Method for producing thallus of seaweed

Abstract

To provide a technique for suppressing the maturation of seaweeds. It is solved by a method for culturing seaweeds, which includes a step of culturing seaweeds under the supply of fine bubbles and a step of recovering the seaweeds.

Description

Technical Field

[0001] The present invention relates to a method for producing thalli of seaweeds and the like.

Background Art

[0002] Many seaweeds have vegetative growth of thalli at low water temperatures, mature at high water temperatures, release spores, and the thalli decrease or disappear. Therefore, in the conventional method of culturing seaweeds on the sea surface, cultivation is carried out only for a short period when the seawater temperature is suitable for the seaweeds to grow vegetatively without maturing.

[0003] In recent years, a method for culturing seaweeds on land that is less affected by seasonal increases in seawater temperature has attracted attention. In the land cultivation method used in Non-Patent Document 1, it is described that by supplying deep seawater at low water temperatures several times a day and cooling the seawater in the culture tank, year-round harvesting of Gracilaria lemaneiformis has become possible. However, in this method, energy and equipment are required for pumping up deep seawater, and the production cost increases. In addition, in summer when the air temperature is high, it is necessary to increase the amount of seawater obtained.

Prior Art Documents

Non-Patent Documents

[0004]

Non-Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0005] An object of the present invention is to provide a technique for suppressing the maturation of seaweeds and obtaining thalli with higher yields.

Means for Solving the Problems

[0006] As a result of intensive research in view of the above problems, the present inventors have found that the maturation of seaweeds can be suppressed by culturing the seaweeds under the supply of fine bubbles. Based on this finding, further research has been carried out, and as a result, the present invention has been completed.

[0007] That is, the present invention includes the following aspects.

[0008] Item 1. A method for producing a thallus of seaweed, comprising a step of culturing the seaweed under the supply of fine bubbles and a step of recovering the thallus of the seaweed. Item 2. The production method according to claim 1, wherein the seaweed is an alga in which somatic cells differentiate into spores. Item 3. The production method according to claim 1 or 2, wherein the seaweed is Sargassum fusiforme, Enteromorpha linza, Enteromorpha prolifera, Enteromorpha clathrata, Monostroma angicava, Cladophora fracta, Cladophora hutchinsiae, Gelidium amansii, Porphyra tenera, Porphyra yezoensis. Item 4. The production method according to any one of claims 1 to 3, wherein the seaweed is Sargassum fusiforme, Cladophora hutchinsiae, Porphyra tenera. Item 5. The production method according to any one of claims 1 to 4, wherein the seaweed is cultured under stirring conditions. Item 6. The production method according to any one of claims 1 to 5, wherein the fine bubbles include fine bubbles having a particle diameter of 500 nm or less. Item 7. The production method according to any one of claims 1 to 6, wherein the fine bubbles are supplied at a supply rate of 0.0001 L / min to 1 L / min per 1 L of the culture solution. Item 8. The production method according to any one of claims 1 to 7, wherein the fine bubbles are generated by a microporous method. Item 9. The thallus of the seaweed obtained by the production method according to any one of claims 1 to 8.

Advantages of the Invention

[0009] According to the present invention, by suppressing the maturation of seaweeds, it is possible to suppress the reduction of the thalli of seaweeds at a temperature suitable for growth and increase the yield.

Brief Description of the Drawings

[0010]

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Modes for Carrying Out the Invention

[0011] In this specification, the expressions "containing" and "comprising" include the concepts of "containing", "comprising", "consisting essentially of", and "consisting only of".

[0012] Method for producing thallus of algae In one aspect, the present invention relates to a method for producing a thallus of seaweed (which may also be referred to as "the production method of the present invention" in this specification), including a step of culturing seaweed under the supply of fine bubbles and a step of recovering the thallus of seaweed. Hereinafter, this will be described.

[0013] The seaweed is not particularly limited as long as it is a multicellular marine alga large enough to be confirmed with the naked eye, and examples include seaweeds such as green algae, red algae, and brown algae. Seaweeds that are raw materials for food and biofuels, for which industrial-scale cultivation is desired, are particularly preferred. If the seaweed grows vegetatively and matures at a temperature of 15 to 25°C, which is the temperature used in industrial aquaculture, the effects of the production method of the present invention can be more effectively exerted. Also, from the perspective of being able to further increase the yield during cultivation by the production method of the present invention, seaweeds in which somatic cells differentiate into spores are more preferred, and examples of such seaweeds include seaweeds belonging to the Ulvophyceae class of the Chlorophyta phylum, the Gracilariaceae class of the Rhodophyta phylum, and the Phaeophyceae class of the Xanthophyta phylum. Examples of seaweeds belonging to the Ulvophyceae class include, for example, Enteromorpha linza, Enteromorpha intestinalis, Enteromorpha compressa, Enteromorpha clathrata, Enteromorpha prolifera, Enteromorpha flexuosa, Enteromorpha linza var. kuroii, Monostroma angicava, Monostroma latissimum, Monostroma nitidum, Monostroma grevillei, Monostroma hariotii, Monostroma lanceolatum, Monostroma angicava var. angustata, Monostroma latissimum var. latissimum, Monostroma nitidum var. nitidum, Monostroma grevillei var. grevillei, Monostroma hariotii var. hariotii, Monostroma lanceolatum var. lanceolatum. Examples of seaweeds belonging to the Gracilariaceae class include, for example, Gracilaria asiatica, Gracilaria chorda. Examples of seaweeds belonging to the Phaeophyceae class include, for example, Undaria pinnatifida, Eisenia bicyclis, Laminaria japonica, Saccharina japonica, Sargassum fusiforme, Ecklonia cava. Preferably, edible seaweeds such as Enteromorpha linza, Enteromorpha intestinalis, Enteromorpha compressa, Enteromorpha clathrata, Monostroma angicava, Monostroma lanceolatum, Monostroma hariotii, Sargassum fusiforme, Ecklonia cava are mentioned. The seaweed may be a single species or a combination of two or more species.

[0014] According to the production method of the present invention, it is possible to obtain thalli of seaweeds with an increased wet weight or volume from spores, germlings, etc. used as starting materials. The wet weight of the obtained thalli of seaweeds is, for example, 10 times or more, 50 times or more, 100 times or more the wet weight at the start of culture, and 150 times or less, 200 times or less, 250 times or less, 300 times or less, 400 times or less, 500 times or less, 1000 times or less, 2000 times or less, 5000 times or less, 10000 times or less, 100000 times or less, 1000000 times or less.

[0015] Maturation means that the thalli of seaweeds come to form and release spores. Generally, seaweeds grow vegetatively with a low water temperature, mature with a high water temperature, release spores, and then the thalli decrease or disappear. Therefore, at the seaweed cultivation site, it is desirable to harvest before maturation occurs and the thalli decrease. Seaweeds such as the class Chlorophyceae, the class Ulvophyceae, and the class Phaeophyceae, when they mature, somatic cells directly differentiate into reproductive cell sacs, and the differentiated somatic cells fall off from the thalli, so the leaf length and the thallus weight decrease. In addition, since the part where the cells have fallen off turns white, the commercial value as a food decreases.

[0016] The evaluation of whether the maturation of algae is suppressed can be carried out by visually or under a microscope checking the presence and number of spores in the culture solution. For seaweeds such as the class Chlorophyceae, the class Ulvophyceae, and the class Phaeophyceae, in addition to the above method, it can be confirmed by whether the wet weight of the thalli after culture or the average daily growth rate during the culture period is increased compared to the control group. The average daily growth rate is an index representing the increase rate of the wet weight of the thalli per day. Taking the initial weight as W0 and the weight after t days as W t it can be calculated by the following formula. Average daily growth rate (%) = [(W t / W0) 1 / t - 1] × 100

[0017] Also, for seaweeds such as the class Chlorophyceae, the class Ulvophyceae, and the class Phaeophyceae, it is also possible to evaluate whether the maturation of algae is suppressed by visually checking the presence or absence of white - colored parts at the tip of the thalli after culture. The less the white - colored parts, the more the maturation is suppressed.

[0018] A spore is a reproductive cell. Examples of spores include zoospores, gametes, zygotes, tetraspores, carpospores, unilocular zoospores, neutral zoospores, and monospores.

[0019] In the production method of the present invention, spores or germlings can be used as starting materials. The spores are not particularly limited as long as they are spores obtained from seaweeds. The spores may be aggregated spores in which the spores are attached to each other. The aggregated spores can be obtained, for example, by the method described in Patent No. 3828359. The germlings can be obtained by transferring aggregated spores finely pulverized to about 5 mm or less in diameter, if necessary, to a culture container and culturing them while aerating to germinate. The spores may be used alone or in combination of two or more species.

[0020] In this specification, microbubbles refer to bubbles with a diameter of less than 100 μm. Microbubbles with a diameter of less than 100 μm are defined as fine bubbles (registered trademark) by the International Organization for Standardization (ISO). Among fine bubbles, those with a diameter of less than 1 to 100 μm are defined as microbubbles, and those with a diameter of less than 1 μm are defined as ultra-fine bubbles (registered trademark) (ISO). Bubbles with a diameter of less than 1 μm are also referred to as nanobubbles. Fine bubbles are characterized in that the bubble surface is charged negatively or positively, attracting an object charged with the opposite polarity and repelling an object charged with the same polarity. The microbubbles used in the production method of the present invention preferably have a diameter of less than 100 μm, 50 μm or less, 20 μm or less, 10 μm or less, 5 μm or less, 1 μm or less, 500 nm or less, 400 nm or less, 300 nm or less, 200 nm or less, or 100 nm or less. By using microbubbles with a small diameter, the damage caused to algal cells during microbubble generation can be reduced. In addition, microbubbles with a diameter of less than 100 μm are characterized by slowly rising, and by supplying gas in the form of microbubbles, it is possible to make the effect of gas molecules in water last longer. Microbubbles with a diameter of less than 1 μm can make the effect of gas molecules in water last even longer, especially by staying in water for a long time. Microbubbles with a diameter of less than 1 μm are transparent to the naked eye because they can transmit visible light. The particle size and number of microbubbles can be measured by methods such as nanoparticle tracking analysis, particle trajectory analysis, laser diffraction / scattering method, dynamic light scattering method, resonant mass measurement method, electrical sensing zone method, dynamic image analysis method, and light blocking method.

[0021] Fine bubbles can be generated using a commonly available fine bubble generator. The method of generating fine bubbles is not particularly limited. Microbubbles can be generated by methods such as the swirling liquid flow type, static mixer type, microporous type, ejector type, Venturi type, pressure dissolution type, cooling dissolution type, or mixed vapor condensation type. Fine bubbles with a diameter of less than 1 μm can be generated by using methods such as the microporous type, static mixer type, swirling liquid flow type, ultrasonic method, or pressure dissolution type. The fine bubbles may be generated in the culture vessel or, for example, in another vessel connected to the culture vessel. In the production method of the present invention, from the viewpoints of simplicity and little impact on algae, it is preferable to generate fine bubbles by the microporous method in the culture vessel. The micropores for aeration are not particularly limited, and for example, non-woven fabric or ceramic can be used. When the fine bubble generator is installed in the culture vessel, the installation location is not particularly limited, and it can be installed, for example, on the bottom surface or side surface.

[0022] The gas used as the generation source of fine bubbles is not particularly limited, and examples include air or a gas containing a gas contained in air such as H2, O2, CO2, Ar, water vapor, etc.

[0023] In the production method of the present invention, it is desirable that the fine bubbles are supplied at a supply rate of 0.0001 to 1 L / min per 1 L of the culture solution volume, more preferably at a supply rate of 0.0005 to 0.5 L / min per 1 L, and even more preferably at a supply rate of 0.001 to 0.2 L / min per 1 L.

[0024] The cultivation is preferably carried out under stirring conditions. The cultivation under stirring conditions is not particularly limited as long as part or all of the culture medium is stirred. For example, it can be carried out by aerating the culture medium with an aeration pump or the like (aeration culture), passing a liquid (for example, the culture medium etc.) through the culture medium with a pump or the like, moving a stirrer (for example, rotating etc.), shaking the culture vessel, etc. During the cultivation, the seaweeds may grow, for example, in a floating state, or may grow in a state where part of them is fixed to the wall surface or a suitable carrier. By carrying out the cultivation under stirring conditions, the oxygen bubbles attached to the surface of the algal thalli by photosynthesis can be removed, and photosynthesis can be carried out efficiently. Also, by flowing the algal thalli aggregates, uniform light irradiation can be performed on each algal thallus, and they can be grown evenly.

[0025] The cultivation temperature is not particularly limited as long as it is suitable for the vegetative growth and maturation of the seaweed. For example, it is 5 to 35 °C, preferably 10 to 30 °C, more preferably 15 to 25 °C.

[0026] The light conditions during cultivation are not particularly limited as long as they are light conditions under which the seaweed can grow. For example, natural light light-dark cycle conditions, artificial light conditions, artificial light-dark cycle conditions, etc. can be mentioned.

[0027] The cultivation period is not particularly limited. However, in one embodiment, according to the production method of the present invention, by culturing at a temperature suitable for the growth of the seaweeds while suppressing maturation, an increase in yield can be achieved earlier. From this viewpoint, in one embodiment, the cultivation period is, for example, 1 day or more, 2 days or more, 3 days or more, 4 days or more, 5 days or more, 6 days or more, 7 days or more, and preferably 60 days or less, 40 days or less, 30 days or less, 25 days or less, 20 days or less, 15 days or less, 10 days or less, 7 days or less.

[0028] The cultivation of the present invention is usually carried out in a container. Also, the cultivation of the present invention is usually static cultivation. The container is not particularly limited as long as it can be used for culturing algae, and one with a suitable volume can be selected according to the scale of cultivation. For example, flasks, beakers, aquariums made of glass or plastic, large circular tanks or connected tanks can be used. The culture container may be installed indoors or outdoors.

[0029] The cultivation of the present invention is carried out in water. The liquid used for cultivation is not particularly limited as long as it is suitable for the growth of seaweed, but it is usually seawater. Seawater is not particularly limited as long as it is suitable for the growth of seaweed, but it is a liquid with a salt concentration of 0% to 5%. Either natural seawater or artificial seawater can be used. Artificial seawater refers to a liquid obtained by artificially adjusting fresh water by adding salts such as sodium chloride, potassium chloride, calcium chloride, magnesium chloride, magnesium sulfate, etc., a pH buffer, and other trace components according to the composition of natural seawater. Also included are seawaters from which salts and other components in natural seawater have been removed and / or those containing active ingredients and the like. Seawater may be sterilized before use.

[0030] If necessary, a nutrient for promoting the growth of seaweed may be added to the seawater. When using artificial seawater, a seaweed morphogenesis inducer may be added. The seaweed morphogenesis inducer refers to a substance produced by microorganisms present in seawater, and various substances reported so far can be used without particular limitation (International Publication No. 2004 / 007510, Japanese Patent Application Laid-Open No. 2003-189,845, etc.). Examples of the seaweed morphogenesis factor include saroysin.

[0031] By the production method of the present invention, it is possible to efficiently grow seaweed vegetatively and increase the yield by suppressing the maturation of seaweed and the accompanying decrease in the algal thallus. Also, in an aquaculture site where it is necessary to keep the water temperature below a certain level, it is possible to reduce the energy required for reducing the water temperature and decrease the production cost.

[0032] In one aspect of the present invention, it is also possible to obtain algal thalli of algae by the production method of the present invention. The obtained algal thalli of algae can be used, for example, as raw materials for foods, active substances such as biofuels and pharmaceuticals, and the like.

Examples

[0033] Hereinafter, the present invention will be described in detail based on examples, but the present invention is not limited by these examples.

[0034] Reference Example 1 After aerating artificial seawater (500 mL) at a pressure of 0.15 MPa and a flow rate of 0.033 L / min for 1 hour using a microporous fine bubble generator (manufactured by Nack Co., Ltd., model FP20 - 70), the particle size and number of bubbles with diameters of 20 to 1000 nm were measured. As a control, the particle size and number were compared with those during normal aeration at the same flow rate using a general diffuser tube. The measurement was performed using ZetaView (manufactured by Particle Metrix Co., Ltd.). The results are shown in Fig. 1A. When aerating using the fine bubble generator, many bubbles with a particle size of less than 100 nm were observed. Also, using the same generator, artificial seawater (2 L with 1 ml of Porphyran Conco added) was treated at a pressure of 0.15 MPa and a total aeration volume of 0.08 L / min (aeration volume from the diffuser tube: aeration volume from the fine bubble generator = 1:1) for 30 minutes, 60 minutes, 120 minutes, and 240 minutes (see Fig. 2(a)), and the particle size and number of bubbles with diameters of 1 to 500 μm were measured. As a control, only normal aeration at 0.08 L / min using a diffuser tube under the same conditions for 240 minutes (see Fig. 2(b)) was compared with the particle size and number of bubbles. The measurement was performed using PartAn SI (manufactured by Microtrac Bell Co., Ltd.). The results are shown in Fig. 1B. By aeration with the fine bubble generator, many bubbles with a particle size of 100 μm or less were generated. Hereinafter, in the examples, fine bubbles were generated using a microporous fine bubble generator (manufactured by Nack Co., Ltd., model FP20 - 70).

[0035] Example 1 The wet weight of the thalli of Sargassum horneri was compared when cultured while supplying fine bubbles in addition to aeration by a general diffuser tube (Fig. 2(a)), or when cultured by aeration with a diffuser tube only (Fig. 2(b)). Five clumps of aggregated thalli of Sargassum horneri strain Kanemura (initial wet weight of thalli: 0.01 g / 5 clumps) were used as test thalli and placed in a medium prepared by adding 1 ml of a nutrient (Porphyran Conco, manufactured by Daiichi Steel Co., Ltd.) to 2 L of sterilized artificial seawater. Under the common culture conditions of a water temperature of 20°C, a light period / dark period of 12 hours / 12 hours, a total aeration volume of 0.06 L / min (aeration volume from the diffuser tube: aeration volume from the fine bubble generator = 1:1), and medium replacement on the 3rd and 5th days, the thalli were cultured for 7 days under each condition of normal aeration only or normal aeration plus fine bubble supply, and the wet weight (g) of the thalli was measured (Fig. 4). Also, the daily growth rate was calculated from the obtained results. As shown in Figs. 3 and 4, Sargassum horneri with fine bubble supply grew larger. The average daily growth rate (%) for 7 days was 1.2 times higher, 113.2% with fine bubbles compared to 92.6% in normal culture. Also, when observing the thalli, when cultured under fine bubble supply, the number of locations (arrows) where spores were released from the tip was small, and the degree of maturity was low (Fig. 5).

[0036] Example 2 Using strains Kanemura, Yoshino, and Matsuzaki of Sargassum horneri, the wet weight of the thalli at 7 days of culture was measured and the daily growth rate was calculated under the same conditions as in Example 1 except that the total aeration volume was 0.04 L / min (aeration volume from the diffuser tube: aeration volume from the fine bubble generator = 1:1). The initial wet weights of the thalli were 0.015 g for strain Kanemura, 0.02 g for strain Yoshino, and 0.027 g / 5 clumps for strain Matsuzaki, respectively. As a result, as shown in Fig. 6, for strains other than strain Kanemura, Sargassum horneri with fine bubble supply also tended to have a higher daily growth rate.

[0037] Example 3 To suppress the turbidity of water by supplying air bubbles from the bottom surface and eliminate the difference in light quantity throughout the container, a microporous generator is installed on the side surface of the culture container as shown in Fig. 7, and the saturated growth light quantity is 500 μmol / m 2 The wet weight (g) of the algal thalli of Sargassum horneri cultured for 8 days at / s was compared with the wet weight (g) of the algal thalli when only aeration with a general air diffuser was performed (Fig. 2(b)). Specifically, the following was done. Five clumps of aggregated algal thalli of Sargassum horneri Akashi strain (starting algal thalli wet weight: 0.01 g / 5 clumps) were used as the test algal thalli and placed in a medium prepared by adding 1 ml of a nutrient agent (Porphyran Conco, manufactured by Daiichi Steel Co., Ltd.) to 2 L of sterilized artificial seawater. The culture was carried out for 8 days under the culture conditions of a water temperature of 20 °C, a light period / dark period = 12 hours / 12 hours, a total aeration volume of 0.08 L / min (aeration volume from the air diffuser: aeration volume from the microporous bubble generator = 1:1), and medium replacement on the 2nd, 4th, and 6th days. The wet weight (g) of the algal thalli was measured and the daily growth rate was calculated. As shown in Fig. 8, even when a microporous bubble generator was installed on the side surface of the container, Sargassum horneri supplied with microporous bubbles grew larger. The average daily growth rate (%) over 8 days was 119.2% with microporous bubbles, which was 1.1 times higher than 107.2% in normal culture. Also, when observing the algal thalli, the part where Sargassum horneri supplied with microporous bubbles released spores from the tip was less, and the degree of maturity was low.

[0038] Example 4 The asexual strain of Cladophora hirohana (starting algal thalli wet weight: 0.017 g / 5 individuals) was used as the test algal thalli and placed in a medium prepared by adding 1 ml of a nutrient agent (Porphyran Conco, manufactured by Daiichi Seitetsu Co., Ltd.) to 2 L of sterilized artificial seawater. Under the common culture conditions of the saturated growth light quantity of Cladophora hirohana being 200 μmol / m 2 / s, a water temperature of 20 °C, a light period / dark period = 12 hours / 12 hours, a total aeration volume of 0.08 L / min (aeration volume from the air diffuser: aeration volume from the microporous bubble generator = 1:1), and medium replacement on the 2nd, 4th, and 6th days, the culture was carried out for 8 days under each condition of only normal aeration (Fig. 2(b)) or in addition to normal aeration, supplying microporous bubbles from the microporous bubble generator installed on the side surface of the container (Fig. 7), and the wet weight (g) of the algal thalli was measured. As shown in Fig. 9, the Undaria pinnatifida with fine bubble supply grew larger and no decrease in wet weight due to maturation was observed. Also, when observing the thallus, the one with fine bubbles had fewer holes and chipped parts at the edges and a lower degree of maturity.

[0039] Example 5 The Nishijo strain of Gracilaria lemaneiformis (initial thallus wet weight 0.19 g / 5 individuals) was used as the co-tested thallus and placed in a medium with 1 ml of nutrient agent (Porphyran Conco, manufactured by Daiichi Netsu Co., Ltd.) added to 2 L of sterilized artificial seawater. Under the common culture conditions of the growth saturation light intensity of Gracilaria lemaneiformis at 300 μmol / m 2 / s, water temperature 20°C, light period / dark period = 12 hours / 12 hours, total aeration volume 0.08 L / min (aeration volume from the diffuser: aeration volume from the fine bubble generator = 1:1), the medium was changed on the 2nd and 4th days, and on the 6th day, it was changed to a medium with 2 ml of Porphyran Conco added to 2 L of sterilized artificial seawater. The thalli were cultured for 8 days under each condition of only normal aeration (Fig. 2(b)) or in addition to normal aeration, fine bubble supply from a fine bubble generator installed on the side of the container (Fig. 7), and the wet weight (g) of the thalli was measured. Also, the daily growth rate was calculated from the obtained results. As shown in Fig. 10, the Gracilaria lemaneiformis with fine bubble supply grew larger. The average daily growth rate (%) over 8 days was about 10% higher at 50.5% with fine bubbles compared to 46.4% in normal culture.

Claims

1. A method for producing thalli of seaweeds, comprising a step of culturing seaweeds under the supply of fine bubbles and a step of recovering the thalli of the seaweeds, wherein the culture temperature in the culturing step is 15°C or higher. A method for producing thalli of seaweeds.

2. The production method according to Claim 1, wherein the seaweeds are seaweeds in which somatic cells differentiate into spores.

3. The production method according to Claim 1, wherein the seaweeds are Sargassum fusiforme, Ulva lactuca, Enteromorpha prolifera, Monostroma nitidum, Gelidium amansii, Porphyra tenera, Porphyra yezoensis, Grateloupia filicina, Sargassum thunbergii, Undaria pinnatifida, Hizikia fusiforme.

4. The production method according to Claim 1, wherein the seaweeds are Sargassum fusiforme, Porphyra yezoensis, Sargassum thunbergii.

5. The production method according to Claim 1, wherein the seaweeds are cultured under stirring conditions.

6. The production method according to Claim 1, wherein the fine bubbles include fine bubbles having a particle diameter of 500 nm or less.

7. The production method according to Claim 1, wherein the fine bubbles are supplied at a supply rate of 0.0001 L / min to 1 L / min per 1 L of the culture solution volume.

8. The production method according to Claim 1, wherein the fine bubbles are generated by a micropore method.

9. The thalli of the seaweeds obtained by the production method according to any one of Claims 1 to 8.

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