KR101199820B1 - The plant cultivating method from soil having a lot of salt - Google Patents

Abstract

The present invention relates to a method for growing a plant in soil containing a large amount of salt.

The method of cultivating a plant in soil containing a large amount of salt of the present invention is to cultivate a plant by foliaring a water-soluble chitosan decomposed product having a molecular weight of 10,000 Daltons or less or by cultivating the root of the plant, thereby maintaining a large amount of residue in the soil. It is characterized by increasing the plant yield by reducing the cell damage of plants caused by salts.

According to the present invention, there is provided a plant cultivation method which can increase the yield of the plant by reducing the damage of plant cells caused by overuse of chemical fertilizer or salt remaining in the reclaimed soil.

Chitosan degradant, foliar fertilization, root irrigation

Description

THE PLANT CULTIVATING METHOD FROM SOIL HAVING A LOT OF SALT

The present invention relates to a method for growing plants in soils containing a large amount of salt, and in particular, to reduce plant cell damage caused by salts remaining in soil such as reclaimed land, which can increase the yield of the plant. It relates to a plant cultivation method.

Salt accumulation in agriculture is an environmental factor that reduces crop productivity and limits the expansion of arable land. It is estimated that salt accumulation soils are more than 950 million hectares worldwide and are increasing every year.

This is equivalent to about 20% of the world's agricultural land, and the cause of salt accumulation in the soil is increasing due to climatic drying, deficient agricultural water and irrigation in irrigation.

Therefore, various solutions to the salt accumulation problem of the soil are being sought, and in particular, the improvement of the seedling method for the improvement of soil, the minimization of damage, and the development of flame resistant crops, etc. have been made.

The majority of crops are neutral crops (glycophyte) that are sensitive to salt accumulation and show growth disturbances and reduced yields.

The fatal effect of salt accumulation on plant growth is known to be due to the following factors.

1) Plant growth is greatly affected by water osmotic pressure due to low osmotic pressure of soil solution, 2) malnutrition, 3) ionic effects that harm plants, and 4) combined effects of all the above factors.

In plants, major metabolic processes, such as photosynthesis, protein synthesis, carbohydrate and fat metabolism, are affected by salting, resulting in growth disturbances and reduced yields.

The most active areas in various research fields to solve salt disorders are the development of salt-tolerant crops and the development of new varieties by traditional breeding and selection of salt-related genes using recent genetic engineering.

In particular, the development of tolerant crops using genetic engineering is possible by studying mechanisms related to the resistance and resistance to salt accumulation and multiplication of plants.

In nature, salt accumulation does not negatively affect all plants, and there are plant halophytes that include these mangroves, shrubs, grasses, and herbs. .

Among the saline-prone plants, in particular, the quince and rice plants are the main plant groups that understand the flame-resistance mechanisms of plants and are essential for the development of saline-tolerant crops.

The main reactions of salt-tolerant plants known to date include active osmotic control, selective absorption / migration of ions, distribution and active toxic ion release, and generation of toxic mitigating substances to resolve the osmotic and ionic imbalances of cells caused by over-salts.

Unlike traditional flame retardant breeding, modern genetic engineering technology has found various factors involved in such a resistance mechanism and introduced them into new crops, enabling the emergence of flame resistant rice, tomatoes, and rapeseed. It raises the possibility.

Apart from the development of flame resistant varieties by introducing recombinant genes using traditional breeding and genetic engineering techniques, there are relatively few studies on materials that prevent or reduce salt damage during plant cultivation.

In hydroponic cultivated roses, 50-100 ppm silicon (Si) reduces sodium chloride (NaCl) interference and reduces oxidative stress, which reduces malondialdehyde (MDA).

The salting mechanism is accompanied by oxidative stress, which increases the production of MDA (malondialdehyde), a product of peroxidation of phospholipids in the cell membrane, and attenuates the selective permeability of the cell membrane.

Therefore, the ideal method to reduce salt damage is likely to be able to inhibit the absorption of toxic ions in the plant outside the plant tissue and simultaneously reduce the oxidative stress through biochemical mechanisms in the plant.

In this respect, naturally derived chitin has several advantages.

Chitin is a polymer material in which N-acetyl-D-glucosamine is polymerized by β- (1-4) bonds. It is a component of crab shells, insect shells, mollusks, and fungal cell walls. Chitosan is usually obtained by deacetylating chitin. .

The industrial use of chitosan has begun to be studied in terms of waste disposal using the chelate properties of chitosan, and the research has been accelerated due to its ecological characteristics, which are not only abundant in nature but also non-toxic and biodegradable.

Moreover, as it is recently discovered that it is a variety of functional materials, research areas and applications are increasing.

The physiological activity of chitin / chitosan also has antimicrobial activity as well as the defense activation of the plant, and the inhibitory effect of chitosan on the growth of pathogenic microorganisms has been confirmed.

Other pathogens ( Rhizopus stolonifer, Botrytis cinerea, Pythium ultimum, Rhizoctonia solani, Fusarium graminearum ) were also found to have inhibitory effects.

As such, chitosan has been actively researched for antimicrobial activity and activation of self-defense function of plants.

In addition, the study of plant metabolism shows that salt stress using 100 mM of salt in pea root causes oxidative stress that accumulates free radicals, and increases the concentration and activity of G6PDH. At the same time, it has been reported that NADPH production is increased in plant cells.

In addition, the antioxidant activity is increased to remove the free radicals induced by salt damage, G6PDH has been reported to play an important role in overcoming the salt damage of peas.

However, although the mechanism and theoretical background of the plant metabolism are still reported, the main ingredient is a natural substance such as chitosan to prevent the damage of cells by inhibiting the absorption of salts toxic to plant cells in salty soils. However, there is no way to grow plants that increase yields.

The present invention to solve the above problems, by using a water-soluble chitosan decomposition product having a molecular weight of 10,000 Daltons or less to reduce the damage of plant cells caused by overuse of chemical fertilizers or salts remaining in reclaimed soil soil to increase the yield of plants The purpose is to provide a method for growing plants.

The method of cultivating a plant in soil containing a large amount of salt of the present invention is to cultivate a plant by foliaring a water-soluble chitosan decomposed product having a molecular weight of 10,000 Daltons or less or by cultivating the root of the plant, thereby maintaining a large amount of residue in the soil. It is characterized by increasing the plant yield by reducing the cell damage of plants caused by salts.

At this time, the chitosan decomposed product is characterized by being prepared by putting chitosan and chitosan degrading enzyme in an acid solution.

In addition, when the plant is grown, the chitosan decomposed product is diluted and prepared in liquid form, or dried and prepared in powder form, and then the amount of chitosan decomposed product is foliarly applied to plants 1 to 5 times by 10 to 500 mg per 3.3 m ² . It is characterized by the irrigation of the root of the plant.

According to the present invention, there is provided a plant cultivation method which can increase the yield of the plant by reducing the salt damage caused by overuse of chemical fertilizer or salt remaining in the reclaimed soil using water soluble chitosan degradants having a molecular weight of 10,000 Daltons or less. .

Chitosan used in the present invention is deproteinized from shellfish, mushrooms, and mollusc bones, and N-acetyl-D-glucosamine monomer obtained by decalcifying N-acetyl-D-glucosamine monomer is polymerized by adding alkali to chitin, which is a polymer polysaccharide β- (1,4). When the ratio of D-glucosamine generated by acetyl group falling off from acetyl-D-glucosamine is 70% or more, this is called chitosan (Korea Food and Drug Administration, Food Additive Code).

That is, chitosan is a polysaccharide composed of 70% or more of D-glucosamine and 30% or less of N-acetyl-D-glucosamine (Fig. 1).

Chitosan has the property of dissolving in organic acid or inorganic acid such as dilute hydrochloric acid, acetic acid, lactic acid, glutamic acid, ascorbic acid, sulfuric acid, phosphoric acid, malic acid, succinic acid, citric acid, fumaric acid, and industrially Has been applied in various ways.

However, chitosan has a very unstable property when various materials are mixed due to its strong cohesion and adsorptive power, and is insoluble in an alkali, so that application is difficult, and thus, chitosan having a low molecular weight is often used instead of chitosan having a high molecular weight.

There are two methods for lowering the molecular weight of chitosan, a chemical method using hydrochloric acid or nitric acid, and a biological method using enzyme.

The effect of a large amount of salt on the plant, in particular, lowers the selective permeation function of the cell membrane of plant cells, resulting in salt damage. Chitosan treatment has the effect of maintaining the original function by protecting the cell membrane from salt damage.

Therefore, the inventors of the present invention have conducted various studies in cultivating plants, and as a result, fertilize low-molecular-weight chitosan debris in the leaves of plants or irrigate them to the roots, resulting in overuse of chemical fertilizers or salts remaining in reclaimed soil. By reducing the harm, the plant can be grown in soils that contain large amounts of salts, thereby finding ways to increase plant yields.

Hereinafter, a method for growing plants in soil containing a large amount of salt of the present invention will be described.

How to grow plants in soils containing large amounts of salt

1. Preparation of Low Molecular Weight Chitosan Degradate

The chitosan decomposition method by enzyme treatment dissolves chitosan and then decomposes it by adding an enzyme.In this case, the chitosan dissolving agent is hydrochloric acid, acetic acid, lactic acid, glutamic acid, ascorbic acid, sulfuric acid, phosphoric acid, malic acid, succinic acid, citric acid, fumaric acid, or fumaric acid. Inorganic acid is used, and the solubilizer can be selected from organic acid or inorganic acid and used alone or in combination of two or more.The amount of acid is added at an equivalent ratio of at least 5% higher than that of chitosan, and then stirred well at 40 to 60 ° C. It is preferable to dissolve.

Chitosan degrading enzyme is prepared by adding 2 to 50 units of chitosan degrading enzyme per 10 g of chitosan to the prepared chitosan solution.

At this time, the difference in the decomposition time depends on the concentration of the chitosan solution, the decomposition temperature, and the amount of the chitosan degrading enzyme, the enzyme decomposition temperature is 30 to 60 ℃ depending on the substrate of the enzyme, the amount of chitosan degrading enzyme is 2-50 units per 10 g of chitosan In general, it is preferable to add 1 unit per 1 g of chitosan.

2. Plant cultivation

Using the chitosan decomposed product prepared above, foliar fertilization is applied to plants 1 to 5 times, or cultivated in relation to the roots of plants.

In addition, after diluting the low molecular weight chitosan degradation product having a glucosamine content of 40 to 80% in liquid form, or drying and preparing it in powder form, the amount of chitosan decomposition product used is sprayed or irrigated by 10 to 500 mg per 3.3㎡. Grow plants.

In addition, the plant is used in one selected from vegetables, grapes, rice, green tea, herbal plants, peppers, tomatoes.

At this time, it can be seen that the salt disturbance is reduced by inhibiting the accumulation of salt in the plant, the yield of the plant is also increased.

Hereinafter, the present invention will be described in detail through Examples and Experimental Examples, but these are not intended to limit the scope of the present invention.

Example 1 Plant Growing Method 1

1. Chitosan Degradation Preparation

8.5 L of water was added to 350 g of 98% acetic acid, and then 840 g of chitosan having a deacetylation degree of 95% was added thereto, followed by complete dissolution while stirring at 45 ° C. for 2 hours to prepare a chitosan solution.

The chitosan solution was prepared by adding 1,000 units of chitosan degrading enzyme (chitosan N.acetylglucosaminohydrolase EC 3,2,1,132; Sigma) and then decomposing at 45 ° C. for 20 hours to dissolve 9.1 liters of chitosan with 7.8% glucosamine content. Prepared.

As a result of analyzing the molecular weight of the prepared chitosan decomposition product using GPC, the highest molecular weight was 9,800 Daltons, and the average molecular weight was 3,900 Daltons, and it was confirmed that the chitosan decomposition product was prepared in the low molecular weight chitosan decomposition product.

2. Plant cultivation

The low molecular weight chitosan decomposed product thus prepared was diluted in water to prepare a liquid form.

40 mg of the low molecular weight chitosan degradant diluent prepared in the above liquid form per 3.3 m 2 of reclaimed soil was cultivated by spraying five times on the leaves of rice plants.

Example 2 Plant Cultivation Method 2

Prepared in the same manner as in Example 1, but diluted 1,500 times the low molecular weight chitosan decomposed product having a glucosamine content of 10% cultivated hot radish by fertilizing 20 mg per 3.3㎡ of soil at the plant house 4 times at 7 days intervals. It was.

Experimental Example 1 Effect Measurement Through Leaves

In order to verify the correlation between the low molecular weight chitosan degradation product and the effect of antioxidant activity to prevent the oxidative stress of plants and the yield enhancement effect, the rice grown in Example 1 was pot (diameter 10 cm, height 10 cm) After sowing, 0.25% NaCl treatment and chitosan digestion were carried out to measure the antioxidant enzyme activity, MDA, Na concentration and growth in the leaves after 14 days.

As a result, it was shown in Table 1 below and FIG.

Na concentration in leaves
Na content in leaf
(mg / g dry weight) MDA
(nmol / mg protein) Guaicol peroxidase (GPX)
nmol / mg
protein / min Plant growth degree
Plant growth
mg / plant Control 1.77 0.48 3721 95.1 Control + Nacl 18.55 0.81 4960 85.4 Polymer Chitosan 9.83 0.61 3695 94.8 100-fold dilution of low molecular weight chitosan 4.55 0.50 3464 99.5 500-fold dilution of low molecular weight chitosan 4.37 0.59 3692 100.4

MDA is a representative indicator that appears when the cell membrane is damaged by ROS, and since GPX uses hydrogen peroxide (H ₂ O ₂ ) as a substrate, an increase in GPX indicates a significant increase in oxidative stress.

Thus, as shown in Table 1, NaCl treatment was confirmed to increase the MDA (Malondialdehyde) and GPX (Guaicol peroxidase) activity to increase the active oxygen species (Reactive Oxygen Species) and cell damage adversely affecting plant growth.

On the other hand, the growth rate of the plants was 17.0%, 5.5% higher than the control + NaCl treatment and chitosan treatment treated with low-molecular chitosan decomposition products.

Therefore, it can be predicted that the low molecular weight chitosan degradant reduced salt disturbance by inhibiting the accumulation of salt in the plant, and also affects the degree of growth of the plant, thereby increasing yield.

Experimental Example 2 Effect of Low Molecular Chitosan Degradation Treatment on Plant Growth in Reclaimed Soil

Experimental results were carried out in the field of Seosan reclaimed soil as in Example 1 to the results expected through Experimental Example 1.

The chemistry of the soil before treatment was shown in Table 2 below, and the salt concentration was 6.2 ds / m (corresponding to 0.40% salt concentration).

The rice varieties used for the test were excellent, and chitosan treatment was sprayed a total of 5 times before planting, 2 times every 15 days after planting, and 1 time after planting. Indicated.

That is, as shown in Figure 3, the NaCl treated group was found to have an externally necrotic phenomenon on the plant leaf tissue, which adversely affects the growth of the plant, but in Example 1 (chitosan lysate treated group) almost no cell damage occurs. The plants were confirmed to grow well without necrosis.

Experimental Example 3 Effect of Low Molecular Chitosan Degradation Treatment in Salt Concentration House

Based on the results of Experiment 3, the vegetable growth test was performed in the soil of facility house where salt accumulation is expected due to excessive use of chemical fertilizer as in Example 2.

The test site was carried out at the facility house in Goyang-si, Gyeonggi-do.

The test crop was yeolmu, cultivated by cultivating the soil irrigation 1 time before sowing, 3 times every 7 days at 4-5 leaves after sowing, and then quantitatively surveying the results are shown in Tables 2, 3 and 3.

Acidity
(pH) Salt concentration, ECe (ds / m) Organic matter
(%) Effective Phosphoric Acid
(mg / kg) Substitution K
cmol + / kg Substitutable Ca
cmol + / kg Substitutable Mg
cmol + / kg Control 6.4 6.2 0.77 51 0.36 1.97 2.74 Example 2 6.5 3.8 3.48 55 2.62 13.0 5.96

As shown in Table 2, when the salt concentration was measured in the heat radish grown in soil containing a large amount of salt, it was confirmed that the salt concentration in the heat radish grown in Example 2 treated with the chitosan decomposed product of the present invention was much less than the control. .

In other words, it was found that the low molecular weight chitosan degradant lowered the salt concentration generated inside the plant by inhibiting the accumulation of salt in the plant.

Leaf
(cm) Leaf width
(cm) ground game
(dog) chlorophyll
(SPAD) Ground load
(g) MDA
(nmol / mg
protein) Control 8.1 3.3 6.4 28.7 5.7 1.3 Example 2 11.4 4.6 7.4 33.3 6.7 0.8

As shown in Table 3 and Figure 3, as a result of investigating the yield and MDA of the plant, it was confirmed that the yield cultivated in Example 2 was increased than the control, MDA was confirmed to decrease, which affects the degree of plant growth It could be seen that the yield can be increased.

1 shows a chitosan structure.

Figure 2: Figure comparing the degree of plant leaf tissue necrosis.

3 shows the yield.

Claims (4)

In the method of growing plants in soil containing a large amount of salt, By cultivating plants with water-soluble chitosan degradation products having a molecular weight of 10,000 Daltons or less, or cultivating them at the roots of plants, they reduce plant cell damage caused by a large amount of salt remaining in the soil to increase plant yield. Is, The chitosan decomposed product is prepared by putting chitosan and chitosan degrading enzyme in an acid solution, When cultivating the plant, the chitosan decomposed product is diluted to prepare in liquid form, or dried to prepare in powder form, and the amount of chitosan decomposed product is foliar fertilized in plants 1 to 5 times by 10 to 500 mg per 3.3 m ² It is characterized by irrigation to the roots, The plant cultivation type is characterized in that it is used for one selected from the Chinese cabbage, radish, jerk, rice, melon, strawberries, tomatoes, How to grow plants in soils containing a large amount of salt. delete delete delete

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