KR101385557B1 - Chitosan and chitooligosaccharide composition for preventing and disease control of plant disease and method for increase of plant growth and disease control of plant by using the same - Google Patents

Abstract

The present invention relates to a chitosan composition for preventing and controlling phytopathogens containing chitosan, chitooligosaccharide, wood vinegar and brown rice vinegar as active ingredients, and to a method for controlling phytopathogens using the same, the composition according to the present invention is a phytopathogen, in particular tomato leaf fungus It is excellent in the control and prevention effect against the germ, the control effect against the phytopathogen is persistent, and is very effective in promoting the growth of plants.

Description

Chitosan and chitooligosaccharide composition for preventing and disease control of plant disease and method for increase of plant growth and disease control of plant by using the chitosan and chitooligosaccharide composition for preventing and controlling phytopathogens same}

The present invention relates to a chitosan and chitooligosaccharide composition for preventing and controlling phytopathogens and a method for controlling the same, more specifically vitamin C, grapefruit extract, to enhance the effective ingredients and effects of chitosan, chitooligosaccharide, wood vinegar and brown rice vinegar, It relates to a chitosan and chitooligosaccharide composition for the prevention and control of plant diseases and garlic plant growth containing garlic extract and onion extract and a method using the same.

Phytopathogens that are problematic in plant cultivation, such as potato and tomato late blight, cucumber, melon, watermelon, strawberry and rose powdery mildew, pepper and apple anthrax, and grass brown patch and large fat germs Mycelium penetrates through the pores and branches of leaves and often occurs when the humidity is high. Among them, tomato leaf fungal disease ( Fulvia fulva ) occurs after a pathogen has incubated for about 14 days after invasion, starting from the lower leaves. Severe outbreaks can cause significant damage to fruit production.

Conventional plant disease control mainly relies on chemicals. Examples of such chemicals include Tolylfluanid WP, Iprohydrate, Thiram + Vinclozolin WP, and Iminoctadin tris (albesilate) WP.

Recently, as the eco-friendly tomato cultivation area is expanded due to the increase in the consumption of eco-friendly agricultural products, a method of using microbial preparations is being progressed as a method for controlling tomato leaf fungus that can replace chemicals. For example, using Bacillus amyloliquefaciens, Cellulosimicrobium cellulans, etc. (Jeonju, Development of Fulvia fulva control microbial pesticides, Dong-A University) , 2006), using rkholderia cepacia (Ki Hyuk Choi, the development of microbial pesticides to control tomato leaf fungus (Fulvia fulva), Dong-A University, 2008).

In this regard, Korean Patent No. 10-0634874 discloses Streptomyces spp. Microorganisms having antifungal activity against fungal bacteria including tomato leaf fungal fungi and methods for controlling plant fungal diseases using the same, and Korean Patent Registration No. 10 -0587447 group Bacillus subtilis EB120 strain (KCTC 10578BP) or a culture, extract or spore thereof, comprising barley powdery mildew, cucumber powdery mildew, rice blast, tomato blight and wheat rust A microbial agent for controlling plant diseases selected from is disclosed.

Meanwhile, as another alternative to chemicals, chitosan and chitooligosaccharides, which are known as one of natural substances, can be used to control tomato leaf fungus through several previous studies. However, there are no studies on tomato leaf fungal diseases using chitosan and chitooligosaccharide.

Chitosan is a substance in which chitin polysaccharide is dissolved in acid, and its use in agriculture is greatly increased.As a result, it is reported that it plays various roles such as antimicrobial activity, soil improvement effect, plant growth promoting action, and self-defense mechanism. And new biomaterials are expected to be developed (Lee et al., 2005).

Chitosan is effective in controlling disease by antibacterial activity against various molds (Hadwiger, 1999; Stossel and Leuba, 1984) and by reducing the incidence of soil disease by soil treatment (Bengamou et al., 1992; El Ghaouth et al., 1992). It is effective in controlling Fusarium disease (Bell et al., 1998), and it also induces disease resistance due to the growth delay of the disease resistant strawberry bacterium (Eikemo et al., 2003). In addition, by increasing the density of non-pathogenic Fusarium in soil, it has the effect of reducing the density of pathogenic Fusarium and delaying its growth (Bell, 1998), it is effective in controlling Chinese cabbage, and also improves the growth of Chinese cabbage (Chang et al., 2002; Et al., 2000). In addition to direct disease control, chitosan is known to be involved in defense mechanisms against many plant pathogens, including stimulating plant chitinases (Barber et al., 1989; Bengamou et al., 1992; El Ghaouth et al., 1992), or inducing pisatin. (Kendra and Hadwiger, 1984), in particular, strengthen the defenses of plant cells, weakening the cell wall invasion of fungi and enhancing the vitality of plant cells (Bengamou et al., 1992). Chitooligosaccharides made of water-soluble chitosan by cutting the sugar chains of chitosan to 2-10 sugar levels have a bactericidal power as chitosan was prepared using organic acids, vitamin C and grapefruit extract and chitonase (Lee et al., 2004).

Chitosan is classified as a substance that can be used for eco-friendly bottle control in the domestic environmental promotion law, but currently it is not registered for bottle control. Chitosan is a polymer polysaccharide compound made by deacetylation of chitin, and is a polymer compound having a general number of single sugars linked to about 500. However, chitosan has a higher antimicrobial effect (Lee et al., 2000). Chitosan, which is currently used in agriculture, is used more frequently for crop growth than for disease control. The disease control effect on chitosan is known as a substance that reduces or prevents disease by inducing defense mechanisms and disease resistance against various plant pathogens including strawberry plague (Bohland et al., 1997; Bengamou et al., 1992; Eikemo et al. , 2003; Kendra and Hadwiger, 1984; Vender et al., 1998).

The effects of chitosan on disease defense mechanisms have been shown to induce the localization of wounds when treated on leaves of wound wheat (Bhaskara Reddy et al., 1999). The disease resistance and control effect prevented strawberry fungus and Rhizopus sp. Infections after harvest, including strawberry blight (Eikemo et al., 2003; El Ghaouth et al., 1992; Reddy et al., 2000; Romanazzi et al., 2000). Direct control of many other diseases is also known (Hadwiger and Beckman, 1980; Stossel, 1984), and they also increase the quality of fruit in the market by extending the life of harvested strawberries (Yoo et al., 1999). ). Chitosan also plays a role in the production of substances that defend against plant diseases (Hirano and Nagao, 1989). Increased phytoalexin production in maize and sprouted peanuts before harvesting, as well as inhibiting Aspergillus flavus mycelial growth and aflatoxin production (Bengamou et al., 1992; Bhaskara et al., 1999). Additional effects of chitosan also include plant growth (Hadwiger et al., 1999) and root development (e.g., 2000).

In this way, the use of chitosan for the control of the bottle is treated with chitosan in powder form directly to the soil like chitin (crab or shrimp shells soil treatment), but the chitosan acid dissolved in water is used to dilute the liquid. Recently, water-soluble chitooligosaccharides made of chitosan using chitinase and acid to cut sugar chains into sugars of less than 10 sugars have been used in farms for disease control and growth, but the effects on disease control are specifically tested. It is not.

Accordingly, the present inventors conducted extensive research to search for substances to solve the above-mentioned problems in environmentally friendly bottle control, and developed chitosan compositions using chitosan and chitooligosaccharides using wood vinegar and brown rice vinegar, and plants. The present invention was completed by assaying the effects and growth effects on the tomato leaf fungal pathogens.

Accordingly, an object of the present invention is a chitosan composition for plant disease prevention and control and plant growth promotion consisting of chitosan, chitooligosaccharide, wood vinegar and brown rice vinegar and vitamin C, grapefruit extract, garlic extract and onion extract used to enhance the effect. To provide.

In addition, another object of the present invention is to provide a plant disease control method using the chitosan composition for plant disease control.

An object of the present invention as described above is to prepare a chitosan composition using chitosan, chitooligosaccharide, wood vinegar and brown rice vinegar as raw materials, and using this to control, preventive effect, control against tomato leaf fungal pathogens The effect of control of the disease according to the lasting effect and concentration of use, while tomato The growth promoting effect was tested by confirming that the chitosan composition was effective in controlling tomato leaf fungal disease while also promoting tomato growth.

The present invention provides a chitosan composition for plant pathogen control and growth promoting chitosan, chitooligosaccharide, wood vinegar and brown rice vinegar as an active ingredient.

In the present invention, the term "phytopathogen" includes potato and tomato late blight, cucumber, melon, watermelon, strawberry and rose powdery mildew, pepper and apple anthrax, grass brown patch and large fat germ. This may include, but is not limited to, various other phytopathogens known in the art.

The chitosan composition for controlling phytopathogens according to the present invention is preferably 1 to 5% by weight of chitosan, 10 to 40% by weight of chitooligosaccharide, 10 to 65% by weight of vinegar and 10 to 65% by weight of vinegar, and 1 to 10% by weight of vitamin C. %, Grapefruit extract 5-30% by weight, garlic extract 10-30% by weight, onion extract 10-30% by weight.

In addition, chitosan composition for controlling phytopathogens according to the present invention 10 to 50% by weight of chitooligosaccharide, 10 to 65% by weight of vinegar and 10 to 65% by weight of vinegar, 1 to 10% by weight of vitamin C, 5 to 30% by weight of grapefruit extract , Garlic extract may be made of 10 to 30% by weight, onion extract 10 to 30% by weight,

In addition, the present invention provides a method for controlling phytopathogens using the chitosan composition for controlling the phytopathogens.

The present invention also provides a method for promoting plant growth using the chitosan composition for controlling phytopathogens.

According to the present invention, it is possible to promote growth and root development of fruit and vegetables such as potatoes, tomatoes, peppers, cucumbers, melons, watermelons and strawberries, and leafy vegetables such as lettuce and cabbage, using the composition. Treatment can also increase the density of soil microbes and promote the growth of turfgrass and cold grass.

In the present invention, the chitosan composition prepared using chitosan and chitooligosaccharides, which are safe for the environment and human beings, was developed as a crop protection agent that can be used in environmentally friendly organic farming and tested its effects. Chitosan is a water-insoluble substance, a polysaccharide whose molecular weight is usually about 7-8 million, but chitosan with a molecular weight of less than 50,000 by breaking down glycoside bonds, which are sugar chains of chitosan, has an antibacterial effect. Chitooligosaccharides (glucoseamine, ai 50-60%) together with vinegar were used to prepare two kinds of liquid chitosan compositions having different composition ratios.

The two chitosan compositions prepared in the present invention had a difference in the content of chitosan and chitooligosaccharide raw material input and the content of wood vinegar and brown rice vinegar, but did not show any difference in the effect of controlling the bottle. In the present invention, the use concentration of chitosan 100 times (10 ml / l) of SH-1-100 and SH-2-100 is more than 200 times (5 ml / l) of SH-1-200 and SH-2-200 Although the control effect was shown to be excellent, the difference in statistical significance (p = 0.05) showed a difference according to the test period, and it is necessary to reconsider the economics of the use concentration. In particular, chitosan is considered to have an effect of inducing disease resistance as the incidence rate is significantly lower than that of no treatment when spraying three times every 7 days before leaf fungal disease (Fig. 1). In addition, even in the packaging with increased leaf fungal disease, the disease control effect was also seen as a single spray in the test that investigated the control rate of the bottle every spray of chitosan (FIG. 3). Chitosan also has a therapeutic effect against leaf fungal disease. It can be seen as. As with other tests, however, the higher the concentration of chitosan used, the more effective the disease control effect was.In the case of SH-1-200 with low chitosan content, the disease control rate decreased as the incidence increased in the package. there was. Although the growth effect of chitosan was conducted in pot tests in greenhouses, it seems to have an effect on growth, and currently, most farmers use irrigation in soil for crop growth, which means that crop growth effect appears. When foliar spraying was over 50 times higher, young leaves collapsed, but at 100 times higher concentration, no weakening symptoms were observed. Therefore, in order to control the environment-friendly tomato leaf fungal disease of the present invention, spraying chitosan mainly in the prevention and early onset of disease is thought to be effective in preventing and treating the disease and help to increase the growth of tomatoes.

Chitosan composition according to the present invention is excellent in the control and prevention effect against phytopathogens, persistent effect on the control of phytopathogens, very effective in promoting the growth of various plants, very useful as a natural control agent for phytopathogen control to be.

1 is a graph showing the results of the tomato leaf fungal disease prevention effect test using the chitosan composition according to the present invention.
Figure 2 is a graph showing the results of the disease control sustaining effect test using the chitosan composition according to the present invention.
Figure 3 is a graph showing the disease control effect test results according to the use concentration using the chitosan composition according to the present invention.

Hereinafter, preferred embodiments of the present invention will be described in more detail with reference to Examples. However, the scope of the present invention is not limited to these examples.

Example  1: Preparation of the Chitosan Composition of the Invention

In this example, chitosan and chitooligosaccharide were dissolved in wood vinegar and brown rice vinegar to prepare a chitosan composition. In the preparation of the chitosan composition, the amount of wood vinegar and brown rice vinegar was adjusted to a range that did not affect the bottle control effect through a preliminary test, and the final prepared chitosan composition was named SH-1 and SH-2, respectively. (See Table 1).

Chitosan, the main raw material used to prepare the chitosan composition of the present invention, used a water-soluble chitosan oligosaccharide having a molecular weight of less than 50,000 and a molecular weight of less than 2,000, and these raw materials were supplied by Mirabiotech. Wood vinegar was purchased from Soochon Forest Co., Ltd. and brown rice vinegar from Millennial Food Co., Ltd.

Preferred mixing amount according to the present invention was prepared as shown in Table 1 below.

designation Formulation Spray amount (mL / L) Active ingredient (mg / kg) SH-1-200 Liquid (2 g of chitosan, 20 g of chitooligosaccharides,
20 ml of wood vinegar, 40 g of brown rice vinegar) 5 Total Glucosamine: 600 SH-1-100 10 Total Glucosamine: 1,200 SH-2-200 Liquid (2 g of chitosan, 20 g of chitooligosaccharides,
40 ml of wood vinegar, 40 g of brown rice vinegar) 5 Total Glucosamine: 600 SH-2-100 10 Total Glucosamine: 1,200

Experimental Example  One : Leaf fungus  Package test for control.

In this Experimental Example, a packaging test for tomato leaf fungal disease control and prevention was conducted using the chitosan composition prepared in Example 1.

The field trial was conducted in a farm house in the spring, and tomatoes (super-dotaerang) were purchased from a nursery company (blue seedlings) and leased farm packages. The chitosan compositions SH-1 and SH-2 were sprayed at the concentrations shown in Table 1.

Experimental Example  1-1: Leaf fungus  Control effect test

Chitosan compositions SH-1 and SH-2 of Example 1, which were developed for the control of eco-friendly leaf fungal diseases in tomato growing houses in winter, were sprayed three times every 7 days at the beginning of the onset to investigate the control effect on leaf fungal diseases. . The incidence was checked after 7 days and the test was performed twice.

 As a result, as shown in Table 2 below, the disease control effect on the chitosan compositions SH-1 and SH-2 showed similar control effects in two tests. Although there was no difference in the control effect between the SH-1 and SH-2 formulations and the treatment concentration in the first trial, in Test 2, there was no difference between the formulations, but there was a significant difference (p = 0.05) between the formulations in the treatment concentration.

Experimental Example  1-2: Leaf fungus  Preventive effect test

In order to investigate the preventive effect of leaf fungal disease, after spraying SH-1 and SH-2 three times every 7 days to tomatoes without leaf fungal disease, as shown in FIG. 1 until 14 days, 28 days, and 35 days Three incidence rates were examined.

As a result, almost no disease occurred in all treatments until 14 days after treatment, but the incidence was lower in all treatments compared to no treatment on day 28, and there was no statistically significant difference (p = 0.05) between SH-1 and SH-2. Did. However, the 35-day survey showed significant differences in treatment periods. The incidence of untreated group was three times higher than that of SH-1 and SH-2. The SH-1-100 and SH-2-100 treatments with high chitosan content showed lower incidence than the SH-1-200 and SH-2-200 treatments with low chitosan content.

Experimental Example  1-3: Bottle  Sustainability Test

Three days after SH-1 and SH-2 spraying at the beginning of tomato leaf fungal disease, 7 days after spraying the preparation to investigate how long the chitosan (SH-1 and SH-2) spraying sustains the suppression of leaf fungal disease (7 days, 14 days, 21 days), the disease control effect was examined for 3 weeks.

As a result, as shown in Figure 2, the treatment of SH-1 and SH-2 treatment was significantly reduced compared to the untreated treatment until 21 days after spraying, the effect of the chitosan composition of the present invention was observed for 21 days continuously, 14 days Since then, the incidence rate has tended to increase significantly. In particular, the treatment of SH-1 and SH-2 showed a higher incidence of disease than the control drug, and the effect of inhibiting the onset was lower. However, the treatment of SH-1-100 and SH-2-100 with high chitosan content Was not significantly different from the control drug, indicating that Chitosan's effect persists.

Experimental Example  1-4: According to the concentration of use Bottle  Effect test

The incidence rate was irradiated before spraying every time while spraying 100 times liquid (SH-1-100) and 200 times liquid (SH-1-200) three times every 7 days on the chitosan composition SH-1 of the present invention (after spraying once, twice After spraying and after 3 sprays) bottle control was converted into%. The incidence of leaf fungal disease is zero incidence and lesion area ratio is 40.1% or more and is divided into 4 and 0 by 4 coefficients. It converted into. The morbidity rate was converted into leaf mold morbidity rate = non-treated morbidity (%) s treated morbidity (%) × 100 / untreated morbidity (%).

As a result, as shown in Fig. 3, SH-1 had a bottle control effect by one spray, and the bottle control effect was high at a high concentration of spray. The 100-fold sprayer (SH-1-100) had higher bottle control effect than the 200-fold sprayer, and the effects of chitosan showed a similar tendency to the control drug. , 200-fold spraying port (SH-1-200) showed a tendency to decrease the bottle control effect when spraying two and three times than the bottle control rate when spraying once.

Experimental Example  2: for tomato growth effect Port test

In this Experimental Example, a pot test for tomato growth effect was performed using the chitosan composition prepared in Example 1.

Experimental Example Tomatoes (Super Taurant) seedlings used were purchased from a nursery company (blue seedlings), and the soil (baroque) and soil were 2: 1 (w / w) in pots (20 cm in diameter × 28 cm in height) in a greenhouse. The mixture was formulated. After seeding, seedlings were grown for 15 days, then sprayed chitosan three times at 7-day intervals and examined for growth after 7 days. In the growth effect investigation, the growth amount (growth) representing the growth level was obtained by subtracting the growth amount before the chitosan spraying and the amount after the three sprayings. Each survey was analyzed statistically using SAS program (Jump 6.1).

As a result, as shown in Table 3, both formulations showed a similar growth promoting effect on the top height, biomass, stem diameter, and root biomass weight of the tomato.

a, amount of growth: length before spraying-length after spraying

b, ±: standard deviation

c, lowest significance

Claims (6)

1 to 5% by weight of chitosan having a weight average molecular weight of less than 50,000, 15 to 40% by weight of chitooligosaccharides having a weight average molecular weight of less than 2,000, 20 to 40% by weight vinegar and 15 to 40% by weight of brown rice vinegar,
The glucosamine, which is an active ingredient generated from the chitosan and chitooligosaccharide, comprises 1,200 mg or more of total glucosamine with respect to 1 kg of the composition.
According to claim 1, wherein the composition further comprises vitamin C, garlic extract and onion extract,
1 to 5% by weight of chitosan, 15 to 30% by weight of chitooligosaccharide, 20 to 40% by weight of vinegar and 15 to 30% by weight of brown rice vinegar, 1 to 10% by weight of vitamin C, 5 to 28% by weight of grapefruit extract, 10 to 10% of garlic extract Leaf fungal disease (Fulvia fulva) bacteria prevention and control composition of tomato, characterized in that it comprises 30% by weight and 10-30% by weight of onion extract.
delete delete Claim 1 or 2 of the tomato leaf fungal disease (Fulvia fulva) at the beginning of the development of tomato leaf fungal disease prevention and control, characterized in that spraying at a daily spread of more than 10 ml / L every 7 days Way.
The method of claim 5, wherein the spraying is performed three times at intervals of 7 days.

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