CN102511391B - Hyacinthus orientalis L. in-vitro rapid propagation method - Google Patents
Hyacinthus orientalis L. in-vitro rapid propagation method Download PDFInfo
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- CN102511391B CN102511391B CN201110404105XA CN201110404105A CN102511391B CN 102511391 B CN102511391 B CN 102511391B CN 201110404105X A CN201110404105X A CN 201110404105XA CN 201110404105 A CN201110404105 A CN 201110404105A CN 102511391 B CN102511391 B CN 102511391B
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Abstract
The invention relates to a hyacinthus orientalis L. in-vitro rapid propagation method, and belongs to the field of biotechnology. The hyacinthus orientalis L. in-vitro rapid propagation method comprises the following steps of 1, selecting hyacinthus orientalis L. seed bulbs growing in an open field for 2 to 3 months, carrying out disinfection, and removing outer scales, 2, taking floret petals, flatwise putting the floret petals on a MS solid medium, and carrying out adventitious bud growth induction culture to obtain tissue blocks containing adventitious buds, 3, longitudinally dividing the tissue blocks containing adventitious buds, and carrying out bud elongation culture by a bud elongation medium, and 4, when length values of the adventitious buds obtained by the step 3 are in a range of 2 to 4 centimeters, cutting the adventitious buds, putting the cut adventitious buds into a rooting medium, and carrying out rooting induction culture to obtain hyacinthus orientalis L. seedlings. The young floret petals of hyacinthus orientalis L. are utilized as explants and directly regenerate a mass of adventitious buds and the adventitious buds further develop into regenerated seedlings. Compared with the existing tissue culture method utilizing scales as explants, the hyacinthus orientalis L. in-vitro rapid propagation method has high regeneration efficiency, a high budding rate of about 100% and a large average amount of buds regenerated by induction of each one explant, wherein the large average amount is about 20.
Description
Technical field
The present invention relates to the method for quickly breeding in a kind of biological technical field, be specifically related to a kind of Hyacinthus orientalis.
Background technology
Hyacinth (Hyacinthus orientalis L.) is Liliaceae Hyacinthus herbaceos perennial, originates in Mediterranean and South Africa, is the flowering bulb in spring that has sight of extensive use all over the world.China in last century the eighties from Dutch introduction of plant, but traditional propagation method is consuming time longer, reproductive efficiency is lower, plants simultaneously the expensive of ball, demand that can not satisfying the market, limited hyacinth plantation at home.Therefore, cultivate by tissue that to carry out hyacinthine Fast-propagation be effective technological means, be conducive to strengthen the popularization of hyacinth in China.
The hyacinth scale is the group training material of the Fast-propagation commonly used the most.Research shows, hyacinth blade and the ovary formation seedling of also can regenerating.Although regenerate take bulb as explant, draw materials relatively convenient, but because bulb carries disease germs seriously, usually cause sterilization not thorough, in cultivation process, contamination phenomenon is very serious repeatedly, increase workload, and not very good take scale as explant regeneration efficiency.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art, a kind of Hyacinthus orientalis is provided.Method regeneration efficiency of the present invention increases, and pollution rate reduces, and by regulating culture medium condition, has improved inductivity, has shortened the regeneration period, and this regenerating system is applicable to a plurality of hyacinth kinds.
The present invention realizes by following technical scheme, and Hyacinthus orientalis of the present invention, comprise the steps: to get hyacinth Xiao Hua petal as explant, the cultivation of regenerate, acquisition hyacinth seedling.
Preferably, described Hyacinthus orientalis, comprise the steps:
(a) get the open country plantation hyacinth kind ball of 2～3 months, sterilization, divest outer scale;
(b) strip the Xiao Hua petal, lie in the induced growth that carries out indefinite bud on the MS solid culture medium and cultivate, obtain the tissue block with the indefinite bud clump;
(c) vertical minute tissue block, carry out the bud elongation and cultivate on the bud elongation medium;
(d) during Elongation of adventitious bud to 2～4cm, after cutting, put into root media and carry out the root induction cultivation, obtain the hyacinth seedling.
Preferably, in step (b), described MS solid culture medium is MS+BA 2.0mg/L+NAA 1.0mg/L, and pH 5.8, sucrose 30g/L, agar 5g/L.
Preferably, in step (b), the condition that described induced growth is cultivated is: 25 ℃ of temperature, illumination: dark phase of 16 photophase/8.
Preferably, in step (c), described bud elongation medium is MS+BA0.5mg/L+NAA1.0mg/L, pH5.8, sucrose 30g/L, agar 5g/L.
Preferably, in step (c), the condition that described bud elongation is cultivated is: 25 ℃ of temperature, illumination: dark phase of 16 photophase/8.
Preferably, in step (d), described root media is: MS+NAA0.5mg/L+ active carbon 1.0g/L, pH 5.8, sucrose 30g/L, agar 5g/L.
Preferably, in step (d), the condition that described root induction is cultivated is: 25 ℃ of temperature, illumination: dark phase of 16 photophase/8.
The present invention has following beneficial effect: method of the present invention utilizes the tender small-flowered sheet of hyacinthine children to be explant, the a large amount of indefinite buds of Direct Regeneration, then obtain regrowth, with the use scale, as explant, organize to cultivate and compare, method regeneration efficiency of the present invention increases, bud ratio is 100% left and right, the bud quantity of average each explant induction regeneration can reach 20 left and right, pollution rate reduces, in cultivation process, not there will be situation about repeatedly polluting, by regulating culture medium condition, can improve inductivity and shorten the regeneration period.
Embodiment
The present invention is described in detail below in conjunction with specific embodiment.Following examples will help those skilled in the art further to understand the present invention, but not limit in any form the present invention.It should be pointed out that to those skilled in the art, without departing from the inventive concept of the premise, can also make some distortion and improvement.These all belong to protection scope of the present invention.
Embodiment 1
Select good hyacinth kind Gypsy queen (Gipsy Queen) in about November, planting in large Tanaka then, take kind of a ball mid-January next year;
Get young tender small-flowered sheet and be explant and carry out bud and induce cultivation, medium is MS+BA1.0mg/L+NAA0.5mg/L, and pH 5.8, sucrose 30g/L, agar 5g/L.Condition of culture is: 25 ℃ of temperature, illumination: dark phase of 16 photophase/8.Bud ratio can reach 70%, and on average the bud quantity of each explant induction regeneration is 8 left and right.
A large amount of bud clumps after 40 days, on the petal sheet, occur, by after the cutting of bud clump, put into the bud elongation medium and cultivate, medium is MS+NAA0.5mg/L, pH5.8, sucrose 30g/L, agar 5g/L.Condition of culture is the same.
After 30 days, bud is elongated to 2-4cm, and cutting bud clump, put into root media and cultivate, and root media is MS+NAA0.5mg/L, and pH 5.8, sucrose 30g/L, agar 5g/L.Condition of culture is the same.After after 20 days, bearing 3-4 bar root, obtain regrowth.After hardening, transplant to little basin or open country, suitably shade a week, but the regrowth normal growth.In cultivation process without pollution condition repeatedly.
Embodiment 2
Select good hyacinth kind Gypsy queen (Gipsy Queen) in about November, planting in large Tanaka then, take kind of a ball mid-January next year;
Get young tender small-flowered sheet and be explant and carry out bud and induce cultivation, medium is MS+BA2.0mg/L+NAA1.0mg/L, and pH 5.8, sucrose 30g/L, agar 5g/L.Condition of culture is: 25 ℃ of temperature, illumination: dark phase of 16 photophase/8.Bud ratio can reach 100%, and on average the bud quantity of each explant induction regeneration is 20 left and right.
A large amount of bud clumps after 30 days, on the petal sheet, occur, by after the cutting of bud clump, put into the bud elongation medium and cultivate, medium is MS+BA0.5mg/L+NAA1.0mg/L, pH5.8, sucrose 30g/L, agar 5g/L.Condition of culture is the same.
After 20 days, treat that bud is elongated to 2-4cm, cutting bud clump, put into root media and cultivate, and root media is MS+NAA0.5mg/L+ active carbon 1.0g/L, and pH 5.8, sucrose 30g/L, agar 5g/L.Condition of culture is the same.After 10 days, after bearing 3-4 bar root, obtain regrowth.After hardening, transplant to little basin or open country, suitably shade a week, but the regrowth normal growth.In cultivation process without pollution condition repeatedly.
Embodiment 3
Select good hyacinth kind Gypsy queen (Gipsy Queen) in about November, planting in large Tanaka then, take kind of a ball mid-January next year;
Get young tender small-flowered sheet and be explant and carry out bud and induce cultivation, medium is MS+BA3.0mg/L+NAA2.0mg/L, and pH 5.8, sucrose 30g/L, agar 5g/L.Condition of culture is: 25 ℃ of temperature, illumination: dark phase of 16 photophase/8.Bud ratio can reach 80%, and on average the bud quantity of each explant induction regeneration is 12 left and right.
A large amount of bud clumps after 35 days, on the petal sheet, occur, by after the cutting of bud clump, put into the bud elongation medium and cultivate, medium is MS+NAA1.0mg/L, pH5.8, sucrose 30g/L, agar 5g/L.Condition of culture is the same.
After 25 days, treat that bud is elongated to 2-4cm, cutting bud clump, put into root media and cultivate, and root media is MS+NAA0.5mg/L+ active carbon 2.0g/L, and pH 5.8, sucrose 30g/L, agar 5g/L.Condition of culture is the same.After 12 days, after bearing 3-4 bar root, obtain regrowth.After hardening, transplant to little basin or open country, suitably shade a week, but the regrowth normal growth.In cultivation process without pollution condition repeatedly.
Embodiment 4
Select good hyacinth kind Annamarie (Anna Marie) in about November, planting in large Tanaka then, take kind of a ball mid-January next year;
Get young tender small-flowered sheet and be explant and carry out bud and induce cultivation, medium is MS+BA2.0mg/L+NAA1.0mg/L, and pH 5.8, sucrose 30g/L, agar 5g/L.Condition of culture is: 25 ℃ of temperature, illumination: dark phase of 16 photophase/8.Bud ratio can reach 100%, and on average the bud quantity of each explant induction regeneration is 22 left and right.
A large amount of bud clumps after 30 days, on the petal sheet, occur, by after the cutting of bud clump, put into the bud elongation medium and cultivate, medium is MS+BA0.5mg/L+NAA1.0mg/L, pH5.8, sucrose 30g/L, agar 5g/L.Condition of culture is the same.
After 20 days, treat that bud is elongated to 2-4cm, cutting bud clump, put into root media and cultivate, and root media is MS+NAA0.5mg/L+ active carbon 1.0g/L, and pH 5.8, sucrose 30g/L, agar 5g/L.Condition of culture is the same.After 10 days, after bearing 3-4 bar root, obtain regrowth.After hardening, transplant to little basin or open country, suitably shade a week, but the regrowth normal growth.In cultivation process without pollution condition repeatedly.
Embodiment 5
Select good hyacinth kind starry sky (Sky Jacket) in about November, planting in large Tanaka then, take kind of a ball mid-January next year;
Get young tender small-flowered sheet and be explant and carry out bud and induce cultivation, medium is MS+BA2.0mg/L+NAA1.0mg/L, and pH 5.8, sucrose 30g/L, agar 5g/L.Condition of culture is: 25 ℃ of temperature, illumination: dark phase of 16 photophase/8.Bud ratio can reach 100%, and on average the bud quantity of each explant induction regeneration is 18 left and right.
A large amount of bud clumps after 30 days, on the petal sheet, occur, by after the cutting of bud clump, put into the bud elongation medium and cultivate, medium is MS+BA0.5mg/L+NAA1.0mg/L, pH5.8, sucrose 30g/L, agar 5g/L.Condition of culture is the same.
After 20 days, treat that bud is elongated to 2-4cm, cutting bud clump, put into root media and cultivate, and root media is MS+NAA0.5mg/L+ active carbon 1.0g/L, and pH 5.8, sucrose 30g/L, agar 5g/L.Condition of culture is the same.After 10 days, after bearing 3-4 bar root, obtain regrowth.After hardening, transplant to little basin or open country, suitably shade a week, but the regrowth normal growth.In cultivation process without pollution condition repeatedly.
In sum, method regeneration efficiency of the present invention increases, and pollution rate reduces, and by regulating culture medium condition, has improved inductivity, has shortened the regeneration period, and this regenerating system is applicable to a plurality of hyacinth kinds.
Claims (4)
1. a Hyacinthus orientalis, is characterized in that, comprises the steps:
(a) get the open country plantation hyacinth kind ball of 2～3 months, sterilization, divest outer scale;
(b) strip the Xiao Hua petal, lie in the induced growth that carries out indefinite bud on the MS solid culture medium and cultivate, obtain the tissue block with the indefinite bud clump;
(c) vertical minute tissue block, carry out the bud elongation and cultivate on the bud elongation medium;
(d) during Elongation of adventitious bud to 2～4cm, after cutting, put into root media and carry out the root induction cultivation, obtain the hyacinth seedling;
In step (b), described MS solid culture medium is MS+BA 2.0mg/L+NAA 1.0mg/L, and pH 5.8, sucrose 30g/L, agar 5g/L;
In step (c), described bud elongation medium is MS+BA0.5mg/L+NAA1.0mg/L, pH5.8, sucrose 30g/L, agar 5g/L;
In step (d), described root media is: MS+NAA0.5mg/L+ active carbon 1.0g/L, pH 5.8, sucrose 30g/L, agar 5g/L.
2. Hyacinthus orientalis as claimed in claim 1, its feature be, in step (b), the condition that described induced growth is cultivated is: 25 ℃ of temperature, illumination: dark phase of 16 photophase/8.
3. Hyacinthus orientalis as claimed in claim 1, its feature be, in step (c), the condition that described bud elongation is cultivated is: 25 ℃ of temperature, illumination: dark phase of 16 photophase/8.
4. Hyacinthus orientalis as claimed in claim 1, its feature be, in step (d), the condition that described root induction is cultivated is: 25 ℃ of temperature, illumination: dark phase of 16 photophase/8.
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| CN103891515B (en) * | 2014-04-15 | 2015-11-04 | 李蓁 | Muscari botryoides seeds of flowering plants method for planting |
| CN104488715B (en) * | 2014-12-19 | 2016-08-03 | 浙江省农业科学院 | A kind of method carrying out wide leaf spring grass bulb induction and plant regeneration by alabastrum |
| CN104521527B (en) * | 2015-01-06 | 2017-01-11 | 云南省农业科学院花卉研究所 | Rapid propagation method for hyacinths |
| CN105210877A (en) * | 2015-10-20 | 2016-01-06 | 韦丽 | A kind of Lilium brownii var viridulum method for quickly breeding |
| CN108834895A (en) * | 2018-07-14 | 2018-11-20 | 沿河后花园农业观光旅游综合开发有限公司 | A kind of lily method for in-vitro rapid propagation |
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