CN101731141A - Hybridization breeding method of undaria pinnatifida and sea tangle - Google Patents
Hybridization breeding method of undaria pinnatifida and sea tangle Download PDFInfo
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Abstract
The invention relates to a method adopting undaria pinnatifida female gametophytes saved and cultured indoors and sea tangle female and male gametophytes to be mutually used as male parents and female parents and cutting the gametophytes. The method comprises the following steps of: mixing according to the mode of the undaria pinnatifida female gametophytes * the sea tangle male gametophytes and the sea tangle female gametophytes * undaria pinnatifida male gametophytes; cutting into algae segments of 200 micrometers by a tissue pounding machine, and then scattering on a vinylon seeding bed; and cultivating the vinylon seeding bed at temperature suitable for the growth and the maturity of the undaria pinnatifida and sea tangle gametophytes, wherein after culturing for 7 days to 10 days, the sea tangle female gametophytes are mature to form oocysts and ovulate ova which are respectively fertilized with sperms released by mature sea tangle and undaria pinnatifida male gametophytes so as to germinate hybrid seedlings. The invention can sufficiently utilize the heteroses of the distant hybridization of undaria pinnatifida and sea tangle; in addition, obtained hybrids have inheritable characteristics of the undaria pinnatifida and the sea tangle, high stress resistance, fast growth and good quality and can greatly increase the cultivation output of the hybrid undaria pinnatifida and sea tangle and enhance the quality of the hybrid undaria pinnatifida and sea tangle.
Description
Technical field:
The present invention relates to the breeding method of a kind of section intermolecular hybrid, especially a kind of simple to operate, with low cost, the hybrid vigour of undaria pinnitafida, sea-tangle distant hybridization can be made full use of, hybridization undaria pinnitafida, the cultivation output of sea-tangle and the undaria pinnitafida and the sea-tangle cross breeding method of quality can be increased substantially.
Background technology:
Undaria pinnitafida (Undaria pinnatifida) belongs to Phaeophyta, brown sub-guiding principle, Laminariales, Alariaceae, undaria, belong to the eurythermic algae, its delicious flavour, nutritious, contain 19 seed amino acids, protein, sugar part and the vitamin that comprise whole essential amino acids, because the content of cellulose of undaria pinnitafida only is about 3% of a dry weight, therefore be easy to by human consumption, be very popular healthy food.
Sea-tangle (Laminaria japonica) belongs to Phaeophyta, brown sub-guiding principle, Laminariales, Laminariaceae, Larminaria in classification, belong to cold water algae.Abundant because of albumen, carbohydrate and vitamin content in the sea-tangle, have hypotensive, reducing blood lipid and oxidation resistant function, particularly polyoses content is higher, has the enhancing body immunity, the effect of prevention and inhibition cancer, medicinal and health care is worth very high, is the marine alga of present domestic consumption amount maximum.
At present, the undaria pinnitafida seed produces and to mainly contain indoor artificial seedling rearing and manually to adopt spore sea area half artificial breeding dual mode, the main spore mode of collecting seedling that adopts in the artificial seedling rearing produces wherein, and the while is also carried out the small-scale gametophyte indoor raising of seedling mode of collecting seedling; Sea-tangle is natural daylight low temperature artificial seedling rearing, mainly adopts the spore mode of collecting seedling; What both genetic breedings all adopted is that intraspecific hybridization or the method for controlling ploidy are carried out the germplasm improvement, does not also have the relevant report about undaria pinnitafida and the breeding of Laminariaceae intermolecular hybrid up to now.
Summary of the invention:
The purpose of this invention is to provide a kind of simple to operately, with low cost, can make full use of the hybrid vigour of undaria pinnitafida, sea-tangle distant hybridization, can increase substantially hybridization undaria pinnitafida, the cultivation output of sea-tangle and the undaria pinnitafida and the sea-tangle cross breeding method of quality.
Technical solution of the present invention is: a kind of undaria pinnitafida and sea-tangle cross breeding method is characterized in that carrying out as follows:
A. in the breeding period of undaria pinnitafida, get the sporophyll of the ripe kind algae maturation of buoyant raft cultivation, behind dry in the shade stimulation, put into the beaker that adds the sterilization seawater, when being light brown, the seawater in the beaker pulls sporophyll out, getting spore fluid with suction pipe moves on in the culture dish that adds culture fluid, it is that 20 ℃, intensity of illumination are to cultivate culture fluid full dose exchange in per 10 days 1 time 1 month under the condition of 2000Lx that culture dish is moved on to temperature;
B. in breeding period, get the ripe kind of buoyant raft cultivation algae in sea-tangle, cut the sporangium part with scissors, adopt with a step same procedure spore is captured in the culture dish, condition of culture, time are with a step;
C. distinguish the egagametophyte and the male gametophyte of a step and b step institute output at microscopically, egagametophyte or male gametophyte are moved on to respectively in the different culture dishes, carry out that undaria pinnitafida is female, male gametophyte, female, the male gametophyte separation and Culture of sea-tangle, cultivated 3 months, group peels from the culture dish bottom surface with the female and male gametophytes algae, move on in the flask, add culture fluid and preserve cultivation, culture fluid full dose exchange in 1 month 1 time;
The undaria pinnitafida oogamete of d. going bail for and depositing cultivation, the undaria pinnitafida male gametophyte, each weight in wet base 5mg of sea-tangle egagametophyte and sea-tangle male gametophyte, according to undaria pinnitafida egagametophyte * sea-tangle male gametophyte, sea-tangle egagametophyte * undaria pinnitafida male gametophyte combination mixes female and male gametophytes, in mixture, use behind the interpolation 300ml sterilization seawater respectively and organize the algae section of agitator chopping to 200 μ m, be sprinkling upon and be covered with on the seedling curtain of coiling diameter 3mm breed with rope, treat that gametophyte is behind adhesion-tight on the vinylon rope, it is 15 ℃ that undaria pinnitafida egagametophyte * sea-tangle male gametophyte combination moves on to temperature, intensity of illumination is 2000Lx, light application time is to cultivate at least 7 days in culture fluid under 12 hours/day conditions, culture fluid full dose exchange in per 3 days 1 time; It is that 10 ℃, intensity of illumination are that 2000Lx, light application time are to cultivate 10 days culture fluid full dose exchange in per 3 days 1 time under the condition at least in 12 hours/day in culture fluid that the combination of sea-tangle egagametophyte * undaria pinnitafida moves on to temperature;
Described culture fluid adds NaNO for being heated in 80 ℃ of nature seawaters after filtering to 1L
3100mg, NaH
2PO
412H
2O 20mg and PI solution 1ml.
The present invention is simple to operate, with low cost, the just contrary hybridization that can realize undaria pinnitafida and sea-tangle (belongs to section's intermolecular hybrid, belong to distant hybridization biologically), compare with crossbreeding technologies such as existing sea-tangle interspecific cross, can make full use of undaria pinnitafida, the hybrid vigour of sea-tangle distant hybridization, the crossbreed that is obtained has undaria pinnitafida, the genetic character of sea-tangle, strong stress resistance, fast growth, quality is good, not only can increase substantially the hybridization undaria pinnitafida, cultivation output and the quality of hybridization sea-tangle, and can and backcross by directive breeding, cultivate undaria pinnitafida, the sea-tangle new varieties.
Embodiment:
A. in the breeding period of undaria pinnitafida, get the sporophyll of the ripe kind algae maturation of buoyant raft cultivation, behind dry in the shade stimulation, put into the beaker that adds the sterilization seawater, diffuse and pull sporophyll when spore fluid is light brown out when having observed a large amount of spores, move on to culture dish (the diameter 9cm that adds culture fluid with the suction pipe spore fluid that takes a morsel, down together), after 2 hours, as seen the culture dish bottom surface has circular sporoblast to adhere to, it is that 20 ℃, intensity of illumination are to cultivate culture fluid full dose exchange in per 10 days 1 time 1 month under the condition of 2000Lx that culture dish is moved on to temperature.
B. in breeding period, get the ripe kind of buoyant raft cultivation algae in sea-tangle, cut the sporangium part with scissors, adopt with a step same procedure spore is captured in the culture dish, condition of culture, time are with a step.
Cultivate after 2 days, sprout the formation female and male gametophytes, cultivate after 1 month the female and male gametophytes algae group of naked eyes visible growth in culture dish attached to the undaria pinnitafida in the culture dish, sea-tangle sporoblast.
C. distinguish the egagametophyte and the male gametophyte of a step and b step institute output at microscopically, sterilization tweezers egagametophyte or male gametophyte are moved on to respectively in the different culture dishes, carry out that undaria pinnitafida is female, andro gamete, female, the male gametophyte separation and Culture of sea-tangle, cultivate after 3 months, female and male gametophytes algae group reaches 0.4～0.6mm, and group peels from the culture dish bottom surface with the female and male gametophytes algae, moves on in the 300ml flask, add culture fluid and preserve cultivation, culture fluid full dose exchange in 1 month 1 time;
The undaria pinnitafida oogamete of d. going bail for and depositing cultivation, the undaria pinnitafida male gametophyte, each weight in wet base 5mg of sea-tangle egagametophyte and sea-tangle male gametophyte, according to undaria pinnitafida egagametophyte * sea-tangle male gametophyte, sea-tangle egagametophyte * undaria pinnitafida male gametophyte combination mixes female and male gametophytes, in mixture, use behind the interpolation 300ml sterilization seawater respectively and organize the algae section of agitator chopping to 200 μ m, be sprinkling upon on the seedling curtain that is covered with coiling diameter 3mm vinylon rope, treat that gametophyte is behind adhesion-tight on the vinylon rope, it is 15 ℃ that undaria pinnitafida egagametophyte * sea-tangle male gametophyte combination moves on to temperature, intensity of illumination is 2000Lx, light application time is to cultivate at least 7 days in culture fluid under 12 hours/day conditions, culture fluid full dose exchange in per 3 days 1 time; It is that 10 ℃, intensity of illumination are that 2000Lx, light application time are to cultivate 10 days culture fluid full dose exchange in per 3 days 1 time under the condition at least in 12 hours/day in culture fluid that the combination of sea-tangle egagametophyte * undaria pinnitafida moves on to temperature.
Undaria pinnitafida egagametophyte * sea-tangle male gametophyte is combined in to cultivate after 7 days and observes, and visible undaria pinnitafida egagametophyte germinates and ripely forms egg capsule and discharge ovum; The sea-tangle male gametophyte is also reached maturity and is formed anthreid and emit sperm, sprouts into undaria pinnitafida and sea-tangle crossbreed seedling after the feritilization of ovum.
Equally, sea-tangle egagametophyte * undaria pinnitafida is combined in cultivates the maturation that germinates after 10 days, and the sea-tangle egagametophyte forms egg capsule and discharges ovum; The undaria pinnitafida male gametophyte forms anthreid and emits sperm, sprouts into sea-tangle, undaria pinnitafida crossbreed seedling after the feritilization of ovum.
A～described culture fluid of d step is same as the prior art, promptly adds the sterilization seawater of nutritive salt, with reference to list of references.
Experiment: when undaria pinnitafida egagametophyte * sea-tangle male gametophyte, sea-tangle egagametophyte * undaria pinnitafida two combined hybrid kind growth of seedling to 200 μ m left and right sides, the seedling curtain of growth being hybridized seedling moves on on the buoyant raft of sea area foster temporarily; Support temporarily through 1 month sea area, two combined hybrid seedling length reach 0.5cm, the vinylon rope of growth seedling is cut into the 3cm segment, is clipped in the cultivation Miao Shengshang of long 8m, diameter 3cm, will cultivate Miao Shengping again and hang over that cultivation is the crossbreed adult on the undaria pinnitafida cultivation buoyant raft with the 35cm spacing.
The result: it is good that Hybrid strong stress resistance, fast growth, quality are educated by institute.
Described culture fluid adds NaNO for being heated in 80 ℃ of nature seawaters after filtering to 1L
3100mg, NaH
2PO
412H
2O 20mg and PI solution 1ml.
PI solution is disclosed in western Ze one person of outstanding talent, thousand former light heros.Algae organon [M] East capital: upright altogether Co., Ltd., 1979:281～293. of publishing
Claims (1)
1. undaria pinnitafida and sea-tangle cross breeding method is characterized in that carrying out as follows:
A. in the breeding period of undaria pinnitafida, get the sporophyll of the ripe kind algae maturation of buoyant raft cultivation, behind dry in the shade stimulation, put into the beaker that adds the sterilization seawater, when being light brown, the seawater in the beaker pulls sporophyll out, getting spore fluid with suction pipe moves on in the culture dish that adds culture fluid, it is that 20 ℃, intensity of illumination are to cultivate culture fluid full dose exchange in per 10 days 1 time 1 month under the condition of 2000Lx that culture dish is moved on to temperature;
B. in breeding period, get the ripe kind of buoyant raft cultivation algae in sea-tangle, cut the sporangium part with scissors, adopt with a step same procedure spore is captured in the culture dish, condition of culture, time are with a step;
C. distinguish the egagametophyte and the male gametophyte of a step and b step institute output at microscopically, egagametophyte or male gametophyte are moved on to respectively in the different culture dishes, carry out that undaria pinnitafida is female, andro gamete, female, the male gametophyte separation and Culture of sea-tangle, cultivated 3 months, group peels from the culture dish bottom surface with the female and male gametophytes algae, move on in the flask, add culture fluid and preserve cultivation, culture fluid full dose exchange in 1 month 1 time;
The undaria pinnitafida oogamete of d. going bail for and depositing cultivation, the undaria pinnitafida male gametophyte, each weight in wet base 5mg of sea-tangle egagametophyte and sea-tangle male gametophyte, according to undaria pinnitafida egagametophyte * sea-tangle male gametophyte, sea-tangle egagametophyte * undaria pinnitafida male gametophyte combination mixes female and male gametophytes, in mixture, use behind the interpolation 300ml sterilization seawater respectively and organize the algae section of agitator chopping to 200 μ m, be sprinkling upon and be covered with on the seedling curtain of coiling diameter 3mm breed with rope, treat that gametophyte is behind adhesion-tight on the rope, it is 15 ℃ that undaria pinnitafida egagametophyte * sea-tangle male gametophyte combination moves on to temperature, intensity of illumination is 2000Lx, light application time is to cultivate at least 7 days in culture fluid under 12 hours/day conditions, culture fluid full dose exchange in per 3 days 1 time; It is that 10 ℃, intensity of illumination are that 2000Lx, light application time are to cultivate 10 days culture fluid full dose exchange in per 3 days 1 time under the condition at least in 12 hours/day in culture fluid that the combination of sea-tangle egagametophyte * undaria pinnitafida moves on to temperature;
Described culture fluid adds NaNO for being heated in 80 ℃ of nature seawaters after filtering to 1L
3100mg, NaH
2PO
412H
2O 20mg and PI solution 1ml.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101946712A (en) * | 2010-09-03 | 2011-01-19 | 中国海洋大学 | Algae breeding method based on intergeneric hybridization |
| CN102917582A (en) * | 2010-07-31 | 2013-02-06 | 中国海洋大学 | Method of breedind Laminaria based on induction of polyploid gametophyte |
| CN103651101A (en) * | 2013-12-06 | 2014-03-26 | 中国海洋大学 | Novel hybrid variety of Saccharinajaponica and Costariacostata |
| CN103704121A (en) * | 2013-12-02 | 2014-04-09 | 中国海洋大学 | Costaria costata gametophyte clone hybridization seedling raising technology |
| CN104094832A (en) * | 2014-06-30 | 2014-10-15 | 中国科学院海洋研究所 | Method for crossbreeding of undaria pinnatifida by using unisexual planospores |
| CN105706893A (en) * | 2016-02-03 | 2016-06-29 | 大连海宝渔业有限公司 | Cloned undaria pinnatifida seedling attachment method |
| CN117296702A (en) * | 2023-11-29 | 2023-12-29 | 山东东方海洋科技股份有限公司 | Cultivation method and application of tetraploid hybrid kelp |
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2009
- 2009-11-30 CN CN2009102202628A patent/CN101731141B/en not_active Expired - Fee Related
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102917582A (en) * | 2010-07-31 | 2013-02-06 | 中国海洋大学 | Method of breedind Laminaria based on induction of polyploid gametophyte |
| CN102917582B (en) * | 2010-07-31 | 2014-09-03 | 中国海洋大学 | Method of breedind Laminaria based on induction of polyploid gametophyte |
| CN101946712A (en) * | 2010-09-03 | 2011-01-19 | 中国海洋大学 | Algae breeding method based on intergeneric hybridization |
| CN103704121A (en) * | 2013-12-02 | 2014-04-09 | 中国海洋大学 | Costaria costata gametophyte clone hybridization seedling raising technology |
| CN103651101A (en) * | 2013-12-06 | 2014-03-26 | 中国海洋大学 | Novel hybrid variety of Saccharinajaponica and Costariacostata |
| CN104094832A (en) * | 2014-06-30 | 2014-10-15 | 中国科学院海洋研究所 | Method for crossbreeding of undaria pinnatifida by using unisexual planospores |
| CN104094832B (en) * | 2014-06-30 | 2017-02-01 | 中国科学院海洋研究所 | Method for crossbreeding of undaria pinnatifida by using unisexual planospores |
| CN105706893A (en) * | 2016-02-03 | 2016-06-29 | 大连海宝渔业有限公司 | Cloned undaria pinnatifida seedling attachment method |
| CN117296702A (en) * | 2023-11-29 | 2023-12-29 | 山东东方海洋科技股份有限公司 | Cultivation method and application of tetraploid hybrid kelp |
| CN117296702B (en) * | 2023-11-29 | 2024-02-02 | 山东东方海洋科技股份有限公司 | Cultivation method and application of tetraploid hybrid kelp |
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