CN107646684A - A kind of breeding method of purpleback murdannia herb and its application - Google Patents
A kind of breeding method of purpleback murdannia herb and its application Download PDFInfo
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- CN107646684A CN107646684A CN201710995256.4A CN201710995256A CN107646684A CN 107646684 A CN107646684 A CN 107646684A CN 201710995256 A CN201710995256 A CN 201710995256A CN 107646684 A CN107646684 A CN 107646684A
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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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- 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
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Abstract
The present invention discloses breeding method and its application of a kind of purpleback murdannia herb.The present invention carries out aseptic seeding after the purpleback murdannia herb capsule that 28~30 days are grown after pollination is plucked；Then obtained protocorm is moved into subculture growth medium, protocorm Stem nematode and sprouting and rooting is induced, using LED as tissue culture light source, with proportion 2~4：6~8 proportioning feux rouges mixing blue lights are cultivated；By obtained plant height up to the sterile transplantation of seedlings of more than 7cm to greenhouse, cultivation management is carried out.Present invention determine that purpleback murdannia herb seed sprout after Subculture in optimal culture condition, including LED light matter and light intensity and processing time, effectively facilitate plant chlorophyll accumulation, stem thickening and root long elongation, strengthening seedling and rooting process is needed not move through after squamous subculture directly to transplant, and save production cost and period.Especially LED promotes purpleback murdannia herb activity in vivo composition flavonoids, aldehydes matter accumulation, significantly improves plant anti-oxidation ability, significant to medicinal plant cultivation.
Description
Technical field
The present invention relates to field of plant growing technology, the breeding method of more particularly to a kind of purpleback murdannia herb and its application.
Background technology
Purpleback murdannia herb (Arundina graminifolia) is orchid family Shi Lan races ground non-hibernating eggs, is filled blood with tune, Qinghuo Jiedu
And other effects, it is one of main material that the dai medicine Dai Nationality hundred solves (hundred solution capsules).Numerous studies find that purpleback murdannia herb extract has very high
Anti-oxidant, antitumor and antiviral activity and lipoid peroxidization resistant, therefore can be treating in food, toadstool, medicine
The diseases such as poison, wherein polyphenol, flavones, saponin(e equal size are in notable dosage effect.In addition, purpleback murdannia herb flower exactly likes Bowring cattleya, flower pattern
It is big and beautiful, it can persistently bloom and environmental suitability is strong whole year, be a kind of excellent potted plant and ground by wild orchid.It is but a large amount of
Artificial excavation, cause purpleback murdannia herb wild resource to reduce increasingly, be listed in country II grade protection plant.Therefore, it is badly in need of establishing
A set of efficiently seedling breeding and planting technology system, meet the market demand, promote purpleback murdannia herb popularization and application and industry development.
During planting management, light quality is to influence its important factor to grow.Light emitting diode
(light-emitting diodes, LED) is the high-quality light source that developed recently gets up, and it can send the monochromatic light of accurate wave spectrum,
Wave-length coverage is narrower, and amplitude is no more than 20nm, and and can controls light intensity by regulation device, can be effectively facilitated Plant Light cooperation
With having wide application prospect.At present have LED light source be used for hybrid cymbidium, iris, Cymbidium lianpan, a beautiful phalaenopsis, chrysanthemum,
The report of the Plant Tissue Breeding such as African Chrysanthemum, common calla, the white palm, tree peony, grape, strawberry, and find that different light medium is planted with comparison
The adjustment effect that thing grows has larger difference.But current research all concentrates on the thing of scale breeding technology relative maturity
Kind, had not been reported for the light quality regulation and control in purpleback murdannia herb planting process.
The content of the invention
The shortcomings that in order to overcome prior art and deficiency, it is an object of the invention to provide a kind of cultivation side of purpleback murdannia herb
Method.This method can effectively facilitate plant growth, improve medicinal active ingredient, shorten growing-seedling period.
Another object of the present invention is to the application for the breeding method for providing above-mentioned purpleback murdannia herb.
The purpose of the present invention is achieved through the following technical solutions：
A kind of breeding method of purpleback murdannia herb, comprises the following steps：Aseptic seeding, squamous subculture, acclimatization and transplantses；
(1) aseptic seeding：Aseptic seeding is carried out after the purpleback murdannia herb capsule that 28~30 days are grown after pollination is plucked；Culture medium
Composition be：Spend precious No. 1 culture medium+10~30g/L+6- of sucrose benzyl purine (6-BA) 0.5~1.5mg/L+ methyl α-naphthyl acetates (NAA)
0.1~0.2mg/L+ activated carbon 0.5~1g/L+ banana puree 10~30g/L+, 6~8g/L of agar, pH=5.6~5.8；Cultivate bar
Part：20~40 μm of o1m of light intensity-2·s-1, 12~16h/d of photoperiod cultures 25~60 days；
(2) squamous subculture：The protocorm that step (1) obtains is moved into subculture growth medium, induction protocorm Stem nematode and
Sprouting and rooting, condition of culture are as follows：Using LED as tissue culture light source, 620~660nm of feux rouges, blue light Blue 450~
480nm, in proportion 2~4：6~8 proportionings, 30~60 μm of o1m of light intensity-2·s-1, 12~16h/d of photoperiod cultures 30~60
My god, improve plant growth rate；The composition of subculture growth medium is：Spend precious No. 1 culture medium+sucrose 10~30g/L+6- benzyls
10~30g/L+ of purine (6-BA) 0.5~3mg/L+ methyl α-naphthyl acetates (NAA) 0.2~0.5mg/L+ activated carbon 0.5~1g/L+ banana purees
6~8g/L of agar, pH=5.6~5.8；
(3) purpleback murdannia herb test tube transplantation of seedlings：The plant height that step (2) is obtained is carried out up to the sterile transplantation of seedlings of more than 7cm to greenhouse
Cultivation management；Cultivation matrix includes：At least one of coconut palm chaff, peat soil, peanut shell, bark and sphagna；Cultivation condition：Transplanting
Humidity 60%~80%, 75% two layers of shade are kept within two weeks afterwards.
Above-mentioned culture is carried out under the conditions of being unless otherwise specified 40~60% in 20~30 DEG C, relative humidity.
Described feux rouges is preferably the feux rouges that wavelength is 630nm.
Sowing culture medium described in step (1) is preferably to spend No. 1 culture medium+6-BA 0.5mg/L+NAA0.2mg/L+ of treasured
Activated carbon 1g/L+ sucrose 30g/L+ banana puree 30g/L+ agar 7g/L, pH=5.6~5.8.
Subculture growth medium described in step (2) is preferably to spend No. 1 culture medium+6-BA1.0mg/L+NAA of treasured
0.5mg/L+ activated carbon 0.5g/L+ sucrose 30g/L+ banana puree 30g/L+ agar 7g/L, pH=5.6~5.8.
Condition of culture described in step (2) is preferably：Using LED as tissue culture light source, blue light and feux rouges 3：1 mixing
Collocation, 45 μm of o1m of light intensity-2·s-1, photoperiod 16h/d cultures 60 days, environment temperature is 24 ± 1 DEG C, relative humidity 50~
60%.
Newborn seedling described in step (2) be preferably after planting protocorm stem length to 2.5~3.5cm newborn seedling.
Cultivation matrix described in step (3) is preferably：Using coconut palm chaff as transplanting medium, environment temperature is 24 ± 1 DEG C, phase
To humidity 60%~80%.
Application of the cultural method of the purpleback murdannia herb in large-scale cultivation purpleback murdannia herb.
The present invention is had the following advantages relative to prior art and effect：
(1) present invention determine that promote during purpleback murdannia herb aseptic seeding seed sprout and Subculture in promote it is new
Sprout the different culture media best configuration of fast-growth, effectively facilitate efficiently quickly purpleback murdannia herb seedling breeding.
(2) present invention determine that purpleback murdannia herb seed sprout after Subculture in optimal culture condition, including LED light matter
With light intensity and processing time, effectively facilitate plant chlorophyll accumulation, stem thickening and root long elongation, after squamous subculture without through
Crossing strengthening seedling and rooting process can directly transplant, and save production cost and period.Especially LED promotes purpleback murdannia herb activity in vivo composition flavones
Class, aldehydes matter accumulation, significantly improve plant anti-oxidation ability, significant to medicinal plant cultivation.
(3) present invention determine that the optimum substrate condition of the sterile transplantation of seedlings of purpleback murdannia herb, transplanting survival rate more than 80%, effectively
Promote purpleback murdannia herb factorial praluction.
(4) in the present invention, using LED lamp as tissue culture light source, there are saving electric resources 30~50%, reduce
Cost, and improve the efficiency of large-scale production.
Brief description of the drawings
Fig. 1 is the influence of different LED light confrontation purpleback murdannia herb tissue-cultured seedling chlorophyll fluorescence parameters.
Embodiment
With reference to embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention are unlimited
In this.
The aseptic seeding medium optimization of embodiment 1
By grown after pollination 28~30 days purpleback murdannia herb capsule harvesting after, 70% alcohol disinfecting 60s, sterilized water wash down after again
Use 0.1%HgCl2Solution disinfection 15min, then with aseptic water washing 3~5 times, finally blot surface Ming Shui with aseptic filter paper, use
Scalpel cuts the capsule disinfected, and takes out seed and is disseminated in the good sowing culture medium of autoclave sterilization.
Culture medium is to spend treasured No. 1 culture medium+agar 7g/L+ sucrose 30g/L as control group, addition 6- benzyl purines (6-BA)
0.5~1.5mg/L, methyl α-naphthyl acetate (NAA) 0.1~0.2mg/L, 0.5~1g/L of activated carbon, 10~30g/L of banana puree are optimization group,
Specific proportioning is as shown in table 1.PH value=5.6~5.8.
Table 1 sows culture medium prescription
| Culture medium prescription | 6-BA(mg/L) | NAA(mg/L) | Activated carbon (g/L) | Banana puree (g/L) |
| CK | 0 | 0 | 0 | 0 |
| 1 | 0.5 | 0.1 | 0.5 | 15 |
| 2 | 0.5 | 0.2 | 1 | 30 |
| 3 | 1.0 | 0.1 | 0.5 | 15 |
| 4 | 1.0 | 0.1 | 1 | 15 |
| 5 | 1.0 | 0.2 | 1 | 30 |
| 6 | 1.5 | 0.1 | 1 | 30 |
Every kind of culture medium prescription sows 10 bottles.Light source is made with Philip (PHILIPS) ordinary white fluorescent tube.Cultivate bar
It is 16h/d, 20 μm of o1m of light intensity the photoperiod that part, which is,-2·s-1, environment temperature is 24 ± 1 DEG C, relative humidity 50~60%.Continuously
Seed starts to expand after cultivating 20 days, and counting germination rate after continuous culture in 40 days, (germination rate=yellow-white or green are expanded
The seed sum of protuberance/pollution-free sowing seed sum × 100%), counted after continuous culture in 60 days newborn height of seedling degree and
The number of blade, it the results are shown in Table 2.
The purpleback murdannia herb of table 2 counts newborn height of seedling degree and the number of blade after continuous culture in 60 days
| Culture medium prescription | Germination rate | Plant height (cm) | The number of blade |
| CK | 42.98 | 2.58 | 2.8 |
| 1 | 62.27 | 2.74 | 2.7 |
| 2 | 72.40 | 3.61 | 3.1 |
| 3 | 59.85 | 2.98 | 2.8 |
| 4 | 63.40 | 3.34 | 3.2 |
| 5 | 68.83 | 2.87 | 3.5 |
| 6 | 62.19 | 3.10 | 3.2 |
Statistical result (table 2) is shown, No. 1 culture medium+6-BA0.5mg/L+ of treasured is spent with formula 2 in Different Optimization combination
NAA 0.2mg/L+ activated carbon 1g/L+ sucrose 30g/L+ banana puree 30g/L+ agar 7g/L (pH=5.6~5.8) sprouting and life
Long best results.
The subculture growth medium optimizing components of embodiment 2
Newly sprouting in culture medium will be sowed, be seeded to progress fast-growth culture in subculture growth medium.Every bottle connects
10 buds of kind, each 10 bottles of processing.Culture medium is to spend treasured No. 1 culture medium+agar 7g/L+ sucrose 30g/L as control group, wherein adding
Add 6- benzyl purines (6-BA) 0.5~3mg/L, methyl α-naphthyl acetate (NAA) 0.2~1.0mg/L, 0.5~1g/L of activated carbon, banana puree 10
~30g/L is optimization group, and specific proportioning is as shown in table 3.PH value=5.6~5.8.
The subculture grown cultures based formulas of table 3
| Culture medium prescription | 6-BA(mg/L) | NAA(mg/L) | Activated carbon (g/L) | Banana puree (g/L) |
| CK | 0 | 0 | 0 | 0 |
| 1 | 0.5 | 0.2 | 0.5 | 10 |
| 2 | 1.0 | 0.2 | 1 | 30 |
| 3 | 1.0 | 0.5 | 0.5 | 30 |
| 4 | 2.0 | 0.2 | 0.5 | 15 |
| 5 | 2.0 | 0.5 | 1 | 30 |
| 6 | 3.0 | 0.2 | 0.5 | 20 |
| 7 | 0.5 | 1.0 | 1 | 30 |
| 8 | 2.0 | 1.0 | 1 | 30 |
Condition of culture is photoperiod 16h/d, 45 μm of o1m of light intensity-2·s-1, environment temperature is 24 ± 1 DEG C, relative humidity
50~60%.Test tube seedling is after 60d is continuously cultivated, the average plant height of detection inoculation bud, individual plant mean number of sheets and fresh weight, Yi Jijie
Seed bud increases bud number newly, and concrete outcome is shown in Table 4.Plant height vernier caliper measurement (is accurate to 0.01mm).
The test tube seedling of table 4 testing result after 60d is continuously cultivated
| Culture medium prescription | Plant height | The number of sheets | Root long | Fresh weight (g) |
| CK | 7.08 | 5.4 | 4.9 | 0.26 |
| 1 | 8.73 | 6.3 | 5.5 | 0.37 |
| 2 | 9.21 | 6.1 | 5.9 | 0.39 |
| 3 | 8.52 | 6.2 | 6.7 | 0.51 |
| 4 | 8.16 | 5.6 | 5.5 | 0.42 |
| 5 | 7.98 | 5.5 | 6.2 | 0.45 |
| 6 | 7.53 | 5.2 | 5.1 | 0.29 |
| 7 | 7.32 | 5.4 | 6.4 | 0.38 |
| 8 | 7.08 | 4.9 | 6.3 | 0.35 |
As a result show, within the specific limits, NAA can promote the growth of under ground portion root.Consider plant height and root long, with
Optimization culture based formulas 3 spends No. 1 culture medium+6-BA1.0mg/L+NAA0.5mg/L+ activated carbon 0.5g/L+ sucrose 30g/L+ of treasured
Banana puree 30g/L+ agar 7g/L (pH=5.6~5.8) rate of rise is very fast, and plant is sturdy, and root long is than control group increase about
50%, fresh weight increase is higher than control group 2 times, elects optimal subculture growth medium as.
The using effect of embodiment 3, culture medium collocation LED
1st, subculture growth medium
Spend precious No. 1 culture medium+6-BA1.0mg/L+NAA0.5mg/L+ activated carbon 0.5g/L+ sucrose 30g/L+ banana puree
30g/L+ agar 7g/L solid medium (pH=5.6~5.8).
2nd, vaccination ways
Purpleback murdannia herb seedling is seeded to Multiplying culture in subculture growth medium after sowing 60 days.10 buds of every bottle of inoculation,
Each 10 bottles of processing.
3rd, condition of culture
Photoperiod is 16h/d, 45 μm of o1m of light intensity-2·s-1, environment temperature is 24 ± 1 DEG C.At Setup Experiments 9
Reason, is specifically shown in Table 5, is control with white fluorescent lamp (WFL), and LED light source processing is warm white (WW), cool white light (CW), remote
Feux rouges (DR), feux rouges (R), blue light (B), red blue proportioning light (R：B=3：1、R：B=2:2、R：B=1：3 use 3R/ individually below
1B, 2R/2B, 1R/3B are represented).In triplicate, intensity of illumination is maintained at 45 μm of olm for each processing-2·s-1, intensity of illumination use
Apogee Instrument measurings.
9 light sources processing that table 5 is set
4th, influences of the LED to plant growth
After purpleback murdannia herb new life seedling is seeded to new culture medium and continuously cultivated 30 days, plant strain growth situation, statistical item are investigated
Including：Plant height, root long, stem are thick, leaf is long, leaf width；Leaf area calculates single leaf leaf area according to formula；With electronic balance claim fresh weight,
(108 DEG C finish 15 minutes dry weight, 80 DEG C of oven for drying 48 hours to constant weight；Soluble sugar content is surveyed with anthrone colorimetry；With examining
Mas bright blue method surveys soluble protein content；Chlorophyll content is surveyed with ethanol acetone method.Experiment is repeated 3 times, with the SPSS sides of progress
Difference is analysed, Ducan testing significance of differences, P<0.05.
Chlorophyll fluorescence parameters using MINI-PAM- II (WALZ, Germany) ultra portable modulated chlorophyll fluorescence instrument, not from
Purpleback murdannia herb seedling leaves after body measurement difference LED light matter is handled 30 days.Actual Photochemical Efficiency Y (II), photochemical quenching coefficient
(qp) directly directly it is measured at room temperature with non-photochemical quenching coefficient (NPQ) value, maximal photochemistry efficiency Fv/Fm values exist
By at least 30 minutes dark processings after determining at room temperature, items measure is repeated 3 times blade.Operating method makes with reference to instrument
Use specification.
The influence that 6 different LED lights of table are verified to the growth of purpleback murdannia herb tissue-cultured seedling
| Light quality ratio | Plant height/mm | Root long/mm | Radical/ | Stem is thick/mm | Leaf width/mm | Dry weight/g | Fresh weight/g |
| T1 | 56.413bc | 44.1567a | 5a | 3.303a | 5.133a | 0.069a | 0.487a |
| T2 | 58.167bc | 36.987b | 7ab | 3.140a | 4.437b | 0.062a | 0.411b |
| T3 | 61.150b | 36.850b | 5ab | 2.297bc | 3.903c | 0.037b | 0.285c |
| T4 | 58.760b | 31.483c | 4b | 1.787de | 3.530d | 0.033b | 0.246d |
| T5 | 71.943a | 21.138f | 3b | 1.730c | 1.990g | 0.026b | 0.223d |
| T6 | 72.783a | 23.590f | 5b | 1.387de | 2.313fg | 0.030b | 0.234d |
| T7 | 57.083bc | 26.947de | 4b | 2.327bc | 2.537ef | 0.031b | 0.221d |
| T8 | 52.377c | 23.947ef | 3b | 1.927cd | 2.213fg | 0.026b | 0.191e |
| CK | 59.373b | 28.843cd | 4b | 2.383b | 2.713e | 0.026b | 0.230d |
The influence of pigment content in the different LED light confrontation purpleback murdannia herb tissue culture seedling leafs of table 7
| Light quality ratio | Chlorophyll a/(mgg-1) | Chlorophyll b/(mgg-1) | Chlorophyll content/(mgg-1) | Chlorophyll a/b |
| T1 | 5.5790b | 2.669b | 8.274b | 2.09b |
| T2 | 7.1460a | 4.328a | 10.7640a | 1.6511a |
| T3 | 4.2310c | 2.49c | 6.7640c | 1.6991c |
| T4 | 3.8840e | 2.116e | 6e | 1.836e |
| T5 | 1.9550g | 1.577g | 3.5330g | 1.2397g |
| T6 | 2.4530b | 2.7070b | 5.1520b | 0.906b |
| T7 | 4.3890cd | 2.433cd | 6.8250c d | 1.804cd |
| T8 | 3.4110f | 1.776f | 5.1880f | 1.921f |
| CK | 3.9340d | 2.337d | 6.2740d | 1.683d |
The influence of the different LED light confrontation purpleback murdannia herb tissue-cultured seedling soluble protein and sugar contents of table 8
| Light quality ratio | Soluble sugar/(mgg-1) | Soluble protein/(mgg-1) |
| T1 | 9.5603f | 2.5910c |
| T2 | 9.4373f | 3.9841a |
| T3 | 10.658e | 3.0260b |
| T4 | 13.407a | 1.7503e |
| T5 | 12.815c | 1.6680e |
| T6 | 13.055b | 1.5082f |
| T7 | 11.317d | 2.4130d |
| T8 | 10.879e | 2.3122.3407d |
| CK | 11.215d | 2.4972.4787cd |
We are by determining plant growth parameter (table 6) and the light such as plant height, root long, stem thick, leaf area, dry weight, fresh weight
The physical signs such as pigment content (table 7), chlorophyll fluorescence parameters (Fig. 1) and soluble sugar, soluble protein content (table 8) are closed,
Analyze the influence of different LED light matter and proportioning processing to purpleback murdannia herb tissue-cultured seedling.Relative to common fluorescent as light source, it has been found that
450~480nm blue light illuminations are remarkably improved Chlorophyll and soluble protein content, promote root growth, and plant is sturdy,
Biomass increase, but suppress purpleback murdannia herb aerial part elongation growth.And 630nm feux rouges and 735nm far-red lights are then led as light source
Plant height is promoted to extend, internal soluble sugar content increase, but be unfavorable for taking root, and internal Chlorophyll synthesis is significantly inhibited, it is raw
Object amount is also accordingly reduced.Red blue light combination can reach Overlay, wherein with blue light and feux rouges 3：1 mix and match treatment effect is most
It is good.Compared to common fluorescent, T2 processing (3B1R) makes purpleback murdannia herb take root number increase by 75%, and biomass increase nearly 80% is soluble
Protein content improves 59.5%, and Chlorophyll content improves 71.6%, chlorophyll fluorescence parameters Fv/Fm, Y (II), qp and NPQ values
There is a rise, plant photosynthetic capability is stronger.
5th, influences of the LED to purpleback murdannia herb active component
Purpleback murdannia herb has very high medical value, and extract has very high anti-oxidant, antitumor and antiviral activity and resisted
Lipid peroxidation, therefore can be treating the diseases such as food, toadstool, drug poisoning, wherein polyphenol, flavones, saponin(e equal size
In notable dosage effect.After purpleback murdannia herb new life seedling is seeded into new culture medium and continuously cultivated 30 days, investigate at different LED light matter
Reason detects to plant activity in vivo composition.The total phenol of plant (TP) content is determined with Folin-Ciocalteu methods；Use NaN02-
AI (N03) 3 colorimetric method for determining plant flavonoids (Flavonoid) content；It is clear with azanol oxidizing process measure plant superoxide anion
Removing solid capacity；With Fenton method measuring plants Hydroxyl radical-scavenging abilities；With ultraviolet absorption method measure plant catalase (CAT),
The content of peroxidase (POD)；With FRAP methods measure plant TAC (T-AOC).
The different LED light matter of table 9 handle the influence to total phenol, flavonoids in plant body
| Light quality ratio | Total phenol (mg/g) | Flavonoids (mg/g) |
| T1 | 19.0097c | 25.8747b |
| T2 | 21.1757a | 31.1291a |
| T3 | 19.2535b | 24.2885c |
| T4 | 16.0270e | 20.5521d |
| T5 | 12.8461g | 17.7453e |
| T6 | 10.0918h | 10.0242h |
| T7 | 18.1483d | 15.2380f |
| T8 | 15.8380f | 14.5463g |
| CK | 18.0903d | 16.0280f |
The different LED light matter of table 10 handle the influence to oxidation resistance in plant body
From result in table 9 and table 10, compared to common fluorescent, T2 processing (3B1R) is to purpleback murdannia herb effective active composition
Accumulation has notable synergistic effect.Flavonoids improves nearly 2 times, and superoxide anion is removed and about 1.5 times of TAC increase, can
As the high-quality light source of plant growth, medicinal active ingredient accumulation is effectively facilitated.
Embodiment 4, transplanting medium optimization
3 months big bottle seedlings of taking root are moved on into warmhouse booth, add 75% to shelter from heat or light for two layers.First choice unscrews bottle cap and is allowed to ventilative,
Bottle cap is opened after adapting it to external environment within two weeks to be placed, rooted seedling is taken out from bottle, cleans the culture medium of root, is used
After 0.1% carbendazim soaks 5~10min, pull out and dry, move into matrix.Irrigated with sprayer, seedling is bonded with matrix.
Ambient humidity is kept within subsequent two weeks to keep humidity 50~60% after seedling survives 80% or so.
Sterile transplantation of seedlings carries out growth indexes measure after 2 months.Survey item has：Survival rate, average plant height, individual plant leaf
Number, increase bud number, fresh weight newly.Plant height vernier caliper measurement (is accurate to 0.01mm).As a result it is as shown in table 11.
The sterile transplantation of seedlings of table 11 carries out growth indexes measurement result after 2 months
| Matrix | Plant height (cm) | The number of sheets | Fresh weight (g) | Survival rate (%) |
| T1 | 13.57 | 6.9 | 0.97 | 86.5 |
| T2 | 11.71 | 6.1 | 0.84 | 70.5 |
| T3 | 12.52 | 6.3 | 1.11 | 80.2 |
| T4 | 13.09 | 6.6 | 1.03 | 78.6 |
| T5 | 13.78 | 6.5 | 1.16 | 73.2 |
| T6 | 10.89 | 6.1 | 1.0 9 | 70 |
| T7 | 11.08 | 6.9 | 1.04 | 71.9 |
| T8 | 14.31 | 7.1 | 1.17 | 73.4 |
| T9 | 12.53 | 7.0 | 1.13 | 82.3 |
It can be seen that purpleback murdannia herb adaptability is stronger, survival rate is up to more than 70%, but different disposal grows feelings in different substrates
Condition differs greatly.Result of the test shows, survival rate highest after coconut palm chaff is transplanted as matrix, after sphagna transplant survival seedling growth compared with
It is good, but to compare coconut palm chaff low as matrix for survival rate, may be relevant with water holding capacity.Integrated comparative is most suitable for the life of purpleback murdannia herb test tube seedling
Long matrix coconut palm chaff.It can be seen that difference, the purpleback murdannia herb that attribute is given birth to for ground come the hot bandwidth for having aerial root with other
Say, the preferable coconut palm chaff cultivation of ventilative and water conservation is advantageous to aseptic seedling and survives and improve its growth rate.
Above-described embodiment is the preferable embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any Spirit Essences without departing from the present invention with made under principle change, modification, replacement, combine, simplification,
Equivalent substitute mode is should be, is included within protection scope of the present invention.
Claims (9)
1. a kind of breeding method of purpleback murdannia herb, it is characterised in that comprise the following steps：Aseptic seeding, squamous subculture, acclimatization and transplantses；
(1) aseptic seeding：Aseptic seeding is carried out after the purpleback murdannia herb capsule that 28~30 days are grown after pollination is plucked；The group of culture medium
Turn into：Spend precious No. 1 culture medium+sucrose 10~30g/L+6-BA, 0.5~1.5mg/L+NAA, 0.1~0.2mg/L+ activated carbons 0.5
~1g/L+ banana puree 10~30g/L+, 6~8g/L of agar, pH=5.6~5.8；Condition of culture：20~40 μm of o1m of light intensity-2·
s-1, 12~16h/d of photoperiod cultures 25~60 days；
(2) squamous subculture：The protocorm that step (1) obtains is moved into subculture growth medium, induction protocorm Stem nematode and quick
Seedling, condition of culture are as follows：Using LED as tissue culture light source, 620~660nm of feux rouges, blue light 450~480nm of Blue, press
Ratio 2~4：6~8 proportionings, 30~60 μm of o1m of light intensity-2·s-1, 12~16h/d of photoperiod cultures 30~60 days, improve and plant
The thing speed of growth；The composition of subculture growth medium is：Spend treasured No. 1 culture medium+10~30g/L+6-BA of sucrose, 0.5~3mg/L
+ NAA 0.2~0.5mg/L+ activated carbon 0.5~1g/L+ banana puree 10~30g/L+, 6~8g/L of agar, pH=5.6~5.8；
(3) purpleback murdannia herb test tube transplantation of seedlings：The plant height that step (2) is obtained is cultivated up to the sterile transplantation of seedlings of more than 7cm to greenhouse
Management；Cultivation matrix includes：At least one of coconut palm chaff, peat soil, peanut shell, bark and sphagna；Cultivation condition：Two after transplanting
Keep humidity 60%~80%, 75% two layers of shade week.
2. the breeding method of purpleback murdannia herb according to claim 1, it is characterised in that：
Step (1), (2), the culture described in (3) are unless otherwise specified 40~60% conditions in 20~30 DEG C, relative humidity
Lower progress.
3. the breeding method of purpleback murdannia herb according to claim 1, it is characterised in that：
Described feux rouges is the feux rouges that wavelength is 630nm.
4. the breeding method of purpleback murdannia herb according to claim 1, it is characterised in that：
Sowing culture medium described in step (1) is to spend precious No. 1 culture medium+6-BA 0.5mg/L+NAA 0.2mg/L+ activated carbon
1g/L+ sucrose 30g/L+ banana puree 30g/L+ agar 7g/L, pH=5.6~5.8.
5. the breeding method of purpleback murdannia herb according to claim 1, it is characterised in that：
Subculture growth medium described in step (2) is to spend precious No. 1 culture medium+6-BA 1.0mg/L+NAA 0.5mg/L+ to live
Property charcoal 0.5g/L+ sucrose 30g/L+ banana puree 30g/L+ agar 7g/L, pH=5.6~5.8.
6. the breeding method of purpleback murdannia herb according to claim 1, it is characterised in that：
Condition of culture described in step (2) is：Using LED as tissue culture light source, 450~480nm of blue light Blue and feux rouges
630nm presses 3：1 mix and match, 45 μm of o1m of light intensity-2·s-1, photoperiod 16h/d cultures 60 days, environment temperature is 24 ± 1 DEG C,
Relative humidity 50~60%.
7. the breeding method of purpleback murdannia herb according to claim 1, it is characterised in that：
Newborn seedling described in step (2) for after planting protocorm stem length to 2.5~3.5cm newborn seedling.
8. the breeding method of purpleback murdannia herb according to claim 1, it is characterised in that：
Cultivation matrix described in step (3) is：Using coconut palm chaff as transplanting medium, environment temperature is 24 ± 1 DEG C, relative humidity
60%~80%.
9. application of the cultural method of any one of claim 1~8 purpleback murdannia herb in large-scale cultivation purpleback murdannia herb.
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| CN109136256A (en) * | 2018-08-01 | 2019-01-04 | 广东省农业科学院环境园艺研究所 | A kind of orchid gene transformation method |
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| CN109601379A (en) * | 2018-12-07 | 2019-04-12 | 广东省农业科学院环境园艺研究所 | A kind of method and its application promoting the development of purpleback murdannia herb seed fast-growth |
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| CN115362936A (en) * | 2022-06-30 | 2022-11-22 | 广东省农业科学院环境园艺研究所 | Cultivation method of hybrid orchid dragon roots |
| CN115644057A (en) * | 2022-09-22 | 2023-01-31 | 广东省农业科学院环境园艺研究所 | Culture medium combination and method for rapid propagation of oncidium glaucescens |
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