promoted (FIG. 2): height of shoots (p = 0.006), weight of shoots (p = 0.003). These results clearly show that the culture or culture filtrate of I. javanica pf185 promotes the growth of tobacco.

1 and 2 showing the results of the experiment, the 14-day-old tobacco seedlings treated with the fungus culture and the culture filtrate mentioned in the experiment method were treated for 5 weeks. PDB or water treated with the same amount was used as a control. Six seedlings per repetition were used and showed the mean and standard deviation based on 2 times. When statistics were compared between controls and growth between treatments (Student's t test, α <0.05), they were provided as substantial P-values, and images of tobacco seedlings are inserted in FIGS. 1 and 2.

(2) Isaria javanica  Confirmation of the direct effect of spores of pf185 on the growth of tobacco plants

In the pot experiment, it was confirmed that the seedlings of the tobacco plants treated with the spore suspension increased in the weight of the shoot (p = 0.02) and the root weight (p = 0.008) (FIG. 3). These results clearly show that spores of I. javanica pf185 promote the growth of tobacco.

Referring to FIG. 3 showing the experimental results in detail, the 14-day-old tobacco seedlings treated with the roots of the fungal spore suspension mentioned in the experimental method were treated for 5 weeks. PDB or water treated with the same amount was used as a control. Six seedlings per repetition were used and showed the mean and standard deviation based on 2 times. When comparing the control and growth statistics between treatments (Student's t test, α <0.05), it was provided as a substantial P-value.

(3) Isaria javanica  Confirmation of indirect tobacco plant growth promoting effect of spores of pf185

The effect of the volatiles produced from pf185 on tobacco using the I plate was confirmed (FIG. 4). Compared to seedlings grown without fungi (control), the growth of tobacco treated with fungal volatiles was greatly accelerated (Tukey's test with alpha value <0.000001). The height of the shoots and the length of the roots were increased by 1.5 times compared to the control group by volatiles; The weight of the shoots and the weight of the roots increased more than twice (Fig. 4). This change was also observed in the leaf chlorophyll (p = 0.000001) and stem diameter. Both lateral roots and root hair formation increased. Therefore, it was found that the volatile material obtained from I. javanica pf185 has the effect of promoting growth with changes in the root structure of tobacco and improvement of biomass.

4, which is the result of this experiment, is data for plants irradiated 2 weeks after treatment in the I plate. Plant growth parameters of tobacco grown without fungi are shown on average. Statistical differences for the response of the control group evaluated by Student's t test were shown as three stars (α <0.001). A photograph of tobacco seedlings and tobacco root hair formation treated with a spore suspension of I. javanica pf185 is shown in FIG. 4. Root hair formation pictures are micrographs.

Claims (8)

Isaria javanica pf185 ( Isaria javanica pf185) at least one selected from the group consisting of peppers, tomatoes, potatoes, bell peppers, goji berries and tobacco, including at least one selected from the group consisting of culture, filtrate and spores of KACC93241 Composition for promoting plant growth.
delete The method according to claim 1, The plant is tobacco ( Nicotiana ), a composition for promoting plant growth.
The method according to claim 1, The growth is a growth, at least one site selected from the group consisting of shoots, roots and stems, plant growth promoting composition.
A method of promoting growth of at least one plant selected from the group consisting of pepper, tomato, potato, bell pepper, goji and tobacco, comprising; spraying the composition of claim 1 onto a plant or soil in which the plant is planted.
delete The method of claim 5, wherein the plant is tobacco ( Nicotiana ).
The method of claim 5, wherein the growth is growth of at least one site selected from the group consisting of shoots, roots, and stems.

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