CN114793876A - Method for quickly culturing porphyra yezoensis protonema nutrient algae filaments - Google Patents
Method for quickly culturing porphyra yezoensis protonema nutrient algae filaments Download PDFInfo
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- CN114793876A CN114793876A CN202210406776.8A CN202210406776A CN114793876A CN 114793876 A CN114793876 A CN 114793876A CN 202210406776 A CN202210406776 A CN 202210406776A CN 114793876 A CN114793876 A CN 114793876A
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- Prior art keywords
- seawater
- filament
- culturing
- nutrient
- porphyra yezoensis
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- 241000206613 Pyropia yezoensis Species 0.000 title claims abstract description 30
- 238000012258 culturing Methods 0.000 title claims abstract description 25
- 241000195493 Cryptophyta Species 0.000 title claims abstract description 21
- 235000015097 nutrients Nutrition 0.000 title claims abstract description 20
- 238000000034 method Methods 0.000 title claims abstract description 16
- 239000013535 sea water Substances 0.000 claims abstract description 28
- 230000001954 sterilising effect Effects 0.000 claims abstract description 10
- 239000012528 membrane Substances 0.000 claims abstract description 7
- 150000003839 salts Chemical class 0.000 claims abstract description 6
- 239000004677 Nylon Substances 0.000 claims abstract description 4
- 230000005484 gravity Effects 0.000 claims abstract description 4
- 229920001778 nylon Polymers 0.000 claims abstract description 4
- 238000001816 cooling Methods 0.000 claims abstract description 3
- 238000001914 filtration Methods 0.000 claims abstract description 3
- 238000011081 inoculation Methods 0.000 claims abstract description 3
- 230000000050 nutritive effect Effects 0.000 claims abstract description 3
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 4
- 239000000741 silica gel Substances 0.000 claims description 4
- 229910002027 silica gel Inorganic materials 0.000 claims description 4
- 239000011521 glass Substances 0.000 claims description 3
- 239000012452 mother liquor Substances 0.000 claims description 2
- 239000011148 porous material Substances 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 abstract description 7
- 206010021033 Hypomenorrhoea Diseases 0.000 abstract description 3
- 230000012010 growth Effects 0.000 description 4
- 238000007789 sealing Methods 0.000 description 4
- 230000001276 controlling effect Effects 0.000 description 3
- 230000016574 developmental growth Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005286 illumination Methods 0.000 description 2
- 239000010413 mother solution Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 230000009105 vegetative growth Effects 0.000 description 2
- 206010042674 Swelling Diseases 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 229910000365 copper sulfate Inorganic materials 0.000 description 1
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 229920001296 polysiloxane Polymers 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G33/00—Cultivation of seaweed or algae
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/04—Electric or magnetic or acoustic treatment of plants for promoting growth
- A01G7/045—Electric or magnetic or acoustic treatment of plants for promoting growth with electric lighting
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Marine Sciences & Fisheries (AREA)
- Biodiversity & Conservation Biology (AREA)
- Botany (AREA)
- Ecology (AREA)
- Forests & Forestry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a method for quickly culturing porphyra yezoensis protonema nutrient algae filaments, which comprises the following steps: (1) sterilizing a transparent culture bottle by a high-temperature sterilization pot to obtain a container for culturing the filaments; (2) filtering seawater with the specific gravity of 1.016-1.018 by using a 200-mesh double-layer nylon bolting silk, sterilizing by using an autoclave, cooling to room temperature, and applying nutritive salt to serve as seawater for culturing porphyra yezoensis filaments; (3) putting the algae filaments into a culture bottle, and adding the seawater treated in the step (2), wherein the inoculation density of the algae filaments in the seawater is 0.60-0.80 g fresh weight/L; (4) air is taken as a gas source, and air is delivered to a culture flask through a filter membrane filter with the aperture of 0.22 mu m, so that the filament is ensured to be suspended in seawater; controlling the culture condition at 19-22 deg.c, light intensity of 2000 Lx-2500 Lx and light period of 14L to 10D; the invention provides sufficient provenance guarantee for meeting the demand of good varieties of porphyra yezoensis in production, has simple and easy operation and low cost, and can guarantee that the filament can grow rapidly in the stage of nutrient algae filament.
Description
Technical Field
The invention relates to a method for quickly culturing porphyra yezoensis protonema nutrient algae filaments, which is used for culturing porphyra yezoensis protonema nutrient algae filaments and belongs to the technical field of germplasm culture.
Background
Porphyra yezoensis is an important economic red alga, with two distinct generations, in which the fronds are the stages of consumption and the filaments are an important source of seedlings in porphyra yezoensis production. The thallus Porphyrae yezoensis is cultured in seawater to obtain filament, which is also called free filament because it is different from filament grown in pearl layer of shell in nature. The porphyra yezoensis free filament has the characteristics of indoor culture and capability of manually regulating and controlling the growth environment, and has become a main source of provenance in production. The porphyra yezoensis free filament body has a vegetative growth stage and a developmental growth stage, wherein the vegetative growth stage is called as a vegetative phycofilament stage, the developmental growth stage is also called as an expansion phycofilament stage, the vegetative phycofilaments can quickly penetrate into a shell nacre layer to grow, the existing observation result does not show that the expanded algae filament can directly penetrate into the shell pearl layer, so in order to meet the requirements of seed sources in production, the culture of the free filament needs to be kept in the state of the nutrient algae filament, which is also the key of the artificial culture of the porphyra yezoensis filament, in the filament culturing method which has been mainly carried out at present, a filament is cultured in a container such as a triangular flask, the growth is carried out under indoor natural light, the control of temperature and photoperiod has no clear standard, it is often the case that the filament develops to the stage of swelling of the algal filaments, which loses its function as a seed source.
Disclosure of Invention
The invention aims to provide a method for quickly culturing porphyra yezoensis protonema nutrient algae filaments, which has the advantages of simple and convenient operation, low cost and good effect, and provides sufficient provenance guarantee for meeting the requirement of good porphyra yezoensis varieties in production.
In order to achieve the purpose, the method for quickly culturing the porphyra yezoensis protonema nutrient algae filaments comprises the following steps:
(1) the transparent culture bottle is sterilized in a high-temperature sterilizer and then used as a container for culturing the filament. The culture bottle is a transparent glass bottle or a triangular bottle, and is soaked in 1% hydrochloric acid for 12h, cleaned, dried at 80 ℃, sterilized in a high-temperature sterilizer and used as a container for culturing the filaments.
(2) Filtering seawater with the specific gravity of 1.016-1.018 by using a 200-mesh double-layer nylon bolting silk, sterilizing by using an autoclave, cooling to room temperature, and applying nutritive salt to serve as seawater for culturing porphyra yezoensis filaments. Specifically, the nutrient salt is applied by adding 5ml of PES mother liquor per liter of seawater every 7 days.
(3) Putting the algae filaments into a culture bottle, and adding the seawater treated in the step (2), wherein the inoculation density of the algae filaments in the seawater is 0.60-0.80 g fresh weight/L;
(4) air is taken as a gas source, air is delivered to the culture bottle through a filter membrane filter with the aperture of 0.22 mu m by a silica gel hose, an adjustable airflow switch is externally connected to an air outlet, and when the adjustable airflow switch is used, bubbles are adjusted to flow out of seawater at the bottom of the culture bottle one by one, so that the filament is ensured to be suspended in the seawater; the culture conditions are controlled to be 19-22 ℃, the light intensity is 2000 Lx-2500 Lx, and the photoperiod is 14L: 10D. Specifically, air was filtered through a 0.22 μm-pore filter by an air pump and filled into the culture flask from the 2 nd hour to the 3 rd hour of irradiation, from the 8 th hour to the 9 th hour of irradiation, and from the 2 nd hour to the 2.5 th hour of darkness.
The invention improves the growth speed of the porphyra yezoensis filament nutrient algae filament by controlling the culture conditions (19-22 ℃, the light intensity is 2000 Lx-2500 Lx, the light period is 14L:10D), sterilizing the seawater, inflating at regular time, applying nutrient salt and other technical measures. The method has strong operability, and the filament can keep the rapid growth in the state of nutrient algae filament, and can provide a large amount of provenances for the production of porphyra yezoensis.
The invention has the beneficial technical effects that:
(1) the method of the invention provides a scientific and standard culture method for the development and utilization of porphyra yezoensis germplasm resources, and also provides sufficient provenance guarantee for meeting the demand of good varieties of porphyra yezoensis in production.
(2) The method is simple and easy to operate, has low cost, and can ensure that the filament can grow rapidly in the nutritional algae filament stage.
Detailed Description
A method for quickly culturing porphyra yezoensis protonema nutrient algae filaments comprises the following steps:
culture bottles: is a 5L transparent glass reagent bottle, is soaked in 1% hydrochloric acid for 12h, is cleaned and dried at 80 ℃, and is used as a container for culturing the filament after being sterilized by a high-temperature sterilization pot.
Seawater treatment: natural seawater with the seawater specific gravity of 1.016-1.018 is filtered by a 200-mesh double-layer nylon bolting silk, sterilized by an autoclave, cooled to room temperature, and then 5ml of PES mother solution is added into each liter of seawater to be used as seawater for culturing porphyra yezoensis filaments.
Temperature control: the temperature of the culture room is controlled to be 19-22 ℃ by using a household air conditioner.
Illumination: the full-wavelength LED lamp is used as a light source, the light intensity is controlled to be 2000 Lx-2500 Lx through the modulatable switch, and the light period is set to be 14L: 10D.
And (3) aeration culture: a centrifugal air pump is selected as air supply equipment, the power of the air pump can be selected to be 1.3kw, air is firstly filtered by 1% copper sulfate solution, air is sent to a culture bottle through a silica gel hose through a filter membrane filter with the aperture of 0.22 mu m, the silica gel hose is connected with an adjustable airflow switch, air is inflated twice in the illumination period, each time is inflated for 1 hour, and the air is inflated for 0.5 hour in the dark period.
Selecting prepared Porphyra yezoensis germplasm Y-0602, weighing 4g fresh and heavy filament, subpackaging into two 5L culture bottles, adding 2.5L sterilized seawater into each bottle, sealing with high temperature resistant tissue culture sealing membrane (filter membrane diameter 30mm), introducing ventilating silicone tube into the culture bottle at the edge of the sealing membrane, and sealing at the interface. Aerating and culturing, controlling the temperature to be 19-22 ℃, the light intensity to be 2000 Lx-2500 Lx, the light cycle to be 14L:10D, and adding 12.5ml of PES mother solution into the seawater every 7 days. After 65 days, 15g of Porphyra yezoensis free filaments were obtained.
The above embodiments do not limit the present invention in any way, and all technical solutions obtained by means of equivalent substitution or equivalent transformation fall within the protection scope of the present invention.
Claims (5)
1. A method for quickly culturing porphyra yezoensis protonema nutrient algae filaments is characterized by comprising the following steps:
(1) sterilizing a transparent culture bottle by a high-temperature sterilizing pot to obtain a container for culturing the filamentous body;
(2) filtering seawater with the specific gravity of 1.016-1.018 by using a 200-mesh double-layer nylon bolting silk, sterilizing by using an autoclave, cooling to room temperature, and applying nutritive salt to serve as seawater for culturing porphyra yezoensis filaments;
(3) putting the algae filaments into a culture bottle, and adding the seawater treated in the step (2), wherein the inoculation density of the algae filaments in the seawater is 0.60-0.80 g fresh weight/L;
(4) air is taken as a gas source, air is delivered to the culture bottle through a filter membrane filter with the aperture of 0.22 mu m by a silica gel hose, an adjustable airflow switch is externally connected to an air outlet, and when the adjustable airflow switch is used, bubbles are adjusted to flow out of seawater at the bottom of the culture bottle one by one, so that the filament is ensured to be suspended in the seawater; the culture conditions are controlled to be 19-22 ℃, the light intensity is 2000 Lx-2500 Lx, and the photoperiod is 14L: 10D.
2. The method for rapidly culturing a porphyra yezoensis filament nutrient algal filament according to claim 1, wherein: the culture bottle is a transparent glass bottle or a triangular flask.
3. The method for rapidly culturing a porphyra yezoensis filament nutrient algal filament according to claim 1, wherein: the nutrient salt is applied by adding 5ml of PES mother liquor per liter of seawater every 7 days.
4. The method for rapidly culturing a porphyra yezoensis filament nutrient algal filament according to claim 1, wherein: air was filtered through a 0.22 μm-pore filter by an air pump and filled into the culture flask at the 2 nd to 3 rd hours of irradiation, at the 8 th to 9 th hours of irradiation, and at the 2 nd to 2.5 th hours of darkness.
5. The method for rapidly culturing a porphyra yezoensis filament nutrient algal filament according to claim 1, wherein: the culture bottle is soaked in 1% hydrochloric acid for 12h, cleaned, dried at 80 ℃, sterilized in a high-temperature sterilization pot and used as a container for culturing the filament.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210406776.8A CN114793876A (en) | 2022-04-18 | 2022-04-18 | Method for quickly culturing porphyra yezoensis protonema nutrient algae filaments |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210406776.8A CN114793876A (en) | 2022-04-18 | 2022-04-18 | Method for quickly culturing porphyra yezoensis protonema nutrient algae filaments |
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| CN114793876A true CN114793876A (en) | 2022-07-29 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210406776.8A Pending CN114793876A (en) | 2022-04-18 | 2022-04-18 | Method for quickly culturing porphyra yezoensis protonema nutrient algae filaments |
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| CN (1) | CN114793876A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115380814A (en) * | 2022-09-28 | 2022-11-25 | 中国农业科学院农业环境与可持续发展研究所 | Porphyra yezoensis plant factory production system and method |
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| CA2546336A1 (en) * | 2003-11-20 | 2005-06-09 | Noritech Seaweed Biotechnologies Ltd | Technology for cultivation of porphyra and other seaweeds in land-based sea water ponds |
| JP2012200189A (en) * | 2011-03-25 | 2012-10-22 | Yamaguchi Prefecture | Method of fixing laver belonging to genus porphyra generating thallus directly from filament to yarn and fixing device using the same |
| CN103907527A (en) * | 2014-03-31 | 2014-07-09 | 江苏省海洋水产研究所 | Porphyra yezoensis germplasm propagation expanding method |
| CN105123491A (en) * | 2015-08-05 | 2015-12-09 | 汕头大学 | Laver free filament factory cultivation method based on function foodstuff development |
| CN105993909A (en) * | 2016-05-13 | 2016-10-12 | 集美大学 | Pyropia haitanensis pure line seedling cultivation method |
-
2022
- 2022-04-18 CN CN202210406776.8A patent/CN114793876A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2546336A1 (en) * | 2003-11-20 | 2005-06-09 | Noritech Seaweed Biotechnologies Ltd | Technology for cultivation of porphyra and other seaweeds in land-based sea water ponds |
| JP2012200189A (en) * | 2011-03-25 | 2012-10-22 | Yamaguchi Prefecture | Method of fixing laver belonging to genus porphyra generating thallus directly from filament to yarn and fixing device using the same |
| CN103907527A (en) * | 2014-03-31 | 2014-07-09 | 江苏省海洋水产研究所 | Porphyra yezoensis germplasm propagation expanding method |
| CN105123491A (en) * | 2015-08-05 | 2015-12-09 | 汕头大学 | Laver free filament factory cultivation method based on function foodstuff development |
| CN105993909A (en) * | 2016-05-13 | 2016-10-12 | 集美大学 | Pyropia haitanensis pure line seedling cultivation method |
Non-Patent Citations (1)
| Title |
|---|
| 孙维华: "紫菜自由丝状体的培育及移植", 中国水产 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115380814A (en) * | 2022-09-28 | 2022-11-25 | 中国农业科学院农业环境与可持续发展研究所 | Porphyra yezoensis plant factory production system and method |
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Application publication date: 20220729 |
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