CN102577942A - Tissue culture method of callistemon hybridus 'golden ball' - Google Patents
Tissue culture method of callistemon hybridus 'golden ball' Download PDFInfo
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- CN102577942A CN102577942A CN2011100020798A CN201110002079A CN102577942A CN 102577942 A CN102577942 A CN 102577942A CN 2011100020798 A CN2011100020798 A CN 2011100020798A CN 201110002079 A CN201110002079 A CN 201110002079A CN 102577942 A CN102577942 A CN 102577942A
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Abstract
The invention discloses a tissue culture method of callistemon hybridus 'golden ball'. The tissue culture method comprises the following steps of: clipping an axillary bud from a parent as an explant, cleaning and sterilizing firstly, and then entering an induction culture, wherein an induction culture medium comprises MS (Murashige Sroog), 0.7-0.9 mg/L of 6-BA (benzyladenine) and 0.1-0.3 mg/L of NAA (Naphthyl Acetic Acid); entering an enrichment culture, wherein an enrichment culture comprises MS, 0.4-0.6 mg/L of 6-BA and 0.1+/-0.05 mg/L of NAA; at last, entering a rooting culture, wherein a rooting culture medium comprises 1/2 MS and 0.1-0.3 mg/L of IBA (indolebutyric acid); the culture conditions are as follows: the illumination intensity is 3000-4500 Lux and the temperature is 26+/-2 DEG C. According to the tissue culture method, the quick industrial breeding is realized, the multiplication multiplying power in 27 days reaches 3.2 times, the rooting rate in 30 days reaches 97%, and the transplanting survival rate is more than 95%. Therefore, the tissue culture method solves the seedling requirement problem for popularizing cultivation of the callistemon hybridus 'golden ball' in a large area.
Description
Technical field
The invention belongs to biological technical field, specifically refer to the method for tissue culture of gold string Qian Liu.
Background technology
Gold string Qian Liu (Callistemon * hybridus ' Golden Ball ') call Wong Kam-po tree, incense with golden color willow, thousand layers of gold, be Myrtaceae Melaleuca aiphyllium.It is the variation new varieties that select the early 1990s, and annual golden yellow of blade or light yellow look are one of most popular in the world at present, that visual effect is best color leaf arbor species.Its property happiness high temperature, sufficient sunshine, be adapted at China's south most of areas popularizing planting.Not only can be used for the flower garden view, road beautifies, and also can be used for transformation of forest phase, simultaneously because these seeds have stronger anti-saline and alkaline, anti-high wind characteristic, also be suitable for beach and artificial marine reclamation land greening, make scape, check winds and fix drifting sand etc.But this kind adopts propagation methods such as conventional sowing, cuttage, press strip, owing to receive the restriction of conditions such as season, weather, the speed of breeding is slow, and genetic variation is big, is difficult to satisfy the growing market demand in a short time.
Plant Tissue Breeding (Tissue culture) is meant the totipotency of utilizing plant cell; In plant corpus, take out part organ, tissue or cell; Physiological environment in the simulating plant body; Under aseptic, proper temperature and certain condition of culture, make it existence, growth, keep the method for its normal configuration and function.
The inventor finds after deliberation, adopts method for tissue culture, both can keep the merit of parent, effectively saves production cost, and shortens breeding cycle, can alleviate the demand and supply contraction of present gold string Qian Liu again, and this case is arisen spontaneously.
Summary of the invention
The present invention aims to provide the method for tissue culture of a kind of gold string Qian Liu, thereby realizes the industrialization fast breeding.
In order to realize the foregoing invention purpose, technical scheme of the present invention is:
The method for tissue culture of gold string Qian Liu; The clip axillalry bud is as explant from the gold string money willow parent with merit; Earlier explant is cleaned, sterilizes; Insert inducing culture then and carry out inducing culture, the inducing culture proportioning is: the conventional minimal medium of MS (Marashige Sroog), 6-BA (6-Bian Ji purine) 0.7～0.9mg/L, NAA (methyl) 0.1～0.3mg/L; Insert proliferated culture medium again and carry out enrichment culture, the proliferated culture medium proportioning is: the conventional minimal medium of MS, 6-BA 0.4～0.6mg/L, NAA 0.1 ± 0.05mg/L; At last, get into the culture of rootage stage, the root media proportioning is: 1/2MS (get 1/2 of macroelement, other trace element remains unchanged, and is as follows), IBA (indolebutyric acid) 0.1～0.3mg/L; Above inducing culture, enrichment culture, culture of rootage condition are: intensity of illumination is 3000～4500Lux, and temperature is 26 ± 2 ℃.
The concrete steps that above-mentioned explant cleans, sterilizes are: earlier with non-phosphide detergent aqueous solution soaking 15 minutes, place under the clear water and washed 30 minutes with current; With alcohol-pickled 15 seconds of concentration of volume percent 75%, aseptic water washing 1 time was used mass percent concentration 0.1%HgCl again
2Solution disinfection 4 minutes, aseptic water washing 5 times.
After above-mentioned explant cleaned, sterilizes, suck dry moisture was cut to the stem section of 2.5～3.5cm, inserted inducing culture then and carried out inducing culture.
The composition of above-mentioned MS medium by routine is:
The MS culture medium prescription:
After adopting said method; Method for tissue culture of the present invention has solved gold string money willow and has adopted conventional method to breed long, problems such as genetic variation is big, survival rate is low, production cost height of garden cycle; Realized the industrialization fast breeding, the propagation multiplying power reached 3.5 times in 27 days, and rooting rate reached more than 97% in 30 days; Transplanting survival rate has solved the root problem of nursery stock demand more than 95% for large tracts of land popularizing planting gold string money willow.
Embodiment
Utilization the present invention carries out the specific embodiment of the tissue culture of gold string Qian Liu.
A, explant are handled and inducing culture
The clip axillalry bud cleans, sterilizes explant earlier as explant from the gold string money willow parent with merit, and concrete steps are with non-phosphide detergent aqueous solution soaking 15 minutes, place under the clear water and wash 30 minutes with current; With alcohol-pickled 15 seconds of concentration of volume percent 75%, aseptic water washing 1 time was used mass percent concentration 0.1%HgCl again
2Solution disinfection 4 minutes; Aseptic water washing 5 times, suck dry moisture are cut to the stem section about 2.5～3.5cm, induce differentiation culture on the inducing culture being inoculated on the superclean bench then; Condition of culture: intensity of illumination is: 3000～4500Lux, temperature is 26 ± 2 ℃.The inducing culture proportioning is: MS, 6-BA 0.7～0.9mg/L, NAA 0.1～0.3mg/L, inductivity reaches 40%.
B, successive transfer culture
At inducing culture about about 30 days, when treating the high 3～4cm of axillalry bud, can get into enrichment culture; Condition of culture is identical with inducing culture; The proliferated culture medium proportioning is: MS, 6-BA 0.4～0.6mg/L, NAA0.1 ± 0.05mg/L, to cultivate 27 days, and the propagation multiplying power reaches 3.2 times.
C, culture of rootage
About 27 days, height of seedling just can carry out culture of rootage when 3～4cm is above at enrichment culture; Condition of culture is identical with above cultivation, and the root media proportioning is: 1/2MS, IBA 0.1～0.3mg/L, about about 30 days of culture of rootage; Rooting rate reaches 97%, and root is about 2～3cm, about the about 4cm of height of seedling; Just can transplant by bottle outlet, be suitable for the requirement that industrialization is produced fully.
D, caption
Table one: different 6-BA and NAA concentration are to the influence of gold string money willow induction frequency, and the highest induction frequency is 40%.
Table two: different 6-BA and NAA concentration is to the influence of gold string money willow propagation multiplying power, and high proliferation multiplying power is 3.2 times.
Table three: different I BA concentration is to the influence of gold string money willow rooting rate, and the highest rooting rate is 97%.
Different 6-BA of table 1. and NAA concentration are to the influence of gold string money willow induction frequency
Different 6-BA of table 2. and NAA concentration are to the influence of gold string money willow propagation multiplying power
Table 3. different I BA concentration is to the influence of gold string money willow rooting rate
Claims (3)
1. the method for tissue culture of gold string Qian Liu is characterized in that:
The clip axillalry bud cleans, sterilizes explant earlier as explant from the gold string money willow parent;
Insert inducing culture then and carry out inducing culture, the inducing culture proportioning is: MS, 6-BA0.7～0.9mg/L, NAA 0.1～0.3mg/L;
Insert proliferated culture medium again and carry out enrichment culture, the proliferated culture medium proportioning is: MS, 6-BA 0.4～0.6mg/L, NAA0.1 ± 0.05mg/L;
At last, get into the culture of rootage stage, the root media proportioning is: 1/2MS, IBA 0.1～0.3mg/L;
The condition of above inducing culture, enrichment culture, culture of rootage is: intensity of illumination is 3000～4500Lux, and temperature is 26 ± 2 ℃.
2. the method for tissue culture of gold string Qian Liu according to claim 1, it is characterized in that: the concrete steps that explant cleans, sterilizes are to soak 15 minutes with detergent solution earlier, place clear water under and wash 30 minutes; The use concentration of volume percent is 75% alcohol-pickled 15 seconds, and aseptic water washing 1 time is used mass percent concentration 0.1%HgCl again
2Solution disinfection 4 minutes, aseptic water washing 5 times.
3. the method for tissue culture of gold string Qian Liu according to claim 1, it is characterized in that: after explant cleaned, sterilizes, suck dry moisture was cut to the stem section of 2.5～3.5cm, inserted inducing culture then and carried out inducing culture.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102960248A (en) * | 2012-11-28 | 2013-03-13 | 巴中市光雾山植物研究所 | Melaleuca bracteata tissue culture and rapid propagation process |
| CN103222426A (en) * | 2013-05-03 | 2013-07-31 | 贵州大学 | Method for establishing Melaleuca bracteata regeneration system |
| CN103733997A (en) * | 2013-12-24 | 2014-04-23 | 镇江山水湾生态农业开发有限公司 | Fast reproduction method for energy willow tissue culture |
| CN103931499A (en) * | 2014-04-16 | 2014-07-23 | 华中农业大学 | Tissue culture rapid propagation method for callistemon rigidus |
| CN105724177A (en) * | 2016-05-14 | 2016-07-06 | 崔子扬 | Cultivating method of callistemon viminalis |
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| CN101032223A (en) * | 2006-08-24 | 2007-09-12 | 厦门涌泉科技有限公司 | Tissue cultivating method of spiceberry melaleuca alternifolia |
| CN101049074A (en) * | 2007-05-14 | 2007-10-10 | 广西玉林市恒汇隆植物香料开发有限公司 | Breeding and reproducing eucalyptus type cajeput with alternate leaves |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101032223A (en) * | 2006-08-24 | 2007-09-12 | 厦门涌泉科技有限公司 | Tissue cultivating method of spiceberry melaleuca alternifolia |
| CN101049074A (en) * | 2007-05-14 | 2007-10-10 | 广西玉林市恒汇隆植物香料开发有限公司 | Breeding and reproducing eucalyptus type cajeput with alternate leaves |
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| 曲芬霞: "影响千层金腋芽及愈伤组织诱导因子初探", 《北方园艺》, no. 8, 30 April 2010 (2010-04-30) * |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102960248A (en) * | 2012-11-28 | 2013-03-13 | 巴中市光雾山植物研究所 | Melaleuca bracteata tissue culture and rapid propagation process |
| CN103222426A (en) * | 2013-05-03 | 2013-07-31 | 贵州大学 | Method for establishing Melaleuca bracteata regeneration system |
| CN103222426B (en) * | 2013-05-03 | 2014-08-20 | 贵州大学 | Method for establishing Melaleuca bracteata regeneration system |
| CN103733997A (en) * | 2013-12-24 | 2014-04-23 | 镇江山水湾生态农业开发有限公司 | Fast reproduction method for energy willow tissue culture |
| CN103931499A (en) * | 2014-04-16 | 2014-07-23 | 华中农业大学 | Tissue culture rapid propagation method for callistemon rigidus |
| CN103931499B (en) * | 2014-04-16 | 2016-07-06 | 华中农业大学 | A kind of quick breeding method for tissue culture of callistemon rigidus |
| CN105724177A (en) * | 2016-05-14 | 2016-07-06 | 崔子扬 | Cultivating method of callistemon viminalis |
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Application publication date: 20120718 |