CN106069753A - A kind of Bougainvillea spectabilis aseptic seeding and method for quickly breeding - Google Patents
A kind of Bougainvillea spectabilis aseptic seeding and method for quickly breeding Download PDFInfo
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- CN106069753A CN106069753A CN201610429327.XA CN201610429327A CN106069753A CN 106069753 A CN106069753 A CN 106069753A CN 201610429327 A CN201610429327 A CN 201610429327A CN 106069753 A CN106069753 A CN 106069753A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
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Abstract
The invention discloses a kind of Bougainvillea spectabilis aseptic seeding and method for quickly breeding, the method includes: carry out aseptic seeding sprout-induction with seed: to accessing bud inducement culture medium after seed disinfection sterilizing, carry out bud inducement cultivation under suitable temperature, light application time, intensity of illumination；Then the root induction again of Shoot propagation expanding propagation is utilized to breed seedling: to sprout the sprouting and access in Shoot propagation culture medium, carry out Shoot propagation successive transfer culture under the conditions of suitable temperature, light application time, intensity of illumination；Sprout after Shoot propagation successive transfer culture accesses root media, carries out rooting seedling under the conditions of suitable temperature, light application time, intensity of illumination.Present invention seed is as breeding outer implant, and simply, pollution rate is low, and the germination percentage of seed reaches 100, and within 30 days, Shoot propagation multiple reaches 5.22 times, rooting rate 95.6 for seed treatment, and the breeding not by limited season time can whole year production of seedling.
Description
Technical field
The invention belongs to field of plant tissue culture technique, particularly relate to a kind of Bougainvillea spectabilis aseptic seeding and Fast-propagation side
Method.
Background technology
Bougainvillea spectabilis (Bougainvillea glabra) is Nyctaginaceae bougainvillea shrub, and its pattern is many, florescence length, flower
Sense prosperity, and barren-resistant, drought-resistant, strong adaptability, the most almost there is introducing and planting in the whole world, is widely used in city trees and shrubs
Greening view and admire with potted plant, be Republic of Zambia's national flower and province of Hainan Province of China spends, Zhuhai City, Shenzhen, Xiamen City, Liuzhou
The city flower in the cities such as city, Sanya, Haikou City.Bougainvillea spectabilis or a kind of multiduty resource plant, contained beet pigments is used
In exploitation natural food colour；The alternative insulin for treating diabetes of the pinitol of isolated and without insulin in its blade and root
The side effect that therapy is brought；Blade and root at least contain 2 kinds of antiviral proteins, various plants virus is had in various degree
Inhibitory action, can be used for develop the efficient biological pesticide of low toxicity, its industrial prospect is boundless.
Bougainvillea spectabilis is usually with being propagated from cuttings, it is difficult to by the kind of cottage propagation, can be come by the planting of layer or grafting
Breeding.Therefore, propagating materials requirement is big, and the repoductive time of some kinds is longer, and appreciable variety is then difficult to Fast-propagation to be pushed away
Wide application.The propagation method of this quasi-tradition may also be subjected to boisterous impact simultaneously.
The collection of current seed and process；The method for disinfection and sterilization of seed；Seed germination and lure grinding of bud culture medium prescription
System；The development of Shoot propagation culture medium prescription；The development of induced bud prescription of rooting medium is the most perfect.
Summary of the invention
It is an object of the invention to provide a kind of Bougainvillea spectabilis aseptic seeding and method for quickly breeding, it is intended to solve current seed
Collection and process；The method for disinfection and sterilization of seed；Seed germination and the development luring bud culture medium prescription；Shoot propagation cultivates basigamy
The development of side；The problem that the development of induced bud prescription of rooting medium is the most perfect.
The present invention is achieved in that a kind of Bougainvillea spectabilis aseptic seeding and method for quickly breeding, this Bougainvillea spectabilis aseptic seeding
Comprise the following steps with method for quickly breeding:
Aseptic seeding sprout-induction is carried out: to accessing in bud inducement culture medium MS+ sucrose after seed disinfection sterilizing with seed,
Bud inducement cultivation is carried out under suitable cultivation temperature, light application time, intensity of illumination；
Then the root induction again of Shoot propagation expanding propagation is utilized to breed seedling: to sprout the sprouting and access Shoot propagation culture medium MS+
In 6-BA+NAA+ sucrose, in suitable cultivation temperature, light application time, under the conditions of intensity of illumination, carry out Shoot propagation successive transfer culture；Bud
Sprout after proliferation and subculture is cultivated accesses in root media MS+NAA+ sucrose, at suitable cultivation temperature, light application time, light
According to carrying out rooting seedling under strength condition.
Further, Bougainvillea spectabilis aseptic seeding specifically includes with method for quickly breeding:
Seed disinfection: choose full Bougainvillea spectabilis seed, is positioned in the vial sterilized on superclean bench, first
With 75% alcohol-pickled 20s, remove ethanol, rinse 3 times with sterilized water, rinse time 1min, the most again with 0.1% every time
HgCl2Sterilization 8min, disinfectant solution should exceed seed about 1, be kept stirred vial with hands, after removing disinfectant solution in disinfecting process
With aseptic water washing 5 times, rinse time 1min every time；
Seed germination and bud inducement are cultivated:
Seed is inoculated in bud inducement culture medium MS+ sucrose, and every bag is accessed 1 seed, in temperature 24-26 DEG C, during illumination
Between 9h/d, under the condition of culture of intensity of illumination 1500lx, carry out seed germination and bud inducement and cultivate, first send out roots after 5d, after 15d
Height of seedling 2～3, grows 1-3 stipes, inductivity 100%；
Shoot propagation successive transfer culture: sprout the segment of the sprouting dissecting knife cutting growth 1.5cm, containing the stem of
Joint, removes blade, is inserted lower end in Shoot propagation culture medium, in temperature 26-28 DEG C, light application time 9h/d, intensity of illumination 1
Cultivate under the condition of culture of 500lx.Cultivating 30d, the growth potential of bud is good, and stem is sturdy, and the height of average bud is by 1.5 increasings when inoculating
Being added to 4.01, >=3 bud quantity that can be used for taking root are many, account for 85.7, proceed to identical cultivation after 25-30d cutting
Subculture multiplication in base；
Root culture: the sprout of high about 3cm is cut insertion root media from base portion, in temperature 26-28 DEG C, illumination
Time 9h/d, root induction under the condition of culture of intensity of illumination 1 500lx, cultivation cycle is 30 days, and rooting rate reaches 95.6%,
Mean elements reaches 6.39, robust plant, dark green leaf color.
Further, need the collection carrying out seed with processing method to be before seed disinfection: in 2 months summers, the last ten-days period gather three
Angle plum seed, be placed in refrigerator preserve standby, reserve temperature 4 DEG C.
Further, bud inducement culture medium is MS+ sucrose 30g/L, pH value 6.0.
Further, Shoot propagation subculture medium is: MS+6-BA1.0mg/L+NAA0.2mg/L+ sucrose 30g/L, pH value
6.0。
Further, root media is: MS+NAA 1.5mg/L+ sucrose 30g/L, pH value 6.0.
Bougainvillea spectabilis prior art mainly uses its branch cutting to breed, the general branch selecting 1-2 raw, is cut into length
The cutting of 12-15cm, cutting and cutting medium are both needed to disinfect, and 20-30 days long new, sprouts after 40 days, and cottage propagation is easy
By season, climatic effect, survival rate 70-95.Although branch cutting molding is very fast, but a cutting breeds a strain only, because of
This need to the amount of kind very big.The Bougainvillea spectabilis tissue culture propagation of prior art mainly with stem section, blade, petiole, bract etc. as outer implant, this
The outer implant of class is all difficult to sterilization and survival rate is low, also needs through callus induction using blade, petiole and bract as outer implant
In the stage, incubation time is longer.The present invention is that seed treatment is simple, and success rate is high, uncontaminated kind as breeding outer implant with seed
Sub-germination percentage reaches 100, and without callus induction, within 30 days, Shoot propagation multiple reaches 5.22 times, rooting rate 95.6, and
Breeding not by limited season time can whole year production of seedling.
The present invention is the tissue culture propagating kind seedling-growing method using seed as outer implant, and its seed treatment is simple, germination percentage
High；Propagating materials is few, and reproduction speed is fast；Condition of culture is controlled, is not subject to seasonal restrictions, can whole year production；Preserve low cost, it is simple to
Plant mass transter；Utilize embryo culture technique to overcome incompatibility and the early stage abortion of hybridization embryo of hybridization, increase new varieties and occur
Probability and speed；
With the method for tissue culture that seed is outer implant, sterilized after non-staining percentage of seedgermination reach 100, pending
After bud, in units of 1 stipes about 1.5 length breedings i.e. can get 5.22 sproutings for 30 days, and cottage propagation branch length is
12-15cm, and a cutting breeds 1 strain, the propagating materials therefore, in terms of the length of propagating materials, needed for tissue culture only
Being about the 1/10 of cottage propagation material, proliferation times is but more than 5 times of cottage propagation, and without being again with new material,
The most monthly can increase with geometry multiple.Cultivating because of whole process and all carry out in culturing room, the condition of culture such as temperature and illumination all can root
Regulate according to needs.Bougainvillea spectabilis mainly cultivates new kind by cross-breeding and two kinds of methods of Bud mutation at present；Great majority three
Angle prunus mume (sieb.) sieb.et zucc. kind ends after spending i.e. nature fallen flowers, is difficult to bear seeds, and seed soil sowing germination percentage is low, and (we natural seeding germination percentage are
0), therefore evolved new breeds certain difficulty by hybridization；Bud mutation is to choose variation part from the plant of excellent bud mutation
Bud or branch, make variation character be continued and fix by vegetative method, through programs such as primary election, final election, final elections
Selection, observe and identify, finally select major clique be bred as new varieties；
The method for tissue culture utilizing aseptically sowing seeds obtains the approach of new varieties appearance to be had: 1, seed sexual propagation
Shape separates the propagating materials that can obtain multiple phenotype；2, the bud mutation material occurred in incubation.More than 10 studied at us
The aseptic seeding propagating materials of individual kind, all there is 2-5 different phenotype in each kind, provides for cultivating new kind
Germplasm materials.
Accompanying drawing explanation
Fig. 1 is Bougainvillea spectabilis aseptic seeding and the method for quickly breeding flow chart of embodiment of the present invention offer.
Detailed description of the invention
In order to make the purpose of the present invention, technical scheme and advantage clearer, below in conjunction with embodiment, to the present invention
It is further elaborated.Should be appreciated that specific embodiment described herein, only in order to explain the present invention, is not used to
Limit the present invention.
Below in conjunction with the accompanying drawings the application principle of the present invention is further described.
As shown in Figure 1: the Bougainvillea spectabilis aseptic seeding that the present invention provides comprises the following steps with method for quickly breeding:
S101: carry out aseptic seeding sprout-induction with seed: to accessing in culture medium MS+ sucrose after seed disinfection sterilizing,
Bud inducement cultivation is carried out under suitable cultivation temperature, light application time, intensity of illumination；
S102: then utilize the root induction again of Shoot propagation expanding propagation to breed seedling: to sprout the sprouting and access culture medium MS+6-
In BA+NAA+ sucrose, in suitable temperature, light application time, under the conditions of intensity of illumination, carry out Shoot propagation successive transfer culture；Shoot propagation continues
Sprout after culture accesses in root media MS+NAA+ sucrose, in suitable temperature, light application time, intensity of illumination condition
Under carry out rooting seedling.
Bougainvillea spectabilis aseptic seeding of the present invention includes with method for quickly breeding concrete steps:
Seed disinfection: choose full Bougainvillea spectabilis seed, is positioned in the vial sterilized on superclean bench, first
With 75% alcohol-pickled 20s, remove ethanol, rinse 3 times with sterilized water, rinse time 1min, the most again with 0.1% every time
HgCl2Sterilization 8min, disinfectant solution should exceed seed about 1, be kept stirred vial with hands therebetween, goes after disinfectant solution with aseptic
Water rinses 5 times, rinses time 1min every time.
Seed germination and bud inducement are cultivated:
Culture medium prescription
MS+ sucrose 30g/L, pH value 6.0.
Condition of culture
Cultivation temperature 24-26 DEG C, light application time 9h/d, intensity of illumination 1500lx.
Inoculation
Seed accesses in culture medium sprouts after 5d, and root first grows, height of seedling 2-3 after 15d, grows 1-3 stipes, inductivity
100%, 30d pollution rate 30%.
Shoot propagation successive transfer culture
Culture medium prescription
MS+6-BA1.0mg/L+NAA0.2mg/L+ sucrose 30g/L, pH value 6.0.
Condition of culture
Cultivation temperature 26-28 DEG C, light application time 9h/d, intensity of illumination 1 500lx.
Inoculation
Sprouting the sprouting dissecting knife to be cut into and be about the single stipes of 1.5cm and access in culture medium and cultivate, 30d breeds
Coefficient reaches 5.22.After 25-30d cutting, proceed to subculture multiplication in identical culture medium later.
Root culture
Culture medium prescription
MS+NAA 1.5mg/L+ sucrose 30g/L, pH value 6.0.
Condition of culture
Cultivation temperature 26-28 DEG C, light application time 9h/d, intensity of illumination 1 500lx.
Inoculation
The sprout cutting high about 3cm accesses in root media, and 14d starts to take root, 30d rooting rate 95.6%, the most often
Strain takes root 6.39, robust plant, and leaf color is dark green.
Further, need to carry out collection and the process of seed before seed disinfection:
Seed collection: in 2 months summers, the last ten-days period gather Bougainvillea spectabilis seed, be placed in refrigerator preserve standby, reserve temperature 4
℃。
Bougainvillea spectabilis is propagated from cuttings easily by season, climatic effect, breeding coefficient low and need to the amount of kind big, the training of existing Bougainvillea spectabilis group is numerous
Growing and mainly use stem section, blade, petiole and bract as outer implant, it is difficult to sterilization and survival rate is low, with blade, petiole and bract
Also needing through the callus induction stage as outer implant, incubation time is longer.The present invention is as breeding outer implant with seed,
Seed treatment is simple, and pollution rate is low, and success rate is high, and uncontaminated percentage of seedgermination reaches 100, it is not necessary to callus induction, and 30
It Shoot propagation multiple reaches 5.22 times, rooting rate 95.6, and the breeding not by limited season time can whole year production of seedling.
The present invention is the tissue culture propagating kind seedling-growing method using seed as outer implant, and its seed treatment is simple, germination percentage
High；Propagating materials is few, and reproduction speed is fast；Condition of culture is controlled, is not subject to seasonal restrictions, can whole year production；Preserve low cost, it is simple to
Plant mass transter；Utilize embryo culture technique to overcome incompatibility and the early stage abortion of hybridization embryo of hybridization, increase new varieties and occur
Probability and speed.
The foregoing is only presently preferred embodiments of the present invention, not in order to limit the present invention, all essences in the present invention
Any amendment, equivalent and the improvement etc. made within god and principle, should be included within the scope of the present invention.
Claims (6)
1. a Bougainvillea spectabilis aseptic seeding and method for quickly breeding, it is characterised in that this Bougainvillea spectabilis aseptic seeding and Fast-propagation
Method comprises the following steps:
Aseptic seeding sprout-induction is carried out: to accessing in bud inducement culture medium MS+ sucrose after seed disinfection sterilizing, suitable with seed
Suitable cultivation temperature, light application time, carry out bud inducement cultivation under intensity of illumination；
Then the root induction again of Shoot propagation expanding propagation is utilized to breed seedling: to sprout the sprouting and access Shoot propagation culture medium MS+6-BA+
In NAA+ sucrose, in suitable cultivation temperature, intensity of illumination, under the conditions of light application time, carry out Shoot propagation successive transfer culture；Shoot propagation
Sprout after successive transfer culture accesses in root media MS+NAA+ sucrose, and in suitable cultivation temperature, intensity of illumination, during illumination
Rooting seedling is carried out under the conditions of between.
2. Bougainvillea spectabilis aseptic seeding as claimed in claim 1 and method for quickly breeding, it is characterised in that Bougainvillea spectabilis aseptic seeding
Specifically include with method for quickly breeding:
Seed disinfection: choose full Bougainvillea spectabilis seed, be positioned on superclean bench in the vial sterilized, first use
75% alcohol-pickled 20s, removes ethanol, rinses 3 times with sterilized water, rinses time 1min every time, uses 0.1%HgCl the most again2
Sterilization 8min, disinfectant solution exceeds seed 1, is kept stirred vial with hands in disinfecting process, uses sterilized water after removing disinfectant solution
Rinse 5 times, rinse time 1min every time；
Seed germination and bud inducement are cultivated:
Seed is inoculated in bud inducement culture medium MS+ sucrose, and every bag is accessed 1 seed, in cultivation temperature 24-26 DEG C, during illumination
Between 9h/d, under conditions of intensity of illumination 1500lx, sprout after 5d, height of seedling 2-3 after 15d, have 1-3 stipes；
Shoot propagation successive transfer culture: sprout the segment of the sprouting dissecting knife cutting growth 1.5cm, containing the stipes of, remove
Remove blade, lower end is inserted in Shoot propagation culture medium, in temperature 26-28 DEG C, light application time 9h/d, the training of intensity of illumination 1500lx
Cultivate under the conditions of Yanging, after 25-30d cutting, proceed to subculture multiplication in identical culture medium；
Root culture: the sprout of high about 3cm is cut insertion root media from base portion, in temperature 26-28 DEG C, light application time
9h/d, root induction under the condition of culture of intensity of illumination 1 500lx, cultivation cycle is 30d.
3. Bougainvillea spectabilis aseptic seeding as claimed in claim 1 and method for quickly breeding, it is characterised in that need before seed disinfection into
The collection of row seed with processing method is: in 2 months summers, the last ten-days period gather Bougainvillea spectabilis seed, be placed in refrigerator preserve standby, storage
Hide temperature 4 DEG C.
4. Bougainvillea spectabilis aseptic seeding as claimed in claim 2 and method for quickly breeding, it is characterised in that bud inducement culture medium is
MS+ sucrose 30g/L, pH value 6.0.
5. Bougainvillea spectabilis aseptic seeding as claimed in claim 2 and method for quickly breeding, it is characterised in that Shoot propagation successive transfer culture
Base is: MS+6-BA1.0mg/L+NAA0.2mg/L+ sucrose 30g/L, pH value 6.0.
6. Bougainvillea spectabilis aseptic seeding as claimed in claim 2 and method for quickly breeding, it is characterised in that root media is:
MS+NAA 1.5mg/L+ sucrose 30g/L, pH value 6.0.
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107896995A (en) * | 2017-12-27 | 2018-04-13 | 西藏自治区农牧科学院蔬菜研究所 | A kind of method for tissue culture of Tibetan medicine material himalayan four-o'clock root in imminent danger |
| CN108207466A (en) * | 2018-01-08 | 2018-06-29 | 陕西省西安植物园 | A kind of method for promoting Bougainvillea spectabilis spray Rapid Rooting |
| CN108522289A (en) * | 2018-06-25 | 2018-09-14 | 济南亚龙生物科技有限公司 | A kind of Bougainvillea spectabilis detoxification method for tissue culture |
| CN108901845A (en) * | 2018-07-12 | 2018-11-30 | 海南省农业科学院热带园艺研究所 | A kind of in-vitro propagation method at vegetation water angle |
| CN109156350A (en) * | 2018-09-27 | 2019-01-08 | 中国科学院华南植物园 | A kind of method of the numerous bud of wind resistance paulownia and root media and promotion wind resistance paulownia Vitro Quick Reproduction |
| CN109258467A (en) * | 2018-10-16 | 2019-01-25 | 广西壮族自治区林业科学研究院 | A method of preventing bougainvillea tissue-cultured seedling yellow |
| CN112673957A (en) * | 2020-12-22 | 2021-04-20 | 中国热带农业科学院热带作物品种资源研究所 | Rapid breeding method of bougainvillea spectabilis tissue culture plants |
| CN114667927A (en) * | 2022-04-24 | 2022-06-28 | 中国热带农业科学院热带作物品种资源研究所 | Method for in vitro induction of bougainvillea spectabilis polyploidy |
| CN115428731A (en) * | 2022-08-11 | 2022-12-06 | 厦门路桥百城建设投资有限公司 | Method for rapidly inducing roots through tide-type in-vitro cutting of bougainvillea spectabilis and application of method |
| CN117561980A (en) * | 2024-01-15 | 2024-02-20 | 海南大学三亚南繁研究院 | Tissue culture medium and culture method for bougainvillea spectabilis test tube flowering |
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- 2016-06-16 CN CN201610429327.XA patent/CN106069753A/en active Pending
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Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107896995A (en) * | 2017-12-27 | 2018-04-13 | 西藏自治区农牧科学院蔬菜研究所 | A kind of method for tissue culture of Tibetan medicine material himalayan four-o'clock root in imminent danger |
| CN108207466A (en) * | 2018-01-08 | 2018-06-29 | 陕西省西安植物园 | A kind of method for promoting Bougainvillea spectabilis spray Rapid Rooting |
| CN108522289A (en) * | 2018-06-25 | 2018-09-14 | 济南亚龙生物科技有限公司 | A kind of Bougainvillea spectabilis detoxification method for tissue culture |
| CN108901845B (en) * | 2018-07-12 | 2021-04-30 | 海南省农业科学院热带园艺研究所 | In-vitro propagation method of plant water horns |
| CN108901845A (en) * | 2018-07-12 | 2018-11-30 | 海南省农业科学院热带园艺研究所 | A kind of in-vitro propagation method at vegetation water angle |
| CN109156350A (en) * | 2018-09-27 | 2019-01-08 | 中国科学院华南植物园 | A kind of method of the numerous bud of wind resistance paulownia and root media and promotion wind resistance paulownia Vitro Quick Reproduction |
| CN109258467A (en) * | 2018-10-16 | 2019-01-25 | 广西壮族自治区林业科学研究院 | A method of preventing bougainvillea tissue-cultured seedling yellow |
| CN112673957A (en) * | 2020-12-22 | 2021-04-20 | 中国热带农业科学院热带作物品种资源研究所 | Rapid breeding method of bougainvillea spectabilis tissue culture plants |
| CN114667927A (en) * | 2022-04-24 | 2022-06-28 | 中国热带农业科学院热带作物品种资源研究所 | Method for in vitro induction of bougainvillea spectabilis polyploidy |
| CN115428731A (en) * | 2022-08-11 | 2022-12-06 | 厦门路桥百城建设投资有限公司 | Method for rapidly inducing roots through tide-type in-vitro cutting of bougainvillea spectabilis and application of method |
| CN115428731B (en) * | 2022-08-11 | 2024-04-19 | 厦门路桥百城建设投资有限公司 | Triangle Mei Chaoxi type in-vitro cutting rapid root induction method and application thereof |
| CN117561980A (en) * | 2024-01-15 | 2024-02-20 | 海南大学三亚南繁研究院 | Tissue culture medium and culture method for bougainvillea spectabilis test tube flowering |
| CN117561980B (en) * | 2024-01-15 | 2024-04-05 | 海南大学三亚南繁研究院 | Tissue culture medium and culture method for bougainvillea spectabilis test tube flowering |
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Application publication date: 20161109 |