CN102823496A - Method for rapidly breeding Plectranthus tomentosa through tissue culture - Google Patents
Method for rapidly breeding Plectranthus tomentosa through tissue culture Download PDFInfo
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- CN102823496A CN102823496A CN2012103437181A CN201210343718A CN102823496A CN 102823496 A CN102823496 A CN 102823496A CN 2012103437181 A CN2012103437181 A CN 2012103437181A CN 201210343718 A CN201210343718 A CN 201210343718A CN 102823496 A CN102823496 A CN 102823496A
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Abstract
The invention discloses a method for rapidly breeding Plectranthus tomentosa through tissue culture, which comprises the following steps of: cutting off explants and sterilizing the explants; primary culture, wherein primary culture media comprise MS, 1.5mg/L 6-BA (6-benzyladenine), 0.2mg/L NAA (naphthylacetic acid), 30g/L cane sugar and 10g/L agar; subculture, wherein subculture media comprise MS, 1.0mg/L 6-BA, 0.2mg/L NAA, 30g/L cane sugar and 10g/L agar; rooting culture, wherein rooting culture media comprise 1/2MS, 0.2mg/L NAA, 30g/L cane sugar and 8g/L agar; and washing away culture media, hardening seedlings and transplanting. The formula of the culture media provided by the invention is simple, the breeding process is simple and convenient to operate, the breeding coefficient is high, the goal of large-scale rapid breeding can be realized and the seedlings after breeding grow vigorously and robustly; attentions are required to be paid to sun screening, cooling and moisture preservation after transplanting, and the survival rate can highly reach 100 percent; the seedlings after seedling hardening and transplanting grow well and the fragrance is pure and strong.
Description
Technical field
The present invention relates to a kind of tissue culture and breed the fragrant method of bumping fast.
Background technology
Bump perfume (or spice) (Plectranthus tomentosa), Labiatae Rabdosia, herbaceos perennial.Grow thickly, multi-branched, complete stool is had fine and closely woven white fine hair.Fleshy leaf, alternately to life, green, oval, there is crenation at the edge.Because of giving out the laudatory title that fragrance is enjoyed " bumping perfume (or spice) " after the touching, sweet because of its thick flavor again, quite like apple aroma, so enjoy " apple is fragrant " good reputation again.
Bump perfume (or spice) and can not only distribute the fragrance that makes us spirit and QI cleaning, and capable of purifying air, environmental protection is with conspicuous characteristics, and market is in one's favour; Bump fragrant suitable potted plantly, should perch or be suspended in indoor, also can make the decoration of several cases, desk; Bump that the spiceleaf sheet can be made tea, wine, strange fragrant tempting, produce refreshing effect to the mind clearing away summerheat, mosquito repellent; Bumping fragrant herb and can refine essence, is a kind of emerging plant of following aromatherapy.
Up to now, less for bumping fragrant research both at home and abroad, only there is the correlation analysis of its volatile component to report.At present, bump fragrant breeding method and mainly contain cuttage and two kinds of sexual propagations: fragrant cuttage is bumped in (1) can become alive, but the natural propagation rate is low; (2) sexual propagation can not keep its merit because of variation easily.
Summary of the invention
The objective of the invention is to solve the deficiency of prior art, provide a kind of culture medium prescription simple, breed simple to operate, the speed of breeding is fast, reproduction coefficient is high, cultivates vigorous, the healthy and strong tissue culture of seedling growth and breeds the fragrant method of bumping fast.
The objective of the invention is to realize through following technical scheme: a kind of tissue culture is bred fast and is bumped fragrant method, and it may further comprise the steps:
S1: cut explant and sterilization;
S2: initial culture: the explant after will sterilizing is cut to the long tender stem segments of 2～3cm, is inoculated in the initial culture base and cultivates, wherein; The initial culture base is MS+6-BA1.5mg/L+NAA0.2mg/L+ sucrose 30g/L+ agar 10g/L, and pH value is 5.8, and intensity of illumination is 2800～3200LX; Light application time is 15～17h/d; Temperature is 23～25 ℃, and the cultivation time is 30～60d, accomplishes and to grow the cluster bud of growing thickly on the stem section of initial culture;
S3: successive transfer culture: cut the bud of growing thickly, be divided into the fritter that diameter is 1.5～2.5cm, be inoculated in the subculture medium and cultivate, subculture medium is MS+6-BA1.0mg/L+NAA0.2mg/L+ sucrose 30g/L+ agar 10g/L, and the cultivation time is 6～8d;
S4: culture of rootage: the budlet in the bud of growing thickly grows to 2cm when above; Transfer and in root media, cultivate; Root media is 1/2MS+NAA0.2mg/L+ sucrose 30g/L+ agar 8g/L, and the cultivation time is 18～22d, obtains the strong fragrant tissue cultivating seedling that bumps of taking root after the culture of rootage;
S5: the refining seedling is transplanted: transplant after washing the medium on the tissue cultivating seedling off.
The step that cuts explant and sterilization described in the step S1 comprises following substep:
(1) gathers the fragrant tender stem segments that bumps of no damage by disease and insect;
(2) remove unnecessary blade, in flowing water, wash 30～40min;
(3) in washing agent, soak 4～6min;
(4) with the flowing water flushing, occur until non-foam;
(5) put on the superclean bench, use concentration is alcohol disinfecting 18～22s of 70%～75%;
(6), put into concentration and be 0.1%～0.15% mercuric chloride, the 4～6min that sterilizes with aseptic water washing 4～6 times;
(7) with behind the aseptic water washing 4～6 times, place on the aseptic paper and dry.
Grafting matrix described in the step S5 is a turfy soil mixing vermiculite, and mixed proportion is 1～2:1.
The invention has the beneficial effects as follows: culture medium prescription is simple, and reproductive process is easy and simple to handle, and reproduction coefficient is high, can breed in a large number fast, and the seedling growth after the breeding is vigorous, stalwartness; Transplant the back as long as note shading, lower the temperature and preserve moisture, survival rate can be up to 100%; Seedling behind the acclimatization and transplants grows fine, and fragrance is pure strong.
Description of drawings
Fig. 1 is for growing the photo figure of the bud of growing thickly on the explant behind the initial culture;
Fig. 2 is for growing the photo figure of the bud of growing thickly in a large number on the explant behind the successive transfer culture;
The photo figure that bump fragrant tissue cultivating seedling of Fig. 3 for obtaining after the culture of rootage;
Fig. 4 is the photo figure that bumps the flourishing root system of fragrant tissue cultivating seedling;
Fig. 5 bumps the photo figure that fragrant training tissue culture seedling is transplanted;
Fig. 6 transplants the outdoor growth conditions photo figure in back for bumping fragrant tissue cultivating seedling.
Embodiment
Below in conjunction with specific embodiment and accompanying drawing technical scheme of the present invention is described in further detail, but protection scope of the present invention is not limited to the following stated.
[embodiment 1] a kind of tissue culture is bred fast and is bumped fragrant method, and it may further comprise the steps:
S1: cut explant and sterilization, it comprises following substep again:
(1) gathers the fragrant tender stem segments that bumps of no damage by disease and insect;
(2) remove unnecessary blade, in flowing water, wash 40min;
(3) in washing agent, soak 4min;
(4) with the flowing water flushing, occur until non-foam;
(5) put on the superclean bench, use concentration is 75% alcohol disinfecting 18s;
(6), put into concentration and be 0.1% the mercuric chloride 6min that sterilizes with aseptic water washing 6 times;
(7) with behind the aseptic water washing 6 times, place on the aseptic paper and dry;
S2: initial culture: the explant after will sterilizing is cut to the long tender stem segments of 2cm, is inoculated in the initial culture base and cultivates, wherein; The initial culture base is MS+6-BA1.5mg/L+NAA0.2mg/L+ sucrose 30g/L+ agar 10g/L, and pH value is 5.8, and intensity of illumination is 3200LX; Light application time is 15h/d, and temperature is 25 ℃, and the cultivation time is 30d; Accomplish to grow the cluster bud of growing thickly on the stem section of initial culture, a stem Duan Shangke induces the sprouting up more than 60;
S3: successive transfer culture: cut the bud of growing thickly; Be divided into the fritter that diameter is 1.5cm; Be inoculated in the subculture medium and cultivate, subculture medium is MS+6-BA1.0mg/L+NAA0.2mg/L+ sucrose 30g/L+ agar 10g/L, and the cultivation time is 6d; After accomplishing successive transfer culture, the quantity of the bud of growing thickly can increase more than 3 times;
S4: culture of rootage: the budlet in the bud of growing thickly grows to 2cm when above; Transfer and in root media, cultivate, root media is 1/2MS+NAA0.2mg/L+ sucrose 30g/L+ agar 8g/L, cultivates behind the 5d it is thus clear that take root; After cultivating 18d, obtain the strong fragrant tissue cultivating seedling that bumps of taking root;
S5: the refining seedling is transplanted: transplant after washing the medium on the tissue cultivating seedling off, grafting matrix is a turfy soil mixing vermiculite, and mixed proportion is 2:1.
[embodiment 2] a kind of tissue culture is bred fast and is bumped fragrant method, and it may further comprise the steps:
S1: cut explant and sterilization, it comprises following substep again:
(1) gathers the fragrant tender stem segments that bumps of no damage by disease and insect;
(2) remove unnecessary blade, in flowing water, wash 30min;
(3) in washing agent, soak 6min;
(4) with the flowing water flushing, occur until non-foam;
(5) put on the superclean bench, use concentration is 70% alcohol disinfecting 22s;
(6), put into concentration and be 0.15% the mercuric chloride 4min that sterilizes with aseptic water washing 4 times;
(7) with behind the aseptic water washing 4 times, place on the aseptic paper and dry;
S2: initial culture: the explant after will sterilizing is cut to the long tender stem segments of 3cm, is inoculated in the initial culture base and cultivates, wherein; The initial culture base is MS+6-BA1.5mg/L+NAA0.2mg/L+ sucrose 30g/L+ agar 10g/L, and pH value is 5.8, and intensity of illumination is 2800LX; Light application time is 17h/d, and temperature is 23 ℃, and the cultivation time is 60d; Accomplish to grow the cluster bud of growing thickly on the stem section of initial culture, a stem Duan Shangke induces the sprouting up more than 60;
S3: successive transfer culture: cut the bud of growing thickly; Be divided into the fritter that diameter is 2.5cm; Be inoculated in the subculture medium and cultivate, subculture medium is MS+6-BA1.0mg/L+NAA0.2mg/L+ sucrose 30g/L+ agar 10g/L, and the cultivation time is 8d; After accomplishing successive transfer culture, the quantity of the bud of growing thickly can increase more than 3 times;
S4: culture of rootage: the budlet in the bud of growing thickly grows to 2cm when above; Transfer and in root media, cultivate, root media is 1/2MS+NAA0.2mg/L+ sucrose 30g/L+ agar 8g/L, cultivates behind the 5d it is thus clear that take root; After cultivating 22d, obtain the strong fragrant tissue cultivating seedling that bumps of taking root;
S5: the refining seedling is transplanted: transplant after washing the medium on the tissue cultivating seedling off, grafting matrix is a turfy soil mixing vermiculite, and mixed proportion is 1:1.
[embodiment 3] a kind of tissue culture is bred fast and is bumped fragrant method, and it may further comprise the steps:
S1: cut explant and sterilization, it comprises following substep again:
(1) gathers the fragrant tender stem segments that bumps of no damage by disease and insect;
(2) remove unnecessary blade, in flowing water, wash 35min;
(3) in washing agent, soak 5min;
(4) with the flowing water flushing, occur until non-foam;
(5) put on the superclean bench, use concentration is 72.5% alcohol disinfecting 20s;
(6), put into concentration and be 0.125% the mercuric chloride 5min that sterilizes with aseptic water washing 5 times;
(7) with behind the aseptic water washing 5 times, place on the aseptic paper and dry;
S2: initial culture: the explant after will sterilizing is cut to the long tender stem segments of 2.5cm, is inoculated in the initial culture base and cultivates, wherein; The initial culture base is MS+6-BA1.5mg/L+NAA0.2mg/L+ sucrose 30g/L+ agar 10g/L, and pH value is 5.8, and intensity of illumination is 3000LX; Light application time is 16h/d, and temperature is 24 ℃, and the cultivation time is 45d; Accomplish to grow the cluster bud of growing thickly on the stem section of initial culture, a stem Duan Shangke induces the sprouting up more than 60;
S3: successive transfer culture: cut the bud of growing thickly; Be divided into the fritter that diameter is 2cm; Be inoculated in the subculture medium and cultivate, subculture medium is MS+6-BA1.0mg/L+NAA0.2mg/L+ sucrose 30g/L+ agar 10g/L, and the cultivation time is 7d; After accomplishing successive transfer culture, the quantity of the bud of growing thickly can increase more than 3 times;
S4: culture of rootage: the budlet in the bud of growing thickly grows to 2cm when above; Transfer and in root media, cultivate, root media is 1/2MS+NAA0.2mg/L+ sucrose 30g/L+ agar 8g/L, cultivates behind the 5d it is thus clear that take root; After cultivating 20d, obtain the strong fragrant tissue cultivating seedling that bumps of taking root;
S5: the refining seedling is transplanted: transplant after washing the medium on the tissue cultivating seedling off, grafting matrix is a turfy soil mixing vermiculite, and mixed proportion is 1.5:1.
[embodiment 4] a kind of tissue culture is bred fast and is bumped fragrant method, and it may further comprise the steps:
S1: cut explant and sterilization, it comprises following substep again:
(1) gathers the fragrant tender stem segments that bumps of no damage by disease and insect;
(2) remove unnecessary blade, in flowing water, wash 30min;
(3) in washing agent, soak 5min;
(4) with the flowing water flushing, occur until non-foam;
(5) put on the superclean bench, use concentration is 75% alcohol disinfecting 20s;
(6), put into concentration and be 0.15% the mercuric chloride 5min that sterilizes with aseptic water washing 5 times;
(7) with behind the aseptic water washing 5 times, place on the aseptic paper and dry;
S2: initial culture: the explant after will sterilizing is cut to the long tender stem segments of 2cm, is inoculated in the initial culture base and cultivates, wherein; The initial culture base is MS+6-BA1.5mg/L+NAA0.2mg/L+ sucrose 30g/L+ agar 10g/L, and pH value is 5.8, and intensity of illumination is 3000LX; Light application time is 16h/d, and temperature is 24 ℃, and the cultivation time is 50d; Accomplish to grow the cluster bud of growing thickly on the stem section of initial culture, a stem Duan Shangke induces the sprouting up more than 60;
S3: successive transfer culture: cut the bud of growing thickly; Be divided into the fritter that diameter is 2cm; Be inoculated in the subculture medium and cultivate, subculture medium is MS+6-BA1.0mg/L+NAA0.2mg/L+ sucrose 30g/L+ agar 10g/L, and the cultivation time is 7d; After accomplishing successive transfer culture, the quantity of the bud of growing thickly can increase more than 3 times;
S4: culture of rootage: the budlet in the bud of growing thickly grows to 2cm when above; Transfer and in root media, cultivate, root media is 1/2MS+NAA0.2mg/L+ sucrose 30g/L+ agar 8g/L, cultivates behind the 5d it is thus clear that take root; After cultivating 20d, obtain the strong fragrant tissue cultivating seedling that bumps of taking root;
S5: the refining seedling is transplanted: transplant after washing the medium on the tissue cultivating seedling off, grafting matrix is a turfy soil mixing vermiculite, and mixed proportion is 2:1.
As shown in Figure 1, accomplish to grow the cluster bud of growing thickly on the stem section of initial culture, a stem Duan Shangke induces the sprouting up more than 60; As shown in Figure 2, behind the completion successive transfer culture, the quantity of the bud of growing thickly can increase more than 3 times; Like Fig. 3, shown in Figure 4, visible taking root behind the culture of rootage 5d behind the cultivation 20d, obtains the strong fragrant tissue cultivating seedling that bumps of taking root; Like Fig. 5, shown in Figure 6, the seedling growth after the refining seedling is transplanted is vigorous, grows fine.
Claims (3)
1. a tissue culture is bred fast and is bumped fragrant method, and it is characterized in that: it may further comprise the steps:
S1: cut explant and sterilization;
S2: initial culture: the explant after will sterilizing is cut to the long tender stem segments of 2～3cm, is inoculated in the initial culture base and cultivates, wherein; The initial culture base is MS+6-BA1.5mg/L+NAA0.2mg/L+ sucrose 30g/L+ agar 10g/L, and pH value is 5.8, and intensity of illumination is 2800～3200LX; Light application time is 15～17h/d; Temperature is 23～25 ℃, and the cultivation time is 30～60d, accomplishes and to grow the cluster bud of growing thickly on the stem section of initial culture;
S3: successive transfer culture: cut the bud of growing thickly, be divided into the fritter that diameter is 1.5～2.5cm, be inoculated in the subculture medium and cultivate, subculture medium is MS+6-BA1.0mg/L+NAA0.2mg/L+ sucrose 30g/L+ agar 10g/L, and the cultivation time is 6～8d;
S4: culture of rootage: the budlet in the bud of growing thickly grows to 2cm when above; Transfer and in root media, cultivate; Root media is 1/2MS+NAA0.2mg/L+ sucrose 30g/L+ agar 8g/L, and the cultivation time is 18～22d, obtains the strong fragrant tissue cultivating seedling that bumps of taking root after the culture of rootage;
S5: the refining seedling is transplanted: transplant after washing the medium on the tissue cultivating seedling off.
2. a kind of tissue culture according to claim 1 is bred fast and bumped fragrant method, it is characterized in that: the step that cuts explant and sterilization described in the step S1 comprises following substep:
(1) gathers the fragrant tender stem segments that bumps of no damage by disease and insect;
(2) remove unnecessary blade, in flowing water, wash 30～40min;
(3) in washing agent, soak 4～6min;
(4) with the flowing water flushing, occur until non-foam;
(5) put on the superclean bench, use concentration is alcohol disinfecting 18～22s of 70%～75%;
(6), put into concentration and be 0.1%～0.15% mercuric chloride, the 4～6min that sterilizes with aseptic water washing 4～6 times;
(7) with behind the aseptic water washing 4～6 times, place on the aseptic paper and dry.
3. a kind of tissue culture according to claim 1 is bred fast and is bumped fragrant method, and it is characterized in that: the grafting matrix described in the step S5 is a turfy soil mixing vermiculite, and mixed proportion is 1～2:1.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103241839A (en) * | 2013-05-27 | 2013-08-14 | 沈阳大学 | Treatment method of treating cadmium contaminated water |
| CN105638463A (en) * | 2015-12-30 | 2016-06-08 | 四川禾木本业农林科技有限公司 | Tissue-culture rapid propagation method for succulents |
| CN110972949A (en) * | 2019-12-19 | 2020-04-10 | 美尚生态景观股份有限公司 | In-bottle rooting method for tissue culture seedlings of rabdosia nervosa |
-
2012
- 2012-09-17 CN CN2012103437181A patent/CN102823496A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| 廖苏梅等: "豆蔻天竺葵的组织培养", 《浙江农业科学》 * |
| 熊伟等: "碰碰香挥发物化学成分分析", 《浙江农林大学学报》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103241839A (en) * | 2013-05-27 | 2013-08-14 | 沈阳大学 | Treatment method of treating cadmium contaminated water |
| CN103241839B (en) * | 2013-05-27 | 2014-12-03 | 沈阳大学 | Treatment method of treating cadmium contaminated water |
| CN105638463A (en) * | 2015-12-30 | 2016-06-08 | 四川禾木本业农林科技有限公司 | Tissue-culture rapid propagation method for succulents |
| CN110972949A (en) * | 2019-12-19 | 2020-04-10 | 美尚生态景观股份有限公司 | In-bottle rooting method for tissue culture seedlings of rabdosia nervosa |
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