CN103270949B - Novel peony tissue culture rooting method - Google Patents
Novel peony tissue culture rooting method Download PDFInfo
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Abstract
The invention discloses a novel peony tissue culture rooting method. Buds of pea green, island kam and yellow peonies are used as explants; under a germfree condition, the buds are disinfected by a three-step disinfection method; tissue culture rooting seedlings are obtained by three steps of constructing a germfree strain, performing subculture multiplication culture and performing root induction culture; and the rooting rates are respectively that the pea green peony is 50.0-66.7 percent, the island kam peony is 33.3-86.7 percent and the yellow peony is 31.4-73.5 percent.
Description
Technical field
The invention belongs to field of plant tissue culture technique, particularly a kind of peony tissue cultivates new method of taking root.
Background technology
Tree peony (
paeonia suffruticosa Andr) for Paeoniaceae (
paeoniaceae) Paeonia (
paeonia) perennial deep-rootedness machaka, leaf alternate, tool long handle.Flower unisexuality or both sexes, top is raw, 4 ~ May of florescence.Follicle radish fruit, cracking time ripe, built-in 5 ~ 15 pieces of seeds, in irregular cycle, brown or black.Tree peony originates in the western Qinling Mountains of China and Daba Mountain one band, existing more than 1600 year of cultivation history, and the tree peony of countries in the world plantation is all introduced a fine variety in China.Tree peony property happiness sunlight, cold-resistant, like cool environment, non-refractory, require high to soil condition, be suitable for growing in loose, fertile, the well-drained sandy soil of semi-wet, tree peony root must be longer, and plant is comparatively large, plants with being suitable for.Peony flowers and is born in spray top, many Dan Sheng, diameter can reach about 20cm, flower pattern has the kind more than ten such as single-lobe type, lotus type, chrysanthemum type, rose type, holder osmanthus type, silk ball-shaped, crown-shaped, gold is ring-like, thousand floor platform pavilion types, Zi Tai pavilion, building type, pattern has the 9 large colour systems such as red, white, powder, Huang, purple, indigo plant, green, black and secondary color, reaches hundreds of varieties.Tree peony belongs to precious flowers, is traditional famous flower of China's special product, and purposes is very extensive, can be used for annual flower, fresh cut-flowers, gardens, courtyard, flower bed, lace and potted plant growth etc., have sight as ornamental flower.The petal edible of tree peony and wine brewing are very distinctive delicious foods.Pharmaceutically, the root skin of tree peony is used as medicine and is called the root bark of tree peony, as emmenagogue and strong dose, has antispastic, eases pain, invigorates blood circulation, the effect such as the loose stasis of blood.Its ornamental value and economic worth considerable.The traditional modes of reproduction of tree peony has seeding method, offshoot and grafting etc., and the reproduction coefficient of seeding method is large, but required time is long, the 3-4 year is generally needed from being seeded into bloom, in addition, Paeonia suffruticosa seed has hibernation feature, and simple flower kind is solid many, half double variety takes second place, double variety gynoecium or the easy lobeization of stamen shaky, seedling variation is large, not easily keeps maternal plant merit, therefore, seeding method is not suitable for the requirement of the double variety Fast-propagation with high ornamental value; The seedling of offshoot or grafting breeding can be bloomed then, but reproduction coefficient is low, and production cost is high, and unit are investment is large, and nursery stock former strain damage by disease and insect infectivity gives sub-strain, and limits by time and season.These problems constrain the breeding of the famous and precious kind of tree peony, promote and the large-scale production of nursery stock, cannot meet the market demand of tree peony commercialization growing in recent years, industrialization and large-scale production.
Plant tissue culture fast breeding technique is major technique and the effective means of accelerating and promote tree peony breeding and breeding, since the research of people's reported first tree peony test-tube plantlet propagation techniques such as Li Yulong in 1984, correlative study is increasing, in existing report, domestic and international researcher mainly concentrates on selection and the process of explant for the research of Section Moutan culturation rapid propagating technology, brownization and vitrified control, the aspects such as medium constituent and environmental factor regulation and control, result of study in the past shows, significant difference between the different cultivars of tree peony, the kind that proliferation times is low under field conditions (factors), under conditions of tissue culture, reproduction coefficient is also very little, breeding complexity under conditions of tissue culture is consistent with the complexity of breeding under normal condition.The key obtaining tree peony tissue culture plant is taken root and acclimatization and transplants, especially rooting technique, to show rooting rate low for the group training rooting technique of current tree peony, quality of rooting is poor, be difficult to obtain the problems such as regeneration plant, become the maximum bottleneck of restriction Section Moutan training technical progress, the key of culture of rootage is medium, conventional minimal medium has MS, LP, improvement MS, 1/2MS and improvement WPM etc., IBA is taken root effectively to some Varieties of Peony, also the report using NAA or IAA root induction is had, rooting efficiency neither be very desirable, in the document retrieved, the rooting rate of " Wei is purple " is about 20%, " Luoyang is red " is 38.6%, " WULONGPENGSHANG " is 33.3%, " water chestnut Zhan dew " is 47.9%, " pea green " does not obtain seedling of taking root, the famous and precious kinds such as Hai Huanghe island brocade train the report of seedling research of taking root there are no group.Existing research all shows, same cultural method and medium be used for different tree peony strain or its rooting efficiency difference of kind quite large, it is impossible for thinking to obtain thus multiple tree peony strain or the desirable group training of kind seedling of taking root simultaneously substantially, therefore, different tree peony strain or kind will obtain group training seedling of taking root and can only select diverse ways and medium respectively.
Summary of the invention
A kind of peony tissue utilizing tissue culture technique to carry out is the object of the present invention is to provide to cultivate new method of taking root.
A kind of peony tissue of the present invention is cultivated new method of taking root and is mainly comprised " foundation of aseptic strain ", " shoot proliferation cultivation " and " root induction cultivation " three links, triacontanol (TA) is added in root induction medium, be equipped with growth hormone hestening rooting, after seedling of taking root is formed, carry out acclimatization and transplants.The foundation of aseptic strain: using tree peony bulbil as explant, aseptically adopt three step sterilizations to bulbil disinfection, then residue scale is peeled off, bulbil is inoculated in inducing culture induction bulbil to sprout, inducing culture for minimal medium, adds 6-BA0.1 ~ 1.0mg/L, NAA0.01 ~ 0.1mg/L, sucrose 30g/L and agar 6g/L with WPM medium, sprouts through 12 ~ 15d bulbil, about 30d adventitious bud formation, aseptic strain builds up; Shoot proliferation is cultivated: indefinite bud is cut, be inoculated in subculture multiplication medium and carry out shoot proliferation cultivation induced synthesis Multiple Buds, again Multiple Buds is divided into the budlet clump of band 1-2 bud, be inoculated on identical medium and proceed shoot proliferation cultivation, subculture multiplication medium take WPM as minimal medium, add 6-BA0.5 ~ 3.0mg/L, NAA0.01 ~ 0.1mg/L, AC0 ~ 2g/L, sucrose 30g/L, agar 6g/L, subculture cycle 30 ~ 45d; Root induction is cultivated: the healthy and strong Multiple Buds chosen in Multiplying culture is inoculated into root media and carries out root induction cultivation, root media take MS as minimal medium, macroelement reduces by half, add IBA1.0 ~ 5.0mg/L, NAA0.1mg/L, TA0.1 ~ 2g/L, sucrose 30 ~ 50g/L, agar 6g/L again, the visible tip of a root of inoculation 10 ~ 18d, continues to cultivate 20d and forms seedling of taking root; Seedling acclimatization and transplants of taking root cultivation matrix used selects perlite and peat composed of rotten mosses mixed-matrix, and transplant initial stage button Small plastic shed moisturizing, raise canopy film after one week gradually and ventilate, after planting, two weeks plantlet in vitro blade obviously grows.
Bulbil disinfects three adopted step sterilizations: the first step with 75% ethanol disinfection 30 ~ 60s, second step with 1% hypochlorite disinfectant 8 ~ 15 min, taking-up aseptic water washing, 3rd step with 0.5% hypochlorite disinfectant 10 ~ 20min, take out clean with aseptic water washing again, constantly shake triangular flask in whole disinfecting process, disinfectant is fully contacted with bulbil.Foundation, the shoot proliferation cultivation of aseptic strain are identical with the condition that three links are cultivated in root induction, are temperature 25 ± 2 DEG C, intensity of illumination 2000Lx, light application time 14h/d.The Varieties of Peony that the present invention selects is for sea is yellow, island is bright and beautiful and pea green.
The inventive method is realized by following steps:
1. the foundation of aseptic strain
Get the pea green of robust growth respectively, island brocade, full bulbil on the paeonia lutea maternal plant of sea, remove the 1-2 layer scale of ragged edge, scrub with suds, then tap water is clean, aseptically bulbil is put into aseptic triangular flask, adopt three step sterilizations to bulbil disinfection: the first step with 75% ethanol disinfection 30 ~ 60s, second step with 1% hypochlorite disinfectant 8 ~ 15 min, taking-up aseptic water washing, 3rd step with 0.5% hypochlorite disinfectant 10 ~ 20min, take out clean with aseptic water washing again, constantly triangular flask is shaken in whole disinfecting process, disinfectant is fully contacted with bulbil, taking out after sterilization is placed on aseptic filter paper, residue scale is divested with aseptic nipper, being inoculated in respectively by the bulbil of 3 kinds on inducing culture induces bulbil to sprout, inducing culture used with WPM medium for minimal medium, add 6-BA0.1 ~ 1.0mg/L, NAA0.01 ~ 0.1mg/L, sucrose 30g/L and agar 6g/L, sprout through 12 ~ 15d bulbil, about 30d forms indefinite bud (base portion has a small amount of callus), aseptic strain is set up, condition of culture is temperature 25 ± 2 DEG C, intensity of illumination 2000Lx, light application time 14h/d.
2. shoot proliferation is cultivated
Take out pea green respectively, island brocade, the yellow indefinite bud formed in sea, cut base portion callus, then access respectively in subculture multiplication medium and cultivate 30 ~ 45d, induced synthesis Multiple Buds, Multiple Buds is divided into again the budlet clump of band 1-2 bud, be inoculated in respectively in identical subculture multiplication medium and carry out shoot proliferation cultivation, time 30 ~ 45d, budlet grows up, base portion has sprouting to sprout the new Multiple Buds of formation, subculture multiplication medium take WPM as minimal medium, add 6-BA0.5 ~ 3.0mg/L, NAA0.01 ~ 0.1mg/L, AC0 ~ 2g/L, sucrose 30g/L, agar 6g/L, condition of culture is temperature 25 ± 2 DEG C, intensity of illumination 2000Lx, light application time 14h/d.
3. root induction is cultivated
Choose pea green, island is bright and beautiful, the Multiple Buds of extra large yellow robust growth; cut into simple bud; be inoculated into respectively on root media and carry out root induction cultivation; root induction medium take MS as minimal medium; macroelement reduces by half; add IBA1.0 ~ 5.0mg/L, NAA0.1mg/L, TA0.1 ~ 2g/L, sucrose 30 ~ 50g/L, agar 6g/L; condition of culture is temperature 25 ± 2 DEG C; intensity of illumination 2000Lx, light application time 14h/d, cultivate 10 ~ 18d; bastem portion expands; the tip of a root breaks through epidermis, continues to cultivate 20d, and seedling of taking root is formed.
4. to take root transplantation of seedlings
By pea green, island is bright and beautiful, the seedling of taking root of extra large paeonia lutea shifts out between cultivation; in greenhouse hardening; first to remain silent hardening, unscrew bottle cap the previous day in transplanting, second day when transplanting; wash away the medium of seedling base portion; plant respectively in nutritive cube, cultivation matrix selects perlite and peat composed of rotten mosses mixed-matrix, transplants initial stage button Small plastic shed moisturizing; raise canopy film after one week gradually to ventilate, after planting, two weeks plantlet in vitro blade obviously grows.
The rooting rate of three kinds of tree peony plantlet in vitro is respectively pea green 30.0% ~ 66.7%, island brocade 33.3% ~ 86.7%, sea yellow 31.4% ~ 73.5%.
The present invention select pea green, island is bright and beautiful and sea yellow be first-class kind in tree peony, " pea green " is one of large famous-object of tree peony four, just opens dark green, yellow green in full bloom, and abloom rate is high, and in evening at florescence, the duration is long, has sight, belongs to rare famous and precious kind; Island brocade is a kind of natural " assorted sample brocade " tree peony seldom, and belong to secondary color class Varieties of Peony, petal is large, has pure red, half red half powder, the alternate isochrome of red bar vermicelli; Sea Huang is referred to as golden tree peony; color is pure golden yellow; the pure of its yellow really belongs to exception; extremely precious; three is all " superfine product " in tree peony, and in the present invention, three kinds of tree peonies all obtain desirable rooting efficiency; sea the highest yellow rooting rate reaches 73.5%, the island the highest rooting rate of brocade reaches 86.7%, pea green tree peony not only finishes not organize training and to take root the record of seedling, and its rooting rate has been up to 66.7%.
Triacontanol (TA) extracts from honeybee, a kind of long-chain saturated fatty alcohol with 30 carbon atoms that purifying obtains, there is the transport accumulation strengthening plant photosynthesis and dry matter, promote the function such as plant establishment and cauline leaf growth, before making the present invention, the application of triacontanol on Section Moutan training is taken root is there are no report, triacontanol is applied to pea green by the present invention first, the group training of island brocade and extra large paeonia lutea is taken root in seedling culture technique, be equipped with growth hormone hestening rooting, obtain good rooting efficiency, group for tree peony is trained seedling research of taking root and is provided technical foundation and technology guarantee, the inventive method is not by time and environmental limit, simple and easy to do, the rooting efficiency of examination tree peony plantlet in vitro is obvious, the tissue-culturing rapid propagation of this method to other kind of tree peony also has reference, there is good using value.
Accompanying drawing explanation
The pea green tree peony Multiple Buds of accompanying drawing 1.
The pea green tree peony of accompanying drawing 2. takes root seedling
Accompanying drawing 3. island brocade tree peony Multiple Buds
Brocade tree peony takes root seedling on accompanying drawing 4. island
The extra large paeonia lutea Multiple Buds of accompanying drawing 5.
The extra large paeonia lutea of accompanying drawing 6. is taken root seedling.
Embodiment
Below in conjunction with embodiment, the present invention is described in detail.
Embodiment 1
1. the foundation of aseptic strain
Get the pea green of robust growth respectively, island brocade, full bulbil on the paeonia lutea maternal plant of sea, remove the 1-2 layer scale of ragged edge, scrub with suds, then tap water is clean, aseptically bulbil is put into aseptic triangular flask, adopt three step sterilizations to bulbil disinfection: the first step with 75% ethanol disinfection 45s, second step with 1% hypochlorite disinfectant 10min, taking-up aseptic water washing, 3rd step with 0.5% hypochlorite disinfectant 15min, take out clean with aseptic water washing again, constantly triangular flask is shaken in whole disinfecting process, disinfectant is fully contacted with bulbil, taking out after sterilization is placed on aseptic filter paper, residue scale is divested with aseptic nipper, being inoculated in respectively by the bulbil of 3 kinds on inducing culture induces bulbil to sprout, inducing culture used with WPM medium for minimal medium, add 6-BA0.8mg/L, NAA0.05mg/L, sucrose 30g/L and agar 6g/L, sprout through 12d bulbil, 30d forms indefinite bud (base portion has a small amount of callus), aseptic strain is set up, condition of culture is temperature 25 ± 2 DEG C, intensity of illumination 2000Lx, light application time 14h/d.
2. shoot proliferation is cultivated
Take out pea green respectively, island brocade, the yellow indefinite bud formed in sea, cut base portion callus, then access respectively in subculture multiplication medium and cultivate 35d, induced synthesis Multiple Buds, Multiple Buds is divided into again the budlet clump of band 1-2 bud, be inoculated in respectively in identical subculture multiplication medium and carry out shoot proliferation cultivation, time 35d, budlet grows up, base portion has sprouting to sprout the new Multiple Buds of formation, pea green, island brocade, the Multiple Buds of sea Huang is shown in accompanying drawing 1 respectively, accompanying drawing 3 and accompanying drawing 5, subculture multiplication medium take WPM as minimal medium, add 6-BA2.0mg/L, NAA0.05mg/L, AC1.0mg/L, sucrose 30g/L, agar 6g/L, condition of culture is temperature 25 ± 2 DEG C, intensity of illumination 2000Lx, light application time 14h/d.
3. root induction is cultivated
Choose pea green, island brocade, the Multiple Buds of the yellow robust growth in sea, cut into simple bud, be inoculated into respectively on root media and carry out root induction cultivation, root induction medium take MS as minimal medium, macroelement reduces by half, add IBA3.0mg/L, NAA0.1mg/L, TA1.0g/L, sucrose 50g/L, agar 6g/L, condition of culture is temperature 25 ± 2 DEG C, intensity of illumination 2000Lx, light application time 14h/d, cultivate 12d, bastem portion expands, the tip of a root breaks through epidermis, continue to cultivate 20d, seedling of taking root is formed, pea green, island brocade, the seedling of taking root of sea Huang is shown in accompanying drawing 2 respectively, accompanying drawing 4 and accompanying drawing 6, pea green rooting rate is 66.7%, the rooting rate of island brocade is 86.7%, the rooting rate of sea Huang is 73.5%.
4. to take root transplantation of seedlings
By pea green, island is bright and beautiful, the seedling of taking root of extra large paeonia lutea shifts out between cultivation; in greenhouse hardening; first to remain silent hardening, unscrew bottle cap the previous day in transplanting, second day when transplanting; wash away the medium of seedling base portion; plant respectively in nutritive cube, cultivation matrix selects perlite and peat composed of rotten mosses mixed-matrix, transplants initial stage button Small plastic shed moisturizing; raise canopy film after one week gradually to ventilate, after planting, two weeks plantlet in vitro blade obviously grows.
Embodiment 2
1. the foundation of aseptic strain
Get the pea green of robust growth respectively, island brocade, full bulbil on the paeonia lutea maternal plant of sea, remove the 1-2 layer scale of ragged edge, scrub with suds, then tap water is clean, aseptically bulbil is put into aseptic triangular flask, adopt three step sterilizations to bulbil disinfection: the first step with 75% ethanol disinfection 60s, second step with 1% hypochlorite disinfectant 8min, taking-up aseptic water washing, 3rd step with 0.5% hypochlorite disinfectant 20min, take out clean with aseptic water washing again, constantly triangular flask is shaken in whole disinfecting process, disinfectant is fully contacted with bulbil, taking out after sterilization is placed on aseptic filter paper, residue scale is divested with aseptic nipper, being inoculated in respectively by the bulbil of 3 kinds on inducing culture induces bulbil to sprout, inducing culture used with WPM medium for minimal medium, add 6-BA1.0mg/L, NAA0.1mg/L, sucrose 30g/L and agar 6g/L, sprout through 12d bulbil, 30d forms indefinite bud (base portion has a small amount of callus), aseptic strain is set up, condition of culture is temperature 25 ± 2 DEG C, intensity of illumination 2000Lx, light application time 14h/d.
2. shoot proliferation is cultivated
Take out pea green respectively, island brocade, the yellow indefinite bud formed in sea, cut base portion callus, then access respectively in subculture multiplication medium and cultivate 30d, induced synthesis Multiple Buds, Multiple Buds is divided into again the budlet clump of band 1-2 bud, be inoculated in respectively in identical subculture multiplication medium and carry out shoot proliferation cultivation, time 30d, budlet grows up, base portion has sprouting to sprout the new Multiple Buds of formation, subculture multiplication medium take WPM as minimal medium, add 6-BA3.0mg/L, NAA0.1mg/L, AC2.0g/L, sucrose 30g/L, agar 6g/L, condition of culture is temperature 25 ± 2 DEG C, intensity of illumination 2000Lx, light application time 14h/d.
3. root induction is cultivated
Choose pea green, island brocade, the Multiple Buds of the yellow robust growth in sea, cut into simple bud, be inoculated into respectively on root media and carry out root induction cultivation, root induction medium take MS as minimal medium, macroelement reduces by half, add IBA1.0mg/L, NAA0.1mg/L, TA2.0g/L, sucrose 40g/L, agar 6g/L, condition of culture is temperature 25 ± 2 DEG C, intensity of illumination 2000Lx, light application time 14h/d, cultivate 18d, bastem portion expands, the tip of a root breaks through epidermis, continue to cultivate 20d, seedling of taking root is formed, pea green rooting rate is 30.0%, the rooting rate of island brocade is 33.3%, the rooting rate of sea Huang is 31.4%.
4. to take root transplantation of seedlings
By pea green, island is bright and beautiful, the seedling of taking root of extra large paeonia lutea shifts out between cultivation; in greenhouse hardening; first to remain silent hardening, unscrew bottle cap the previous day in transplanting, second day when transplanting; wash away the medium of seedling base portion; plant respectively in nutritive cube, cultivation matrix selects perlite and peat composed of rotten mosses mixed-matrix, transplants initial stage button Small plastic shed moisturizing; raise canopy film after one week gradually to ventilate, after planting, two weeks plantlet in vitro blade obviously grows.
Embodiment 3
1. the foundation of aseptic strain
Get the pea green of robust growth respectively, island brocade, full bulbil on the paeonia lutea maternal plant of sea, remove the 1-2 layer scale of ragged edge, scrub with suds, then tap water is clean, aseptically bulbil is put into aseptic triangular flask, adopt three step sterilizations to bulbil disinfection: the first step with 75% ethanol disinfection 30s, second step with 1% hypochlorite disinfectant 15 min, taking-up aseptic water washing, 3rd step with 0.5% hypochlorite disinfectant 10min, take out clean with aseptic water washing again, constantly triangular flask is shaken in whole disinfecting process, disinfectant is fully contacted with bulbil, taking out after sterilization is placed on aseptic filter paper, residue scale is divested with aseptic nipper, being inoculated in respectively by the bulbil of 3 kinds on inducing culture induces bulbil to sprout, inducing culture used with WPM medium for minimal medium, add 6-BA0.1mg/L, NAA0.01mg/L, sucrose 30g/L and agar 6g/L, sprout through 15d bulbil, 30d forms indefinite bud (base portion has a small amount of callus), aseptic strain is set up, condition of culture is temperature 25 ± 2 DEG C, intensity of illumination 2000Lx, light application time 14h/d.
2. shoot proliferation is cultivated
Take out pea green respectively, island brocade, the yellow indefinite bud formed in sea, cut base portion callus, then access respectively in subculture multiplication medium and cultivate 45d, induced synthesis Multiple Buds, Multiple Buds is divided into again the budlet clump of band 1-2 bud, be inoculated in respectively in identical subculture multiplication medium and carry out shoot proliferation cultivation, time 45d, budlet grows up, base portion has sprouting to sprout the new Multiple Buds of formation, subculture multiplication medium take WPM as minimal medium, add 6-BA0.5mg/L, NAA0.01mg/L, AC0g/L, sucrose 30g/L, agar 6g/L, condition of culture is temperature 25 ± 2 DEG C, intensity of illumination 2000Lx, light application time 14h/d.
3. root induction is cultivated
Choose pea green, island brocade, the Multiple Buds of the yellow robust growth in sea, cut into simple bud, be inoculated into respectively on root media and carry out root induction cultivation, root induction medium take MS as minimal medium, macroelement reduces by half, add IBA5.0mg/L, NAA0.1mg/L, TA0.1g/L, sucrose 30g/L, agar 6g/L, condition of culture is temperature 25 ± 2 DEG C, intensity of illumination 2000Lx, light application time 14h/d, cultivate 10d, bastem portion expands, the tip of a root breaks through epidermis, continue to cultivate 20d, seedling of taking root is formed, pea green rooting rate is 35.7%, the rooting rate of island brocade is 42.0%, the rooting rate of sea Huang is 37.5%.
4. to take root transplantation of seedlings
By pea green, island is bright and beautiful, the seedling of taking root of extra large paeonia lutea shifts out between cultivation; in greenhouse hardening; first to remain silent hardening, unscrew bottle cap the previous day in transplanting, second day when transplanting; wash away the medium of seedling base portion; plant respectively in nutritive cube, cultivation matrix selects perlite and peat composed of rotten mosses mixed-matrix, transplants initial stage button Small plastic shed moisturizing; raise canopy film after one week gradually to ventilate, after planting, two weeks plantlet in vitro blade obviously grows.
Claims (1)
1. a peony tissue cultivates rooting method, it is characterized in that by " foundation of aseptic strain ", " shoot proliferation cultivation " and " root induction cultivation " three link compositions, triacontanol (TA) is added in root induction medium, be equipped with growth hormone hestening rooting, the foundation of aseptic strain: using tree peony bulbil as explant, aseptically adopt three step sterilization disinfections, bulbil is inoculated in inducing culture induction bulbil and sprouts, inducing culture with WPM medium for minimal medium, add 6-BA0.1 ~ 1.0mg/L, NAA0.01 ~ 0.1mg/L, sucrose 30g/L and agar 6g/L, adventitious bud formation, aseptic strain builds up, shoot proliferation is cultivated: indefinite bud is cut, be inoculated in proliferated culture medium induced synthesis Multiple Buds, again Multiple Buds is divided into the budlet clump of band 1 ~ 2 bud, be inoculated on identical medium and proceed shoot proliferation cultivation, proliferated culture medium is minimal medium with WPM, adds 6-BA0.5 ~ 3.0mg/L, NAA0.01 ~ 0.1mg/L, AC0 ~ 2g/L, sucrose 30g/L, agar 6g/L, root induction is cultivated: the healthy and strong Multiple Buds chosen in Multiplying culture is inoculated into root media and carries out root induction cultivation, formation is taken root seedling, root media take MS as minimal medium, macroelement reduces by half, then adds IBA1.0 ~ 5.0mg/L, NAA0.1mg/L, TA0.1 ~ 2g/L, sucrose 30 ~ 50g/L, agar 6g/L, bulbil disinfects three adopted step sterilizations: the first step with 75% ethanol disinfection 30 ~ 60s, second step with 1% hypochlorite disinfectant 8 ~ 15 min, taking-up aseptic water washing, 3rd step with 0.5% hypochlorite disinfectant 10 ~ 20min, take out clean with aseptic water washing again, constantly shake triangular flask in whole disinfecting process, disinfectant is fully contacted with bulbil, foundation, the shoot proliferation cultivation of aseptic strain are identical with the condition that three links are cultivated in root induction, are temperature 25 ± 2 DEG C, intensity of illumination 2000Lx, light application time 14h/d, the Varieties of Peony selected is for sea is yellow, island is bright and beautiful and pea green.
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| CN104855286A (en) * | 2015-04-22 | 2015-08-26 | 胡进耀 | Tongling peony tissue culture and rapid propagation seedling raising technical method |
| CN105532477A (en) * | 2016-02-04 | 2016-05-04 | 河南农业大学 | Peony rooting culture method |
| CN106538395B (en) * | 2017-01-18 | 2018-11-13 | 西藏农牧学院 | A method of improving endangered plants Tibet Paeonia ludlowii planting percent and seedling percent |
| CN111727879B (en) * | 2019-07-11 | 2021-10-15 | 中国科学院植物研究所 | Culture medium and method for oil peony propagation |
| CN111165356B (en) * | 2020-02-28 | 2021-01-05 | 上海培林生物科技有限公司 | Tissue culture propagation method of peony |
| CN113875596B (en) * | 2021-10-26 | 2022-10-28 | 河南省农业科学院园艺研究所 | Processing method for relieving terminal bud dormancy of peony tissue culture seedlings |
| CN116602212A (en) * | 2023-06-06 | 2023-08-18 | 北京市农林科学院 | Paeonia plant in-vitro culture and proliferation method and special culture medium |
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| JP2011142903A (en) * | 2009-12-15 | 2011-07-28 | Kinki Univ | Method for raising seedlings of peony |
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| CN102084812A (en) * | 2009-12-08 | 2011-06-08 | 上海市农业科学院 | Tissue culture and bud induction method for peony |
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