CN104488715A - Method for carrying out bulb induction and plant regeneration of wide-leaf albuca namaquensis by flower buds - Google Patents
Method for carrying out bulb induction and plant regeneration of wide-leaf albuca namaquensis by flower buds Download PDFInfo
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- CN104488715A CN104488715A CN201410797136.XA CN201410797136A CN104488715A CN 104488715 A CN104488715 A CN 104488715A CN 201410797136 A CN201410797136 A CN 201410797136A CN 104488715 A CN104488715 A CN 104488715A
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Abstract
The invention discloses a method for carrying out bulb induction and plant regeneration of wide-leaf albuca namaquensis by flower buds. The method comprises the following specific steps: (1) preparation of a culture medium including a basic culture medium and components of the culture medium at all stages of tissue culture; (2) selection and disinfection of explants; (3) induction and multiplication of bulbs; (4) rooting of strong seedlings; and (5) transplantation. The method disclosed by the invention has the beneficial effects that the direct induction and the plant regeneration of the bulbs are carried out on flower buds of the wide-leaf albuca namaquensis by utilizing a plant-tissue culture technology, and the procedure of tissue-culture operation is simplified, so that a great quantity of high-quality strong seedlings with consistent genetic characters can be obtained in short time, simultaneously, the defects of female-parent scarcity and slowness of conventional multiplication methods are overcome, and the method has the positive significance for both protecting germplasm resources of the albuca namaquensis and industrializing seedling production.
Description
Technical field
The present invention relates to plant tissue culture technique, be specifically related to a kind of method of being carried out wide leaf spring grass bulb induce and plant regeneration by bud.
Background technology
Wide leaf spring grass (Albuca concordiana) views and admires succulent for hyacinthaceae wide leaf spring grass belongs to perennial, originate in South Africa, its zonate frond distortion is spiraled, the simple and unsophisticated uniqueness of spherical bulb, and pattern is pure and fresh simple and elegant, it is precious spring grass product kind.Wide leaf spring grass poor growth under field conditions (factors), reproduction rate is lower; Simultaneously international succulent research organization (TheInternational Organization for Succulent Plant Study) predatoryly gathers the threat of even becoming extinct to protect wild succulent to exempt from; formulate " Rules of Conduct of succulent cultivation knowledge and gatherer " in the seventies; the succulent that significantly limit China introduces a fine variety work; make commercially available the holding at high price of current wide leaf spring grass, its promotion and application are restricted.But; the Fast-propagation utilizing plant tissue culture technique to carry out wide leaf spring grass can obtain a large amount of high quality seedling consistent with maternal plant genetic background in a short time, and this is for meeting the domestic and international market demand for wide leaf spring grass, protecting rare wide leaf spring grass germ plasm resource to have great importance.But, also do not set up the report of spring grass tissue cultures before this, more do not induced the examples of many successful of wide leaf spring grass bulb and plant regeneration by bud.
Summary of the invention
The object of the invention is to the deficiency overcoming prior art existence, and a kind of method of being carried out wide leaf spring grass bulb induce and plant regeneration by bud is provided.
The object of the invention is to have come by following technical solution.This method of being carried out wide leaf spring grass bulb induce and plant regeneration by bud, is mainly comprised the steps:
1), the preparation of medium:
(1) minimal medium: MS medium, wherein sucrose 20 ~ 30g/L, agar 5 ~ 9g/L, pH=5.8;
(2) inducing culture: MS+6-BA 3 ~ 6mg/L+NAA0.2 ~ 1.0mg/L;
(3) proliferated culture medium: MS+6-BA 0.5 ~ 1.0mg/L+NAA0.05 ~ 0.2mg/L;
(4) strengthening seedling and rooting medium: MS+6-BA 0.01 ~ 0.05mg/L+NAA0.05 ~ 0.2mg/L;
2) the choosing and sterilization of explant: the immature inflorescence getting wide leaf spring grass, by for subsequent use after bud cleaning and sterilizing;
3) induction of bulb and propagation: by step 2) disinfect after bud be aseptically inoculated on inducing culture, after 1 ~ 2 month, directly induce bulb, bulb be transferred on proliferated culture medium and carry out Multiplying culture;
4) strengthening seedling and rooting is cultivated: by step 3) bulb is transferred to after being aseptically divided into individual plant on strengthening seedling and rooting medium, cultivates and can become the strong sprout with 1cm young root after 20 ~ 40 days;
5) transplant: by step 4) take root after strong sprout takes out from blake bottle and clean afterwards transplanting and cultivate to matrix.
Further, the present invention is in described step 1) in, the component that described medium comprises minimal medium and each stage medium of group training is specially:
(1) minimal medium: MS medium, wherein sucrose 30g/L, agar 7.5g/L, pH=5.8;
(2) inducing culture: MS+6-BA 4.0mg/L+NAA 0.5mg/L;
(3) proliferated culture medium: MS+6-BA 0.5mg/L+NAA 0.05mg/L;
(4) strengthening seedling and rooting medium: MS+6-BA 0.01mg/L+NAA 0.1mg/L.
Further, the present invention is in described step 2) in, described disinfects the outsourcing sheet being to remove closed bud on inflorescence, again bud is cut one by one together with part scape, after first soaking 30 ~ 60min with saturated washing powder solution, clean with tap water again, then successively volume ratio be 70% alcohol and effective chlorine density be soak 0.5 ~ 1min and 4 ~ 7min respectively in the liquor natrii hypochloritis of 1%, finally use aseptic water washing 3 ~ 5 times;
Further, the present invention is in described step 3) in, described in disinfect after bud, be first inoculated in after its base portion cut wound on inducing culture with scalpel, and make Wound contact to medium; Cultivate and directly can induce bulb from bud wound in 1 ~ 2 month;
Further, the present invention is in described step 4) in, described bulb is transferred to after being divided into individual plant on strengthening seedling and rooting medium and cultivates after 20 ~ 40 days and can directly become the strong sprout with 1cm young root.
Beneficial effect of the present invention is: utilize the bud of plant tissue culture technique to wide leaf spring grass to carry out direct induction and the plant regeneration of bulb; and carry out strong sprout and culture of rootage simultaneously; simplify group training operation sequence; the high-quality strong sprout that a large amount of genetic character is consistent can be obtained within a short period of time; overcome the shortcoming that female parent is rare and Sterile culture method is slow, all there is positive effect to the seeling industry of protection germ plasm resource and batch production.
Embodiment
Below by embodiment, the present invention is further elaborated, and help is understood the present invention by embodiment better, but the present invention is not limited only to following embodiment.
The invention provides a kind of method of being carried out wide leaf spring grass bulb induce and plant regeneration by bud, the steps include:
1), the preparation of medium
(1) minimal medium: MS medium, wherein sucrose 20 ~ 30g/L, agar 5 ~ 9g/L, pH=5.8;
(2) inducing culture: MS+6-BA 4 ~ 8mg/L+NAA0.05 ~ 0.5mg/L;
(3) differential medium: MS+6-BA 0.05 ~ 0.5mg/L+NAA 0.05 ~ 0.5mg/L;
(4) proliferated culture medium: MS+6-BA 0.5 ~ 1.0mg/L+NAA 0.05 ~ 0.2mg/L;
(5) strong seedling culture base: MS+6-BA 0.05 ~ 0.1mg/L+NAA 0.05 ~ 0.1mg/L;
2), the choosing and sterilizing of explant
Get the immature inflorescence of wide leaf spring grass, remove the outsourcing sheet of closed bud on inflorescence, again bud is cut one by one together with part scape, after first soaking 30 ~ 60min with saturated washing powder solution, clean with tap water again, then successively volume ratio be 70% alcohol and effective chlorine density be soak 0.5 ~ 1min and 4 ~ 7min respectively in the liquor natrii hypochloritis of 1%, finally use aseptic water washing 3 ~ 5 times;
3), the induction of bulb and Multiplying culture
Bud after disinfecting aseptically first is inoculated in after its base portion cut wound on inducing culture with scalpel, and makes Wound contact to medium.Cultivated after 1 ~ 2 month 1 ~ 2 month can from bud wound direct induction bulb, bulb is transferred on proliferated culture medium and carries out Multiplying culture; Cultivation temperature is 23 ± 2 DEG C, intensity of illumination is 30 ~ 60 μm of olm
-2s
-1, light application time is 8 ~ 10 hours/day;
4), strengthening seedling and rooting is cultivated
Be transferred to cultivation on strengthening seedling and rooting medium after bulb being divided into individual plant and can form the strong sprout with 1cm young root after 20 ~ 40 days; Cultivation temperature is 23 ± 2 DEG C, intensity of illumination is 30 ~ 80 μm of olm
-2s
-1, light application time is 8 ~ 10 hours/day;
5), transplant
Strong sprout after taking root is taken out from blake bottle and transplants after cleaning and cultivate to matrix, transplanting survival rate about 90%.
Embodiment
The invention provides a kind of method of being carried out wide leaf spring grass bulb induce and plant regeneration by bud, the steps include:
1), the preparation of medium
(1) minimal medium: MS medium, wherein sucrose 30g/L, agar 7.5g/L, pH=5.8;
(2) inducing culture: MS+6-BA 4.0mg/L+NAA 0.5mg/L;
(3) proliferated culture medium: MS+6-BA 0.5mg/L+NAA 0.05mg/L;
(4) strengthening seedling and rooting medium: MS+6-BA 0.01mg/L+NAA 0.1mg/L.
2), the choosing and sterilizing of explant
Get the immature inflorescence of wide leaf spring grass, remove the outsourcing sheet of closed bud on inflorescence, again bud is cut one by one together with part scape, after first soaking 60min with saturated washing powder solution, clean with tap water again, then successively volume ratio be 70% alcohol and effective chlorine density be soak 1min and 6min respectively in the liquor natrii hypochloritis of 1%, finally use aseptic water washing 5 times; 3), the induction of bulb and Multiplying culture
Bud after disinfecting aseptically first is inoculated in after its base portion cut wound on inducing culture with scalpel, and makes Wound contact to medium.Cultivated after 1 ~ 2 month and directly can induce bulb from bud wound in 1 ~ 2 month, bulb is transferred on proliferated culture medium and carries out Multiplying culture; Cultivation temperature is 23 ± 2 DEG C, intensity of illumination is 30 ~ 60 μm of olm
-2s
-1, light application time is 8 ~ 10 hours/day;
4), strengthening seedling and rooting is cultivated
Be transferred to after bulb being divided into individual plant on strengthening seedling and rooting medium and cultivate after 20 ~ 40 days and can directly become the strong sprout with 1cm young root; Cultivation temperature is 23 ± 2 DEG C, intensity of illumination is 30 ~ 80 μm of olm
-2s
-1, light application time is 8 ~ 10 hours/day;
5), transplant
Taken out from blake bottle the strong sprout after taking root and transplant after cleaning and cultivate to matrix, transplanting survival rate is greater than 90%.
Finally it should be noted that, except the present embodiment, the present invention can also have other embodiment and distortion, and what more than enumerate is only specific embodiments of the invention.All distortion that all those of ordinary skill in the art can directly derive from content disclosed by the invention or associate, all should think protection scope of the present invention.
Claims (5)
1. carried out a method for wide leaf spring grass bulb induce and plant regeneration by bud, it is characterized in that: the method comprises the steps:
1), the preparation of medium:
(1) minimal medium: MS medium, wherein sucrose 20 ~ 30g/L, agar 5 ~ 9g/L, pH=5.8;
(2) inducing culture: MS+6-BA 3 ~ 6mg/L+NAA0.2 ~ 1.0mg/L;
(3) proliferated culture medium: MS+6-BA 0.5 ~ 1.0mg/L+NAA0.05 ~ 0.2mg/L;
(4) strengthening seedling and rooting medium: MS+6-BA 0.01 ~ 0.05mg/L+NAA0.05 ~ 0.2mg/L;
2) the choosing and sterilization of explant: the immature inflorescence getting wide leaf spring grass, by for subsequent use after bud cleaning and sterilizing;
3) induction of bulb and propagation: by step 2) disinfect after bud be aseptically inoculated on inducing culture, after 1 ~ 2 month, directly induce bulb, bulb be transferred on proliferated culture medium and carry out Multiplying culture;
4) strengthening seedling and rooting is cultivated: by step 3) bulb is transferred to after being aseptically divided into individual plant on strengthening seedling and rooting medium, cultivates and can become the strong sprout with 1cm young root after 20 ~ 40 days;
5) transplant: by step 4) take root after strong sprout takes out from blake bottle and clean afterwards transplanting and cultivate to matrix.
2. method of being carried out wide leaf spring grass bulb induce and plant regeneration by bud according to claim 1, be is characterized in that: in described step 1) in, the component that described medium comprises minimal medium and each stage medium of group training is specially:
(1) minimal medium: MS medium, wherein sucrose 30g/L, agar 7.5g/L, pH=5.8;
(2) inducing culture: MS+6-BA 4.0mg/L+NAA 0.5mg/L;
(3) proliferated culture medium: MS+6-BA 0.5mg/L+NAA 0.05mg/L;
(4) strengthening seedling and rooting medium: MS+6-BA 0.01mg/L+NAA 0.1mg/L.
3. method of being carried out wide leaf spring grass bulb induce and plant regeneration by bud according to claim 1, it is characterized in that: in described step 2) in, described disinfecting is the outsourcing sheet removing closed bud on inflorescence, again bud is cut one by one together with part scape, after first soaking 30 ~ 60min with saturated washing powder solution, clean with tap water again, then successively volume ratio be 70% alcohol and effective chlorine density be soak 0.5 ~ 1min and 4 ~ 7min respectively in the liquor natrii hypochloritis of 1%, finally use aseptic water washing 3 ~ 5 times.
4. method of being carried out wide leaf spring grass bulb induce and plant regeneration by bud according to claim 1, it is characterized in that: in described step 3) in, described disinfect after bud, first be inoculated in after its base portion cut wound on inducing culture with scalpel, and make Wound contact to medium; Cultivate and directly induce bulb in 1 ~ 2 month.
5. method of being carried out wide leaf spring grass bulb induce and plant regeneration by bud according to claim 1, it is characterized in that: in described step 4) in, described bulb is transferred to after being divided into individual plant on strengthening seedling and rooting medium and cultivates after 20 ~ 40 days and can directly become the strong sprout with 1cm young root.
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| CN201410797136.XA CN104488715B (en) | 2014-12-19 | 2014-12-19 | A kind of method carrying out wide leaf spring grass bulb induction and plant regeneration by alabastrum |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107810854A (en) * | 2017-11-27 | 2018-03-20 | 北京农学院 | Lithops pseudotruncatella cultured in vitro and fast numerous method |
| CN108617510A (en) * | 2018-05-10 | 2018-10-09 | 漳州市农业科学研究所 | A method of improving wide leaf spring grass tissue-cultured seedling transplanting survival rate |
| CN108901857A (en) * | 2018-09-19 | 2018-11-30 | 闽卉（福建）园艺有限公司 | A kind of tissue culture propagation method of spring grass |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102119664A (en) * | 2011-01-21 | 2011-07-13 | 江苏九久环境科技有限公司 | Method for culturing muscari botryoides mill with plant tissues |
| CN102511391A (en) * | 2011-12-07 | 2012-06-27 | 上海交通大学 | Hyacinthus orientalis L. in-vitro rapid propagation method |
| USPP22954P2 (en) * | 2011-09-23 | 2012-08-14 | Zuidgeest Honselersdijk v.o.f. | Albuca plant named ‘Frizzle Sizzle’ |
| CN103125394A (en) * | 2013-03-13 | 2013-06-05 | 浙江省农业科学院 | Method for establishing tissue culture regeneration system of tylecodon paniculatus |
-
2014
- 2014-12-19 CN CN201410797136.XA patent/CN104488715B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102119664A (en) * | 2011-01-21 | 2011-07-13 | 江苏九久环境科技有限公司 | Method for culturing muscari botryoides mill with plant tissues |
| USPP22954P2 (en) * | 2011-09-23 | 2012-08-14 | Zuidgeest Honselersdijk v.o.f. | Albuca plant named ‘Frizzle Sizzle’ |
| CN102511391A (en) * | 2011-12-07 | 2012-06-27 | 上海交通大学 | Hyacinthus orientalis L. in-vitro rapid propagation method |
| CN103125394A (en) * | 2013-03-13 | 2013-06-05 | 浙江省农业科学院 | Method for establishing tissue culture regeneration system of tylecodon paniculatus |
Non-Patent Citations (2)
| Title |
|---|
| G.D. ASCOUGH等: "Micropropagation of Albuca bracteata and A. nelsonii — Indigenous ornamentals with medicinal value", 《SOUTH AFRICAN JOURNAL OF BOTANY》, vol. 76, 31 December 2010 (2010-12-31), pages 579 - 584, XP027094012 * |
| 兑宝峰: "宽叶弹簧草栽培技术", 《中国花卉报》, 8 February 2014 (2014-02-08), pages 1 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107810854A (en) * | 2017-11-27 | 2018-03-20 | 北京农学院 | Lithops pseudotruncatella cultured in vitro and fast numerous method |
| CN108617510A (en) * | 2018-05-10 | 2018-10-09 | 漳州市农业科学研究所 | A method of improving wide leaf spring grass tissue-cultured seedling transplanting survival rate |
| CN108617510B (en) * | 2018-05-10 | 2021-06-29 | 漳州市农业科学研究所 | Method for improving transplanting survival rate of tissue culture seedlings of broad-leaf spring grass |
| CN108901857A (en) * | 2018-09-19 | 2018-11-30 | 闽卉（福建）园艺有限公司 | A kind of tissue culture propagation method of spring grass |
| CN108901857B (en) * | 2018-09-19 | 2021-08-24 | 闽卉（福建）园艺有限公司 | Tissue culture propagation method of pogostemon |
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