CN103843662B - A kind of promote Herba Dendrobii tissue cultured seedling strong sprout and the method taken root - Google Patents
A kind of promote Herba Dendrobii tissue cultured seedling strong sprout and the method taken root Download PDFInfo
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Abstract
The invention discloses and a kind of promote Herba Dendrobii tissue cultured seedling strong sprout and the method taken root, comprise the steps: 1) Herba Dendrobii tissue cultured seedling is proceeded to illumination cultivation in strong sprout and defined medium of taking root, strong sprout root induction；2) tissue cultured seedling taken root can be transplanted after seedling exercising；3) by the Herba Dendrobii tissue cultured seedling Transplantation of Regenerated Plantlets taken root in the cultivation matrix that bark mixes with Vermiculitum.The present invention is simple to operate, easy, and strong sprout is fast, and average every strain plant height is high 3 4cm than matched group, and diameter stem is thick 2 3mm than matched group, fresh weight, than matched group weight 3 times, is taken root early, the formation seeing root in 45 days, rooting rate is high, and within 30 days, rooting rate reaches 100%, and average every strain takes root 78；Transplanting survival rate is high, can reach 95%.The present invention can effectively shorten the root induction time, and root system development is good, robust plant；Transplanting survival rate is high, and the cultivation to other the orchid family tissue cultured seedlinies simultaneously also has important references and is worth.
Description
Technical field
The invention belongs to plant biotechnology field, be specifically related to the tissue culture of Herba Dendrobii, especially relate to
And a kind of promote Herba Dendrobii tissue cultured seedling strong sprout and the method taken root.
Background technology
Herba Dendrobii (D.orchids) is the orchid family Dendrobium (Dendrobium) herbaceos perennial,
It is mainly used for viewing and admiring with medicinal.Herba Dendrobii is four your name internationally famous, most ornamental value
One of blue, have of a great variety, pattern is gorgeous, and attitude is graceful, spends many and close, and florescence length etc. is many
Good characteristic, can make potted plant and cut-flower and apply, the intensive commodity production of suitable China most area,
Have a extensive future.The rare Chinese medicine that Herba Dendrobii Ye Shi China is traditional, as famous Herba Dendrobii,
Herba Dendrobii, Dendrobium nobile etc., all have long applicating history in China.Modern pharmacological research shows,
Herba Dendrobii have antitumor, defying age, enhancing body immunity, expansion blood vessel and anti-platelet aggregation,
The health-care effects such as blood circulation promoting, enhancing heart vigor, have high Development volue.
Owing to Herba Dendrobii economic worth is high, wild resource is excessively excavated extremely serious, and in August, 1998 issues
" the Chinese Plants Red Data Book " of cloth almost will all list protection name in by China all of dendrobium plant
Record.Ecological environment is required strict by Herba Dendrobii, and percentage of seedgermination is extremely low, poor growth, natural renovation
Difficulty, far from meeting demand.Application tissue culture carries out Herba Dendrobii Fast-propagation, for solving money
Source is in short supply and to realize industrialization development significant.
The tissue culture of orchid starts from the sixties in 20th century, and Morel uses Cymbidium ensifolium (L.) Sw. stem apex,
Cultivating in KC culture medium containing the basic element of cell division, shoot apical meristem expands and forms class protocorm
Stem, and differentiate root and leaf, obtain the virus-free plantlet of Cymbidium ensifolium (L.) Sw. first.Wimber is to Morel's
Method improves, and uses the method for agitated submerged culture to be greatly accelerated the growth rate of protocorm,
Can obtain a large amount of regeneration plant in a short time, hereafter tissue culture technique has obtained extensively in orchid production
General application.
Strong seedling culture is to improve important measures of transplanting survival rate, and rooting efficiency also directly influences
Productivity effect.The strong sprout of orchid with take root largely with culture medium a great number of elements and trace unit
Element component, plant growth regulator kind relevant with the natural extract of concentration, interpolation, this stage
The kind that it is critical only that strict screening hormone and concentration.Minimal medium add certain density
The hormones such as NAA, IAA, IBA contribute to rooting of vitro seedling, but strong sprout, effect was the most notable.Tang Gui
The result of study of perfume etc. shows, NAA contributes to taking root of test tube Seedling, but NAA concentration is 0.5mg/L
Time, mean elements and root length value are maximum.Meng Aidong etc. utilize tissue culture technique to have studied paclobutrazol pair
The impact that Herba Dendrobii the growth of plants is cultivated, result shows, adding paclobutrazol in MS culture medium can
It is obviously promoted the growth of test tube Seedling, beneficially rooting of vitro seedling, improves transplanting survival rate；But, many
After effect azoles process, test tube Seedling is downgraded, internode shortens, leaf diminishes, stem slightly increases；The number of blade with compare nothing
Significant difference, leaf length and leaf width are substantially reduced；Root length reduces, and radical dramatically increases, and root slightly increases not
Substantially.
Liu Hua etc. think, the culture medium of optimum Herba Dendrobii the growth of plants is B5 or l/2MS base
Basal culture medium adds the NAA of 2mg/L, Fructus Musae extract 10%, sucrose 2%.Strong plantlets and rootage is fitted
Suitable culture medium also has N6+10% bananas juice, 1/2MS+IBA 0.1mg/L, 1/2MS+10% Fructus Musae
Juice+0.5%AC and improvement N6+0.2mg/LNAA+10% bananas juice etc..
Knowable to document is reported, root media is usually added into bananas juice, Sucus Cocois etc. natural attached
Adding thing can hestening rooting strong sprout.But the report such as Lv Xiuli thinks sky such as interpolation 10% Sucus Cocois, bananas juice etc.
So extract, does not has facilitation to Herba Cymbidii Goeringii tissue cultured seedling late growing stage, can cause yellow on the contrary and enter
One step is dead.We are orchids such as hybrid cymbidium, thousand generation orchids, Herba Dendrobii, hook lip Herba Dendrobiis
Group training with the addition of bananas juice, potato juice, it has been found that natural extract promotes the product of brownization on the contrary
Raw, root growth is bad, have impact on the growth of plant.
Additionally, due to the extracting method of natural extract, the Maturity extracting material and physiological status are not
With, purity and the valid density of extract are widely different, and experimental result all can produce impact.Natural
Extract is every secondary existing with existing extraction, it is impossible to preserves for a long time, operates cumbersome.At root media
Middle addition activated carbon, the beneficially growth of Herba Dendrobii test tube Seedling, can make root system look more sturdy.
Add activated carbon in the medium and be favorably improved the survival rate of test tube Seedling, but to the radical of Seedling and root length
Affect less.
Therefore the plant growth regulator that can promote hestening rooting again in strong sprout that selection is more suitable for is still
Necessary.
Summary of the invention
It is an object of the invention to provide and a kind of promote Herba Dendrobii tissue cultured seedling strong sprout and the method taken root, make
Herba Dendrobii tissue cultured seedling is taken root soon in incubation, and rooting rate is high, well developed root system, and root is sturdy, regeneration
Plant strain growth is healthy and strong and transplanting survival rate is high.
To achieve these goals, technical scheme is as follows:
A kind of promote Herba Dendrobii tissue cultured seedling strong sprout and the method taken root, comprise the steps:
1) Herba Dendrobii tissue cultured seedling is proceeded to illumination cultivation in strengthening seedling and rooting culture medium, root induction；Described
The formula of strengthening seedling and rooting culture medium be MS minimal medium+hymexazol HMI and/or sugar alcohol chelating compound
Fertilizer+agar powder+sucrose.
2) the Herba Dendrobii tissue cultured seedling taken root in strengthening seedling and rooting culture medium is transplanted after seedling exercising.
3) by the Herba Dendrobii tissue cultured seedling Transplantation of Regenerated Plantlets taken root in weight ratio be the bark of 1-3:1-3
In the cultivation matrix mixed with Vermiculitum, moisturizing of spraying water, temperature 25 ± 2 DEG C, relative humidity 70-85%.
Wherein, above-mentioned steps 1) in, described Herba Dendrobii tissue cultured seedling is the conventional organization using this area
Cultural method, utilizes the organ (such as root, bud, seed, protocorm etc.) that Vitro Plant is cultivated in nothing
Under the conditions of bacterium and suitable synthetic medium and illumination temperature etc. are artificial, the tissue training of inducing culture gained
Supporting vegetative seedling, Herba Dendrobii tissue cultured seedling herein is unrooted.
Preferably, above-mentioned steps 1) in, the formula of described strengthening seedling and rooting culture medium is that MS trains substantially
Support base+1～10mg/L hymexazol HMI and/or 1000 times of-8000 times of+5-7g/L of sugar alcohol chelate compound fertilizer
Agar powder+20-30g/L sucrose.
Preferably, above-mentioned steps 1) in, the formula of described strengthening seedling and rooting culture medium is that MS trains substantially
Support base+5mg/L hymexazol HMI and/or 1000 times of+5-7g/L agar powders of sugar alcohol chelate compound fertilizer
+ 20-30g/L sucrose.
Further, above-mentioned steps 1) in, excellent 12 elements of described sugar alcohol chelate compound fertilizer preferred Huo Shang Australia
Liquid composite fertilizer.
Above-mentioned steps 1) in, Herba Dendrobii tissue cultured seedling is aseptically cut seedling, proceeds to strong sprout
In root media, illumination cultivation condition is: cultivation temperature is 25 DEG C ± 2 DEG C, and light application time is 12-16
Hour/day, intensity of illumination is 1000-2000lux.
Above-mentioned steps 2) in, seedling is general in strengthening seedling and rooting culture medium to be cultivated 4-5 week, now children
Seedling robust growth, stem is sturdy, and leaf color is dark green, well developed root system, is gradually opened lid in culturing room,
Seedling is removed bottle in one week by seedling exercising.
The excellent 12 element liquid composite fertilizers of Huo Shang Australia that the present invention is used in the medium are Australia Ai Erfu
The 12 element liquid composite fertilizers (broad spectrum foliar) of company's research and development in 1991.
The hymexazol that the present invention is used in the medium, chemical name is 3-hydroxy-5-methyl base isoxazole,
English name is Hymemzo1 (being abbreviated as HMI, hereafter repeat no more), is that absorption is killed in one
Microbial inoculum and seed disinfectant.It is under suitable concentration, right in addition to having good bactericidal action
The growth of various crop has regulation effect.Hymexazol imposes on rice seedling bed, can promote root growth, increases
Add tiller, improve seedling quality；Germination percentage can be improved with hymexazol leaching seed rice, increase plant weights,
Process soil with 70% hymexazol wettable powder, increase production Oryza glutinosa 12%；Additionally, hymexazol is to Ma Ling
Potato, Bulbus Allii Cepae, Fructus Fragariae Ananssae, Radix Betae, Semen Pisi sativi all have promotion growth and production-increasing function.Have no that it is at stone before this
The report of application in Hu Lan tissue culture.
Plant desired nutritional element either a great number of elements N P and K or trace element sulfur in culture medium
The existences such as magnesium zinc boron and concentration proportioning are all the biggest to plant strong sprout and Rooting effect.Sugar alcohol material
Mineral nutrient fast transportation in plant phloem can be carried as other nutrient carriers such as boron.
Sugar alcohol chelating chemical fertilizer is the good chelating agent of the nutrients such as middle and trace element, can be with multiple nutrients thing
Matter combines and forms stable complex, has the advantage that
1. it is that the currently the only mineral nutrient that can carry carries out the material of fast transportation in phloem；
2. be the natural extract in plant phloem juice, nontoxic, to plant, human body without any damage
Wound；
3. molecular weight is low, it is easy to absorbed by blade, and easily degrading in entering into plant body, it is foster to discharge
Point, consume energy low；
4. being stable existence form with liquid, especially in alkaline solution, dissolubility is higher.Due to tough
It is alkaline environment in skin zone, major part metal class mineral nutrient dissolubility and mobility under alkaline environment
The most poor, and sugar alcohol complex more can embody it and can carry the advantage that mineral nutrient moves at phloem；
5. can improve the resistance of plant, on the one hand, sugar alcohol is the weight participating in intracellular penetration regulation
Wanting material, plant is under the environment stresses such as salt damage, arid, waterflooding, and sugar alcohol can be oozed by regulation cell
Property makes plant adapt to adverse growth thoroughly；On the other hand, sugar alcohol can improve the resistance to active oxygen, keeps away
Exempt from the plant-active oxygen injury caused due to reasons such as ultraviolet day bright, arid, disease, anoxias.
Yet there are no sugar alcohol chelate compound fertilizer report of application in the training of orchid group.
The present invention utilizes the MS culture medium containing HMI and/or sugar alcohol chelate compound fertilizer to Herba Dendrobii group
Seedlings cultivating carries out root induction, drastically increases rooting rate, and transplanting survival rate also significantly improves.
Compared with prior art, there is advantages that
1. utilizing the root media root induction of the present invention, its rooting rate, up to 100%, is transplanted into
Motility rate is up to 95%.
2. the present invention can effectively shorten the root induction time, the generation just seeing root in 4-5 days, and root system
Sturdy, root is many, average 7-8 the root of each plant.
3. due to the fact that the certain density HMI of use and/or sugar alcohol chelate compound fertilizer, promote life
While root, improve the plant quick absorption to nutrient, plant strain growth is fast and healthy and strong, and stem is thick, leaf
Thickness, leaf color is dark green, reaches the purpose in strong sprout.
Operational approach the most of the present invention is simple, low cost.Be not added with other growth regulators and other
Different natural additament, such as Rhizoma Solani tuber osi, bananas juice, Sucus Mali pumilae, coconut juice etc., and the feelings of activated carbon
Under condition, i.e. can reach take root fast, rooting rate height, plant strain growth stalwartness, the effect that survival rate is high, joint
About cost, simplifies operation sequence.
Accompanying drawing explanation
Fig. 1 is the upgrowth situation of the embodiment of the present invention 1 plant.
Fig. 2 is the plant that takes root of the embodiment of the present invention 1.
Fig. 3 is the plant of the embodiment of the present invention 1.
Fig. 4 is the plant that takes root of the embodiment of the present invention 2.
Fig. 5 is the upgrowth situation of the embodiment of the present invention 2 plant.
Fig. 6 is the plant (left) plant (right) comparison diagram with embodiment 1 of the embodiment of the present invention 3.
Detailed description of the invention
Below in conjunction with accompanying drawing and specific embodiment, technical scheme is described in further detail.
Embodiment 1
The candidum tissue culturing seedling strengthening seedling and rooting cultural method of the present embodiment, specifically comprises the following steps that
1) take and come from the Herba Dendrobii unrooted tissue cultured seedling high for 2cm of the outer implant of stem section, under aseptic condition
Cut seedling, proceed to the culture bottle equipped with strengthening seedling and rooting culture medium is cultivated, culture bottle is placed in illumination
Under the conditions of cultivate, cultivation temperature is 25 DEG C ± 2 DEG C, and light application time is 16 hours/every day, and illumination is strong
Degree is 1000lux；The formula of Rooting and hardening-off culture base is MS minimal medium+5mg/L HMI+5g/L
Agar powder+30g/L sucrose.
Cultivate the generation just having root for 4-5 days, add up rooting rate after cultivating one month and reach 100% (see Fig. 1),
Average every strain is taken root 6-8 root, root length plant height average out to 7.1-7.8cm (see Fig. 2), Ye Senong
Green, leaf is big and thick, the average 0.47-0.5cm of diameter stem (see Fig. 3), average every strain fresh weight 1.4-1.6g.
2) seedling cultivates 4-5 week in strengthening seedling and rooting culture medium, and now seedling is healthy and strong, well developed root system,
Progressively open bottle cap seedling exercising one week, seedling is removed bottle, rinse the culture medium of root with clear water, transplant
In the cultivation matrix that bark mixes with Vermiculitum 1:1, moisturizing of spraying water.Temperature 25 DEG C ± 2 DEG C, relatively
Humidity 70%.The survival rate of plant of cultivation is high, reaches 95%.Plant strain growth is healthy and strong.
Embodiment 2
The candidum tissue culturing seedling strengthening seedling and rooting cultural method of the present embodiment, specifically comprises the following steps that
1) take and come from the Herba Dendrobii unrooted tissue cultured seedling high for 2cm of the outer implant of stem section, under aseptic condition
Cut seedling, proceed to the culture bottle equipped with strengthening seedling and rooting culture medium is cultivated, culture bottle is placed in illumination
Under the conditions of cultivate, cultivation temperature is 25 DEG C ± 2 DEG C, and light application time is 16 hours/day, intensity of illumination
For 1000lux；The formula of Rooting and hardening-off culture base is excellent 12 elements of MS minimal medium+Huo Shang Australia
1000 times of+5g/L agar powder+30g/L sucrose of liquid composite fertilizer；
Cultivate the generation having root for 15 days, add up rooting rate after cultivating 45 days and reach 100%, average every strain
Taking root 7-8 root, root is sturdy, neat (see Fig. 4), plant height average out to 8.1-8.8cm, leaf color
Dark green (see Fig. 5), leaf is big and thick, the average 0.5-0.6cm of diameter stem, average every strain fresh weight 1.7-1.8g.
2) seedling cultivates 4-5 week in strengthening seedling and rooting culture medium, and now seedling is healthy and strong, well developed root system,
Progressively open bottle cap seedling exercising one week, seedling is removed bottle, rinse the culture medium of root with clear water, transplant
In the cultivation matrix that bark mixes with Vermiculitum 1:1, moisturizing of spraying water.Temperature 25 DEG C ± 2 DEG C, relatively
Humidity 70%.The survival rate of plant of cultivation is high, reaches 95%.Plant strain growth is healthy and strong.
Embodiment 3
The candidum tissue culturing seedling strengthening seedling and rooting cultural method of the present embodiment, specifically comprises the following steps that
1) take and come from the Herba Dendrobii unrooted tissue cultured seedling high for 2cm of the outer implant of stem section, under aseptic condition
Cut seedling, proceed to the culture bottle equipped with strengthening seedling and rooting culture medium is cultivated, culture bottle is placed in illumination
Under the conditions of cultivate, cultivation temperature is 25 DEG C ± 2 DEG C, and light application time is 16 hours/day, intensity of illumination
For 1000lux；The formula of Rooting and hardening-off culture base is MS minimal medium+5mg/L HMI+ Huo Shang
1000 times of+5g/L agar powder+30g/L sucrose of the excellent 12 element liquid composite fertilizer of Australia；
Cultivating the generation having root for 5 days, add up rooting rate and reach 100% after cultivating 25 days, average every strain is raw
Root 7-8 root, root is sturdy, and neatly, plant strain growth gesture is better than the process (see Fig. 6) only adding HMI,
Leaf color is dark green, and leaf is big and thick.
2) seedling cultivates 4-5 week in strengthening seedling and rooting culture medium, and now seedling is healthy and strong, well developed root system,
Progressively open bottle cap seedling exercising one week, seedling is removed bottle, rinse the culture medium of root with clear water, transplant
In the cultivation matrix that bark mixes with Vermiculitum 1:1, moisturizing of spraying water.Temperature 25 DEG C ± 2 DEG C, relatively
Humidity 70%.The survival rate of plant of cultivation is high, reaches 95%.Plant strain growth is healthy and strong.
Embodiment 4 matched group
1) take and come from the outer implant of stem section, Herba Dendrobii unrooted tissue cultured seedling high for 2cm, aseptic condition
Under cut seedling, proceed to equipped be not added with in the culture bottle of any hormone MS culture medium cultivate, will cultivate
Bottle is placed under illumination condition cultivation, and cultivation temperature is 25 DEG C ± 2 DEG C, and light application time is 16 hours/day,
Intensity of illumination is 1000lux；The formula of culture medium is MS culture medium+5g/L agar powder+30g/L sugarcane
Sugar；
Cultivate the generation having root for 20 days, add up rooting rate after cultivating one month and reach 20%, average every strain
Taking root 3, plant height average out to 3.4cm, plant is tiny, the average 0.2cm of diameter stem, averagely
Every strain fresh weight 0.5g.
2) seedling cultivates 4-5 week in the medium, progressively opens bottle cap seedling exercising one week, is removed by seedling
Bottle, rinses the culture medium of root, is transplanted to the cultivation matrix that bark mixes with Vermiculitum 1:1 with clear water
In, moisturizing of spraying water.Temperature 25 DEG C ± 2 DEG C, relative humidity 70%.The survival rate of plant of cultivation is only
40%.Plant growing way is slow.
It is last it should be noted that, above example is only in order to illustrate technical scheme rather than limit
System, although the present invention being described in detail with reference to preferred embodiment, the ordinary skill people of this area
Member should be appreciated that and can modify the technical scheme of invention or equivalent, without deviating from this
The spirit and scope of inventive technique scheme, it all should be contained in scope of the presently claimed invention.
Claims (4)
1. promote Herba Dendrobii tissue cultured seedling strong sprout and the method taken root, comprise the steps:
1) Herba Dendrobii tissue cultured seedling is proceeded to illumination cultivation in strengthening seedling and rooting culture medium, root induction；Described
The formula of strengthening seedling and rooting culture medium be MS minimal medium+1～10mg/L hymexazol HMI and/
Or 1000 times-8000 times+5-7g/L agar powder+20-30g/L sucrose of sugar alcohol chelate compound fertilizer；Its
In, described sugar alcohol chelate compound fertilizer is the excellent 12 element liquid composite fertilizers of Huo Shang Australia；
2) the Herba Dendrobii tissue cultured seedling taken root in strengthening seedling and rooting culture medium is transplanted after seedling exercising；
3) cultivation that the Herba Dendrobii tissue cultured seedling Transplantation of Regenerated Plantlets taken root is mixed with Vermiculitum in bark
In substrate, moisturizing of spraying water, temperature 25 ± 2 DEG C, relative humidity 70-85%.
2. promoting Herba Dendrobii tissue cultured seedling strong sprout and the method taken root as claimed in claim 1, its feature exists
In, the formula of described strengthening seedling and rooting culture medium is MS minimal medium+5mg/L hymexazol HMI
And/or 1000 times of+5-7g/L agar powder+20-30g/L sucrose of sugar alcohol chelate compound fertilizer.
3. promoting Herba Dendrobii tissue cultured seedling strong sprout and the method taken root as claimed in claim 1, its feature exists
In, step 1) in illumination cultivation condition be: cultivation temperature is 25 DEG C ± 2 DEG C, and light application time is
12-16 hour/day, intensity of illumination was 1000-2000lux.
4. promoting Herba Dendrobii tissue cultured seedling strong sprout and the method taken root as claimed in claim 1, its feature exists
In, step 3) described in the cultivation matrix that mixes with Vermiculitum of bark both weight ratios be 1-3:
1-3。
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| CN103843662A (en) | 2014-06-11 |
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