CN103621402B - The induction method of a kind of sargassun regeneration plant - Google Patents
The induction method of a kind of sargassun regeneration plant Download PDFInfo
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- CN103621402B CN103621402B CN201310525496.XA CN201310525496A CN103621402B CN 103621402 B CN103621402 B CN 103621402B CN 201310525496 A CN201310525496 A CN 201310525496A CN 103621402 B CN103621402 B CN 103621402B
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- 238000000034 method Methods 0.000 title claims abstract description 35
- 230000006698 induction Effects 0.000 title claims abstract description 12
- 230000008929 regeneration Effects 0.000 title claims abstract description 12
- 238000011069 regeneration method Methods 0.000 title claims abstract description 12
- 230000001954 sterilising effect Effects 0.000 claims abstract description 20
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 20
- 229920001817 Agar Polymers 0.000 claims abstract description 18
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims abstract description 18
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims abstract description 18
- 229930006000 Sucrose Natural products 0.000 claims abstract description 18
- 239000008272 agar Substances 0.000 claims abstract description 18
- 239000005720 sucrose Substances 0.000 claims abstract description 18
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000007844 bleaching agent Substances 0.000 claims abstract description 8
- 239000000460 chlorine Substances 0.000 claims abstract description 8
- 229910052801 chlorine Inorganic materials 0.000 claims abstract description 8
- 229960002523 mercuric chloride Drugs 0.000 claims abstract description 8
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 claims abstract description 8
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 claims abstract description 8
- 229920000036 polyvinylpyrrolidone Polymers 0.000 claims abstract description 8
- 230000015572 biosynthetic process Effects 0.000 claims description 14
- 238000005286 illumination Methods 0.000 claims description 7
- 239000013535 sea water Substances 0.000 claims description 7
- 239000007787 solid Substances 0.000 claims description 7
- 238000011146 sterile filtration Methods 0.000 claims description 7
- 239000005556 hormone Substances 0.000 claims description 2
- 229940088597 hormone Drugs 0.000 claims description 2
- 230000008569 process Effects 0.000 abstract description 7
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- 238000004161 plant tissue culture Methods 0.000 description 2
- 241000143060 Americamysis bahia Species 0.000 description 1
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 238000009360 aquaculture Methods 0.000 description 1
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The present invention discloses the induction method of a kind of sargassun regeneration plant, comprises and prepares aseptic explant and be inoculated on inducing culture by explant evoked callus and formed, and inducing temperature is 15-20 DEG C, light intensity is 4000-5000Lx, and the photoperiod is 8h; The polyvidone process 3min of sargassun rhizoid in 0.2%, then sterilize 5min in the chlorine bleach liquor of 3%, is in the mercuric chloride of 0.1% to sterilize that 3min can obtain the aseptic explant survived; 1L? do you PES substratum add ZT? 0.1-1mg, IAA1-2mg, xitix 1-3g, sucrose 30g, agar 7g, be conducive to sargassun to form bud after autoclave sterilization 10-20min.
Description
Technical field
The present invention relates to field of plant tissue culture technique, it is specifically related to a kind of method of sargassun tissue culture.
Background technology
Sargassun is a kind of large-scale economic brown alga, and frond length is maximum reaches more than 7m, and abundant Biological resources are contained in the natural algae field that sargassun is formed, be fish, Shrimp waste good breed place, be one of ecosystem that fecundity of the sea is the highest. Sargassun is rich in the trace element of carbohydrate, lipid, protein, VITAMIN, mineral substance and multiple needed by human, and its exploitation prospect is very wide. Sargassun is a kind of perennial marine alga, owing to the speed of growth is fast, biomass big and Ecosystem Service important in shallow-sea ecology environment, can be used as one of important structure species of " blue carbon converges ", the reconstruction of algae field, ecosystem environment remediation, artificial fish shelter and sea ranch, there is huge economic benefit, ecological benefits and social benefit. The germ plasm resource of sargassun is deficienter, biological engineering is carried out at present mainly through sexual propagation approach, repoductive time is long, and need a large amount of kind algaes as the source adopting spore, the commercial aquaculture of current sargassun is not still popularized, main using natural resources as germplasm origin, can not originate for the kind algae that biological engineering provides stable, sufficient.
Plant tissue culture technique not only can breed a large amount of callus or bud of growing thickly within the short period of time, and regeneration plant and original plant there is no genetic diversity; And callus can form new callus by shoot proliferation, callus can also induced synthesis bud or root thus form new plant, the breeding coefficient of this kind of propagation method is very high, it is possible to reach the effect got twice the result with half the effort, and is the most effective plant cultivation method. And the tissue culture technique of sargassun is also immature at present, what also do not have that successful method can be stable forms a large amount of callus or bud of growing thickly.
Summary of the invention
For the defect that prior art exists, technical problem to be solved by this invention is to provide the induction method of a kind of sargassun regeneration plant, is reached the object of asexual breeding by the approach inducing sargassun to form bud on rhizoid.
In order to solve the problems of the technologies described above, the technical scheme of the present invention is as follows:
The induction method of a kind of sargassun regeneration plant, comprise the steps: using the rhizoid of sargassun as explant, clean after totally, in the polyvidone process 3min of 0.2%, then sterilize 5min in the chlorine bleach liquor of 3%, is in the mercuric chloride of 0.1% to sterilize 3min; Explant sterile filtration seawater after sterilization is cleaned after totally and it is cut into the segment that length is 0.3-0.5mm, segment is inoculated in the solid PES substratum adding hormone, in intensity of illumination to be 4000-5000Lx, light application time be 8h, temperature are 15-20 DEG C when, cultivates 20-30d to Buds formation.
Further, the configuration method of described substratum is, adds ZT0.1-1mg, IAA1-2mg, xitix 1-3g, sucrose 30g, agar 7g, autoclave sterilization 20min in 1LPES substratum.
Further, the configuration method of described substratum is, adds ZT0.1mg, IAA1mg, xitix 1g, sucrose 30g, agar 7g, autoclave sterilization 20min in 1LPES substratum.
Further, the configuration method of described substratum is, adds ZT1mg, IAA2mg, xitix 3g, sucrose 30g, agar 7g, autoclave sterilization 20min in 1LPES substratum.
Further, the configuration method of described substratum is, adds ZT0.5mg, IAA1mg, xitix 2g, sucrose 30g, agar 7g, autoclave sterilization 20min in 1LPES substratum.
Further, the configuration method of described substratum is, adds ZT1mg, IAA1mg, xitix 3g, sucrose 30g, agar 7g, autoclave sterilization 20min in 1LPES substratum.
Further, the configuration method of described substratum is, adds ZT0.5mg, IAA1.5mg, xitix 3g, sucrose 30g, agar 7g, autoclave sterilization 20min in 1LPES substratum.
The invention has the beneficial effects as follows:
First successfully using sargassun rhizoid as explant induction Buds formation, and the inductivity of bud can reach 42%, and bud can be grown further and be formed sargassun plant, is the germ plasm resource that sargassun algae field provides abundant.
Embodiment
Set forth the useful effect of the present invention by the following examples further:
Embodiment 1:
Using the rhizoid of sargassun as explant, clean clean after, in the polyvidone process 3min of 0.2%, then sterilize 5min in the chlorine bleach liquor of 3%, is in the mercuric chloride of 0.1% to sterilize 3min; Explant sterile filtration seawater after sterilization is cleaned after totally and it is cut into the segment that length is 0.3-0.5mm; Segment is inoculated in solid PES substratum, 1LPES substratum adds ZT0.1mg, IAA1mg, xitix 1g, sucrose 30g, agar 7g; Be 4000-5000Lx, light application time in intensity of illumination it is that 8h, temperature continue when being 18 DEG C to cultivate 28d to Buds formation; Inoculating 100 pieces of rhizoid strippings and slicings altogether, have and have Buds formation on 20 pieces of explants in 100 block organization's blocks, the inductivity of bud is 20%.
Embodiment 2:
Using the rhizoid of sargassun as explant, clean clean after, in the polyvidone process 3min of 0.2%, then sterilize 5min in the chlorine bleach liquor of 3%, is in the mercuric chloride of 0.1% to sterilize 3min; Explant sterile filtration seawater after sterilization is cleaned after totally and it is cut into the segment that length is 0.3-0.5mm; Segment is inoculated in solid PES substratum, 1LPES substratum adds ZT1mg, IAA2mg, xitix 3g, sucrose 30g, agar 7g; Be 4000-5000Lx, light application time in intensity of illumination it is that 8h, temperature continue when being 18 DEG C to cultivate 28d to Buds formation; Inoculating 100 pieces of rhizoid strippings and slicings altogether, have and have Buds formation on 40 pieces of explants in 100 block organization's blocks, the inductivity of bud is 42%.
Embodiment 3:
Using the rhizoid of sargassun as explant, clean clean after, in the polyvidone process 3min of 0.2%, then sterilize 5min in the chlorine bleach liquor of 3%, is in the mercuric chloride of 0.1% to sterilize 3min; Explant sterile filtration seawater after sterilization is cleaned after totally and it is cut into the segment that length is 0.3-0.5mm; Segment is inoculated in solid PES substratum, 1LPES substratum adds ZT0.5mg, IAA1mg, xitix 2g, sucrose 30g, agar 7g; Be 4000-5000Lx, light application time in intensity of illumination it is that 8h, temperature continue when being 18 DEG C to cultivate 28d to Buds formation; Inoculating 100 pieces of rhizoid strippings and slicings altogether, have and have Buds formation on 30 pieces of explants in 100 block organization's blocks, the inductivity of bud is 30%.
Embodiment 4:
Using the rhizoid of sargassun as explant, clean clean after, in the polyvidone process 3min of 0.2%, then sterilize 5min in the chlorine bleach liquor of 3%, is in the mercuric chloride of 0.1% to sterilize 3min; Explant sterile filtration seawater after sterilization is cleaned after totally and it is cut into the segment that length is 0.3-0.5mm; Segment is inoculated in solid PES substratum, 1LPES substratum adds ZT1mg, IAA1mg, xitix 3g, sucrose 30g, agar 7g; Be 4000-5000Lx, light application time in intensity of illumination it is that 8h, temperature continue when being 18 DEG C to cultivate 28d to Buds formation; Inoculating 100 pieces of rhizoid strippings and slicings altogether, have and have Buds formation on 38 pieces of explants in 100 block organization's blocks, the inductivity of bud is 38%.
Embodiment 5:
Using the rhizoid of sargassun as explant, clean clean after, in the polyvidone process 3min of 0.2%, then sterilize 5min in the chlorine bleach liquor of 3%, is in the mercuric chloride of 0.1% to sterilize 3min; Explant sterile filtration seawater after sterilization is cleaned after totally and it is cut into the segment that length is 0.3-0.5mm; Segment is inoculated in solid PES substratum, 1LPES substratum adds ZT0.5mg, IAA1.5mg, xitix 3g, sucrose 30g, agar 7g; Be 4000-5000Lx, light application time in intensity of illumination it is that 8h, temperature continue when being 18 DEG C to cultivate 28d to Buds formation; Inoculating 100 pieces of rhizoid strippings and slicings altogether, have and have Buds formation on 42 pieces of explants in 100 block organization's blocks, the inductivity of bud is 42%.
Above-mentioned example just for technical conceive and the technology feature of the present invention are described, can not limit the scope of the invention with this. All equivalent transformations of doing according to the essence of the present invention or modification, all should be encompassed within protection scope of the present invention.
Claims (6)
1. the induction method of a sargassun regeneration plant, it is characterised in that, comprise the steps: using the rhizoid of sargassun as explant, clean after totally, processing 3min in the polyvidone of 0.2%, then sterilize 5min in the chlorine bleach liquor of 3%, and sterilize 3min in the mercuric chloride of 0.1%; Explant sterile filtration seawater after sterilization is cleaned after totally and it is cut into the segment that length is 0.3-0.5mm, segment is inoculated in the solid PES substratum adding hormone, in intensity of illumination to be 4000-5000Lx, light application time be 8h, temperature are 15-20 DEG C when, cultivates 20-30d to Buds formation;
The configuration method of above-mentioned substratum is, adds ZT0.1-1mg, IAA1-2mg, xitix 1-3g, sucrose 30g, agar 7g, autoclave sterilization 20min in 1LPES substratum.
2. the induction method of sargassun regeneration plant according to claim 1, it is characterized in that, the configuration method of described substratum is, adds ZT0.1mg, IAA1mg, xitix 1g, sucrose 30g, agar 7g, autoclave sterilization 20min in 1LPES substratum.
3. the induction method of sargassun regeneration plant according to claim 1, it is characterized in that, the configuration method of described substratum is, adds ZT1mg, IAA2mg, xitix 3g, sucrose 30g, agar 7g, autoclave sterilization 20min in 1LPES substratum.
4. the induction method of sargassun regeneration plant according to claim 1, it is characterized in that, the configuration method of described substratum is, adds ZT0.5mg, IAA1mg, xitix 2g, sucrose 30g, agar 7g, autoclave sterilization 20min in 1LPES substratum.
5. the induction method of sargassun regeneration plant according to claim 1, it is characterized in that, the configuration method of described substratum is, adds ZT1mg, IAA1mg, xitix 3g, sucrose 30g, agar 7g, autoclave sterilization 20min in 1LPES substratum.
6. the induction method of sargassun regeneration plant according to claim 1, it is characterized in that, the configuration method of described substratum is, adds ZT0.5mg, IAA1.5mg, xitix 3g, sucrose 30g, agar 7g, autoclave sterilization 20min in 1LPES substratum.
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| CN108849502A (en) * | 2018-06-12 | 2018-11-23 | 金华市胤宏农业科技有限公司 | A kind of cultural method of bladder-wrack |
| CN109197593B (en) * | 2018-10-29 | 2021-11-09 | 杭州园泰生物科技有限公司 | Rapid breeding method of gulfweed |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1373633A (en) * | 2000-08-31 | 2002-10-09 | 科学与工业研究委员会 | Improved process for cultivation of algae |
| CN103141371A (en) * | 2013-04-08 | 2013-06-12 | 广东海洋大学 | Transplanting and cultivating method for gulfweed on rock |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| JP2006129833A (en) * | 2004-11-09 | 2006-05-25 | Japan Science & Technology Agency | Method for culturing sargassum |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1373633A (en) * | 2000-08-31 | 2002-10-09 | 科学与工业研究委员会 | Improved process for cultivation of algae |
| CN103141371A (en) * | 2013-04-08 | 2013-06-12 | 广东海洋大学 | Transplanting and cultivating method for gulfweed on rock |
Non-Patent Citations (1)
| Title |
|---|
| Physiological differences in the growth of Sargassum horneri between the germling and adult stages;Han Gil Choi;《J Appl Phycol》;20081231;第20卷;第729-735页 * |
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