JPS5995828A - Laver samplng method - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for collecting seaweed seedlings in seaweed cultivation.

In seaweed aquaculture, the seaweed seedling method of culturing shell filaments and obtaining shell spores to attach to the seaweed net is an important task that determines the success or failure of seaweed aquaculture production.

Conventionally, the glue seedling method involves artificially perforating the shells of oysters, scallops, and other shells with caracospores released from protoalgae collected as mother algae from fishing grounds in a land aquarium. A method of growing for a long period of time until the autumn cool season when the chospores are released while periodically managing them under specific conditions, and a method of growing free filaments with fixed cultivar characteristics and genetic traits by shredding them with a mixer etc. A method of culturing shell filaments using free filament transplantation has been carried out, in which shell filaments are cultured by drilling sections into shells in the same manner as carospores.

However, these methods require a long culture period after transplantation into the shell, and require large land aquarium facilities for culturing.
Adjustment of irradiation light dose, irradiation time, culture solution temperature and specific gravity, cleaning to remove foreign algae that adhere to the shell surface and inhibit the growth of shell filaments, exchange of culture solution, etc. during the cold autumn season. This method has the drawback that it imposes a large economic burden and management effort on cultivators over a long period of time until spore collection.

The present inventors have conducted various studies to solve the above-mentioned economic and management disadvantages associated with the conventional glue seedling method, and as a result, they have developed the method of culturing cholaspores formed in shell filaments through pure culture. The mature chlospores are exposed by exposing them to an artificial culture solution by treating them at high temperature for a long period of time, separating the sporangium branches from the surface of the shell filament, and culturing them under substrate-free conditions to multiply as the sporangium branches. The present invention was completed after discovering that it can be obtained in large quantities in a short period of time.

That is, in the present invention, a pure cultured shell filament is cultured to make the shell sporangia branches extend, and then the bare part is separated and cut into pieces, and after being further cultured and multiplied, a low temperature and short day This is a method for collecting seedlings using glue, which is characterized by processing to release shell spores.

The present invention will be explained in detail below.

First, shell filaments that have been pure cultured using a known method are treated in an artificial culture solution at high temperature for a long period of time. Specifically, high temperature 24~
Static culture was performed for 21 to 30 days at 28°C, light irradiation amount (hereinafter referred to as "illuminance") to the shell filament surface, 1000 to 3000 lux, and 16 hours of light (8 hours of darkness) per day. The sporangia branches in the shell are extended into the artificial culture medium.

Next, the chosporangium branches exposed in this artificial culture solution are separated pure from the surface of the shell filament, and the separated chospore branches are shredded with a mixer (12,000 revolutions, 20 seconds), and further added to the artificial culture solution. Inside temperature 22~28℃, illuminance 1000~30
00lux, light period 13-16 hours per day (dark period 11-16 hours)
8 hours), the culture container is gently shaken 4 to 6 times per day, and the culture is statically cultured in a substrate-free condition to proliferate as cholosporangia branches. The cholaspore branches thus obtained are subjected to a known low-temperature, short-day treatment (temperature 18-20°C, illuminance 4000-5000 lux, 10 hours light/14 hours dark per day) as complementary culture for ripening. After approximately 4 days of aerated culture), the spores immediately enter meiosis, release a large amount of chospores, settle on the glue net, and germinate.

The composition of the artificial culture solution used in the present invention is city water 1000%
Sodium chloride 24g, potassium chloride 0.7g per ml
, magnesium sulfate 8g, calcium chloride 0.37g,
Sodium nitrate 0.32, sodium dihydrogen phosphate 0.
025g, sodium bicarbonate 0.336g, vitamin B12 0.02mg, phytic acid 0.5mg "Klewat-32" (trade name, manufactured by Teikoku Kagaku Sangyo Co., Ltd., metal ion sequestering agent) 20mg, "Klewat-N" 20mg
The pH of the solution is adjusted to 7.9 to 8.2.

Furthermore, the type of seaweed that can be applied to the present invention may be any type as long as it can be normally cultivated, but representative examples thereof include Asakusa nori, Narawasu sabinori, Futamatasu sabinori, Susabi nori, and Obasakusanori. , Malva Asakusanori, etc.

In the present invention, pure cultured shell filaments obtained by known methods are used, but in order to obtain the best effect, it is preferable to culture them under specific conditions for a certain period of time. As a culture solution, the above-mentioned artificial culture solution of the inventors, artificial seawater with a known composition, or even natural seawater are all possible, but sterilized culture from which foreign algae such as diatoms and green algae have been removed is acceptable. After perforating the shell with caracospores or free filaments using the liquid, the temperature is 15-22°C, the illuminance is 1000-3000 lux, and the light period is 12-13 hours (dark period 12-11 hours) for 5 to 5 hours. Shell filaments cultured for 7 months have well-grown filaments in the shell, and some that have matured to the point of releasing shell spores through low-temperature, short-day treatment are observed.

One feature of the present invention is that the shell filament in which fully ripened shell sporangia branches are formed in the shell through pure culture is cultured by high temperature and long day treatment. The growth of the sporangium branches is promoted and extends significantly, showing a growth rate of 10 to 20 times higher than in conventional culture methods.

The separated sporangium branches can be mechanically shredded without any trouble using a mixer, etc., and by such mechanical shredding, they are formed into a shape suitable for later static culture, for example, 100 to 500μ, and are physically The reproductive growth of the sporangium branches is accelerated by the stimulation, which promotes growth and proliferation in the next stationary culture under substrate-free conditions.

By static culture under no-substrate conditions, the strip-shaped conch sporangium branches are multiplied, but by gently shaking the shell sporangium several times a day during culture, the shell sporangium branches are stimulated and elongate. While growing, the cell contents are enriched and a large number of mature shell sporangia are formed in about 1 to 1.5 months.

The conch sporangia branches that have multiplied in large quantities in this way are known to be
By short-day treatment, it transitions to meiosis, releases a large amount of chospores, settles on the glue net, and germinates.

The glue seedling collecting method according to the present invention has the following advantages compared to conventional methods.

1) It is possible to culture large quantities of chospores.

2) Extensive land equipment made of concrete blocks or polyvinyl chloride, which is required in conventional culture of shell filaments, is not required.

3) There is no need for cleaning work to remove attached diatoms etc. from the surface of shell filaments, and for collecting and transporting a large amount of seawater required for water change.

4) The labor and economic burden of seedling management on cultivators will be greatly reduced.

Next, the present invention will be explained in more detail with reference to Examples.

Example 1 A pure culture was carried out for about 6 months using a known method (culture water temperature 20-22°C, light period 13 hours, illuminance 1500-2000 lux), and a species of Asakusa [Asakusa nori, scientific name: Porphyra tenella] with well-grown filaments in its shell was cultivated. porphyra te
nera) ] one shell filament (approx. 20 cm2),
Place the artificial culture solution in a 1L glass incubator with the lid on, and incubate at a high temperature of 26-27°C, under long-day conditions with a light period of 16 hours (dark period: 8 hours) and an illuminance of 1500-2000 lux for 2 hours.
After culturing for one day, the shell sporangia branches in the shell are allowed to extend into the artificial culture medium.

Next, about 0.5 g of wet weight of the bare shell sporangium branches was scraped off from the surface of the shell filament, and the pieces were cut into pieces (100 to 500 μm) using a mixer (12,000 revolutions, 20 seconds).
) was prepared and placed in a 1 liter glass incubator together with the artificial culture solution, and incubated at 22-24°C with a light period of 14 hours (dark period of 10 hours).
The cells are left to stand for one month at an illuminance of 1,000 to 1,500 lux (the incubator is gently shaken 4 to 6 times per day), and grown as sporangia branches. Usually one shell filament (20cm2)
It is possible to proliferate shell sporangia branches with a wet weight of about 2 g. Next, 1 g (wet weight) of the proliferated sporangium branches was placed in a 1 liter round flask with a bottom neck along with an artificial culture solution, and 1 g was added as a substrate for the growth of the sporangia.
Once a day at a fixed time, insert nylon thread (manufactured by Toray Industries, Inc., using the registered trademark "Amilan Net Yarn No. 4") and aerate and stir (
The pH of the culture solution is controlled at 7.9 to 8.2 by mixing carbon dioxide gas with aeration air. ) and temperature 18℃
, light period 10 hours (dark period 14 hours), illuminance 5000lux
Seedlings were collected for 8 days. Once a day, the nylon threads with seedlings taken out from the round flask with a bottom neck were placed in an artificial culture medium in a 4-liter transparent acrylic rectangular aquarium in order to measure the number of spores that had settled on the nylon threads. Aerate and stir at a temperature of 18℃.
The cells were cultured at a light period of 12 hours and an illuminance of 8000 lux. Culture 4
After a day, the lobule that had grown epiphytically on the nylon thread was measured using a microscope.

The results are shown in Table 1.

Shell spore release from 1 g was confirmed from the 4th day after the low-temperature, short-day treatment, peaked on the 6th day, and decreased rapidly thereafter. 8
Approximately 38 million shell spores were released during the one-day seedling collection period, and this amount is equivalent to approximately 7 to 8 sheets of glue net, assuming that 100 shell spores are attached to each 1 cm of glue net. This is equivalent to 50% of the total amount, which indicates that it can be used as seedlings for collecting seedlings at the production level.

Comparative Example One shell filament (approximately 20 cm2) of the same species of Asakusa as that used in Example 1 was grown for 500 m together with an artificial culture solution.
Pour into a 1-volume glass beaker and add nylon thread once a day at the same time as in Example 1 and aerate and stir (for pH, mix carbon dioxide gas with aeration air to adjust the pH of the culture solution to 7.9 to 8.2. ), and seedlings were harvested for 8 days at a temperature of 18°C, a light period of 10 hours (dark period of 14 hours), and an illuminance of 5000 lux.

Furthermore, the collected nylon threads on which the callus spores were attached were cultured under the same conditions as in Example 1, and after 4 days of culture, the lobules that had grown on the nylon threads were measured.

The results are shown in Table 2.

Examples 2 to 4 Various types of sporangia branches grown by the same method as in Example 1 were treated indoors at low temperature and short days for 4 days, and then used as seedlings for seedling collection using the burrow method outdoors. Served as. On the 4th day after seedlings were introduced, the bag was removed and the number of shell spores attached to the expanded glue net was measured. The results are shown in Table 3.

As shown in Table 3, in this seedling method (outdoor) at the production level, there are slight differences in the number of seedlings for each variety, but it is possible to collect seedlings on one glue net with 1 g of sporangium branches. This shows that the seeds and seedlings produced by the present inventors can be fully utilized in the present zubo-type seedling collection method at the production level.

Examples 5 to 6 Various shell sporangia branches grown by the same method as in Example 1 were treated indoors at low temperatures and for short days, and then shelled into the tank of a waterwheel-type seedling device used on land. Sporangium branches were introduced, seedlings were collected continuously for 4 days using the usual method, and Table 4 shows the number of glue nets on which the shell spores were attached to the appropriate density required for glue cultivation.

As shown in Table 4, when seedlings are harvested on land at the production level using the waterwheel seedling method, 1 g of sporangium branches is used to produce approximately 1.5 to 2 g of glue net.
The fact that a single sheet of seedlings could be taken shows that the seeds and seedlings produced by the present inventors can be fully utilized in the water wheel type seedling collecting method at the production level.

Claims (1)

[Claims] 1) Cultivate pure cultured shell filaments to extend shell sporangia branches, then separate and shred the exposed parts, further culture and propagate, and then treat at low temperature and short days. A glue seedling collection method characterized by releasing shell spores.