CN104686342A - Asexual and rapid propagation technology for styrax tonkinensis - Google Patents
Asexual and rapid propagation technology for styrax tonkinensis Download PDFInfo
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- CN104686342A CN104686342A CN201510085932.5A CN201510085932A CN104686342A CN 104686342 A CN104686342 A CN 104686342A CN 201510085932 A CN201510085932 A CN 201510085932A CN 104686342 A CN104686342 A CN 104686342A
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Abstract
The invention discloses an asexual and rapid propagation technology for styrax tonkinensis, and relates to a seedling growing method for obtaining stable-property seedlings through a plant tissue culture technology of styrax tonkinensis. Sterile seedlings are obtained by utilizing sterile sowing, the stem segment with axillary buds is taken as an explant, rapid propagation of the styrax tonkinensis seedlings is realized through processes of propagation culture, rooting culture, seedling hardening, transplanting, and the like, the problem that the high-quality tree seedlings which are integrated with a medicinal purpose, an oil purpose, a material purpose and an ornamental value are absent is solved, and practical significance is provided for developing the wild plant with an extremely high ornamental value and an edible value.
Description
Technical field
The present invention relates to the method for Plant Tissue Breeding in agricultural biotechnologies, specifically, relate to a kind of styrax tonkinensis Craib ex Hart Fast Asexual Propagation Technique.
Background technology
Styrax tonkinensis Craib ex Hart (
styrax tonkinensis) have another name called Styrax tonkinensis, be Styracaceae Styraax deciduous tree, be integrate medicinal, oil, material with and the fine tree species viewed and admired.Mainly be distributed in Perenniporia martius height above sea level in the sparse woods of 100 ~ 2000m, in the ground natural forest such as Yunnan Province of China, Guizhou, Guangdong, Guangxi, Fujian, Hunan and Jiangxi, have fragmentary distribution.
Styrax is the resin of Styrax plant secretion, containing more balsamic acid, have refresh oneself, invigorate blood circulation, the effect such as pain relieving.In styrax tonkinensis Craib ex Hart resin, styrax content can reach more than 25%, is therefore a kind of valuable pharmaceutical material.Styrax tonkinensis Craib ex Hart timber is diffuse porous wood, and trunk is logical straight, and compact structure, material is soft, can make splint, furniture and sheet material.Styrax tonkinensis Craib ex Hart seed oil is called " white flower oil ", and main component is oleic acid, linoleic acid, palmitic acid and arachidic acid, and wherein unsaturated fatty acid total content reaches 85.4%, is only second to tea oil, olive oil and sesame oil, is a kind of broad-spectrum edible oil.In addition, styrax tonkinensis Craib ex Hart or a kind of excellent ornamental tree species.Current south China, just at development scaleization plantation styrax tonkinensis Craib ex Hart, lacks high-quality and spends the bottleneck that seeds seedling is its large-scale planting of restriction in vain.But current styrax tonkinensis Craib ex Hart mainly adopts planting seed, cottage propagation, root turion modes of reproduction to carry out nursery, and efficiency is low.Therefore, the styrax tonkinensis Craib ex Hart group Fast Asexual Propagation Technique setting up complete set is necessary very much.
Summary of the invention
The object of the present invention is to provide out a kind of styrax tonkinensis Craib ex Hart Fast Asexual Propagation Technique, logical the present invention utilizes aseptic seeding to obtain aseptic seedling, get its stem segment with axillary bud as explant, achieved the Fast-propagation of styrax tonkinensis Craib ex Hart seedling by Multiplying culture, culture of rootage, hardening and transplanting and other steps, thus achieve object of the present invention.
A kind of styrax tonkinensis Craib ex Hart Fast Asexual Propagation Technique of the present invention, comprises the following steps:
(1) fruit harvesting: the fruit of gathering then of styrax tonkinensis Craib ex Hart fine individual plant of gathering, Fruit dries in the shade after regaining and is stored in 4 DEG C of refrigerators after sloughing pericarp for subsequent use.
(2) aseptic seeding: the kind benevolence of styrax tonkinensis Craib ex Hart is first used after 75% ethanol disinfection 5 ~ 30s with aseptic washing 3 ~ 5 times, again with 0.1% mercuric chloride solution sterilization 10 ~ 25min, in superclean bench, carry out seed germination cultivation by being inoculated on MS medium by embryo after the endosperm excision outside kind benevolence with after aseptic water washing 4 ~ 6 times.
(3) Multiplying culture: the bud of aseptic seedling step (1) obtained is cut into 1.5 ~ 2.0cm length, every section of bud that at least one, band is healthy and strong, be incorporated on proliferated culture medium and carry out Multiplying culture, first full light culture 7 ~ 14 days under 25 ~ 28 DEG C of conditions after inoculation, then illumination every day is placed in 12 ~ 16 hours, intensity of illumination is 1500 ~ 2500lx, and being placed in cultivation temperature is cultivate under the condition of 25 ~ 28 DEG C, and switching in 35 ~ 40 days once.
(4) culture of rootage: the bud of step (1) or (2) aseptic seedling is cut into 1.5 ~ 2.0cm length, every section of bud that at least one, band is healthy and strong is cut to be inoculated on root media and is carried out culture of rootage, first full light culture 7 ~ 14 days under 25 ~ 28 DEG C of conditions after inoculation, then illumination every day is placed in 14 ~ 16 hours, intensity of illumination is 2000 ~ 3000lx, and cultivation temperature is be cultured under the condition of 25 ~ 28 DEG C to take root.
(5) acclimatization and transplants: the rooting tube plantlet decap of height about 8 ~ 10cm was placed in natural lighting lower refining seedling after 5 ~ 7 days, test-tube plantlet is taken out from blake bottle, wash root medium off, to plant in the matrix be mixed into by peat soil and yellow sand mud and to be colonizated in large Tanaka.
Proliferated culture medium described in above-mentioned steps (3) is: MS+6-BA0.3 ~ 0.9mg/L+NAA0.2 ~ 0.4mg/L+ sucrose 15 ~ 30g/L+ agar 3.5 ~ 6.0g/L, pH is 5.4 ~ 5.8.
Root media described in above-mentioned steps (4) is: MS+NAA0.2 ~ 0.5mg/L+IBA0.2 ~ 0.5mg/L+ sucrose 15 ~ 30g/L+ agar 3.5 ~ 6.0g/L, pH is 5.4 ~ 5.8.
Compared with prior art advantage of the present invention is: the present invention stablizes the seedling-cultivating method of seedling by plant tissue culture technique acquired character.Aseptic seeding is utilized to obtain aseptic seedling, get its stem segment with axillary bud as explant, the Fast-propagation of styrax tonkinensis Craib ex Hart seedling is achieved by Multiplying culture, culture of rootage, hardening and transplanting and other steps, solve the problem of the fine tree species seedling shortage that this integrates medicinal, oil is used, material is used and view and admire, to abundant this wild plant with high in ornamental value and edibility of exploitation, there is realistic meaning.
Embodiment
Following examples further illustrate of the present invention, is not limitation of the present invention.
Embodiment 1:
(1) fruit harvesting: the fruit of gathering then of styrax tonkinensis Craib ex Hart fine individual plant of gathering, Fruit dries in the shade after regaining and is stored in 4 DEG C of refrigerators after sloughing pericarp for subsequent use.
(2) aseptic seeding: the kind benevolence of styrax tonkinensis Craib ex Hart is first used after 75% ethanol disinfection 10s with aseptic washing 3 times, again with 0.1% mercuric chloride solution sterilization 10min, in superclean bench, carry out seed germination cultivation by being inoculated on MS medium by embryo after the endosperm excision outside kind benevolence with after aseptic water washing 4 times.Pollution rate is low to moderate less than 5%, and survival rate can reach 92%.
(3) Multiplying culture: the bud of aseptic seedling step (1) obtained is cut into 1.5 ~ 2.0cm length, every section of bud that at least one, band is healthy and strong, be incorporated on proliferated culture medium and carry out Multiplying culture, first full light culture 7 days under 25 DEG C of conditions after inoculation, then illumination every day is placed in 12 hours, intensity of illumination is 1500lx, and being placed in cultivation temperature is cultivate under the condition of 25 DEG C, and switching in 35 days once.Cultivate after 1 month, the average production height of bud is 5.02cm, and growth coefficient is 4.8, and plant strain growth is good.Described proliferated culture medium is: MS+6-BA0.3mg/L+NAA0.2mg/L+ sucrose 15g/L+ agar 3.5g/L, pH is 5.8.
(4) culture of rootage: the bud of step (1) or (2) aseptic seedling is cut into 1.5 ~ 2.0cm length, every section of bud that at least one, band is healthy and strong is cut to be inoculated on root media and is carried out culture of rootage, first full light culture 7 days under 25 DEG C of conditions after inoculation, then illumination every day is placed in 14 hours, intensity of illumination is 2000lx, and cultivation temperature is be cultured under the condition of 25 DEG C to take root.Cultivate after 35 days, average root is raw reaches 6.0cm, well developed root system, and rooting rate reaches more than 86%.Described root media is: MS+NAA0.2mg/L+IBA0.2mg/L+ sucrose 15g/L+ agar 3.5g/L, pH is 5.8.
(5) acclimatization and transplants: the rooting tube plantlet decap of height about 8 ~ 10cm was placed in natural lighting lower refining seedling after 5 ~ 7 days, test-tube plantlet is taken out from blake bottle, wash root medium off, to plant in the matrix be mixed into by peat soil and yellow sand mud and to be colonizated in large Tanaka, transplanting into survival rate 91%.
Embodiment 2:
(1) fruit harvesting: the fruit of gathering then of styrax tonkinensis Craib ex Hart fine individual plant of gathering, Fruit dries in the shade after regaining and is stored in 4 DEG C of refrigerators after sloughing pericarp for subsequent use.
(2) aseptic seeding: the kind benevolence of styrax tonkinensis Craib ex Hart is first used after 75% ethanol disinfection 13s with aseptic washing 5 times, again with 0.1% mercuric chloride solution sterilization 15min, in superclean bench, carry out seed germination cultivation by being inoculated on MS medium by embryo after the endosperm excision outside kind benevolence with after aseptic water washing 4 times.Pollution rate is low to moderate less than 3%, and survival rate can reach 90%.
(3) Multiplying culture: the bud of aseptic seedling step (1) obtained is cut into 1.5 ~ 2.0cm length, every section of bud that at least one, band is healthy and strong, be incorporated on proliferated culture medium and carry out Multiplying culture, first full light culture 7 days under 25 DEG C of conditions after inoculation, then illumination every day is placed in 13 hours, intensity of illumination is 2000lx, and being placed in cultivation temperature is cultivate under the condition of 25 DEG C, and switching in 38 days once.Cultivate after 35 days, the average production height of bud is 4.92cm, and growth coefficient is 5.7, and plant strain growth is good.Described proliferated culture medium is: MS+6-BA0.5mg/L+NAA0.4mg/L+ sucrose 15g/L+ agar 3.5g/L, pH is 5.8.
(4) culture of rootage: the bud of step (1) or (2) aseptic seedling is cut into 1.5 ~ 2.0cm length, every section of bud that at least one, band is healthy and strong is cut to be inoculated on root media and is carried out culture of rootage, first full light culture 7 days under 25 DEG C of conditions after inoculation, then illumination every day is placed in 14 hours, intensity of illumination is 2000lx, and cultivation temperature is be cultured under the condition of 25 DEG C to take root.Cultivate after 38 days, average root is raw reaches 5.6cm, well developed root system, and rooting rate reaches more than 88%.Described root media is: MS+NAA0.3mg/L+IBA0.6mg/L+ sucrose 15g/L+ agar 3.5g/L, pH is 5.8.
(5) acclimatization and transplants: the rooting tube plantlet decap of height about 8 ~ 10cm was placed in natural lighting lower refining seedling after 5 ~ 7 days, test-tube plantlet is taken out from blake bottle, wash root medium off, to plant in the matrix be mixed into by peat soil and yellow sand mud and to be colonizated in large Tanaka, transplanting into survival rate 94%.
Claims (3)
1. a styrax tonkinensis Craib ex Hart Fast Asexual Propagation Technique, is characterized in that comprising the following steps:
(1) fruit harvesting: the fruit of gathering then of styrax tonkinensis Craib ex Hart fine individual plant of gathering, Fruit dries in the shade after regaining and is stored in 4 DEG C of refrigerators after sloughing pericarp for subsequent use;
(2) aseptic seeding: the kind benevolence of styrax tonkinensis Craib ex Hart is first used after 75% ethanol disinfection 5 ~ 30s with aseptic washing 3 ~ 5 times, again with 0.1% mercuric chloride solution sterilization 10 ~ 25min, in workbench, carry out seed germination cultivation by being inoculated on MS medium by embryo after the endosperm excision outside kind benevolence with after aseptic water washing 4 ~ 6 times;
(3) Multiplying culture: the bud of aseptic seedling step (1) obtained is cut into 1.5 ~ 2.0cm length, every section of bud that at least one, band is healthy and strong, be incorporated on proliferated culture medium and carry out Multiplying culture, first full light culture 7 ~ 14 days under 25 ~ 28 DEG C of conditions after inoculation, then illumination every day is placed in 12 ~ 16 hours, intensity of illumination is 1500 ~ 2500lx, and being placed in cultivation temperature is cultivate under the condition of 25 ~ 28 DEG C, and switching in 35 ~ 40 days once;
(4) culture of rootage: the bud of step (1) or (2) aseptic seedling is cut into 1.5 ~ 2.0cm length, every section of bud that at least one, band is healthy and strong is cut to be inoculated on root media and is carried out culture of rootage, first full light culture 7 ~ 14 days under 25 ~ 28 DEG C of conditions after inoculation, then illumination every day is placed in 14 ~ 16 hours, intensity of illumination is 2000 ~ 3000lx, and cultivation temperature is be cultured under the condition of 25 ~ 28 DEG C to take root;
(5) acclimatization and transplants: the rooting tube plantlet decap of height about 8 ~ 10cm was placed in natural lighting lower refining seedling after 5 ~ 7 days, test-tube plantlet is taken out from blake bottle, wash root medium off, to plant in the matrix be mixed into by peat soil and yellow sand mud and to be colonizated in large Tanaka.
2. a kind of styrax tonkinensis Craib ex Hart Fast Asexual Propagation Technique according to claim 1, it is characterized in that the proliferated culture medium described in step (3) is: MS+6-BA0.3 ~ 0.9mg/L+NAA0.2 ~ 0.4mg/L+ sucrose 15 ~ 30g/L+ agar 3.5 ~ 6.0g/L, pH is 5.4 ~ 5.8.
3. a kind of styrax tonkinensis Craib ex Hart Fast Asexual Propagation Technique according to claim 1, it is characterized in that the root media described in step (4) is: MS+NAA0.2 ~ 0.5mg/L+IBA0.2 ~ 0.5mg/L+ sucrose 15 ~ 30g/L+ agar 3.5 ~ 6.0g/L, pH is 5.4 ~ 5.8.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106106145A (en) * | 2016-06-12 | 2016-11-16 | 青岛农业大学 | A kind of method for in-vitro rapid propagation of snowball embryo Seedling stem section |
| CN109329047A (en) * | 2018-08-30 | 2019-02-15 | 江苏省中国科学院植物研究所 | A method of improving beautiful tinkling of pieces of jade flower tissue culture vessel seedling efficiency |
| CN113598048A (en) * | 2021-08-06 | 2021-11-05 | 苏州回文生物种业科技有限公司 | Rooting and seedling raising method for wild jasmine |
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| CN102726146A (en) * | 2012-07-18 | 2012-10-17 | 喻方圆 | Method for breaking dormancy of styrax tonkinensis seeds |
| CN102860204A (en) * | 2012-09-18 | 2013-01-09 | 王继华 | Cuttage propagation method for styrax tonkinensis |
| CN103918430A (en) * | 2014-03-25 | 2014-07-16 | 浙江省林业科学研究院 | Styrax hypoglauca cuttage propagation method |
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2015
- 2015-02-22 CN CN201510085932.5A patent/CN104686342A/en active Pending
Patent Citations (3)
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|---|---|---|---|---|
| CN102726146A (en) * | 2012-07-18 | 2012-10-17 | 喻方圆 | Method for breaking dormancy of styrax tonkinensis seeds |
| CN102860204A (en) * | 2012-09-18 | 2013-01-09 | 王继华 | Cuttage propagation method for styrax tonkinensis |
| CN103918430A (en) * | 2014-03-25 | 2014-07-16 | 浙江省林业科学研究院 | Styrax hypoglauca cuttage propagation method |
Non-Patent Citations (2)
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| HATICE DEMİRAY ET.AL.,: "Identification of benzoin obtained from calli of Styrax officinalis by HPLC", 《TURKISH JOURNAL OF BOTANY》 * |
| 张亮亮: "白花树无性快繁研究", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106106145A (en) * | 2016-06-12 | 2016-11-16 | 青岛农业大学 | A kind of method for in-vitro rapid propagation of snowball embryo Seedling stem section |
| CN109329047A (en) * | 2018-08-30 | 2019-02-15 | 江苏省中国科学院植物研究所 | A method of improving beautiful tinkling of pieces of jade flower tissue culture vessel seedling efficiency |
| CN109329047B (en) * | 2018-08-30 | 2021-06-04 | 江苏省中国科学院植物研究所 | Method for improving seedling efficiency of tissue culture container of Helianthus tuberosus |
| CN113598048A (en) * | 2021-08-06 | 2021-11-05 | 苏州回文生物种业科技有限公司 | Rooting and seedling raising method for wild jasmine |
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