JPH0791169B2 - Plant growth promoter - Google Patents

Description

TECHNICAL FIELD The present invention relates to a plant growth promoter. More specifically, it relates to a plant growth promoter containing peroxidase as an active ingredient.

[Conventional technology]

Heretofore, plant hormones such as auxins, diberylenes, cytokinins, and ethylene have been generally known as plant growth regulators. These phytohormones can bring various effects to plants in trace amounts, but on the other hand,
If it is given in excess, it has a bad effect, and there is a problem that the type of hormone and the concentration to be applied must be strictly determined depending on the growth stage of the plant, the type of plant, and the surrounding environment.

[Problems to be Solved by the Invention]

The problem to be solved by the present invention is to solve the above-mentioned drawbacks of conventional promoters for promoting plant growth, among conventional plant growth regulators.

[Means for Solving the Problems]

In order to solve the above-mentioned problems, the present inventors have conducted extensive studies as a plant growth promoter to find out an excellent one, and as a result, found that peroxidase has a plant growth promoter. The invention was completed.

[Operation and Configuration of Invention]

Peroxidase is widely distributed in plant tissues, animal tissues, blood and the like, and it has been conventionally confirmed that some microorganisms secrete it. Among them, peroxidase existing in plant tissue has been studied for a long time, and many reports have been made on its physiological role. They are summarized as follows.

(1) It has been reported that the activity of peroxidase contained in tissues increases when a plant body is infected with a pathogenic bacterium. As a specific example, Phytophthora citrophthora
In tissues of navel orange infected with so-called rot such as Diplodia natalensis, enzyme activities such as peroxidase, cellulase, catalase, polygalacturonase are increased, and tomato leaves infected with tomato mosaic virus It can be mentioned that the activity of peroxidase increases with an increase in the content of carbohydrates and reducing sugars, and that in soybean tissues infected with soybean mosaic virus, the peroxidase activity increases at the latter stage of infection.

(2) It has been reported that the higher the resistance to plant pathogens, the higher the activity of peroxidase. As a specific example, there are specific peroxidases and proteins in pigeon pea that is resistant to incompetent mosaic disease, and in paddy rice, varieties with high resistance to blast and low varieties are used. It can be mentioned that when the rice blast fungus is inoculated, the increase rate of peroxidase activity in the tissue of the former is more rapid than that of the latter.

(3) It has been reported that peroxidase is involved in the secretion and regulation of plant hormones. As a specific example, when cypress is treated with Sumithion (insecticide),
In non-sensitive cypress, peroxidase is not activated but rather inhibited by sumithione, which suppresses endogenous ethylene production and defoliation. Treating rice leaves with auxins and ethylene increases peroxidase activity. It can be mentioned that the disease is suppressed and that the activity of peroxidase and polyphenol oxidase in the pericarp is increased by treating lacey with a trace amount of ethylene.

(4) It has been reported that peroxidase is involved in plant differentiation and growth, and the indole acetic acid oxidative activity of peroxidase is expressed as plant root elongation and differentiation progress. is there.

Although various reports have been made on the behavior of peroxidase in the plant composition as described above, no report has been made on the reaction of the plant when peroxidase is directly given to the plant or its seed. Then, the present inventors have found that peroxidase has a plant growth promoting action as a result of externally applying peroxidase to plants and their seeds and observing the reaction, and completed the present invention.

The term "plant growth promoting action" as used herein refers to an action of broadly promoting plant growth, and examples thereof include plant growth promotion, germination root promotion, fruit enlargement promotion, and grain yield increase.

Next, the peroxidase according to the present invention will be described. Peroxidase is a heme protein containing trivalent iron ions and is an enzyme that catalyzes the oxidation of various compounds in the presence of hydrogen peroxide and organic peroxide. For example, Chance explains the mechanism of action of horseradish peroxidase.

POD ＋ H ₂ O ₂ ← → Complex I Complex I ＋ AH → Complex II ＋ A Complex II ＋ AH → POD ＋ A ＋ 2H ₂ O Where POD: peroxidase AH: reduced donor A: oxidized donor Peroxidase is thus present in the presence of peroxide. It is an enzyme that catalyzes the oxidation reaction of various compounds. Next, in order to confirm that this plant growth promoting action is due to peroxidase, the plant inactivated peroxidase by heat treatment and the one not subjected to heat treatment were given to the plants, and their plant growth promoting action was observed. Was observed. as a result,
The former did not have a growth promoting action, and the latter had a growth promoting action.

The peroxidase according to the present invention is extracted and purified from a material containing peroxidase and used.
Naturally, the amount and quality of impurities contained together with peroxidase vary depending on the degree of purification. We investigated how the impurities affect the plant growth promoting action of peroxidase. That is, samples containing peroxidases having different degrees of purification were prepared and their plant growth promoting action was observed. As a result, the activity of rice clam peroxidase was 10 ⁶ / g unit or more, and the RZ value was 0.1.
It was observed in the above samples. It should be noted that the same was observed with substances other than rice hull peroxidase, for example, peroxidase from horseradish and microorganisms.

Here, the activity (U) of peroxidase and the RZ value will be described.

For the activity of peroxidase, guaiacol and hydrogen peroxide are used as substrates, and the increase in absorbance at 470 nm due to an enzymatic reaction is measured. The amount of enzyme that increases the absorbance by 0.001 per minute at 30 ° C was expressed as 1U (unit).

RZ value is the ratio of absorbance at 275 nm and 403 nm (A
_403nm / A _275nm ). This value represents the ratio of the heme content to the protein content in the peroxidase solution and is an index of the degree of purification of peroxidase. The value for pure horseradish peroxidase is reported to be 3.04.

Even when actually using peroxidase in the fields of agriculture and horticulture, it is advantageous that the peroxidase is purified and concentrated. For example, it can be easily transported to the site of use and can be stored in a non-harsh place where the environmental temperature and humidity do not rise excessively.

As described above, since peroxidase is widely present in plant tissues, it can be extracted from plant tissues regardless of the type of plant. However, in order to widely utilize the growth promoting action of peroxidase in agricultural and horticultural fields, peroxidase must be inexpensively extracted and purified. Considering this point, the condition for extracting peroxidase is that it contains a large amount of peroxidase,
It can be said that it is easily available, that it exists in a large amount, that the material is inexpensive. Examples thereof include horseradish, Japanese radish, daikon radish, fig, rice husk and the like, and other microorganism-derived ones can also be used. However, the peroxidase according to the present invention is not limited to these types.

As described above, the peroxidase in the present invention is produced by extracting and purifying from plant tissue or microorganism.

In the present invention, the produced peroxidase may be used as it is, or it may be mixed with a suitable carrier such as liquid, clay, talc or kaolin dissolved in a solvent such as water to prepare a powder or granule. It can also be used.

The agent of the present invention is used by foliage spraying or seed soaking, and for cuttings, by soaking cuttings or applying powder or paste to the part where rooting is expected. Further, the agent of the present invention can be used by adding a surfactant so that the agent can easily adhere to and penetrate the leaf surface in foliage application. It can also be mixed with liquid fertilizers, fungicides, insecticides and plant hormones.

The time of use of the agent of the present invention is preferably from the vegetative period to the reproductive period, but is not particularly limited.

Next, the amount of the agent of the present invention used in plants will be described.

In order to show the usage amount of the agent of the present invention, the peroxidase activity value and the concentration used in the agent of the present invention are displayed. As a result, the peroxidase activity at the time of use can be calculated, and the required amount can be given to the plant at a preferable concentration.

The amount of the present invention agents, plant type and growth stage may vary upon usage or use time, as an example 10 ⁸ U /
The amount of the agent of the present invention having g and RZ = 0.1 will be described. When spraying the foliage of plants with an aqueous solution, the concentration of this drug should be 1 to
2000ppn (Peroxidase activity 10 ² to 2 × 10 ⁵ U / m
l), preferably 10-1500 ppm (peroxidase activity 10
^{3 to} 1.5 × 10 ⁵ U / ml). In seed soaking, the concentration of this agent is 1 to 2000 ppm, preferably 10 to 1500 ppm (peroxidase activity 10 ^{3 to} 1.5 × 10 ⁵ U / ml). The soaking time of seeds is 1 to 72 hours, preferably 5 to 48 hours. When used for cuttings, the concentration used for cutting soaking, powder or paste application is 1 to 2000 ppm, preferably 1
0 to 10,000 ppm (peroxidase activity of 10 ³ to 10 ⁶ U /
and the soaking time is 1 to 48 hours, preferably 3 to 24 hours. However, when powder or paste is applied, cutting is done immediately after application.

〔The invention's effect〕

The effect of using the agent of the present invention is promotion of plant growth. By applying the foliar agent of the present invention to leafy vegetables such as Japanese mustard spinach and salad, the growth of both the foliage and the root is promoted, early harvesting is possible, and the yield is increased. For fruit vegetables such as tomatoes, cucumbers, eggplants and bell peppers, and shells such as rice and wheat, seedlings can be cultivated and foliage sprayed to grow healthy seedlings, and the yield can be increased.

〔Example〕

 The present invention will be described below with reference to examples.

Example 1 Salad vegetables were sown on March 15 in a 1/4000 are Wagner pot containing commercially available flower soil. March 20, 3
The peroxidase aqueous solution was thoroughly sprayed on the foliage twice on the 27th of each month. Further, distilled water was sprinkled in the same manner without treatment. The growth was investigated on May 1, leaving 10 well-collected individuals. The fertilizer is chemical fertilizer per pot on March 28, N: 0.4g, P:
0.4g, K: 0.3g was given.

The peroxidase used is as follows.

・ Chive moth peroxidase Extracted and produced from rice husk Enzyme activity 10 ⁸ U / g, RZ value 0.1 ・ Horse horseradish peroxidase Sigma made and produced from horseradish extract Enzyme activity 1.5 × 10 ⁹ U / g, RZ value 0.3 ・ Microbial peroxidase Suntory Produced from culture fluid of filamentous fungus Arthomyces ramosus Enzyme activity 4.8 × 10 ⁸ U / g, RZ value 2.5 Perfoliate application of peroxidase increased dry matter weight in both foliage and root.

Example 2 Komatsuna was sown on March 31 in a 1/4000 arel Wagner pot containing vermiculite. On April 7th and April 17th, the aqueous solution of peroxidase was thoroughly sprayed on the foliage.
Distilled water was sprayed as no treatment. The growth condition was investigated on May 19, leaving 15 well-collected individuals. The fertilizer is 4
On the 10th of each month, N: 0.4g, P: 0.4g, K: 0.
Giving 3g. Also, on May 9, liquid fertilizer N: 0.1g, P: 0.1
g, K: 0.1 g was topdressed.

The peroxidase used is as follows.

・ Chicken peroxidase Enzyme activity 10 ⁶ U / g, RZ value 0.1 ・ Horse horseradish peroxidase The same results as in Example 1 are shown in Table 2. Similar to horseradish peroxidase, which has a high activity, chaff perilla oxidase, which has a low peroxidase activity, can be sprayed on the foliage,
Increased grain weight in both roots.

Example 3 Tomato seedlings (cultivar Momotaro) that had been seeded for 80 days after planting were planted on March 8 in a 1/5000 arel Wagner pot containing flower soil. After planting, on March 10, March 18, April 7, and April 11, the peroxidase aqueous solution was sprayed four times on the leaves and fruits. Untreated and sprayed with distilled water. And May 9, 12th, 15th, 18th, 22nd, 25th, 29th, 31st
The fruits were harvested and surveyed on Sunday, June 5, 9, 15, 22, and 26th. In addition, 1 ward has 4 pots. The fertilizer was a chemical fertilizer and N: 0.8g, P: 0.8g, and K: 0.6g were given four times on March 9, March 28, April 14, and May 12. The peroxidase used is the same as in Example 1. The results are shown in Table 3. An increase in the yield of fruits and the average fruit weight per one was observed by the foliar application of peroxidase.

Example 4 Cucumber seeds (variety: Kinsei Yotsuba cucumber) were dipped in an aqueous solution of peroxidase or untreated in distilled water for 2 days,
Immediately, three bees having a diameter of 9 cm and containing vermiculite were sowed. The seeding date was June 1st. One individual in a good germination state was left, and 5 pots were prepared in 1 area. Fertilizer June 13
N: 0.16 g, P: 0.16 g, and K: 0.12 g were given per one day.
Then, on July 13, the growth condition was investigated. The peroxidase used is the same as in Example 1. The results are shown in Table 4. By dipping the seeds in peroxidase, the weight of grain increased in both the foliage and roots.

Example 5 The cut end of the green branch of metasequoia was immersed in an aqueous solution of peroxidase, or untreated and then soaked in distilled water for 8 hours, and immediately cut into Kanuma soil for cutting. (August 25). The rooting condition was investigated on August 24, 60 days later. There are 20 wards in one ward, and a two-district system.

Rice chaff peroxidase (activity value 10 ⁸ units / g
An RZ value of 0.1) was used. The results are shown in Table 5. Rooting was promoted by the peroxidase immersion treatment.

Example 6 On May 15, paddy rice (cultivar Sasanishiki) was sown in a kneaded granular soil O. And on June 15, Kumiai granular soil O
It was transplanted to a 1/5000 ares pot containing. May 24
On June 1, June 21, the peroxidase solution was added to the foliage, and undiluted distilled water was applied to the leaves. And 10
On the 5th of each month, the above-ground portion was cut and the yield was investigated. There were 3 pots in 1 ward. The kumiai granular soil O used contained N: 0.6g, P: 1.2g, K: 1.1g per 1.7kg, and the amount of fertilizer component per pot was N: 1.2g, P: 2.2. The soil was used so that it would be g. Also, on August 1, 1 with chemical fertilizer
N: 0.3g, P: 0.08g, K: 0.3g was given per pot. The peroxidase used is the same as in Example 1. The results are shown in Table 6. Yield was increased by foliar application of peroxidase.

Example 7 Paddy rice (cultivar Sasanishiki) fir was immersed in a peroxidase solution or in distilled water for 2 days without treatment. Then, on May 15, it was transplanted to a pot of 1/5000 ares containing Kumiai granular soil O. Then, on October 5, the above-ground portion was cut and the yield was investigated. The fertilization is the same as in Example 6. The peroxidase used is the same as in Example 1. The results are shown in Table 7. The yield was increased by soaking the seed fir in the aqueous solution of peroxidase.

Example 8 Threshed rice seeds (Sasanishiki) were thoroughly washed with water and then 0.3
% Sodium hypozincnate solution for 30 minutes for sterilization.
After washing well with sterilized water, put 50 tablets each in a dish that has been dried and covered with filter paper, and filter and sterilize various enzyme solutions (rice mosquito peroxidase, horseradish peroxidase, microbial peroxidase) for each concentration (100ppm, 10pp
m, 1 ppm) enzyme solution was added in 10 ml increments. As control, sterilized water and 10 ppm enzyme solution boiled for 30 minutes were used.

Germination was carried out by incubating at 30 ° C. for 72 hours, and the lengths of extended shoots and roots were measured. The results are shown in Table 8.

The activity values of each enzyme used were: Rice mosquito peroxidase 5.7 × 10 ⁹ U / g, RZ value 1.59 ・ Horseradish peroxidase 1.9 × 10 ⁹ U / g, RZ value 0.8 ・ Microbial peroxidase 4.8 × 10 ⁸ U The value was / g and the RZ value was 2.5.

Since the test date and time for each enzyme was different, a control section was provided in each test section.

None of the enzyme solutions whose peroxidase activity was inactivated by heat treatment showed a growth promoting action, whereas the enzyme solutions not subjected to heat treatment showed a growth promoting action.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Aso 12-46 Sannomachi, Tsuruoka City, Yamagata Prefecture (72) Inventor Naoichi Sakoda 1-23-24, Sumiyoshihoncho, Higashinada-ku, Kobe City, Hyogo Prefecture (56) References JP 47-1305 (JP, A)

Claims (1)

[Claims] 1. A plant growth promoter containing peroxidase as an active ingredient in a range in which the action of ethylene does not substantially appear.