CN104904594A - Anthurium adndraeanum pollen culture method - Google Patents
Anthurium adndraeanum pollen culture method Download PDFInfo
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- CN104904594A CN104904594A CN201510253722.2A CN201510253722A CN104904594A CN 104904594 A CN104904594 A CN 104904594A CN 201510253722 A CN201510253722 A CN 201510253722A CN 104904594 A CN104904594 A CN 104904594A
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Abstract
The invention relates to an anthurium adndraeanum pollen culture method. The method comprises the following steps: an anther at a mid-late uninucleate stage is adopted and is inoculated to a callus induction medium, and callus is obtained by culturing; the callus is inoculated to a differentiation medium, such that test tube seedlings are obtained; the test tube seedlings are inoculated to a rooting medium, and screening is carried out, such that single plants are obtained. With the above culturing manner, an anthurium adndraeanum pollen callus induction rate reaches 88%, a rooting rate reaches 94%, and a transplanting survival rate reaches 90%.
Description
Technical field
The present invention relates to a kind of method for plant tissue culture, be specifically related to a kind of red palm pollen cultures method.
Background technology
The red palm (Anthurium adndraeanum) is Araeceae Anthurium perennial evergreen herbaceous plant, can make potted flower and cut-flower and use, and is one of most popular rare flower in the world today, has very high ornamental value and economic worth.
The red palm is important tropical cut-flower, spadix, and its Buddhist flame petal is very large, plump tool wax, and color and luster has red, powder, white, green, double-colored etc.It is bright in colour, and appearance is peculiar, applied range, and economic worth is high, is current global evolution is fast, demand is larger top grade torrid zone cut-flower and pot flowers.Red palm original producton location tropical rain forest, the available seminal propagation of the red palm, but it is long to enter flowering time.Division propagation is the major way that the red palm was bred in the past.Red palm plant base portion grows suction bud, and producing after root system can plant division, and can divide 3-4 strain every year, reproduction coefficient is lower, is difficult to the seedling met needed for large-scale production.Present red palm seeling industry carries out the Fast-propagation of seedling mainly through tissue cultures, namely the clone technology of the red palm.Like this can within the shorter time high quality seedling of production neat and consistent, the needs that supply is produced.Produce red palm seedling by tissue culture technique and mainly contain foundation, Multiplying culture, the strengthening seedling and rooting of regenerating system, sport technique segment of transplanting and to nurse young plants in hothouses etc.
Along with the development of plant tissue culture technique, plant cell has, and " totipotency " is widely confirmed, therefore, pollen is cultivated by vitro method under in vitro condition, the development pathway of artificial change microspore, makes its Development of Gametophytes approach stop, turns to sporophyte to grow, occurred by organ or embryo occur approach, form complete plant.Also there is not the report cultivated about red palm in-vitro pollen at present.
Summary of the invention
The present invention is directed to current problem, a kind of red palm pollen cultures method is provided.
For solving the problems of the technologies described above, the technical scheme that the present invention adopts is: provide a kind of red palm pollen cultures method, comprise the following steps,
1) pollen getting mid-late uninucleate stage is inoculated into callus inducing medium, cultivate and obtain callus, described callus inducing medium: WPM+NAA 0.3 ~ 0.5mg/L+6-BA 0.2 ~ 0.4mg/L+2.4-D 0.4 ~ 0.7mg/L+ gibberellin 2.2 ~ 2.5mg/L+agar 1500 ~ 1700mg/L+ sucrose 600 ~ 800mg/L;
2) inoculate differential medium, cultivate and obtain test tube seedling, described differential medium is WPM+6-BA 0.4 ~ 0.6mg/L+NAA 0.1 ~ 0.3mg/L+ ZT 0.6 ~ 1mg/L+ 600 ~ 800mg/L sucrose;
3) inoculate root media, screening obtains monomer plant, and described root media is that 1/2WPM+IBA 0.1 ~ 0.3mg/L+NAA0.6 ~ 0.8mg/L+ agar 600 ~ 800mg/L+ sucrose 1400 ~ 1600mg/L+ quality of activated carbon is than 0.1% ~ 0.3%.
Preferably, described callus inducing medium: WPM+NAA 0.4mg/L+6-BA 0.3mg/L+2.4-D 0.6mg/L+ gibberellin 2.4mg/L+agar 1600mg/L+ sucrose 700mg/L.
Preferably, described differential medium is WPM+6-BA 0.5mg/L+NAA 0.2mg/L+ ZT 0.8mg/L+ 700mg/L sucrose.
Preferably, described root media is that 1/2WPM+IBA 0.2mg/L+NAA0.7mg/L+ agar 700mg/L+ sucrose 1500mg/L+ quality of activated carbon is than 0.2%.
Preferably, condition of culture: the method that condition of culture adopts illumination cultivation and light culture to combine, illumination cultivation light source adopts fluorescent lamp, and in culturing room, day temperature is 28 DEG C-30 DEG C, illumination 12h, intensity of illumination 15 μm of ol.m
-2.s
-1-20 μm of ol.m
-2.s
-1, temperature controls at 16 DEG C-18 DEG C.
Preferably, also comprise seedling replanting: until red palm seedling healthy and strong and have 2-3 sheet true leaf, 3-5cm height time, open wide bottleneck hardening 2-3d, then medium subsidiary on red palm seedling root system thoroughly washed, be just transplanted in the flowerpot in greenhouse and heel in.
The invention has the beneficial effects as follows: the present invention adopts red palm pollen to be explant, carries out the induction of pollen callus, adopt the method alternate culture that illumination cultivation and light culture combine, obtain pollen plant.Medium callus proliferation of the present invention is obvious, easily forms pollen plant.Key have found suitable cultural method, basal medium is revised as WPM, the concentration of corresponding adjustment hormone simultaneously, cultivate with the in vitro pollen of method of the present invention to the red palm, there is pollen germination and become that the embryo time is short, doubling etticiency is high, output is large and be easier to obtain the advantages such as excellent variation breeding intermediate materials, at aspects such as breeding material purification and rejuvenation, Germplasm enhancement, polyploid breeding and the researchs of pollen development process, there is larger practical value.
Embodiment
Below preferred embodiment of the present invention is described in detail, can be easier to make advantages and features of the invention be readily appreciated by one skilled in the art, thus more explicit defining is made to protection scope of the present invention.
The embodiment of the present invention comprises:
Embodiment 1:
A kind of red palm pollen cultures method, comprises the following steps,
1) pollen 1000 getting mid-late uninucleate stage is inoculated into callus inducing medium, cultivate and obtain callus, described callus inducing medium: WPM+NAA 0.3mg/L+6-BA 0.2mg/L+2.4-D 0.4mg/L+ gibberellin 2.2mg/L+agar 1500mg/L+ sucrose 600mg/L;
2) inoculate differential medium, cultivate and obtain test tube seedling, described differential medium is WPM+6-BA 0.4mg/L+NAA 0.1mg/L+ ZT 0.6mg/L+ 600mg/L sucrose;
3) inoculate root media, screening obtains monomer plant, and described root media is that 1/2WPM+IBA 0.1mg/L+NAA0.6mg/L+ agar 600mg/L+ sucrose 1400mg/L+ quality of activated carbon is than 0.1%.
The method that condition of culture adopts illumination cultivation and light culture to combine, illumination cultivation light source adopts fluorescent lamp, and in culturing room, day temperature is 28 DEG C DEG C, illumination 12h, intensity of illumination 15 μm of ol.m
-2.s
-1, temperature controls at 16 DEG C.
Healthy and strong and when having 2 true leaves, 3cm height, open wide bottleneck hardening 2d, then medium subsidiary on red palm seedling root system is thoroughly washed, be just transplanted in the flowerpot in greenhouse and heel in until red palm seedling.
Embodiment 2:
A kind of red palm pollen cultures method, comprises the following steps,
1) pollen getting mid-late uninucleate stage is inoculated into callus inducing medium, cultivate and obtain callus, described callus inducing medium: WPM+NAA0.5mg/L+6-BA0.4mg/L+2.4-D 0.7mg/L+ gibberellin 2.5mg/L+agar 1700mg/L+ sucrose 800mg/L;
2) inoculate differential medium, cultivate and obtain test tube seedling, described differential medium is WPM+6-BA 0.6mg/L+NAA 0.3mg/L+ ZT 1mg/L+ 800mg/L sucrose;
3) inoculate root media, screening obtains monomer plant, and described root media is that 1/2WPM+IBA 0.3mg/L+NAA 0.8mg/L+ agar 800mg/L+ sucrose 1600mg/L+ quality of activated carbon is than 0.3%.
The method that condition of culture adopts illumination cultivation and light culture to combine, illumination cultivation light source adopts fluorescent lamp, and in culturing room, day temperature is 30 DEG C, illumination 12h, intensity of illumination 20 μm of ol.m
-2.s
-1, temperature controls at 18 DEG C.
Healthy and strong and when having 3 true leaves, 5cm height, open wide bottleneck hardening 3d, then medium subsidiary on red palm seedling root system is thoroughly washed, be just transplanted in the flowerpot in greenhouse and heel in until red palm seedling.
Embodiment 3:
A kind of red palm pollen cultures method, comprises the following steps,
1) pollen getting mid-late uninucleate stage is inoculated into callus inducing medium, cultivate and obtain callus, described callus inducing medium: WPM+NAA 0.4mg/L+6-BA 0.3mg/L+2.4-D 0.6mg/L+ gibberellin 2.4mg/L+agar 1600mg/L+ sucrose 700mg/L;
2) inoculate differential medium, cultivate and obtain test tube seedling, described differential medium is WPM+6-BA 0.5mg/L+NAA 0.2mg/L+ ZT 0.8mg/L+ 700mg/L sucrose;
3) inoculate root media, screening obtains monomer plant, and described root media is that 1/2WPM+IBA 0.2mg/L+NAA0.7mg/L+ agar 700mg/L+ sucrose 1500mg/L+ quality of activated carbon is than 0.2%.
The method that condition of culture adopts illumination cultivation and light culture to combine, illumination cultivation light source adopts fluorescent lamp, and in culturing room, day temperature is 29 DEG C DEG C, illumination 12h, intensity of illumination 17 μm of ol.m
-2.s
-1, temperature controls at 17 DEG C DEG C.
Healthy and strong and when having 3 true leaves, 5cm height, open wide bottleneck hardening 3d, then medium subsidiary on red palm seedling root system is thoroughly washed, be just transplanted in the flowerpot in greenhouse and heel in until red palm seedling.
Average result through above-mentioned cultivation is that the inductivity of callus reaches 88%, and rooting rate reaches 94%, and transplanting survival rate reaches 90%.
The foregoing is only embodiments of the invention; not thereby the scope of the claims of the present invention is limited; every utilize description of the present invention to do equivalent structure or equivalent flow process conversion; or be directly or indirectly used in other relevant technical fields, be all in like manner included in scope of patent protection of the present invention.
Claims (6)
1. a red palm pollen cultures method, is characterized in that,
1) pollen getting mid-late uninucleate stage is inoculated into callus inducing medium, cultivate and obtain callus, described callus inducing medium: WPM+NAA 0.3 ~ 0.5mg/L+6-BA 0.2 ~ 0.4mg/L+2.4-D 0.4 ~ 0.7mg/L+ gibberellin 2.2 ~ 2.5mg/L+agar 1500 ~ 1700mg/L+ sucrose 600 ~ 800mg/L;
2) inoculate differential medium, cultivate and obtain test tube seedling, described differential medium is WPM+6-BA 0.4 ~ 0.6mg/L+NAA 0.1 ~ 0.3mg/L+ ZT 0.6 ~ 1mg/L+ 600 ~ 800mg/L sucrose;
3) inoculate root media, screening obtains monomer plant, and described root media is that 1/2WPM+IBA 0.1 ~ 0.3mg/L+NAA0.6 ~ 0.8mg/L+ agar 600 ~ 800mg/L+ sucrose 1400 ~ 1600mg/L+ quality of activated carbon is than 0.1% ~ 0.3%.
2. red palm pollen cultures method according to claim 1, is characterized in that, described callus inducing medium: WPM+NAA 0.4mg/L+6-BA 0.3mg/L+2.4-D 0.6mg/L+ gibberellin 2.4mg/L+agar 1600mg/L+ sucrose 700mg/L.
3. red palm pollen cultures method according to claim 1 and 2, is characterized in that, described differential medium is WPM+6-BA 0.5mg/L+NAA 0.2mg/L+ ZT 0.8mg/L+ 700mg/L sucrose.
4. red palm pollen cultures method as claimed in claim 1 or 2, is characterized in that, described root media is that 1/2WPM+IBA 0.2mg/L+NAA0.7mg/L+ agar 700mg/L+ sucrose 1500mg/L+ quality of activated carbon is than 0.2%.
5. red palm pollen cultures method as claimed in claim 1 or 2, is characterized in that, condition of culture: the method that condition of culture adopts illumination cultivation and light culture to combine, illumination cultivation light source adopts fluorescent lamp, in culturing room, day temperature is 28 DEG C-30 DEG C, illumination 12h, intensity of illumination 15 μm of ol.m
-2.s
-1-20 μm of ol.m
-2.s
-1, temperature controls at 16 DEG C-18 DEG C.
6. red palm pollen cultures method as claimed in claim 1 or 2, it is characterized in that, also comprise seedling replanting: until red palm seedling healthy and strong and have 2-3 sheet true leaf, 3-5cm height time, open wide bottleneck hardening 2-3d, then medium subsidiary on red palm seedling root system is thoroughly washed, be just transplanted in the flowerpot in greenhouse and heel in.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109197588A (en) * | 2018-08-28 | 2019-01-15 | 江苏高航农业科技有限公司 | A kind of red palm poison-removing method based on Anther Culture |
| CN112352680A (en) * | 2020-11-23 | 2021-02-12 | 锡山区先锋家庭农场 | Seedling-raising and rapid rooting method for anthurium tissue culture propagation |
Citations (2)
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| US20110232189A1 (en) * | 2004-06-10 | 2011-09-29 | Visser 's-Gravendeel Holding B.V. | Container for cultivating biological materials |
| CN104137777A (en) * | 2014-07-29 | 2014-11-12 | 浙江省萧山棉麻研究所 | Method for obtaining anthurium haplobionts |
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- 2015-05-19 CN CN201510253722.2A patent/CN104904594B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110232189A1 (en) * | 2004-06-10 | 2011-09-29 | Visser 's-Gravendeel Holding B.V. | Container for cultivating biological materials |
| CN104137777A (en) * | 2014-07-29 | 2014-11-12 | 浙江省萧山棉麻研究所 | Method for obtaining anthurium haplobionts |
Non-Patent Citations (3)
| Title |
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| ÇIMEN ATAK,ET AL.: "MICROPROPAGATION OF ANTHURIUM ANDRAEANUM FROM LEAF EXPLANTS", 《PAK. J. BOT.》 * |
| 杜宝贵等: "红掌花药培养", 《生物技术通报》 * |
| 束晓春等: "红掌组织培养研究", 《江苏农业科学》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109197588A (en) * | 2018-08-28 | 2019-01-15 | 江苏高航农业科技有限公司 | A kind of red palm poison-removing method based on Anther Culture |
| CN112352680A (en) * | 2020-11-23 | 2021-02-12 | 锡山区先锋家庭农场 | Seedling-raising and rapid rooting method for anthurium tissue culture propagation |
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