CN104642141B - Gerbera adventitious bud induction and plant regeneration method by using root as explant - Google Patents
Gerbera adventitious bud induction and plant regeneration method by using root as explant Download PDFInfo
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- CN104642141B CN104642141B CN201510110573.4A CN201510110573A CN104642141B CN 104642141 B CN104642141 B CN 104642141B CN 201510110573 A CN201510110573 A CN 201510110573A CN 104642141 B CN104642141 B CN 104642141B
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Abstract
The invention relates to a plant tissue culture propagation technology, more specifically, relates to a gerbera adventitious bud induction and plant regeneration method by using a root as an explant. The method is characterized by comprising the following steps: by using degraded gerbera root as the explant, proliferating and rooting culturing to obtain gerbera rooted seedling. The gerbera rooted seedling is stable in inheritable character, simple in culture process, easy to root, short in culture and propagation period, and easy to survive after being transplanted.
Description
Technical field
The present invention relates to plant tissue culture reproduction technique, more particularly, to a kind of African Chrysanthemum with root as explant
Induce aerosor and plant regeneration method.
Background technology
African Chrysanthemum (gerbera jamesoniiBolus), another name African daisy, heronsbill, Herba Erigerontiss etc., are Compositae nail-like boil
Grass belongs to perennial evergreen perennial root herbage flower, originates in Africa, is one of big cut-flower in the world 5.African Chrysanthemum rich color, flower is large
Greatly, be artistic flower arrangement ideal material, there is important ornamental value, under suitable cultivation condition can bloom in the anniversary, can make
For city ornamental flower and potted plant decoration.But because African Chrysanthemum is cross-pollinatd plant, seedling variation is big, leads to bennet to become
Carefully, deliquescing, flower pattern diminish etc. it is impossible to keep breeding characteristic, and division propagation method breeding coefficient is low can not meet market need
Ask, therefore on International Flower market, the African Chrysanthemum industry of flowers and plants has been used up tissue culture technique asexual propagation and obtains breeding seedling.
China is in the popularization stage of African Chrysanthemum, and the research and development of the high breeding African Chrysanthemum kind of quality are external monopolization technology,
The seed of African Chrysanthemum breeding initial species of domestic flowers company and seedling are all introduced from external, and cost is very high.Meanwhile, domestic
, after the successive transfer culture proliferation of propagation in excessive generation, can quality deterioration by generation existing in the breeding seedling that flowers company high price is bought
As, typical phenomenon is exactly that the flower pattern of African Chrysanthemum diminishes, directly result in produce in enormous quantities African Chrysanthemum out not up to standard it is impossible to pin
Sell, cause flowers company must just ask an exorbitant fare for every 2 ~ 3 years and introduce new breeding seedling from external, cost is high.Understand state
After the interior main flowers company producing African Chrysanthemum and research unit, the phenomenon that after this many propagation from generation to generation, flower pattern is degenerated is very general
Time, domestic African Chrysanthemum this problem of industry urgent need to resolve.
Many researcheres are compared research to the culture effect of African Chrysanthemum Different Organs explant material, from flower
Calyx, holder, blade, stem section, petal, ovary, multiple explant such as bennet, alabastrum, stem apex obtains regeneration plant, its culture
Formula has been announced.Number of patent application is 200810025775.9, entitled " in-vitro high-frequency regeneration replication method of African chrysanthemum "
Patent of invention, discloses a kind of Induce aerosor method using African Chrysanthemum test tube seedling petiole as explant；Number of patent application is
200810014604.6th, entitled " a kind of tissue culture and rapid propagation method for Gerbera jamesonii Bolus ", discloses one kind with African Chrysanthemum holder as explant
Induce aerosor method.But in numerous isolated culture report, using root segment as the African Chrysanthemum tissue culture of explant it is still
Blank.
It is aerial partss organ in view of organs such as floral organ, stem, leaves, there is obvious maturation effect, age effect
And position effect, by these effects, it is very universal phenomenon that the plant that asexual propagation is cultivated out degenerates.And root system is not
There is phasic development, be juvenile forever, the nursery stock being obtained for parent material breeding with root, it is obtained in that such as seminal propagation
Same juvenile of nursery stock, thus plant of realizing failing return young effect of rejuvenation, be the ideal material recovering plant degenerativ character.
Content of the invention
The aerial partss organ such as floral organ, Miao Ye is mainly adopted to plant as present in explant induction for prior art
Strain degradation phenomena, the present invention proposes a kind of African Chrysanthemum Induce aerosor with root as explant and plant regeneration method.
The African Chrysanthemum Induce aerosor with root as explant of the present invention and plant regeneration method are it is characterised in that press following
Step is implemented:
(1) acquisition of explant: with the aseptic bottle seedling of rudimentary African Chrysanthemum as original material, by the nothing of high about 3 ~ 4 cm
Vaccine is cut into individual plant Seedling, is inoculated in culture medium, and culture medium prescription is 1/2ms+iba 0.2 ~ 1.0 mg/l, and ph 5.8 ~
6.2, sucrose 20 ~ 30 g/l, agar 4 ~ 7 g/l；Cultivation temperature is 21 ± 2 DEG C, with intensity of illumination for 1600 ~ 1800
Illumination 12 h/d of lux；After growth 14 ~ 20 d, when root extends to 1 ~ 2 cm, intercept the root segment conduct of 0.5 ~ 1.5cm
Explant material；
(2) Primary culture: after the root segment of intercepting is rinsed well under distilled water, be inoculated in adventitious bud induction culture base
In, the formula of culture medium is ms+6-ba 0.5 ~ 1.5 mg/l+naa 0.1 ~ 1.0 mg/l, ph 5.8 ~ 6.2, sugarcane
Sugared 20 ~ 30 g/l, agar 4 ~ 7 g/l；Cultivation temperature is 21 ± 2 DEG C, with the light for 1600 ~ 1800 lux for the intensity of illumination
According to 12 h/d, cultivate 25 ~ 30 d；
(3) enrichment culture: preferable 1 ~ 2 cm adventitious bud of growing way that root segment base portion is obtained is transferred to proliferated culture medium
On, the formula of increment culture medium is: ms+6-ba 0.5 ~ 2.0 mg/l+naa 0.05 ~ 0.2 mg/l, ph 5.8 ~
6.2, sucrose 20 ~ 30 g/l, agar 4 ~ 7 g/l；Cultivation temperature is 21 ± 2 DEG C, with intensity of illumination for 1600 ~ 1800
Illumination 12 h/d of lux, cultivates 25 ~ 30 d, forms the high Multiple Buds of 2 ~ 3cm；The Multiple Buds of formation can be cut into list
Bud, is placed on proliferated culture medium and carries out enrichment culture；
(4) root culture: when breeding obtained Multiple Buds plant and grow to 2 ~ 3cm, the bud of subculture is cut into 2 ~ 4
Piece leaf is one plant of simple bud, is inoculated on root media, and prescription of rooting medium is: 1/2ms+iba 0.2 ~ 1.0
Mg/l, adds 20 ~ 30 g/l sucrose, 4 ~ 7 g/l agar；Cultivation temperature is 21 ± 2 DEG C, with intensity of illumination for 1600 ~
Illumination 12 h/d of 1800 lux；After culture 25 ~ 30 d, a length of 1.5 ~ 2.5 cm of root, obtain African Chrysanthemum and take root Seedling.
(5) seedling exercising and transplanting: after the root of root to be generated is up to 2 cm, the indoor seedling exercising 2 ~ 4 days being 25 DEG C in room temperature, take
Birth root, cleans the culture medium attaching in root, and being transplanted to substrate composition afterwards is laterite: fertile soil: perlite=1:1:
In 1 sand table, after transplanting note control greenhouse in temperature between 25 ~ 30 DEG C, humidity between 75 ~ 80%, 10 ~
Survive after 15 days.
The present invention has the prominent advantages that: the African Chrysanthemum aseptic bottle seedling root segment not up to standard of the degeneration being diminished with flower pattern is outer
Implant material, rinses through sterile distilled water and both can inoculate, and decreases the link of explant sterilization；Root segment with aseptic bottle seedling is
Explant, has and draws materials conveniently, be not subject to seasonal restrictions, and proliferative induction coefficient is high, by inducing seedling with root segment for explant, increases
Grow coefficient and reach 3.6 ~ 5.1 times, and the tissue culture cycle be 3 months, far below other with floral organ official rank as explant material 6 ~
Tissue culture cycle of 7 months, tissue culture cost savings 1/2nd.African Chrysanthemum plant regeneration method provided by the present invention has something lost
Transmissibility shape is stable, and incubation is simple, easily takes root, and cultivates expanding propagation cycle is short, transplants the advantage easily surviving.Thus, the present invention
For the asexual propagation accelerating African Chrysanthemum breeding, there is positive role, the character simultaneously also having reached for degeneration African Chrysanthemum kind is extensive
The purpose that multiple and recycling lays the foundation.
Specific embodiment
Embodiment 1:, the Induce aerosor of African Chrysanthemum and plant regeneration are in Southwest Forestry University's forestry institute's laboratory and Green
Greenhouse is carried out, and the time of implementing is 2013.
(1) acquisition of explant: the aseptic bottle seedling (the entitled solar wind of kind) with African Chrysanthemum filigree as Explant Propagation is
Original material, the degradable material that this material diminishes for flower pattern is it is impossible to up to standard sold.On superclean bench, by high by about 3 ~
The aseptic seedling of 4 cm is cut into individual plant Seedling, is inoculated in and draws materials in order to take root in the culture medium of 1/2ms+iba 0.5 mg/l.Wherein,
Ph 5.8, sucrose 20 g/l, agar 4 g/l.Condition of culture: cultivation temperature is 21 ± 2 DEG C, with intensity of illumination for 1600 ~ 1800
Illumination 12 h/d of lux.After growing 15 d, root is elongated to certain length, and the root segment intercepting 0.8 cm length is as explant material.
(2) Primary culture: after the root segment of intercepting being rinsed well under distilled water in superclean bench, be inoculated in adventitious bud
In inducing culture.The formula of culture medium is: ms+6-ba 1.0 mg/l+naa 0.5 mg/l, ph 5.8, sucrose 20
G/l, agar 4 g/l.Condition of culture: cultivation temperature is 21 ± 2 DEG C, with the illumination 12 for 1600 ~ 1800 lux for the intensity of illumination
H/d, cultivates 25 d.
(3) enrichment culture: preferable 1 ~ 2 cm adventitious bud of growing way that root segment base portion is obtained is transferred to proliferated culture medium
On, the formula of proliferated culture medium is: ms+6-ba 1.0 mg/l+naa 0.1 mg/l, ph 5.8, sucrose 30 g/l, fine jade
Fat 4 g/l.Condition of culture: cultivation temperature is 21 ± 2 DEG C, illumination 12 h/d with intensity of illumination for 1600 ~ 1800 lux, training
Support 25 d, form the high Multiple Buds of 2 ~ 3cm.Repeatedly the Multiple Buds of formation can be cut into simple bud, be placed on proliferated culture medium
Carry out enrichment culture, African Chrysanthemum can be made to carry out the expanding propagation of rapid, high volume, its increment multiple is 5.07 times.
(4) root culture: when breeding obtained Multiple Buds plant and grow to 2 ~ 3cm, the bud of subculture is cut into 2 ~ 4
Piece leaf is one plant of simple bud, is inoculated on root media, and prescription of rooting medium is: 1/2ms+iba 0.5 mg/l,
Additional 20 g/l sucrose, 4 g/l agar.Condition of culture: cultivation temperature is 21 ± 2 DEG C, with intensity of illumination for 1600 ~ 1800
Illumination 12 h/d of lux.After cultivating 25 d, a length of 1.5 ~ 2.5 cm of root, obtain African Chrysanthemum and take root Seedling.
(5) seedling exercising and transplanting: after the root of root to be generated is up to 2 cm, bottle cap, seedling exercising 2 are opened in the interior being 25 DEG C in room temperature
After ~ 4 days, take out Seedling of taking root, clean the culture medium attaching in root, be transplanted to substrate composition afterwards and be laterite: fertile soil: be precious
In the sand table of Zhu Yan=1:1:1, note after transplanting controlling the temperature in greenhouse between 25 ~ 30 DEG C, humidity is 75 ~ 80%
Between, survive after 10 ~ 15 days, move in the nutrient bag of a diameter of 10 cm of rim of a cup, the same sand table of transplanting medium proportioning.Pass through again
Maintenance 2 months, when height of seedling reaches 10-15 cm, can sell.
Claims (1)
1. the African Chrysanthemum Induce aerosor with root as explant and plant regeneration method are it is characterised in that follow these steps to implement:
(1) acquisition of explant: with the aseptic bottle seedling of rudimentary African Chrysanthemum as original material, by the aseptic seedling of high about 3 ~ 4 cm
It is cut into individual plant Seedling, is inoculated in culture medium, culture medium prescription is ms+6-ba 1.0 mg/l+naa 0.1 mg/l, ph
5.8, sucrose 20 ~ 30 g/l, agar 4 ~ 7 g/l；Cultivation temperature is 21 ± 2 DEG C, with intensity of illumination for 1600 ~ 1800
Illumination 12 h/d of lux；After growth 14 ~ 20 d, when root extends to 1 ~ 2 cm, intercept the root segment conduct of 0.5 ~ 1.5cm
Explant material；
(2) Primary culture: after the root segment of intercepting is rinsed well under distilled water, be inoculated in adventitious bud induction culture base, training
The formula of foster base is ms+6-ba 0.5 ~ 1.5 mg/l+naa 0.1 ~ 1.0 mg/l, ph 5.8 ~ 6.2, sucrose 20 ~
30 g/l, agar 4 ~ 7 g/l；Cultivation temperature is 21 ± 2 DEG C, illumination 12 h/ with intensity of illumination for 1600 ~ 1800 lux
D, cultivates 25 ~ 30 d；
(3) enrichment culture: preferable 1 ~ 2 cm adventitious bud of growing way that root segment base portion is obtained is transferred on proliferated culture medium, increases
The formula growing culture medium is: ms+6-ba 0.5 ~ 2.0 mg/l+naa 0.05 ~ 0.2 mg/l, ph 5.8 ~ 6.2, sugarcane
Sugared 20 ~ 30 g/l, agar 4 ~ 7 g/l；Cultivation temperature is 21 ± 2 DEG C, with the light for 1600 ~ 1800 lux for the intensity of illumination
According to 12 h/d, cultivate 25 ~ 30 d, form the high Multiple Buds of 2 ~ 3cm；The Multiple Buds of formation are cut into simple bud, are placed on propagation
Enrichment culture is carried out on culture medium；
(4) root culture: when breeding obtained Multiple Buds plant and grow to 2 ~ 3cm, the bud of subculture is cut into 2 ~ 4 leaves
The simple bud for one plant for the son, is inoculated on root media, prescription of rooting medium is: 1/2ms+iba 0.2 ~ 1.0 mg/
L, adds 20 ~ 30 g/l sucrose, 4 ~ 7 g/l agar；Cultivation temperature is 21 ± 2 DEG C, with intensity of illumination for 1600 ~ 1800
Illumination 12 h/d of lux；After culture 25 ~ 30 d, a length of 1.5 ~ 2.5 cm of root, obtain African Chrysanthemum and take root Seedling；
(5) seedling exercising and transplanting: after the root of root to be generated is up to 2 cm, the indoor seedling exercising 2 ~ 4 days being 25 DEG C in room temperature, takes out and gives birth to
Root, cleans the culture medium attaching in root, is transplanted to matrix volume afterwards than for laterite: fertile soil: perlite=1:1:1
Sand table in, after transplanting note control greenhouse in temperature between 25 ~ 30 DEG C, humidity between 75 ~ 80%, 10 ~ 15
Survive after it.
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| CN104920048B (en) * | 2015-06-30 | 2017-06-16 | 云南省农业科学院花卉研究所 | A kind of hardening transition method for improving tobacco K326 tissue-cultured seedling transplanting survival rates |
| CN105104203B (en) * | 2015-09-10 | 2017-06-23 | 郝毅 | A kind of efficient expanding propagation method of African Chrysanthemum virus-elimination seedlingses |
| CN106332784A (en) * | 2016-11-10 | 2017-01-18 | 福建农林大学 | Open gerbera jamesonii tissue culture method |
| CN109362524B (en) * | 2018-11-07 | 2020-11-17 | 中国科学院昆明植物研究所海盐工程技术中心 | Cultivation method of new gerbera jamesonii variety |
| CN112753393B (en) * | 2020-12-31 | 2022-04-22 | 华南农业大学 | Broussonetia papyrifera root propagation method |
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Inventor after: He Chengzhong Inventor after: Li Dan Inventor after: Zhao Yanming Inventor after: Xu Changhui Inventor after: Wu Yingying Inventor after: Xin Peiyao Inventor after: Zhang Bin Inventor before: He Chengzhong Inventor before: Zhao Yanming Inventor before: Li Dan Inventor before: Xu Changhui Inventor before: Wu Yingying Inventor before: Xin Peiyao Inventor before: Zhang Bin |
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