CN106489748A - A kind of quick breeding by group culture method of wintergreen - Google Patents

A kind of quick breeding by group culture method of wintergreen Download PDF

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Publication number
CN106489748A
CN106489748A CN201611231227.2A CN201611231227A CN106489748A CN 106489748 A CN106489748 A CN 106489748A CN 201611231227 A CN201611231227 A CN 201611231227A CN 106489748 A CN106489748 A CN 106489748A
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culture
wpm
agar
sucrose
wintergreen
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CN106489748B (en
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解玮佳
李世峰
彭绿春
王继华
宋杰
张露
蔡艳飞
李树发
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Flower Research Institute of YAAS
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01HNEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/005Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01HNEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/001Culture apparatus for tissue culture
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01HNEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/008Methods for regeneration to complete plants

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  • Life Sciences & Earth Sciences (AREA)
  • Developmental Biology & Embryology (AREA)
  • Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • Botany (AREA)
  • Environmental Sciences (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

Abstract

The invention provides a kind of quick breeding by group culture method of wintergreen.The method includes disinfecting for seed, the acquisition of aseptic seedling, the acquisition of Multiple Buds and root culture.Using the inventive method, the Multiple Buds quantity that simple bud induces is more, about 38, inducing clumping bud rate is high, inductivity is 83.33 100%, and sprout robust growth, individual plant may be up to 3 5cm, breeding coefficient is high, up to 4.12 5.58, rooting rate is high for breeding coefficient, and rooting rate is up to 75.00 97.20%, breeding cycle is short, 90 120 days breeding cycles.The tissue cultured seedling for breeding is healthy and strong, well developed root system, and per plant takes root 3 10, and 0.92 1.26cm of average root length, its tissue cultured seedling transplanting survival rate reach more than 95%, and after transplanting, growing way is strong.

Description

A kind of quick breeding by group culture method of wintergreen
Technical field
The present invention relates to field of plant tissue culture technique, and in particular to a kind of quick breeding by group culture side of wintergreen Method.
Background technology
The branch and leaf of Gaultheria (Gaultheria Kalm ex Linn.) most of species contains aromatic oil, is light industry The natural type fine chemical material of industry and medical industry.The platymiscium is bloomed milky, and corolla is bell and sagging, and fruit is red, white Or blue, and fruiting period is longer, is precious ornamental plants in garden again.As the resource plant that Important Economic is worth, Gaultheria Plant has larger potentiality to be exploited.China has 35 kinds of Gaultheria plant, accounts for the nearly a quarter of world's species amount, wherein 14 kinds is peculiar.However, at present domestic for the monoid that this germ plasm resource is abundant, economic worth and potentiality to be exploited are higher, only There are the basic researchs such as typoiogical classification, species identification, origin and evolution, compound separation and Extraction, and to the monoid germ plasm resource Introduction and acclimatization and protective development research also in the lag phase, to its tissue culture propagation in terms of research also there is not been reported.For Accelerate rational exploitation and utilization China's abundant Gaultheria resource, carry out the platymiscium group culturation rapid propagating technology have highly important Meaning.
The wintergreen (Gaultheria procumbens L.) being related in the present invention is one in Gaultheria The species of great sight, calls round-leaved wintergreen, is the short evergreen shrubss of 10-15cm, tool Herba pyrolae japonicae fragrance, floral white, The capsule that fruit is split for berry shape, 5 slit bamboo or chopped woods, contains and after spending expands and become in the calyx of meat, red, and sight is high, into For very fast-selling fruit effect plant on International Flower market.Nearly 2 years, with the rise of domestic households gardening, wintergreen by The concern of domestic production of flowers and plants business is gradually received.But, due to the careful such as grit of the kind of wintergreen, in conventional seed planting nursery There are problems that sowing inconvenience, emergence rate are low and conserve, and can be limited by season and resource, be difficult in the short time Carry out extensive nursery work.For this purpose, developing a kind of tissue culture quick propagation plantation technology of suitable wintergreen, effectively can solve Nursery amount certainly in actual production is low and slow-paced problem, to promote China's Gaultheria plant resourceses reasonable development and guarantor Shield.
Content of the invention
For wintergreen nursery emergence rate in effectively solving actual production is low and slow-paced technical problem, the present invention There is provided that a kind of cultivation cycle is short, be not subject to seasonal restrictions and wintergreen that survival rate is high quick breeding by group culture method, with Wintergreen industry development and the protective development of the platymiscium resource is promoted to utilize.
Technical scheme is as follows:
1st, a kind of quick breeding by group culture method of wintergreen, comprises the following steps:
(1) wintergreen seed is disinfected
Aseptically, by the wintergreen seed that gives birth to then in the H that mass fraction is 25%2O2Soak in solution 20min, blots the surface of the seed moisture with aseptic filter paper, standby;
(2) acquisition of aseptic seedling
Aseptically, will sow on seed germination medium through the seed that step (1) disinfects, cultivate 30- 40 days, aseptic seedling of the height for 2-3cm is obtained, condition of culture is:25 DEG C ± 2 DEG C of temperature, illumination 12h d-1, illuminance is 4000lx;The seed germination medium is:WPM+ agar 6g/L+ sucrose 30g/L, pH5.4;
(3) acquisition of Multiple Buds
Aseptically, stem with bud is cut in the aseptic seedling that step (2) is obtained accesses inducing clumping bud culture medium, Culture 30-40 days, obtains Multiple Buds, and condition of culture is identical with the condition of culture described in step (2), and described inducing clumping bud is trained Foster base is:WPM+ZT 1.0~3.0mg/L+IBA 0.1mg/L+ agar 6g/L+ sucrose 30g/L, pH5.4;
(4) root culture
The Multiple Buds that step (3) is obtained access root media, cultivate -40 days 30 days, obtain band and take root in strain, condition of culture Identical with the condition of culture described in step (2), described root media is:0.5~1.5mg/L+NAA of WPM+IBA 0.2~ 0.75mg/L+ agar 6g/L+ sucrose 30g/L, pH5.4.
2. the quick breeding by group culture method of a kind of wintergreen according to technical scheme 1, described in step (3) Inducing clumping bud culture medium is:WPM+ZT 2.0mg/L+IBA 0.1mg/L+ agar 6g/L+ sucrose 30g/L, pH5.4;Step (4) root media described in is:WPM+IBA 1.0mg/L+NAA 0.50mg/L+ agar 6g/L+ sucrose 30g/L, pH5.4.
3. the quick breeding by group culture method of a kind of wintergreen according to technical scheme 1 or 2, step are cut in (3) The length of the stem with bud for taking is 1~2cm.
4. the special culture media of the quick breeding by group culture of one group of wintergreen, described special culture media is by following seed Germination medium, inducing clumping bud culture medium and root media composition:
The seed germination medium is:WPM+ agar 6g/L+ sucrose 30g/L, pH5.4;
Described inducing clumping bud culture medium is:WPM+ZT 1.0~3.0mg/L+IBA 0.1mg/L+ agar 6g/L+ sugarcanes Sugared 30g/L, pH5.4;
Described root media is:WPM+IBA 0.5~1.5mg/L+NAA, 0.2~0.75mg/L+ agar 6g/L+ sugarcanes Sugared 30g/L, pH5.4.
5. the special culture media of the quick breeding by group culture of one group of wintergreen according to technical scheme 4, described Special culture media is made up of following seed germination medium, inducing clumping bud culture medium and root media:
The seed germination medium is:WPM+ agar 6g/L+ sucrose 30g/L, pH5.4;
Described inducing clumping bud culture medium is:WPM+ZT 2.0mg/L+IBA 0.1mg/L+ agar 6g/L+ sucrose 30g/ L, pH5.4;
Described root media is:WPM+IBA 1.0mg/L+NAA 0.50mg/L+ agar 6g/L+ sucrose 30g/L, pH5.4.
The beneficial effects of the present invention is:
The present invention is studied for the group culturation rapid propagating technology of Gaultheria plant wintergreen first, is proposed a kind of flat The quick breeding by group culture method and special culture medium of paving Gaultheria.Using the inventive method, the Multiple Buds that simple bud induces Quantity is more, about 3-8, and inducing clumping bud rate is high, and inductivity is 83.33-100%, and sprout robust growth, individual plant may be up to 3- 5cm, breeding coefficient are high, and breeding coefficient reaches 4.12-5.58, and rooting rate is high, and rooting rate is up to 75.00-97.20%, breeding cycle Short, 90-120 days breeding cycle.The tissue cultured seedling for breeding is healthy and strong, well developed root system, per plant of 3-10 bar of taking root, the long 0.92- of average root 1.26cm, its tissue cultured seedling transplanting survival rate reach more than 95%, and after transplanting, growing way is strong.
Description of the drawings
Fig. 1:The stem with bud of wintergreen accesses inducing clumping bud culture medium.
Fig. 2:The wintergreen Multiple Buds that tissue culture is obtained.
Fig. 3:The root induction culture of wintergreen.
Fig. 4:The Seedling of taking root that wintergreen tissue culture is obtained.
Specific embodiment
Following examples are conventional plant tissue culture method without specified otherwise, and in each example, test material used is city Sell.Each embodiment is respectively provided with three repetitions, results averaged.
Culture medium preparation method of the preparation of each embodiment culture medium according to culture medium prescription routinely plant tissue culture Prepare.
Embodiment 1
(1) wintergreen seed is disinfected
Aseptically, by the wintergreen seed that gives birth to then in the H that mass fraction is 25%2O2Soak in solution 20min, blots the surface of the seed moisture with aseptic filter paper, standby.
(2) acquisition of aseptic seedling
Aseptically, will sow on seed germination medium through the seed that step (1) disinfects, culture 35 My god, aseptic seedling of the height for 2-3cm is obtained, condition of culture is:25 DEG C ± 2 DEG C of temperature, illumination 12h d-1, illuminance is 4000lx;The seed germination medium is:WPM+ agar 6g/L+ sucrose 30g/L, pH5.4.
(3) acquisition of Multiple Buds
Aseptically, the stem with bud that length is 1~2cm is cut in the aseptic seedling that step (2) is obtained accesses clump Sprout inducing culture, cultivate 35 days, obtain Multiple Buds, condition of culture is identical with the condition of culture described in step (2), described Inducing clumping bud culture medium is:WPM+ZT 1.0mg/L+IBA 0.1mg/L+ agar 6g/L+ sucrose 30g/L, pH5.4.
(4) root culture
The Multiple Buds that step (3) is obtained access root media, cultivate 35 days, obtain band and take root in strain, condition of culture and step Suddenly the condition of culture described in (2) is identical, and described root media is:WPM+IBA 0.5mg/L+NAA 0.2mg/L+ agar 6g/L+ sucrose 30g/L, pH5.4.
, with embodiment 3 in addition to measure listed by table 1 is different, remaining measure is same as Example 1, repeats no more for embodiment 2.
1 embodiment 2- embodiment of table, 3 each step and the difference of embodiment 1
The reproductive effect of embodiment 1-3 is shown in Table 2.
The reproductive effect of 2 embodiment 1- embodiment 3 of table
The Multiple Buds quantity that the inventive method simple bud induces is more, about 3-8, and inducing clumping bud rate is high, and inductivity is 83.33-100%, sprout robust growth, individual plant may be up to 3-5cm, and breeding coefficient is high, and breeding coefficient is taken root up to 4.12-5.58 Rate is high, and, up to 75.00-97.20%, the breeding cycle is short for rooting rate, 90-120 days breeding cycle.The tissue cultured seedling for breeding is healthy and strong, Well developed root system, per plant of 3-10 bar of taking root, the long 0.92-1.26cm of average root, its tissue cultured seedling transplanting survival rate reach more than 95%, move After cultivation, growing way is strong.

Claims (5)

1. a kind of quick breeding by group culture method of wintergreen, it is characterised in that comprise the following steps:
(1) wintergreen seed is disinfected
Aseptically, by the wintergreen seed that gives birth to then in the H that mass fraction is 25%2O2Soak in solution 20min, blots the surface of the seed moisture with aseptic filter paper, standby;
(2) acquisition of aseptic seedling
Aseptically, will sow on seed germination medium through the seed that step (1) disinfects, cultivate 30-40 days, Aseptic seedling of the height for 2-3cm is obtained, condition of culture is:25 DEG C ± 2 DEG C of temperature, illumination 12h d-1, illuminance is 4000lx; The seed germination medium is:WPM+ agar 6g/L+ sucrose 30g/L, pH5.4;
(3) acquisition of Multiple Buds
Aseptically, stem with bud is cut in the aseptic seedling that step (2) is obtained accesses inducing clumping bud culture medium, culture 30-40 days, Multiple Buds are obtained, condition of culture is identical with the condition of culture described in step (2), described inducing clumping bud culture medium For:WPM+ZT 1.0~3.0mg/L+IBA0.1mg/L+ agar 6g/L+ sucrose 30g/L, pH5.4;
(4) root culture
The Multiple Buds that step (3) is obtained access root media, cultivate -40 days 30 days, obtain band and take root in strain, condition of culture and step Suddenly the condition of culture described in (2) is identical, and described root media is:0.5~1.5mg/L+NAA of WPM+IBA 0.2~ 0.75mg/L+ agar 6g/L+ sucrose 30g/L, pH5.4.
2. the quick breeding by group culture method of a kind of wintergreen according to claims 1, it is characterised in that:Step (3) the inducing clumping bud culture medium described in is:WPM+ZT 2.0mg/L+IBA 0.1mg/L+ agar 6g/L+ sucrose 30g/L, pH5.4;Root media described in step (4) is:WPM+IBA 1.0mg/L+NAA 0.50mg/L+ agar 6g/L+ sucrose 30g/L, pH5.4.
3. the quick breeding by group culture method of a kind of wintergreen according to claims 1 or 2, it is characterised in that:Step The length of the stem with bud for cutting in (3) suddenly is 1~2cm.
4. the special culture media of the quick breeding by group culture of one group of wintergreen, it is characterised in that described special culture media by Following seed germination medium, inducing clumping bud culture medium and root media composition:
The seed germination medium is:WPM+ agar 6g/L+ sucrose 30g/L, pH5.4;
Described inducing clumping bud culture medium is:WPM+ZT 1.0~3.0mg/L+IBA 0.1mg/L+ agar 6g/L+ sucrose 30g/L, pH5.4;
Described root media is:WPM+IBA 0.5~1.5mg/L+NAA, 0.2~0.75mg/L+ agar 6g/L+ sucrose 30g/L, pH5.4.
5. the special culture media of the quick breeding by group culture of one group of wintergreen according to claim 4, it is characterised in that Described special culture media is made up of following seed germination medium, inducing clumping bud culture medium and root media:
The seed germination medium is:WPM+ agar 6g/L+ sucrose 30g/L, pH5.4;
Described inducing clumping bud culture medium is:WPM+ZT 2.0mg/L+IBA 0.1mg/L+ agar 6g/L+ sucrose 30g/L, pH5.4;
Described root media is:WPM+IBA 1.0mg/L+NAA 0.50mg/L+ agar 6g/L+ sucrose 30g/L, pH5.4.
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Publication number Priority date Publication date Assignee Title
CN109006326A (en) * 2018-08-20 2018-12-18 潘国长 A kind of artificial method for planting of spice all over the mountain
CN110476815B (en) * 2019-09-27 2021-06-29 四川七彩林科股份有限公司 Tissue culture and rapid propagation method of Ligumu
CN116548314A (en) * 2023-07-07 2023-08-08 云南卉兴农业有限公司 Method for tissue culture and rapid propagation of flat white bead clone

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109006326A (en) * 2018-08-20 2018-12-18 潘国长 A kind of artificial method for planting of spice all over the mountain
CN110476815B (en) * 2019-09-27 2021-06-29 四川七彩林科股份有限公司 Tissue culture and rapid propagation method of Ligumu
CN116548314A (en) * 2023-07-07 2023-08-08 云南卉兴农业有限公司 Method for tissue culture and rapid propagation of flat white bead clone
CN116548314B (en) * 2023-07-07 2023-09-12 云南卉兴农业有限公司 Method for tissue culture and rapid propagation of flat white bead clone

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