JP6301703B2 - Hydroponics method and strains used for hydroponics - Google Patents

Description

  The present invention relates to a hydroponic cultivation of vegetables.

In soil cultivation of crops using soil, there are a wide variety of soil microorganisms and bacteria that are generated and coexist by humus in the soil, and these microorganisms and bacteria can skillfully regulate the environment essential for crop growth. We are using.
However, some of the microorganisms and bacteria in the soil can cause crop diseases, so these measures are necessary. In soil cultivation, it is necessary to consider the problem of continuous cropping caused by continuous cropping of specific types of crops.

  On the other hand, hydroponics (hydroponic cultivation), which is soilless cultivation without using soil, is performed for soil cultivation of crops using soil. Hydroponic cultivation has advantages such as year-round production, production on unsuitable land due to soil, and avoidance of continuous cropping failures. On the other hand, since hydroponics does not have the buffering power of the soil, the antibacterial action against pathogenic bacteria does not work, so the types and amounts of agricultural chemicals and fertilizers tend to increase.

  For example, Patent Document 1 describes that tomato hydroponics is performed using a specific rhizosphere microorganism.

JP-A-6-31826

  In hydroponic cultivation that does not have the buffering power of the soil, it is desirable to cultivate vegetables that complete a single crop in a short span because there is a risk of a balance of pesticides and fertilizers and pathogens.

  Accordingly, an object of the present invention is to provide a hydroponics method capable of promoting the growth of a specific vegetable and a strain used for the hydroponics.

  As a result of earnest examination of the method for hydroponics of specific vegetables, the present inventors added Bacillus amyloliquefaciens, Pantoea agglomerans and Pseudomonas fluorescens to the culture broth of the hydroponic culture, It was found that can be promoted.

That is, in order to achieve the above object, the following inventions [1] to [6] are provided.
[1] A hydroponic method for growing the vegetable by adding at least one of Bacillus amyloliquefaciens, Pantoea agglomerans and Pseudomonas fluorescens to promote the growth of the vegetable , the Bacillus -Amiloliquefaciens is a strain indicated by any of the accession numbers NITE P-01825 and NITE P-01826, and the Pantoea agglomerans is a strain indicated by the accession number NITE P-01823, and the Pseudomonas A hydroponic cultivation method in which the fluorescens is a strain indicated by the accession number NITE P-01824 .
[2] The hydroponics method according to [1], wherein the vegetable is a leaf vegetable.
[3] The hydroponics method according to [2], wherein the leafy vegetables are selected from the group of bees and lettuce.
[4] Bacillus amyloliquefaciens, which is a strain represented by any of the accession numbers NITE P-01825 and NITE P-01826 used for promoting growth in the hydroponic cultivation of vegetables.
[5] Pantoea agglomerans, which is a strain indicated by the accession number NITE P-01823 used for promoting growth in hydroponic cultivation of vegetables.
[6] Pseudomonas fluorescens, which is a strain indicated by the accession number NITE P-01824 used for promoting growth in hydroponic cultivation of vegetables.

The existence of bacteria having a growth-promoting effect on plants as plant growth-promoting rhizosphere bacteria (PGPR) has been recognized in rhizosphere bacteria living in the rhizosphere, which is a soil environment in which plants grow. In the present invention, in hydroponic cultivation, PGPR which is Bacillus amyloliquefaciens (a strain indicated by any of accession numbers NITE P-01825 and NITE P-01826) , Pantoea agglomerans (accession number NITE P-01823) At least one of Pseudomonas fluorescens ( strain indicated by accession number NITE P-01824) is added to the culture solution. Growth can be promoted by using such a culture solution for hydroponics of vegetables, especially leafy vegetables. As the leaf vegetables, for example, bees, lettuce, etc. are preferably used. In Examples to be described later, it was confirmed that the growth of these leaf vegetables was promoted (increase in fresh weight on the ground).

  Therefore, in this invention, the growth promotion effect can be acquired with a simple method by introduce | transducing a specific microorganism into a culture solution in the nourishing culture of vegetables.

Hereinafter, embodiments of the present invention will be described with reference to the drawings.
In the present invention, in the hydroponic cultivation of vegetables, at least one of PGPR, Bacillus amyloliquefaciens, Pantoea agglomerans and Pseudomonas fluorescens is added to the culture solution.

(Hydroculture)
Hydroponic cultivation is a cultivation method that provides nutrient nutrients necessary for plant growth as liquid fertilizer. A culture solution containing liquid fertilizer is supplied into a cultivation tank having a desired scale, and a part of the plant body is transferred to the culture solution. Cultivate in a state immersed in The cultivation tank is usually installed in a closed space where temperature and humidity and lighting can be controlled separately, and the plant (vegetables) is cultivated while circulating the culture solution supplied to the cultivation tank. However, the present invention is not limited to such an embodiment.

The mode of hydroponics is not particularly limited, and methods such as hydroponics, spray plowing, and solid medium plowing can be applied.
Hydroponic methods include circulating submerged hydroponic method, non-circular submerged submerged hydroponic method, vented submerged submerged hydroponic method, liquid level vertical submerged submerged hydroponic method, capillary hydroponic method and NFT (Nutrient Film) Technique: Thin-film hydroponics method may be applied.
A spray hydroponic method may be applied as the spray plowing method.
As the solid medium cultivation method, an inorganic medium cultivation method, an organic medium cultivation method, or the like may be applied. As the medium in the inorganic medium cultivation method, rubble, sand, rice bran charcoal, vermiculite, perlite, rock wool and the like are used. As the medium in the organic medium cultivation method, natural organic substances such as bark, coconut husk, peat moss, sawdust and rice husk and organic compounds such as polyurethane, polyphenol and vinylon are used. In particular, it is preferable to perform an inorganic medium cultivation method using rock wool from the viewpoint of improving the retention of the culture medium and improving the gas phase rate.

  The culture solution used for the hydroponics is not particularly limited as long as the nutrient (essential element) required by the plant to be cultivated is dissolved in water in a mode and concentration suitable for absorption. The nutrient is usually present in an ionic state, and has a large amount element and a small amount element.

Major elements include nitrogen (N: nitrate (NO ₃ -N) and ammonia (NH ₄ -N), and in nutrient solution cultivation, nitrate is the main nitrogen source), phosphorus (P: phosphate ( PO ₄ -P)), potassium (K), calcium (Ca), magnesium (Mg), sulfur (S: sulfate (SO ₄ -S)) and the like.

  Trace elements include iron (F), manganese (Mn), boron (B), copper (Cu), zinc (Zn), molybdenum (Mo), silicon (Si), chlorine (Cl), aluminum (Al), Examples include sodium (Na) and nickel (Ni).

  Such nutrients are prepared to an appropriate concentration, pH, and EC (electric conductivity) and used as a culture solution. In hydroponics, culture solution management that controls these factors of the culture solution is important. Furthermore, in hydroponic cultivation, environmental control that controls environmental factors such as temperature, humidity, and light conditions is also important.

  What is necessary is just to set the conditions of the factor mentioned above suitably according to the kind of vegetable to grow. For example, pH is 5.0 to 7.0, EC is 0.5 to 5.0 mS / cm (25 ° C.), air temperature is 10 to 30 ° C., humidity is 50 to 70%, and light conditions are 8 known artificial light sources. Although it may be controlled to irradiate for about 12 hours, it is not limited to these ranges.

(Rhizosphere bacteria)
The rhizosphere is an area that is affected by the roots of the plant, and the rhizosphere bacteria are bacteria that live in the rhizosphere, which is the soil environment in which the plants grow. In the present invention, as a rhizosphere bacterium, a bacterium having a growth promoting effect on a plant as a plant growth promoting rhizosphere bacterium (PGPR), a Bacillus bacterium, Bacillus amyloliquefaciens, Pantoea bacterium Pantoea agglomerans and Pseudomonas fluorescens can be used.

  Bacillus amyloliquefaciens is known as an antagonistic microorganism against pathogens of plant diseases and is an aerobic bacterium often found in soil samples. As the Bacillus amyloliquefaciens, for example, a strain (# 111) represented by the accession number NITE P-01825 and a strain (# 167) represented by NITE P-01826 can be used.

Bacteria other than Bacillus are known as antagonistic microorganisms against pathogens of plant diseases like Pantoea agglomerans and Bacillus amyloliquefaciens, which are endemic in wheat, and are often found in soil samples. The aerobic bacterium Pseudomonas fluorescens that can be used can be used. For Pantoea agglomerans, for example, a strain (# 74) represented by accession number NITE P-01823, and for Pseudomonas fluorescens, for example, a strain (# 78) represented by accession number NITE P-01824 can be used. In the present invention, these bacteria may be used alone or in combination.
All the strains to which the above-mentioned deposit numbers are attached have been deposited on March 12, 2014 at the National Institute for Product Evaluation and Technology (NITE) Patent Microorganism Deposit Center (NPMD).
Moreover, in this invention, the mutant strain or derivative strain of the strain mentioned above may be sufficient, and it can be used as long as it has the effect | action mentioned later by using such a strain for hydroponics.

  For the culture of the above-mentioned strain, media such as TSB (Trypticase Soy Broth, manufactured by BBL) and TSA (Trypticase Soy Agar, manufactured by BBL) can be used, but are not limited thereto. In addition, for washing and recovery (resuspension) of cells after culture, sterilized water or PBS (phosphate buffered saline) can be used after centrifugation, but is not limited thereto. It is not a thing.

(Leaf vegetables)
In the present invention, leaf vegetables are preferably applied as vegetables. Leaf vegetables refer to vegetables that mainly use the leaf portion as food. In the present invention, leaf vegetables are not particularly limited. Mizuna, mugwort, etc. can be applied.

  In the present invention, in hydroponic cultivation, at least one of PGPR, Bacillus amyloliquefaciens, Pantoea agglomerans, Pseudomonas fluorescens, is added to the culture solution. Growth can be promoted by using such a culture solution for hydroponics of vegetables, especially leaf vegetables.

Examples of the present invention will be described.
Hydroponic cultivation of honey bees (white stems, three leaves, Kansai, Takii seedlings) and Sunny lettuce (Takii seedlings) using a non-circulating submerged hydroponic system with normal hydroponic cultivation equipment (water field, manufactured by Zenpo) Went.

Mitsuba seeds were immersed in a 17% (W / V) aqueous sodium chloride solution for 30 minutes to remove immature seeds and impurities, then washed with water, immersed in running water overnight, and drained. Thereafter, it was stored at 4 ° C. for 2 weeks and used after pre-treatment. Sunny lettuce seeds were used as they were. Both Mitsuba and Sunny lettuce are cultivated with water only during the period from sowing to the start of development of cotyledons, and thereafter, until the planting, Otsuka Chemical A formulation (pH 6.0, electric conductivity 1.2 mS / It was adjusted to cm (20 ° C.) and cultivated at a capacity of 500 mL).
Mitsuba seedlings and sunny lettuce seedlings were respectively planted on the 14th day after sowing. As the culture solution, Otsuka Chemical A formulation (pH 6.0, electric conductivity adjusted to 2.3 mS / cm (20 ° C.), capacity 10 L) was used.

Two types of Bacillus amyloliquefaciens (# 111 strain (Accession No. NITE P-01825) and # 167 strain (Accession No. NITE P-01826)) were each 10 ⁵ cfu / Culture systems added at a concentration of mL were constructed (Examples 1 and 2). In addition, Pantoea agglomerans # 74 strain (Accession No. NITE P-01823) (Example 3)), Pseudomonas fluorescens # 78 strain (Accession No. NITE P-01824) (Example 4: A culture system was constructed in which only Sunny lettuce was added at a concentration of 10 ⁵ cfu / mL.
The strains mentioned above were obtained by inoculating each strain in a TSB liquid medium, shaking culture at 30 ° C. for 16 hours at 140 rpm, culturing, centrifuging, washing with PBS, and resuspension three times. It was used.

  Hydroponics is carried out under light conditions of 8000 Lx for 13 hours (20 ° C.), dark periods of 11 hours (15 ° C.), and humidity of about 50% RH in both light and dark periods. Was cultivated for 41 days after planting and Sunny lettuce was cultivated for 45 days after planting. For each example, Mitsuba grows a total of 60 of 12 x 5 urethane blocks, Sunny lettuce grows a total of 5 of 1 x 5 urethane blocks, and the plant height of Mitsuba is the average of the maximum length of each urethane block The value was the total value of the seedlings of all five urethane blocks for the above-ground and root fresh weights of bees and sunny lettuce.

In addition, as Example 5, cultivated honey bees (white stems, three leaves, Kansai, Takii seedlings) in a container using a plant culture container (Incuttis, manufactured by Biomedical Science Co., Ltd.) System). The seeds that have been drained and pre-treated are used for vermiculite sterilized by autoclaving (121 ° C, 15 minutes), and the period from sowing to the start of the development of cotyledons is cultivated only with water. As a culture solution, Otsuka Chemical A formulation (pH 6.0, electric conductivity adjusted to 2.3 mS / cm (20 ° C.), 100 mL) was added to vermiculite.
A culture system was constructed in which Pseudomonas fluorescens strain # 78 (accession number NITE P-01824) was added at a concentration of 10 ⁵ cfu / mL to the culture solution thus prepared at the 14th day after sowing. did.

  Hydroponic cultivation is performed at a light period of 10000 Lx for 16 hours (20 ° C.), a dark period of 8 hours (20 ° C.), and a humidity of about 55% RH in both the light and dark periods. Cultivated for 39 days. Nine honey bees are cultivated, and the plant height, the above-ground part and the root fresh weight are the average values of all the seedlings.

  In Examples 1 to 5, after the hydroponics, all seedlings were harvested, and the plant height, the above-ground fresh weight, and the root fresh weight were measured. The results are shown in Table 1. The numerical values shown in Table 1 are the values when the plant height, the above-ground fresh weight, and the root fresh weight in the control (honey bees and sunny lettuce cultivated without adding any microorganisms having a growth-promoting effect to the culture solution) are set to 1.0. Relative values are shown.

  As a result, in Example 1-5, the height of the ground part of the edible part was 1.1 to 1.4, and the fresh part of the ground part was 1.2 to 1.5. . Moreover, in the sunny lettuce, in Examples 1-4, the value of 1.4-1.8 was obtained for the above-ground fresh weight.

  Therefore, growth can be promoted by adding any of PGPR Bacillus amyloliquefaciens, Pantoea agglomerans and Pseudomonas fluorescens to the culture solution in the hydroponic culture of honey bees and sunny lettuce. It was.

  The present invention can be used for hydroponics of vegetables.

NITE P-01825
NITE P-01826
NITE P-01823
NITE P-01824

Claims (6)

Bacillus amyloliquefaciens that promotes the growth of vegetables, Pantoea agglomerans and Pseudomonas fluorescens are added to the culture solution to cultivate the vegetables ,
The Bacillus amyloliquefaciens is a strain indicated by any of the accession numbers NITE P-01825 and NITE P-01826, and the Pantoea agglomerans is a strain indicated by the accession number NITE P-01823, A hydroponic cultivation method in which Pseudomonas fluorescens is a strain represented by an accession number NITE P-01824.   The hydroponic cultivation method according to claim 1, wherein the vegetable is a leaf vegetable.   The hydroponic cultivation method according to claim 2, wherein the leafy vegetables are selected from the group of honey bees and lettuce. Bacillus amyloliquefaciens, which is a strain represented by any of the accession numbers NITE P-01825 and NITE P-01826 used for the promotion of growth in hydroponic cultivation of vegetables. Pantoea agglomerans which is a strain indicated by the accession number NITE P-01823 used for the promotion of growth in hydroponic cultivation of vegetables. Pseudomonas fluorescens which is a strain shown by the accession number NITE P-01824 used for the promotion of growth in hydroponic cultivation of vegetables.