CN109122425B - Method for inducing Pacific eupatorium odoratum to produce resting eggs - Google Patents
Method for inducing Pacific eupatorium odoratum to produce resting eggs Download PDFInfo
- Publication number
- CN109122425B CN109122425B CN201811105423.4A CN201811105423A CN109122425B CN 109122425 B CN109122425 B CN 109122425B CN 201811105423 A CN201811105423 A CN 201811105423A CN 109122425 B CN109122425 B CN 109122425B
- Authority
- CN
- China
- Prior art keywords
- culture solution
- salinity
- eggs
- temperature
- culturing
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/20—Culture of aquatic animals of zooplankton, e.g. water fleas or Rotatoria
Abstract
The invention discloses a method for inducing Pacific daphnia to lay resting eggs, which comprises the following steps: culturing with culture medium to obtain culture solution of Platymonas subcordiformis and Skeletonema costatum, mixing microalgae culture solution with sterilized seawater and attractant prepared from abscisic acid and acetylcholine to obtain culture solution of Eudaphnia magna; and (3) breeding the adult daphnia magna in a culture solution, and performing combined stimulation by adopting illumination, temperature and salinity to obtain the resting eggs of the daphnia magna. The method for inducing the Pacific daphnia magna to produce the resting eggs has the advantages of strong operability, high repeatability, high resting egg occurrence rate and high induction efficiency, fully utilizes the measure of combined stimulation of illumination, temperature and salinity, and utilizes the progressive decline type temperature-illumination stimulation method to obtain more resting eggs of the Pacific daphnia magna with more sufficient energy storage.
Description
Technical Field
The invention relates to the technical field of aquatic organisms, in particular to a method for inducing Pacific daphnia to produce resting eggs.
Background
Copepods belonging to the phylum Arthropoda, Crustacea and the subclass copepoda are important components of zooplankton, which are small crustaceans with a body length of less than 3mm, and are distributed in oceans, fresh water or brackish water for planktonic and parasitic life. It is one of the initial feeds for most fish larvae, and therefore has great significance in food nets for aquatic ecosystems and aquaculture. Pacific daphnia is a species belonging to the order Choriophyceae, the family Daphtaleidae, the genus Daphtales, and is one of the very common warm-water coastal copepods species in low-salt regions. Pacific Eudaphnia hybrida mainly appears in summer and autumn, and begins to lay dormant eggs by the end of autumn, and the dormant eggs sink into the sea bottom and are buried in sea mud and continuously hatch to supplement the population.
Copepods are a key group of the ecosystem of water and play an important role in material circulation, energy flow and information transfer. The dormant eggs are an important survival strategy of the copepods, play an important role in resisting adverse environments, maintaining population continuation and the like, and have very important significance for seasonal dynamics and species continuation of copepods. The dormant eggs are the products of offspring continued by sexual reproduction of copepods under adverse environmental conditions, have the characteristics of resisting external adverse environments, being easy to diffuse, generating genetic diversity through recombination and the like, are also important seed sources for intensive culture of zooplankton, and are propagated in a large amount in a short time through rapid parthenogenesis of copepods, so that the copepods are used as fresh and alive baits to play an extremely important role in the production of offspring seeds of aquatic economic animals.
The generation of dormant eggs, the induction of the formation of the dormant eggs and the timely hatching and utilization of the dormant eggs are also very significant problems in the aquaculture industry. Copepods have many applications in aquaculture due to their high content of certain essential fatty acids, particularly EPA and DHA, and are often used as starter feed for some commercial fish. With the deep research and the development of scientific technology, a large number of nauplii obtained through the dormant eggs of the copepods are gradually valued and utilized by people, and the large-scale culture of the dormant eggs plays an important role in modern mariculture and has wide application prospect.
However, the problems of the structure of the resting eggs and the like are limited by the fact that the resting eggs can be collected in the sea only in one season within one year, and researches show that the rhythmicity of the production of the resting eggs by the Pacific daphnia speciosa is mainly the influence of external environmental stimuli such as temperature, light period, environmental salinity and the like or animal behaviors such as sexual selection, parasitic behaviors, spawning behaviors and the like on the prior representation of the current representation, so that how to induce the production of a large number of resting eggs is a difficult problem to solve urgently.
Disclosure of Invention
The invention aims to provide a method for inducing the Pacific daphnia to lay dormant eggs, which has the advantages of strong operability, high repeatability, high incidence rate of the dormant eggs and high induction efficiency.
Aiming at the problems mentioned in the background technology, the invention adopts the technical scheme that:
a method for inducing Pacific Eupatorium Adenophorum to lay dormant eggs is implemented by subjecting adult Eupatorium Adenophorum to combined stimulation of progressively decreasing illumination, temperature and salinity to obtain and collect dormant eggs. The method is simple and convenient, the measures of combined stimulation of illumination, temperature and salinity are fully utilized to complete high-efficiency induction, the progressive descending temperature-illumination stimulation method can be utilized to stimulate the sexual reproduction of the parent of the real daphnia magna, the higher incidence rate of dormant eggs is obtained, and the aim of quickly and efficiently inducing and producing the dormant eggs is fulfilled.
Preferably, the specific steps of the combined stimulation of the progressive decline type illumination, the temperature and the salinity are as follows:
step 1: adjusting the salinity of the culture solution to be 2.8-3.1%, and temporarily culturing for 3-5 days under the conditions that the temperature is 24-27 ℃, the illumination intensity is 2500-3000 lux, and the light-dark time is 16h:8 h;
step 2: adjusting the salinity of the culture solution to 3.3-3.6%, and culturing for 23-26 h under the conditions that the temperature is 18-22 ℃, the illumination intensity is 1800-2000 lux, and the light-dark time is 12h:12 h;
and step 3: adjusting the salinity of the culture solution to 3.3-3.6%, and culturing for 23-26 h under the conditions that the temperature is 15-18 ℃, the illumination intensity is 1800-2000 lux, and the light-dark time is 12h:12 h;
and 4, step 4: adjusting the salinity of the culture solution to 3.3-3.6%, and culturing at 12-16 deg.C, illumination intensity of 1800-2000 lux, and light-dark time of 12h:12h until the generation of resting eggs. The abiotic factors such as illumination, temperature, water environment salinity, humidity and food have certain induction effect on the reproduction of zooplankton and the production of resting eggs, for example, at higher temperature, the adult daphnia pseudostreda can accelerate the development, shorten the time required by sexual maturity, accelerate and promote the growth and reproduction of the daphnia pseudostreda, and at lower temperature, the amphoterism of copepods can be induced; other abiotic factors also have a significant effect on the reproductive capacity and resting egg incidence of daphnia magna.
Preferably, the population density of the adult daphnia pseudobrevicaulis in the culture solution is 0.06-1.6 per ml. The population density is an important factor for keeping the population stable by self-regulation of animals, the collision rate between real daphnia magna is increased along with the increase of the population density, the probability of generating dormant eggs is obviously increased, and the adequate control of the population density can avoid insufficient food and space crowding caused by overlarge population density, thereby avoiding reduction of the reproductive capacity.
Preferably, the collection of resting eggs is performed by using a bolting silk with a pore diameter of 45-50 μm.
Preferably, the culture medium used for inducing the pacific eupatorium to lay the resting eggs contains an attractant consisting of abscisic acid and acetylcholine. Abscisic acid and acetylcholine form a highly active state after being stimulated by illumination, alpha and beta unsaturated ketone structures contained in molecules of abscisic acid and acetylcholine are easily combined with hydrogen atoms of amino acids in protein and further combined with a diapause hormone receptor on an oocyte membrane, so that the activity of trehalase on the oocyte membrane is improved, the trehalase in maternal blood is decomposed into glucose, the glucose enters the oocyte through the cell membrane, a large amount of glycogen is synthesized in the oocyte and is stored, and the laid fertilized eggs enter a dormant state at the initial embryo stage before the differentiation of embryo dermis is completed, so that the incidence rate of the dormant eggs is further improved, and meanwhile, more sufficient energy can be stored for the dormant eggs.
Further preferably, the optimal technical scheme is that the weight ratio of the abscisic acid to the acetylcholine in the attractant is 2-2.5: 1.5.
Further preferably, the culture solution for inducing the pacific eupatorium to lay the resting eggs also comprises a culture solution of Platymonas subcordiformis and Skeletonema costatum, and the preparation steps are as follows: inoculating Platymonas subcordiformis and skeletonema costatum to an MS liquid culture medium, adjusting the pH of the culture medium to 7.5-8.5, adjusting the salinity to 15-35 per mill, and culturing at the temperature of 20-30 and the illumination intensity of 5000-10000 lux. The unicellular algae not only has proper size and can be ingested by zooplankton, but also can be digested and absorbed quickly in animal bodies, and contains rich nutrition, thereby being beneficial to the growth and development of animals.
Further preferably, the culture solution for inducing the Pacific Dolichos pelagi to lay the resting eggs comprises the following raw materials in parts by weight: 5-10 parts of culture solution of Platymonas subcordiformis and Skeletonema costatum, 20-25 parts of sterilized seawater and 0.25-0.55 part of attractant.
And further preferably, the sterilized seawater is obtained by filtering floating sand and impurities from natural seawater by using a silk screen with the aperture of 35-45 mu m, sterilizing and continuously inflating for 20-25 hours.
Compared with the prior art, the invention has the advantages that: 1) the method for inducing the Pacific daphnia variegata to lay the resting eggs effectively solves the problem of collecting the resting eggs of the Pacific daphnia variegata, is not limited by the collection of the open sea area and the rhythmicity of biological egg laying, and can obtain more resting eggs with more sufficient energy storage; 2) in the process of inducing the Pacific real daphnia to lay the resting eggs, measures of combined stimulation of illumination, temperature and salinity are utilized to complete high-efficiency induction, and the progressive descending temperature-illumination stimulation method is utilized to stimulate the sexual reproduction of the mother generation of the real daphnia to obtain higher resting egg occurrence rate, so that the aim of quickly and efficiently inducing the resting eggs to be produced is fulfilled; 3) the method for inducing the Pacific eudaphnia to lay the resting eggs is simple, convenient and fast, has strong operability, repeatability, high resting egg occurrence rate and high induction efficiency,
a higher proportion of resting eggs can be obtained.
Detailed Description
The scheme of the invention is further illustrated by the following examples:
example 1:
a method for inducing Pacific Eupatorium odoratum to produce resting eggs is characterized by that the adult Eupatorium odoratum cultured by 2 generations of domestication and culture and having good activity is progressively undergone the processes of reduced illumination, temp. and salinity combined stimulation to obtain and collect resting eggs. The method is simple, convenient and fast, has strong operability, fully utilizes the measures of combined stimulation of illumination, temperature and salinity to complete high-efficiency induction, and utilizes the progressive descending temperature-illumination stimulation method to stimulate the sexual reproduction of the hymexana brevifolia offspring, thereby obtaining higher incidence rate of dormant eggs and realizing the purpose of quickly and efficiently inducing and producing the dormant eggs.
The specific steps of the progressive decline type illumination, temperature and salinity combined stimulation are as follows:
step 1: adjusting the salinity of the culture solution to 2.8%, and temporarily culturing for 3d under the conditions of 24 deg.C, illumination intensity of 2500lux, and light and dark time of 16h:8 h;
step 2: adjusting the salinity of the culture solution to 3.3%, and culturing for 23h under the conditions of 18 deg.C, 1800lux illumination intensity, 12h light and shade time;
and step 3: adjusting the salinity of the culture solution to 3.3%, and culturing for 23h under the conditions of 15 deg.C, 1800lux illumination intensity, and 12h light and shade time;
and 4, step 4: adjusting salinity of the culture solution to 3.3%, and culturing at 12 deg.C, illumination intensity of 1800lux, and light-dark time of 12h:12h until dormant eggs are produced. The abiotic factors such as illumination, temperature, water environment salinity, humidity and food have certain induction effect on the reproduction of zooplankton and the production of resting eggs, for example, at higher temperature, the adult daphnia pseudostreda can accelerate the development, shorten the time required by sexual maturity, accelerate and promote the growth and reproduction of the daphnia pseudostreda, and at lower temperature, the amphoterism of copepods can be induced; other abiotic factors also have a significant effect on the reproductive capacity and resting egg incidence of daphnia magna.
The population density of adult daphnia pseudobrevica in the culture solution is 0.06 pieces/ml. The population density is an important factor for keeping the population stable by self-regulation of animals, the collision rate between real daphnia magna is increased along with the increase of the population density, the probability of generating dormant eggs is obviously increased, and the adequate control of the population density can avoid insufficient food and space crowding caused by overlarge population density, thereby avoiding reduction of the reproductive capacity.
The resting eggs were collected using a silk screen with a pore size of 45 μm.
The culture solution used for inducing the Pacific Eupatorium Adenophorum to lay the resting eggs contains an attractant consisting of abscisic acid and acetylcholine, the weight ratio of the abscisic acid to the acetylcholine in the attractant is 2:1.5, the abscisic acid and the acetylcholine form a highly active state after being stimulated by illumination, alpha and beta unsaturated ketone structures contained in molecules of the attractant are easily combined with hydrogen atoms of amino acids in proteins and further combined with a diapause hormone receptor on an oocyte membrane, the activity of trehalase on the oocyte membrane is improved, so that the trehalase in maternal blood is decomposed into glucose, the glucose enters the oocyte through the membrane, a large amount of glycogen is synthesized in the oocyte and stored, the laid fertilized eggs enter the resting state at the early embryo stage before the differentiation of embryo dermis is completed, and the occurrence rate of the resting eggs is further improved, more sufficient energy can be stored for resting eggs.
The culture solution for inducing the Pacific eupatorium Adenophora to lay the resting eggs also comprises a culture solution of Platymonas subcordiformis and Skeletonema costatum, and the preparation steps are as follows: inoculating Platymonas subcordiformis and Skeletonema costatum to MS liquid culture medium, adjusting pH of the culture medium to 7.5, adjusting salinity to 15 ‰, and culturing at 20 deg.C and illumination intensity of 5000 lux. The unicellular algae not only has proper size and can be ingested by zooplankton, but also can be digested and absorbed quickly in animal bodies, and contains rich nutrition, thereby being beneficial to the growth and development of animals.
The culture solution for inducing the Pacific eupatorium odoratum to lay the resting eggs comprises the following raw materials in parts by weight: 5 parts of culture solution of Platymonas subcordiformis and Skeletonema costatum, 20 parts of sterilized seawater and 0.25 part of a trapping agent.
The sterilized seawater is prepared by filtering natural seawater with 35 μm mesh screen, sterilizing, and continuously aerating for 20 hr.
Example 2:
a method for inducing Pacific Dolichos latipes to produce resting eggs, wherein the step of inducing Pacific Dolichos latipes to produce resting eggs comprises the following steps: placing the adult daphnia magna which is domesticated and cultured for 2 generations and has good activity into a culture solution, leading the population density of the adult daphnia magna in the culture solution to be 0.26/ml, and obtaining and collecting the resting eggs by using bolting silk with the aperture of 50 mu m through the measures of progressive descending illumination, temperature and salinity combined stimulation during the culture.
The specific steps of the progressive decline type illumination, temperature and salinity combined stimulation are as follows:
step 1: adjusting the salinity of the culture solution to 3.0%, and temporarily culturing for 4d under the conditions of 27 deg.C, illumination intensity of 3000lux, and light and dark time of 16h:8 h;
step 2: adjusting the salinity of the culture solution to 3.6%, and culturing for 25h under the conditions of 20 deg.C, 1900lux illumination intensity, and 12h light and shade time;
and step 3: adjusting the salinity of the culture solution to 3.6%, and culturing for 25h under the conditions of 16 deg.C, 1900lux illumination intensity, and 12h light and shade time;
and 4, step 4: adjusting salinity of the culture solution to 3.6%, and culturing at 14 deg.C under illumination intensity of 1900lux and light-dark time of 12h:12h until the generation of resting eggs.
The culture solution for inducing the Pacific eupatorium odoratum to lay the resting eggs comprises the following raw materials in parts by weight: 8 parts of culture solution of Platymonas subcordiformis and Skeletonema costatum, 25 parts of sterilized seawater and 0.50 part of a lure promoter, wherein the weight ratio of abscisic acid to acetylcholine in the lure promoter is 2.5: 1.5.
The preparation method of the culture solution of Platymonas subcordiformis and Skeletonema costatum comprises the following steps: inoculating Platymonas subcordiformis and Skeletonema costatum to MS liquid culture medium, adjusting pH of the culture medium to 8.5, and salinity to 30 ‰, and culturing at 25 deg.C and illumination intensity of 10000 lux.
The sterilized seawater is prepared by filtering natural seawater with 40 μm mesh screen, sterilizing, and continuously aerating for 22 hr.
Example 3:
a method for inducing Pacific Dolichos latipes to produce resting eggs, wherein the step of inducing Pacific Dolichos latipes to produce resting eggs comprises the following steps: placing the adult daphnia magna which is domesticated and cultured for 2 generations and has good activity into a culture solution, leading the population density of the adult daphnia magna in the culture solution to be 0.8/ml, and obtaining and collecting the resting eggs by using bolting silk with the aperture of 48 mu m through the measures of progressive descending illumination, temperature and salinity combined stimulation during the culture.
The specific steps of the progressive decline type illumination, temperature and salinity combined stimulation are as follows:
step 1: adjusting the salinity of the culture solution to 3.0%, and temporarily culturing for 3d under the conditions of 25 ℃, illumination intensity of 2600lux and light and dark time of 16h:8 h;
step 2: adjusting the salinity of the culture solution to 3.5%, and culturing for 24h under the conditions of 22 deg.C, illumination intensity of 2000lux, and light-dark time of 12h:12 h;
and step 3: adjusting the salinity of the culture solution to 3.5%, and culturing for 24h under the conditions of 18 deg.C, illumination intensity of 2000lux, and light-dark time of 12h:12 h;
and 4, step 4: adjusting the salinity of the culture solution to 3.5%, and culturing at 16 deg.C under illumination intensity of 2000lux and light-dark time of 12h:12h until the generation of resting eggs.
The culture solution for inducing the Pacific eupatorium odoratum to lay the resting eggs comprises the following raw materials in parts by weight: 10 parts of culture solution of Platymonas subcordiformis and Skeletonema costatum, 22 parts of sterilized seawater and 0.35 part of a lure promoter, wherein the weight ratio of abscisic acid to acetylcholine in the lure promoter is 2.3: 1.5.
The preparation method of the culture solution of Platymonas subcordiformis and Skeletonema costatum comprises the following steps: inoculating Platymonas subcordiformis and Skeletonema costatum to MS liquid culture medium, adjusting pH to 8.0 and salinity to 25 ‰, and culturing at 28 deg.C and illumination intensity of 8000 lux.
The sterilized seawater is prepared by filtering natural seawater with 45 μm mesh screen, sterilizing, and continuously charging air for 24 hr.
Example 4:
a method for inducing Pacific Eupatorium Adenophorum to lay resting eggs further optimizes the scheme as follows:
when the salinity of the culture solution for inducing the pacific eupatorium to produce the resting eggs is adjusted in the step 4, 0.065 percent by weight of glutamine is added into the culture solution, the weight ratio of L-glutamine to D-glutamine in the glutamine is 5:0.8, at the moment, microalgae in the culture solution is eaten by the eupatorium for a long time and is at a level with lower content of microalgae, the eupatorium is in a hungry state in a short time, so that the permeability of intestinal mucosa of the eupatorium is increased, the glutamine in the proportion is added into the culture solution, so that the intestinal tract can be stimulated to maintain a mucosal structure, the activity of intestinal tract cells is enhanced, the secretion of insulin and testosterone is increased, the sexual reproduction of adults is promoted, meanwhile, the coupling can be formed with antifreeze protein and opsin, the parents are stimulated to quickly reach the diapause induction sensitive period in the closed-loop period, more diapause hormone is released, and external bad stimulation signals received by the parents are converted into breeding signals, directly acts on the offspring, thereby improving the induction efficiency and stimulating the generation of more and more stable dormant eggs.
The steps of inducing the Pacific eudaphnia to lay the resting eggs are as follows: placing the adult daphnia magna which is domesticated and cultured for 2 generations and has good activity into a culture solution, leading the population density of the adult daphnia magna in the culture solution to be 0.8/ml, and obtaining and collecting the resting eggs by using bolting silk with the aperture of 48 mu m through the measures of progressive descending illumination, temperature and salinity combined stimulation during the culture.
The specific steps of the progressive decline type illumination, temperature and salinity combined stimulation are as follows:
step 1: adjusting the salinity of the culture solution to 3.0%, and temporarily culturing for 3d under the conditions of 25 ℃, illumination intensity of 2600lux and light and dark time of 16h:8 h;
step 2: adjusting the salinity of the culture solution to 3.5%, and culturing for 24h under the conditions of 22 deg.C, illumination intensity of 2000lux, and light-dark time of 12h:12 h;
and step 3: adjusting the salinity of the culture solution to 3.5%, and culturing for 24h under the conditions of 18 deg.C, illumination intensity of 2000lux, and light-dark time of 12h:12 h;
and 4, step 4: adjusting the salinity of the culture solution to 3.5%, and culturing at 16 deg.C under illumination intensity of 2000lux and light-dark time of 12h:12h until the generation of resting eggs.
The culture solution for inducing the Pacific eupatorium odoratum to lay the resting eggs comprises the following raw materials in parts by weight: 10 parts of culture solution of Platymonas subcordiformis and Skeletonema costatum, 22 parts of sterilized seawater and 0.35 part of a lure promoter, wherein the weight ratio of abscisic acid to acetylcholine in the lure promoter is 2.3: 1.5.
The preparation method of the culture solution of Platymonas subcordiformis and Skeletonema costatum comprises the following steps: inoculating Platymonas subcordiformis and Skeletonema costatum to MS liquid culture medium, adjusting pH to 8.0 and salinity to 25 ‰, and culturing at 28 deg.C and illumination intensity of 8000 lux.
The sterilized seawater is prepared by filtering natural seawater with 45 μm mesh screen, sterilizing, and continuously charging air for 24 hr.
Comparative example:
a method for inducing Pacific Dolichos latipes to produce resting eggs, wherein the step of inducing Pacific Dolichos latipes to produce resting eggs comprises the following steps: placing the adult daphnia magna which is domesticated and cultured for 2 generations and has good activity into a culture solution, leading the population density of the adult daphnia magna in the culture solution to be 0.8/ml, and obtaining and collecting the resting eggs by using bolting silk with the aperture of 48 mu m through the measures of progressive descending illumination, temperature and salinity combined stimulation during the culture.
The specific steps of the progressive decline type illumination, temperature and salinity combined stimulation are as follows:
step 1: adjusting the salinity of the culture solution to 3.0%, and temporarily culturing for 3d under the conditions of 25 ℃, illumination intensity of 2600lux and light and dark time of 16h:8 h;
step 2: adjusting the salinity of the culture solution to 3.5%, and culturing for 24h under the conditions of 22 deg.C, illumination intensity of 2000lux, and light-dark time of 12h:12 h;
and step 3: adjusting the salinity of the culture solution to 3.5%, and culturing for 24h under the conditions of 18 deg.C, illumination intensity of 2000lux, and light-dark time of 12h:12 h;
and 4, step 4: adjusting the salinity of the culture solution to 3.5%, and culturing at 16 deg.C under illumination intensity of 2000lux and light-dark time of 12h:12h until the generation of resting eggs.
The culture solution for inducing the Pacific eupatorium odoratum to lay the resting eggs comprises the following raw materials in parts by weight: 10 parts of culture solution of Platymonas subcordiformis and Skeletonema costatum, and 22 parts of sterilized seawater.
In this comparative example, the culture solution for inducing the pacific daphnia magna to produce resting eggs was prepared in the same manner as in example 3, except that no attractant was added.
Example 5:
induced production test of resting eggs of Pacific Eupatorium odoratum
Selecting the adult daphnia magna which is domesticated and cultured for 2 generations and has good vigor, respectively placing the adult daphnia magna in 5 culture ponds according to the conditions of the embodiments 1-4 and the comparative example, simultaneously setting a group of blank groups, and culturing by adopting the conditions of common illumination, salinity and temperature. During the incubation period, the conditions of each test were strictly controlled, and when the number of breeding parents was detected to be less than 2, the test was ended. The experimental results are statistically as follows.
TABLE 1 Pacific Eupatorium Adenophorum resting egg induced production test
| Rate of occurrence of males/%) | Resting egg incidence/%) | Normal egg occurrence/%) | |
| Example 1 | 40.7 | 64.5 | 35.5 |
| Example 2 | 40.6 | 64.7 | 35.3 |
| Example 3 | 42.7 | 65.3 | 34.7 |
| Example 4 | 43.3 | 65.5 | 34.5 |
| Comparative example | 37.8 | 57.3 | 42.7 |
| Blank group | 33.3 | 50.5 | 49.5 |
From the above table, the emergence rate of the male bodies of the true daphnia magna obtained in the examples 1-4 is obviously higher than that of the comparative example, especially 13.7% of the true daphnia magna obtained in the examples 3 and 4, and is also obviously higher than that of the blank group by 29.1%; the difference of the dormant egg incidence rate of the examples 1-4 is not large, and the average is higher than that of the comparative example 7.7 percent and is higher than that of the blank group by 15 percent; the experimental result shows that the incidence rate of the dormant eggs produced in the embodiment of the invention is obviously stimulated by the combination of the gradually-decreasing illumination, the temperature and the salinity, the incidence rate of the dormant eggs is obviously improved, particularly the embodiment 3 and the embodiment 4 are obviously influenced by the attractant, and the method can complete the induction of the dormant eggs of the daphnia magna by the measures of the combined stimulation of the illumination, the temperature and the salinity, obtain higher incidence rate of the dormant eggs, obtain more dormant eggs with more sufficient energy storage and realize the purpose of quickly and efficiently inducing and producing the dormant eggs.
The conventional operations in the operation steps of the present invention are well known to those skilled in the art and will not be described herein.
The embodiments described above are intended to illustrate the technical solutions of the present invention in detail, and it should be understood that the above-mentioned embodiments are only specific embodiments of the present invention, and are not intended to limit the present invention, and any modification, supplement or similar substitution made within the scope of the principles of the present invention should be included in the protection scope of the present invention.
Claims (8)
1. A method for inducing Pacific Eupatorium Adenophorum to lay resting eggs is characterized in that: obtaining and collecting resting eggs by carrying out progressive descending type illumination, temperature and salinity combined stimulation on adult daphnia magna;
the specific steps of the progressive decline type illumination, temperature and salinity combined stimulation are as follows:
step 1: adjusting the salinity of the culture solution to be 2.8-3.1%, and temporarily culturing for 3-5 days under the conditions that the temperature is 24-27 ℃, the illumination intensity is 2500-3000 lux, and the light-dark time is 16h:8 h;
step 2: adjusting the salinity of the culture solution to 3.3-3.6%, and culturing for 23-26 h under the conditions that the temperature is 18-22 ℃, the illumination intensity is 1800-2000 lux, and the light-dark time is 12h:12 h;
and step 3: adjusting the salinity of the culture solution to 3.3-3.6%, and culturing for 23-26 h under the conditions that the temperature is 15-18 ℃, the illumination intensity is 1800-2000 lux, and the light-dark time is 12h:12 h;
and 4, step 4: adjusting the salinity of the culture solution to 3.3-3.6%, and culturing under the conditions that the temperature is 12-16 ℃, the illumination intensity is 1800-2000 lux, and the light and shade time is 12h:12h until dormant eggs are generated;
the culture solution contains a promoting agent consisting of abscisic acid and acetylcholine; the weight ratio of the abscisic acid to the acetylcholine in the attractant is 2-2.5: 1.5;
the culture solution comprises the following raw materials in parts by weight: 5-10 parts of culture solution of Platymonas subcordiformis and Skeletonema costatum, 20-25 parts of sterilized seawater and 0.25-0.55 part of attractant.
2. The method of claim 1, wherein the method comprises the steps of: the population density of the adult daphnia pseudobrevicaulis in a culture solution is 0.06-1.6 per ml.
3. The method of claim 1, wherein the method comprises the steps of: the collection of the resting eggs is carried out by using bolting silk with the aperture of 45-50 mu m.
4. The method of claim 1, wherein the method comprises the steps of: the culture solution for inducing the Pacific eupatorium to lay the resting eggs also comprises a culture solution of Platymonas subcordiformis and Skeletonema costatum.
5. The method of claim 1, wherein the method comprises the steps of: the preparation conditions of the culture solution of the Platymonas subcordiformis and the Skeletonema costatum are as follows: an MS liquid culture medium is adopted, the pH value of the culture medium is 7.5-8.5, the salinity is 15-35 per mill, the culture temperature is 20-30 ℃, and the illumination intensity is 5000-10000 lux.
6. The method of claim 1, wherein the method comprises the steps of: the sterilized seawater is obtained by filtering floating sand and impurities from natural seawater by using a silk screen with the aperture of 35-45 mu m, sterilizing and continuously inflating for 20-25 h.
7. The method of claim 1, wherein the method comprises the steps of: the specific steps of the progressive decline type illumination, temperature and salinity combined stimulation are as follows:
step 1: adjusting the salinity of the culture solution to 3.0%, and temporarily culturing for 3d under the conditions of 25 ℃, illumination intensity of 2600lux and light and dark time of 16h:8 h;
step 2: adjusting the salinity of the culture solution to 3.5%, and culturing for 24h under the conditions of 22 deg.C, illumination intensity of 2000lux, and light-dark time of 12h:12 h;
and step 3: adjusting the salinity of the culture solution to 3.5%, and culturing for 24h under the conditions of 18 deg.C, illumination intensity of 2000lux, and light-dark time of 12h:12 h;
and 4, step 4: adjusting the salinity of the culture solution to 3.5%, and culturing at 16 deg.C under illumination intensity of 2000lux and light-dark time of 12h:12h until the generation of resting eggs.
8. The method of claim 1, wherein the method comprises the steps of: the culture solution comprises the following raw materials in parts by weight: 10 parts of culture solution of Platymonas subcordiformis and Skeletonema costatum, 22 parts of sterilized seawater and 0.35 part of a lure promoter, wherein the weight ratio of abscisic acid to acetylcholine in the lure promoter is 2.3: 1.5.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811105423.4A CN109122425B (en) | 2018-09-21 | 2018-09-21 | Method for inducing Pacific eupatorium odoratum to produce resting eggs |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811105423.4A CN109122425B (en) | 2018-09-21 | 2018-09-21 | Method for inducing Pacific eupatorium odoratum to produce resting eggs |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109122425A CN109122425A (en) | 2019-01-04 |
| CN109122425B true CN109122425B (en) | 2021-06-08 |
Family
ID=64823107
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811105423.4A Active CN109122425B (en) | 2018-09-21 | 2018-09-21 | Method for inducing Pacific eupatorium odoratum to produce resting eggs |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109122425B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106577409A (en) * | 2016-12-21 | 2017-04-26 | 厦门大学 | Method for inducing copepoda to produce diapause eggs |
-
2018
- 2018-09-21 CN CN201811105423.4A patent/CN109122425B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106577409A (en) * | 2016-12-21 | 2017-04-26 | 厦门大学 | Method for inducing copepoda to produce diapause eggs |
Non-Patent Citations (3)
| Title |
|---|
| 几种单胞藻对太平洋真宽水蚤生物学特性的影响;夏立萍等;《浙江海洋大学学报(自然科学版)》;20170731;第36卷(第4期);第308-314页 * |
| 河口近海桡足类休眠卵生态学研究进展;王庆等;《应用生态学报》;20150531;第26卷(第7期);第2213-2224页 * |
| 海洋桡足类滞育卵在海水养殖业中的应用及其前景;刘光兴等;《青岛海洋大学学报》;20031231;第33卷(第6期);第901-906页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109122425A (en) | 2019-01-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102210278B (en) | Method for domesticating and breeding wild hippocampus | |
| CN1745725A (en) | Artificial culture of shrimp parents | |
| Andriantahina et al. | Comparison of reproductive performance and offspring quality of domesticated Pacific white shrimp, Litopenaeus vannamei | |
| CN111226843A (en) | Special artemia indoor culture method for seawater fish and shrimp seedling culture | |
| CN106614125A (en) | Breeding method of hybrid bream | |
| CN110100770B (en) | Artificial propagation method of siganus oramin | |
| CN101444192B (en) | Sebastiscus marmoratus industrialized fry breeding method | |
| CN102919186B (en) | Artificial breeding method for sillago sihama | |
| CN103053476B (en) | Cultivating method for obligate parthenogenesis rotifera population | |
| CN105557584A (en) | All-seawater larval rearing and indoor industrialized culture method for trachidermus fasciatus heckel | |
| Ajah | Mass culture of Rotifera(Brachionus quadridentatus[Hermann, 1783]) using three different algal species | |
| Li et al. | The success of yellow catfish aquaculture in China: from rare wild fish to popular farmed fish | |
| CN109122425B (en) | Method for inducing Pacific eupatorium odoratum to produce resting eggs | |
| CN114766397B (en) | Cultivation method of bellied sea horse | |
| CN109892261B (en) | Artificial breeding method of Babuyan jellyfishes | |
| CN104273073A (en) | Breeding method for bay purple hybrid golden yellow adductor muscle scallops | |
| CN110622892B (en) | Water-flowing type production promoting method for oplegnathus fasciatus and juvenile fish culturing method | |
| Vasudevan et al. | Intensive cultivation of the calanoid copepod Oithona rigida for mariculture purpose | |
| KR101476032B1 (en) | Embryo production method for mackerel | |
| CN108283163A (en) | Continuous cultural method in a kind of big artemia room of obligate ovoviviparity breeding | |
| CN110352887A (en) | A kind of saline-alkaline pond phytoplankton-artemia-litopenaeus vannamei ecological cultivation method | |
| Holt et al. | Advances in cobia research in Texas | |
| CN106900600B (en) | Enhanced cultivation method for improving fertility of sepiella maindroni | |
| CN110684666A (en) | Selenium-containing copepods culture method | |
| CN108902055B (en) | Method for inducing Pacific spiny daphnia to produce resting eggs |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |