CN103733988A - Gelidiella acerosa callus induction method - Google Patents
Gelidiella acerosa callus induction method Download PDFInfo
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- CN103733988A CN103733988A CN201310584618.2A CN201310584618A CN103733988A CN 103733988 A CN103733988 A CN 103733988A CN 201310584618 A CN201310584618 A CN 201310584618A CN 103733988 A CN103733988 A CN 103733988A
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Abstract
The invention discloses a Gelidiella acerosa callus induction method. The method comprises the steps of explant temporary rearing, explant disinfection, inoculation induction and callus propagation or storage. A povidone iodine and sodium hypochlorite cooperation use method is adopted to disinfect Gelidiella acerosa explants in order to obtain a large amount of aseptic explants, the Gelidiella acerosa explants are induced, 75% of the explants can form calluses, the calluses can further propagate, and a large amount of high-quality media obtained after the propagation can further differentiate into regeneration buds or plants, and the calluses can also be stored as a Gelidiella acerosa germplasm resource.
Description
Technical field
The present invention relates to field of plant tissue culture technique, be specifically related to a kind of abductive approach of flore dish callus.
Background technology
Flore dish (
gelidiella acerosa(Forsskal)) be one of the important raw and processed materials of industrial production agar product.The agar gel output of flore dish is very high, and the thallophytic output of every square centimeter can reach 600g, and so high agar yield is more favored in the raw material as extraction agar with flore dish people.Every year, from the 200-300 ton flore dried vegetable material of Natural Population results, being used for native country agar produces, due to biotechnological industries rise at home, demand to agar is increasing, cause the use of other agar marine algas, also caused the excessive collection bringing in order to meet the growing market demand of flore dish.The limitation because flore dish distributes, intrinsic Growth and reproduction is slow, therefore recovers and protects the effort of overdeveloped flore dish nature algae bed suppressed.Therefore,, in order to develop the large-scale cultivation culture technique of flore dish in feasible native country, in coastal marine site, carried out and on artificial substratum, cultivated flore dish.
Summary of the invention
The defect existing for prior art, technical problem to be solved by this invention is to provide a kind of abductive approach of flore dish callus, for the vegetative propagation of flore dish and screening fast-growth strain provide sufficient germ plasm resource, improve the economic outlook of flore dish integral production power and cultivation field.
In order to achieve the above object, the present invention adopts following technical scheme: a kind of abductive approach of flore dish callus, comprises the steps:
(1) supporting temporarily of explant: select healthy, subramose plant to cultivate as tissue, brush away by hand except epiphyte and other microorganisms; Flore dish thallus is cut into the long segment of 4-5cm, and by segment in adding Ge0
2the gas filling bottle of PES culture fluid in cultivate 1 week, cultivation temperature remains on 20-22 ° of C, light intensity is white fluorescent lamp 2000-3000Lx, light application time is 12h;
(2) explant sterilization: by flore dish segment 1% the povidone iodine immersion 1-5min that cultivates 1 week in step 1, then soak 1-5min with 1% liquor natrii hypochloritis, aseptic seawater flushing is clean;
(3) inoculation induction: the explant after sterilization in step 3 is placed in to aseptic operating platform, be cut into the inoculation section of 3-5mm, inoculation section is inoculated in to the formation of evoked callus in solid PES inducing culture, condition of culture is for dark, temperature remain on 20-22 ° of C entirely;
(4) callus propagation or storage: the callus obtaining in step 3 is as propagation or as germ plasm resource storage.
Further, the Ge0 described in step 1
2concentration be 10mg/L.
Further, the formula of the induction of the solid PES described in step 3 culture medium is in 1LPES medium, to add sucrose 15-20g, agar 5-8g, IAA1-3mg, KT0.5-2mg.
Further, the formula of the callus proliferated culture medium described in step 4 is, in 1LPES medium, add sucrose 20-40g, agar 5-8g, IAA 0.5-1.5mg, KT 0.2-1mg, propagation condition of culture is: cultivation temperature remains on 23-25 ° of C, light intensity is white fluorescent lamp 2000-3000Lx, and light application time is 12h.
Further, it is in 1LPES medium, to add sucrose 10-15g, KT0.1-0.5mg that described callus stores medium, and described condition of storage is 12-15 ° of C, and light intensity is white fluorescent lamp 500-800Lx, and light application time is 12h.
The invention has the beneficial effects as follows:
The present invention adopts the method that povidone iodine and clorox are used in conjunction with to carry out disinfection to flore dish explant, can obtain a large amount of aseptic explants, adopt the present invention to induce flore dish explant, there is 75% explant can form callus, callus not only can further be bred, a large amount of high-quality medium of propagation can further be divided into regeneration bud or plant, and callus can also store the germ plasm resource as flore dish.
Embodiment
Further set forth by the following examples beneficial effect of the present invention:
Embodiment 1:
An abductive approach for flore dish callus, comprises the steps:
(1) supporting temporarily of explant: select healthy, subramose plant to cultivate as tissue, brush away by hand except epiphyte and other microorganisms; Flore dish thallus is cut into the long segment of 4-5cm, and in adding concentration, is 10mg/L Ge0 by segment
2the gas filling bottle of PES culture fluid in cultivate 1 week, cultivation temperature remains on 20-22 ° of C, light intensity is white fluorescent lamp 2000-3000Lx, light application time is 12h;
(2) explant sterilization: by flore dish segment 1% the povidone iodine immersion 2min that cultivates 1 week in step 1, then soak 2min with 1% liquor natrii hypochloritis, aseptic seawater flushing is clean;
(3) inoculation induction: the explant after sterilization in step 3 is placed in to aseptic operating platform, be cut into the inoculation section of 3-5mm, inoculation section is inoculated in to the formation of evoked callus in solid PES inducing culture, condition of culture is for dark, temperature remain on 20-22 ° of C entirely; The formula of solid PES induction culture medium is in 1LPES medium, to add sucrose 18g, agar 6g, IAA 2mg, KT 1mg; Inoculate altogether 100 inoculation sections, wherein have 75 to form callus, callus induction rate is 75%;
(4) callus propagation or storage: the callus obtaining in step 3 is as propagation or as germ plasm resource storage, the formula of proliferated culture medium is, in 1LPES medium, add sucrose 35g, agar 7g, IAA 1mg, KT 0.5mg, propagation condition of culture is: cultivation temperature remains on 23-25 ° of C, light intensity is white fluorescent lamp 2000-3000Lx, and light application time is 12h; Storing medium is in 1LPES medium, to add sucrose 12g, KT0.3mg, and described condition of storage is 12-15 ° of C, and light intensity is white fluorescent lamp 500-800Lx, and light application time is 12h.
Embodiment 2:
Method is with embodiment 1, and difference is that the formula of solid PES induction culture medium is in 1LPES medium, to add sucrose 18g, agar 6g, IAA 3mg, KT 2mg; Inoculate altogether 100 inoculation sections, wherein have 70 to form callus, callus induction rate is 70%.
Embodiment 3:
Method is with embodiment 1, and difference is that the formula of solid PES induction culture medium is in 1LPES medium, to add sucrose 18g, agar 6g, IAA 1mg, KT 0.5mg; Inoculate altogether 100 inoculation sections, wherein have 68 to form callus, callus induction rate is 68%.
Embodiment 4:
Method is with embodiment 1, and difference is that the formula of solid PES induction culture medium is in 1LPES medium, to add sucrose 18g, agar 6g, IAA 1mg, KT 2mg; Inoculate altogether 100 inoculation sections, wherein have 60 to form callus, callus induction rate is 60%.
Above-mentioned example just, for explanation technical conceive of the present invention and technical characterstic, can not limit the scope of the invention with this.Equivalent transformation or modification that all essence according to the present invention is done, within all should being encompassed in protection scope of the present invention.
Claims (5)
1. an abductive approach for flore dish callus, is characterized in that, comprises the steps:
(1) supporting temporarily of explant: select healthy, subramose plant to cultivate as tissue, brush away by hand except epiphyte and other microorganisms; Flore dish thallus is cut into the long segment of 4-5cm, and by segment in adding Ge0
2the gas filling bottle of PES culture fluid in cultivate 1 week, cultivation temperature remains on 20-22 ° of C, light intensity is white fluorescent lamp 2000-3000Lx, light application time is 12h;
(2) explant sterilization: by flore dish segment 1% the povidone iodine immersion 1-5min that cultivates 1 week in step 1, then soak 1-5min with 1% liquor natrii hypochloritis, aseptic seawater flushing is clean;
(3) inoculation induction: the explant after sterilization in step 3 is placed in to aseptic operating platform, be cut into the inoculation section of 3-5mm, inoculation section is inoculated in to the formation of evoked callus in solid PES inducing culture, condition of culture is for dark, temperature remain on 20-22 ° of C entirely;
(4) callus propagation or storage: the callus obtaining in step 3 is as propagation or as germ plasm resource storage.
2. abductive approach according to claim 1, is characterized in that, the Ge0 described in step 1
2concentration be 10mg/L.
3. abductive approach according to claim 1, is characterized in that, the formula of the solid PES induction culture medium described in step 3 is in 1LPES medium, to add sucrose 15-20g, agar 5-8g, IAA1-3mg, KT0.5-2mg.
4. abductive approach according to claim 1, it is characterized in that, the formula of the callus proliferated culture medium described in step 4 is, in 1LPES medium, add sucrose 20-40g, agar 5-8g, IAA 0.5-1.5mg, KT 0.2-1mg, propagation condition of culture is: cultivation temperature remains on 23-25 ° of C, light intensity is white fluorescent lamp 2000-3000Lx, and light application time is 12h.
5. abductive approach according to claim 1, it is characterized in that, it is in 1LPES medium, to add sucrose 10-15g, KT0.1-0.5mg that described callus stores medium, and described condition of storage is 12-15 ° of C, light intensity is white fluorescent lamp 500-800Lx, and light application time is 12h.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201310584618.2A CN103733988A (en) | 2013-11-20 | 2013-11-20 | Gelidiella acerosa callus induction method |
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| CN201310584618.2A CN103733988A (en) | 2013-11-20 | 2013-11-20 | Gelidiella acerosa callus induction method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2016140318A (en) * | 2015-02-03 | 2016-08-08 | 住友ゴム工業株式会社 | Recovering method of rubber tree, proliferation method of rubber tree, induction method of shoot, growth method of shoot, root method of shoot and conditioning method for infant plant |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002017707A1 (en) * | 2000-08-31 | 2002-03-07 | Council Of Scientific And Industrial Research | An improved process for cultivation of algae |
| CN101731147A (en) * | 2009-12-18 | 2010-06-16 | 中国海洋大学 | Gelidium amansii Lamx. culture method by tissue culture breeding |
| CN102986529A (en) * | 2012-11-25 | 2013-03-27 | 溧阳市天目湖保健品有限公司 | Tissue culture method of gelidium amansii |
-
2013
- 2013-11-20 CN CN201310584618.2A patent/CN103733988A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002017707A1 (en) * | 2000-08-31 | 2002-03-07 | Council Of Scientific And Industrial Research | An improved process for cultivation of algae |
| CN101731147A (en) * | 2009-12-18 | 2010-06-16 | 中国海洋大学 | Gelidium amansii Lamx. culture method by tissue culture breeding |
| CN102986529A (en) * | 2012-11-25 | 2013-03-27 | 溧阳市天目湖保健品有限公司 | Tissue culture method of gelidium amansii |
Non-Patent Citations (2)
| Title |
|---|
| G. RAJAKRISHNA KUMAR ET.AL.,: "Tissue culture and regeneration of thallus from callus of Gelidiella acerosa (Gelidiaies, Rhodophyta)", 《PHYCOLOGIA》, vol. 43, no. 5, 30 September 2004 (2004-09-30) * |
| 姜红霞等: "红藻育种研究进展", 《海洋科学》, vol. 27, no. 6, 31 December 2003 (2003-12-31), pages 25 - 30 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2016140318A (en) * | 2015-02-03 | 2016-08-08 | 住友ゴム工業株式会社 | Recovering method of rubber tree, proliferation method of rubber tree, induction method of shoot, growth method of shoot, root method of shoot and conditioning method for infant plant |
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Application publication date: 20140423 |