CN105454042A - Method for inducing somatic embryo from zostera marina embryo - Google Patents
Method for inducing somatic embryo from zostera marina embryo Download PDFInfo
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- CN105454042A CN105454042A CN201410463126.2A CN201410463126A CN105454042A CN 105454042 A CN105454042 A CN 105454042A CN 201410463126 A CN201410463126 A CN 201410463126A CN 105454042 A CN105454042 A CN 105454042A
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Abstract
The present invention relates to a method for inducing somatic embryo from zostera marina embryo. The method comprises that zostera marina embryo is adopted as an explant, sterile embryo is peeled from fruits or seeds, and is inoculated into a somatic embryo induction culture medium, induced culture is performed at a temperature of 15-25 DEG C under a dark or weak illumination condition, transfer inoculation is performed every two weeks, and small enation is generated on the embryo after 60 days so as to form the somatic embryo, wherein the callus stage does not exist during the process, and the somatic embryo induction culture medium is a solid culture medium adopting a MS culture medium as a basic culture medium and added with an organic additive, a carbon source, plant auxin and a sea salt solid medium. According to the present invention, the callus induction culture stage does not exist, and the zostera marina somatic embryo is directly induced from the embryo within the short time, such that the possible approach is provided for the achievement of the zostera marina sterile tissue culture seedling regeneration and the establishment of the tissue culture rapid propagation system establishing, and the foundation is established for the successful seaweed tissue culture.
Description
Technical field
Plant Tissue Breeding research field belonging to the present invention, the present invention relates to a kind of with the abductive approach of the Zostera marina body embryo blast that is explant.
Background technology
Zostera marina (
zosteramarinal.) be the sociales of northern China coastal (Shandong, Hebei and Liaoning) sea grass bed; there is extremely important Ecosystem Service and economic worth: it is not only the good place that the marine economy biologies such as many economic fishs are perched, breed and hidden oneself; these halobiontic young and ovum are played a protective role; it can also form numerous humus; for the marine animal such as plankton, immature fishes and shrimps provides excellent natural bait; as in Jiaodong Peninsula and Liaodong Peninsula, Zostera marina sea grass bed is the best natural habitat of this famous-brand and high-quality marine products of sea cucumber.
In nearly two ten years, China's Zostera marina sea grass bed ecosystem has suffered serious destruction, area sharply reduces, therefore, the reparation of Zostera marina sea grass bed has in recent years become a content indispensable in the reparation of northern China marine ecosystems, its grassland needs a large amount of seedlings in repairing, and utilizing plant tissue culture technique to turn out aseptic seedling in indoor is the effective ways alleviating seedling shortage in current Zostera marina reparation transplanting.
Somatic embryo (also known as embryoid) is an important intermediate link of aseptic seedling regeneration in Plant Tissue Breeding, and its successful acquisitions can be aseptic seedling and regenerate and establish important basis.The present invention for explant, induces its somatic embryo by tissue culture technique with the embryo of Zostera marina, adopts the Patents of this technical scheme and document not to be reported at present.
Summary of the invention
The object of this invention is to provide a kind of abductive approach of Zostera marina somatic embryo, with the embryo of Zostera marina for explant, effectively can obtain somatic embryo, for the Fast-propagation of Zostera marina and the regeneration of aseptic plantlet in vitro provide important technical support.
The technical solution adopted in the present invention is:
The abductive approach of a kind of Zostera marina somatic embryo of the present invention is the explant that is somatic embryos with the embryo of Zostera marina.
Concrete steps of the present invention are as follows:
1, the choosing of explant: the fruits and seeds gathering Zostera marina, disinfection in superclean bench after cleaning, obtains aseptic fruits and seeds, peels off pericarp or seed coat, takes out aseptic embryo;
2, the Fiber differentiation of somatic embryo: aseptic embryo is inoculated in medium, 15-25 DEG C, cultivate under dark or low light condition.Once, after 60d, on visible embryo, adularescent kick occurs, can confirm as somatic embryo after about 75d, period does not experience callus phase in every first quarter moon switching.Inducing culture is that salinity is 20-35 ‰ and with the addition of the MS solid culture medium of organic additive, carbon source, plant hormone.
In the present invention, institute's fruit picking is real in being close to mellow fruit, and seed is healthy mature seed.
In the present invention, preferred culture medium is that MS medium adds caseinhydrolysate 500mg/L, sucrose 60g/L, PSK0.001mg/L, agar 5g/L, sea salt 20g/L.
In the present invention, preferred condition of culture is 18-20 DEG C, light culture.
The invention has the advantages that: grass wrack seeds can be preserved for a long time, therefore with the embryo in it for explant time, the acquisition time of explant is longer; And fruit and seed are easy to surface sterilization, therefore surface sterilization effect promotes greatly, and the ratio obtaining aseptic embryo also improves greatly.The present invention successfully induces somatic embryo from Zostera marina embryo, for the realization of zostera aseptic plantlet in vitro regeneration provides another possible approaches, also for the success of sea grass tissue cultures is laid a good foundation.
Embodiment
Disclose the present invention with most preferred embodiment below, so it is not intended to limiting the invention, and all equivalences done according to the embodiment of the present invention are replaced and amendment, all belongs in the spiritual and the scope of the claims that defined by claim of invention of the present invention.
embodiment:an abductive approach for Zostera marina somatic embryo, implementation step comprises:
1) the choosing of explant: gather its healthy spathe inflorescence when Zostera marina fruit is close to maturation, peel off the bract outside spathe inflorescence, only stay the inside inflorescence and on fruit, hydrogen peroxide soaking disinfection 20min with 15% in superclean bench after cleaning, use sterilizing seawater flushing 3-5 time again, obtain aseptic inflorescence, take off the fruit on inflorescence one by one, cut pericarp, take out aseptic embryo;
2) Fiber differentiation of somatic embryo: aseptically aseptic embryo is inoculated in inducing culture, whole incubation 18-20 DEG C, carry out under dark condition.Every first quarter moon switching once.Inducing culture is medium based on MS medium, adds caseinhydrolysate 500mg/L, sucrose 60g/L, PSK0.001mg/L, agar 5g/L, sea salt 20g/L.After cultivating 60d, on visible embryo, adularescent kick occurs, can confirm as somatic embryo after about 75d, period does not experience callus phase.Somatic embryo inducement rate is 15.58%.
Claims (5)
1. an abductive approach for Zostera marina somatic embryo, is characterized in that, is the explant that is somatic embryos with the embryo of Zostera marina.
2. abductive approach as claimed in claim 1, it is characterized in that, step is as follows:
1) the choosing of explant: the fruits and seeds gathering Zostera marina, disinfection in superclean bench after cleaning, obtains aseptic fruits and seeds, peels off pericarp or seed coat, takes out aseptic embryo as explant;
2) Fiber differentiation of somatic embryo: aseptic embryo is inoculated in medium, 15-25 DEG C, cultivate under dark or low light condition, every first quarter moon switching once, adularescent kick appearance on visible embryo after 60d, can confirm as somatic embryo after about 75d, period does not experience callus phase; Inducing culture is that salinity is 20-35 ‰ and with the addition of the MS solid culture medium of organic additive, carbon source, plant hormone.
3. method for inducing and cultivating as claimed in claim 2, is characterized in that, institute's fruit picking is real in being close to mellow fruit, and seed is healthy mature seed.
4. method for inducing and cultivating as claimed in claim 2, is characterized in that, preferred culture medium is that MS medium adds caseinhydrolysate 500mg/L, sucrose 60g/L, PSK0.001mg/L, agar 5g/L, sea salt 20g/L.
5. method for inducing and cultivating as claimed in claim 2, it is characterized in that, preferred condition of culture is 18-20 DEG C, dark culturing.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410463126.2A CN105454042B (en) | 2014-09-12 | 2014-09-12 | A kind of method from Zostera marina embryonal induction somatic embryo |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201410463126.2A CN105454042B (en) | 2014-09-12 | 2014-09-12 | A kind of method from Zostera marina embryonal induction somatic embryo |
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| CN105454042A true CN105454042A (en) | 2016-04-06 |
| CN105454042B CN105454042B (en) | 2017-10-03 |
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| CN201410463126.2A Expired - Fee Related CN105454042B (en) | 2014-09-12 | 2014-09-12 | A kind of method from Zostera marina embryonal induction somatic embryo |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101904304A (en) * | 2010-07-16 | 2010-12-08 | 中国海洋大学 | Cultivating method of zostera aseptic seedlings |
| CN102577699A (en) * | 2012-01-01 | 2012-07-18 | 山东东方海洋科技股份有限公司 | Method for cultivating zostera marina aseptic seedlings |
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- 2014-09-12 CN CN201410463126.2A patent/CN105454042B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101904304A (en) * | 2010-07-16 | 2010-12-08 | 中国海洋大学 | Cultivating method of zostera aseptic seedlings |
| CN102577699A (en) * | 2012-01-01 | 2012-07-18 | 山东东方海洋科技股份有限公司 | Method for cultivating zostera marina aseptic seedlings |
Non-Patent Citations (1)
| Title |
|---|
| 吴茜: "大叶藻愈伤组织体系的建立及优化", 《中国优秀硕士学位论文全文数据库农业科技辑》 * |
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