CN106416972A - Method for quickly cultivating bletilla striata seedlings - Google Patents
Method for quickly cultivating bletilla striata seedlings Download PDFInfo
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- CN106416972A CN106416972A CN201610803670.6A CN201610803670A CN106416972A CN 106416972 A CN106416972 A CN 106416972A CN 201610803670 A CN201610803670 A CN 201610803670A CN 106416972 A CN106416972 A CN 106416972A
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- bletillae
- rhizoma bletillae
- pseudobulbus bletillae
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G31/00—Soilless cultivation, e.g. hydroponics
Abstract
The invention discloses a method for quickly cultivating bletilla striata seedlings. The method comprises the steps of performing germination cultivation on bletilla striata seeds, performing protocorm differentiation cultivation and performing bulb induction cultivation and domestication to obtain the bletilla striata seedlings. Compared with the prior art, the method provided by the invention has the advantages of high germination rate, short cultivation time, high regularity and low aberration rate, and has a good market prospect.
Description
Technical field
The invention belongs to field of plant growing technology is and in particular to a kind of method of fast culture Pseudobulbus Bletillae (Rhizoma Bletillae) seedling.
Background technology
The Pseudobulbus Bletillae (Rhizoma Bletillae) is orchid family draft bulbous plant, also known as Rhizoma Bletillae, Lian Jicao, Gan Gen, gives free of charge, and plant is high 18-60 centimetre.The Pseudobulbus Bletillae (Rhizoma Bletillae)
Nascent pseudobulb be spheroidal, growth to a certain extent just formed V-shaped bulk pseudobulb.4-6 piece of leaf, long and narrow circular or drape over one's shoulders
Aciculiform, long 8-29 centimetre, wide 1.5-4 centimetre, tip is tapering, and base portion is received narrow one-tenth sheath and embraced stem.The nice and cool weather of Pseudobulbus Bletillae (Rhizoma Bletillae) happiness and humic
The abundant and well-drained sandy loam of matter, also can cultivate half light, but arid, high temperature, blade can be made to wither yellow.Late March is sprouted
Bud, ground die back after frost, the pollen of the Pseudobulbus Bletillae (Rhizoma Bletillae) is difficult to scatter in bulk, so being not highly advantageous in pollination.The Pseudobulbus Bletillae (Rhizoma Bletillae)
Seed is superfine little, and like powder, shape does not have endosperm, the embryo of the ateliosis of only several cellularities, and seed germination rate is extremely low,
And natural propagation is difficult, so wild resource is sparser, is a kind of protection plant of Precious, Rare, Endangered of China.
The Pseudobulbus Bletillae (Rhizoma Bletillae) belongs to nine species, and originate in China just has four kinds, and the Pseudobulbus Bletillae (Rhizoma Bletillae) all has in China Yangtze river basin and areas to the south
Distribution, growth district is wider.Due to for many years predation formula excavation, add habitat day deteriorate so that the quantity of wild bletilla striata drastically
Reduce, endangered edge.
The main chemical compositions of the Pseudobulbus Bletillae (Rhizoma Bletillae) are bibenzyl, luxuriant and rich with fragrance class and its derivant, additionally, also containing a small amount of volatile oil, mucus
Matter, bletilla mannan and starch (30.5%), glucose (1.5%) etc..Recent years, the chemical constitution study to Pseudobulbus Bletillae (Rhizoma Bletillae)
It is concentrated mainly on bibenzyl, dihydro phenanthrene class and Lian Fei class compound, and therefrom isolation identification goes out a series of new bibenzyls, dihydro
Phenanthrene and connection phenanthrene compound.
The Pseudobulbus Bletillae (Rhizoma Bletillae) has the effect of astringing to arrest bleeding, heat-clearing and toxic substances removing, detumescence and promoting granulation.With the development of China's Chinese medicine, in vain
Splendid achnatherum range of application clinically constantly expands, and the medication demand of the Pseudobulbus Bletillae (Rhizoma Bletillae) sharply increases.But bletilla striata seeds are under field conditions (factors) very
Difficult sprouting and growth, the cultivation of seedling is more difficult, only leans on division propagation, and the cycle is long, and reproductive efficiency is low, and consumes kind of an amount
Greatly, it is difficult to meet the needs of a large amount of cultivations.The Pseudobulbus Bletillae (Rhizoma Bletillae) is faced with the stem reality that resource is difficult to meet market.
Content of the invention
The technical problem to be solved, is to provide a kind of method of fast culture Pseudobulbus Bletillae (Rhizoma Bletillae) seedling.
For solving above-mentioned technical problem, it is raw material that the present invention chooses bletilla striata seeds, using MS substrate or the culture of 1/2MS substrate
Method carries out the mode of sowing that suspends, and promotes the sprouting of bletilla striata seeds and takes root, specific technical scheme is as follows：
A kind of method of fast culture Pseudobulbus Bletillae (Rhizoma Bletillae) seedling, it comprises the following steps：
(1) Pseudobulbus Bletillae (Rhizoma Bletillae) capsule is cleaned with clear water, and detergent solution soaks 10min, then is cleaned with scour without sediment motion, in ultra-clean work
Adopt 75% ethanol disinfection 30s on platform, then sterilized 10～15min with mercuric chloride solution, with aseptic water washing 3～4 times, obtain
The Pseudobulbus Bletillae (Rhizoma Bletillae) is aseptic to mould fruit.
(2) aseptic the moulding of the Pseudobulbus Bletillae (Rhizoma Bletillae) of step (1) gained is really cut open with dissecting knife on super-clean bench, takes out seed, is inoculated in kind
Son is sprouted on fluid medium, under conditions of 22～25 DEG C, 80～120 turns/min, cultivates 40d, is filtrated to get Pseudobulbus Bletillae (Rhizoma Bletillae) protocorm
Stem；
(3) Pseudobulbus Bletillae (Rhizoma Bletillae) protocorm of gained in step (2) is inoculated in protocorm differentiation culture medium, in 25～28 DEG C, light
Cultivate 60d according under conditions of intensity 1000～4000lux, light application time 11～13h/d, obtain Pseudobulbus Bletillae (Rhizoma Bletillae) seedling；
(4) Pseudobulbus Bletillae (Rhizoma Bletillae) seedling of gained in step (3) is inoculated in bulb inducing culture, in 25～28 DEG C, illuminance
Cultivate 60d under conditions of 1000～4000lux, light application time 11～13h/d, choose base portion and form the white of diameter 4～7mm bulb
Splendid achnatherum；
(5) after the Pseudobulbus Bletillae (Rhizoma Bletillae) chosen in step (4) being cleaned with water, it is displaced on the seedbed being covered with domesticating and cultivating substrate, hide
Cloudy 80%, after taming 6 months, gained Pseudobulbus Bletillae (Rhizoma Bletillae) seedling can carry out field-transplanting.
Wherein, in step (1), described detergent is normal domestic use liquid detergent or detergent；Described mercuric chloride is water-soluble
In liquid, the mass fraction of mercuric chloride is 0.1%～0.15%.
Wherein, in step (2), the group that described seed sprouting fluid medium is is divided into：1/2MS+1.0mg/L 6-BA (or
Claim " 6- benzylaminopurine ", be a kind of " basic element of cell division ")+6.8-7.5g/L agar, Ph5.6-5.8.
Wherein, the condition of culture in step (2) is：Under the conditions of 22～25 DEG C, front 7d gives light culture, and illumination is strong afterwards
Spend for 1000～4000lux, preferably 1500-2500lux, particularly preferred 2000lux, light application time 11～13h/d, preferably 12h/
d.
Wherein, in step (3), the group of described protocorm differentiation culture medium is divided into：MS+0.1mg/L NAA (or claim naphthalene second
Acid)+1.0mg/L 6-BA+7.5%wt bananas juice+6.8-7.5g/L agar, Ph5.6-5.8, the preferred 1500- of intensity of illumination
2500lux, particularly preferred 2000lux, the preferred 12h/d of light application time.
Wherein, in step (4), the group of described bulb inducing culture is divided into：1/2MS culture medium+1.0mg/L NAA+
7.5%wt bananas juice+6.8～7.5g/L agar, pH 5.6～5.8, the preferred 1500-2500lux of intensity of illumination, particularly preferably
2000lux, the preferred 12h/d of light application time；
Wherein, in step (5), described domesticating and cultivating substrate compares 1: 1: 0.5 by rural area soil, peat soil and Vermiculitum with weight
Composition.
Beneficial effect：Compared with prior art, the invention has the advantages that：
1st, after the Pseudobulbus Bletillae (Rhizoma Bletillae) moulds fruit sterilization, it is placed on superclean bench to be cut open with dissecting knife and moulds fruit, take out seed, be inoculated into step
(2) on fluid medium in, using 1/2MS minimal medium and be equipped with auxin 6-BA 1.0mg/L, it is 80～120 in rotating speed
Turn/min under the conditions of, first light culture, then illumination cultivated, and compared with existing bletilla striata seeds germination medium, inoculates speed
Improve 2-3 times, and germination rate is high (more than 85%), regularity is good, aberration rate is less than 0.8%.
2nd, in the protocorm differentiation culture medium described in step (3), using MS (a great number of elements) and be equipped with auxin NAA
0.1mg/L and 6-BA 1.0mg/L, compared with traditional culture technique, the method more preferably can promote protocorm differentiation shape faster
Become Pseudobulbus Bletillae (Rhizoma Bletillae) seedling.
3rd, in the bulb inducing culture described in step (4), using 1/2MS (a great number of elements halves) and be equipped with auxin
NAA1.0mg/L and 7.5% bananas juice, compared with traditional culture technique, the method more preferably can promote Pseudobulbus Bletillae (Rhizoma Bletillae) seedling faster
Base portion expands, and forms bulb, the bulb of bletilla tissue culture seedlings can be made in 60d to expand diameter 4～7mm.
Specific embodiment
According to following embodiments, the present invention may be better understood.However, it is as it will be easily appreciated by one skilled in the art that real
Apply the content described by example and be merely to illustrate the present invention, and should not be also without limitation on basis described in detail in claims
Invention.
Embodiment 1：A kind of method of fast culture Pseudobulbus Bletillae (Rhizoma Bletillae) seedling, the method comprises the following steps：
(1) September, after the clip Pseudobulbus Bletillae (Rhizoma Bletillae) artificial pollination 120d mould fruit, clean 10min with detergent, then rinsed with flowing water
10min, brings transfer room into, uses the alcohol-pickled 45～60s of 75v/v%, then be positioned over 0.1% mercuric chloride water on superclean bench
Soak in solution；
(2) aseptic the moulding of the Pseudobulbus Bletillae (Rhizoma Bletillae) that step (1) obtains really is cut open with dissecting knife on superclean bench, take out seed, connect
Plant and sprout on fluid medium in seed, under conditions of 22 DEG C, 120 turns/min, cultivate 15d, seed starts to sprout, after 40d,
By micro- sem observation, the protocorm of about 88% Seed Development straight stem 0.7～1mm, aberration rate is about 0.75%, leaches protocorm
Stem is standby；Wherein, illumination condition is：Front 7d gives light culture, and illuminance is 1000lux, light application time 12h/d later；Described
Fluid medium consist of：1/2MS+1.0mg/L 6-BA+6.8g/L agar, Ph5.6；
(3) Pseudobulbus Bletillae (Rhizoma Bletillae) protocorm obtaining step (2) is placed in protocorm differentiation culture medium, in 25 DEG C, illuminance
Cultivate 60d under conditions of 1000lux, light application time 12h/d, obtain 5cm high left and right Pseudobulbus Bletillae (Rhizoma Bletillae) seedling；Wherein, described protocorm
Division culture medium consists of：MS+0.1mg/L NAA+1.0mg/L 6-BA+7.5%wt bananas juice+6.8g/L agar, Ph5.6-
5.8；
(4) Pseudobulbus Bletillae (Rhizoma Bletillae) seedling obtaining in step (3) is inoculated in bulb inducing culture, in 28 DEG C, illuminance
60d is cultivated, the base portion of the Pseudobulbus Bletillae (Rhizoma Bletillae) expands formation bulb, measured, the squama of formation under conditions of 1500lux, light application time 12h/d
Stem a diameter of 4～7mm mostly, average diameter is 5.81mm, at this moment can carry out rooting culture, and wherein, described bulb induction is trained
Support basis set becoming：1/2MS culture medium+1.0mg/L NAA+7.5%wt bananas juice+7.5g/L agar, pH 5.6；
(5) seedling obtaining step (4) taking-up clear water in bottle washes off after culture medium, and dislocation is covered with domestication and plants
On the seedbed of training substrate, and moisturizing of sheltering from heat or light, after taming 6 months, gained Pseudobulbus Bletillae (Rhizoma Bletillae) seedling can carry out field-transplanting.
Embodiment 2：A kind of method of fast culture Pseudobulbus Bletillae (Rhizoma Bletillae) seedling, the method comprises the following steps：
(1) September, after the clip Pseudobulbus Bletillae (Rhizoma Bletillae) artificial pollination 120d mould fruit, clean 10min with detergent, then rinsed with flowing water
10min, brings transfer room into, uses the alcohol-pickled 45～60s of 75v/v%, then be positioned over 0.15% mercuric chloride on superclean bench
Soak in aqueous solution；
(2) aseptic the moulding of the Pseudobulbus Bletillae (Rhizoma Bletillae) that step (1) obtains really is cut open with dissecting knife on superclean bench, take out seed, connect
Plant and sprout on fluid medium in seed, under conditions of 25 DEG C, 120 turns/min, cultivate 15d, seed starts to sprout, after 40d,
By micro- sem observation, the protocorm of about 90% Seed Development straight stem 0.7～1mm, aberration rate is about 0.76%, leaches protocorm
Stem is standby；Wherein, illumination condition is：Front 7d gives light culture, and illuminance is 1500lux, light application time 12h/d later；Described
Fluid medium consist of：1/2MS+1.0mg/L 6-BA+7.5g/L agar, Ph5.6；
(3) Pseudobulbus Bletillae (Rhizoma Bletillae) protocorm obtaining step (2) is placed in protocorm differentiation culture medium, in 25 DEG C, illuminance
Cultivate 60d under conditions of 2000lux, light application time 12h/d, obtain 5cm high left and right Pseudobulbus Bletillae (Rhizoma Bletillae) seedling；Wherein, described protocorm
Division culture medium consists of：MS+0.1mg/L NAA+1.0mg/L 6-BA+7.5%wt bananas juice+6.8g/L agar, Ph 5.8；
(4) Pseudobulbus Bletillae (Rhizoma Bletillae) seedling obtaining in step (3) is inoculated in bulb inducing culture, in 28 DEG C, illuminance
60d is cultivated, the base portion of the Pseudobulbus Bletillae (Rhizoma Bletillae) expands formation bulb, measured, the squama of formation under conditions of 2000lux, light application time 12h/d
Stem a diameter of 4～7mm mostly, average diameter is 5.85mm, at this moment can carry out rooting culture, and wherein, described bulb induction is trained
Support basis set becoming：1/2MS culture medium+1.0mg/L NAA+7.5%wt bananas juice+6.8g/L agar, pH 5.6；
(5) seedling obtaining step (4) taking-up clear water in bottle washes off after culture medium, and dislocation is covered with domestication and plants
On the seedbed of training substrate, and moisturizing of sheltering from heat or light, after taming 6 months, gained Pseudobulbus Bletillae (Rhizoma Bletillae) seedling can carry out field-transplanting.
By embodiment, the surface present invention uses different cultures in bletilla striata seeds sprouting, inoculation, differentiation, induction period
Base, achieves good effect it is adaptable to the popularization and application of Pseudobulbus Bletillae (Rhizoma Bletillae) seedling large-scale production.
Claims (7)
1. a kind of method of fast culture Pseudobulbus Bletillae (Rhizoma Bletillae) seedling, it comprises the following steps；
(1) Pseudobulbus Bletillae (Rhizoma Bletillae) capsule is cleaned with clear water, and detergent solution soaks 10min, then is cleaned with scour without sediment motion, on superclean bench
Using 75% ethanol disinfection 30s, then sterilized 10～15min with mercuric chloride solution, with aseptic water washing 3～4 times, obtain the Pseudobulbus Bletillae (Rhizoma Bletillae)
Aseptic mould fruit.
(2) aseptic the moulding of the Pseudobulbus Bletillae (Rhizoma Bletillae) of step (1) gained is really cut open with dissecting knife on super-clean bench, takes out seed, is inoculated in seed and sprouts
Send out on fluid medium, under conditions of 22～25 DEG C, 80～120 turns/min, cultivate 40d, be filtrated to get Pseudobulbus Bletillae (Rhizoma Bletillae) protocorm；
(3) Pseudobulbus Bletillae (Rhizoma Bletillae) protocorm of gained in step (2) is inoculated in protocorm differentiation culture medium, 25～28 DEG C, illumination strong
Cultivate 60d under conditions of degree 1000～4000lux, light application time 11～13h/d, obtain Pseudobulbus Bletillae (Rhizoma Bletillae) seedling；
(4) Pseudobulbus Bletillae (Rhizoma Bletillae) seedling of gained in step (3) is inoculated in bulb inducing culture, in 25～28 DEG C, illuminance 1000
Cultivate 60d under conditions of～1500lux, light application time 11～13h/d, choose the Pseudobulbus Bletillae (Rhizoma Bletillae) that base portion forms diameter 4～7mm bulb；
(5), after the Pseudobulbus Bletillae (Rhizoma Bletillae) chosen in step (4) being cleaned with water, it is displaced on the seedbed being covered with domesticating and cultivating substrate, shelters from heat or light
80%, after taming 6 months, gained Pseudobulbus Bletillae (Rhizoma Bletillae) seedling can carry out field-transplanting.
2. the method for fast culture Pseudobulbus Bletillae (Rhizoma Bletillae) seedling according to claim 1 is it is characterised in that in step (1), described goes
Dirty agent is normal domestic use liquid detergent；In described mercuric chloride solution, the mass fraction of mercuric chloride is 0.1%～0.15%.
3. the method for fast culture Pseudobulbus Bletillae (Rhizoma Bletillae) seedling according to claim 1 is it is characterised in that in step (2), described liquid
The group that body culture medium is is divided into：1/2MS+1.0mg/L 6-BA+6.8-7.5g/L agar, Ph5.6-5.8.
4. the method for fast culture Pseudobulbus Bletillae (Rhizoma Bletillae) seedling according to claim 1 is it is characterised in that culture bar in step (2)
Part is：Under the conditions of 22～25 DEG C, front 7d gives light culture, and intensity of illumination is 1000～4000lux afterwards, and light application time 11～
13h/d.
5. the method for fast culture Pseudobulbus Bletillae (Rhizoma Bletillae) seedling according to claim 1 is it is characterised in that in step (3), described is former
The group of bulb division culture medium is divided into：MS+0.1mg/L NAA+1.0mg/L 6-BA+7.5%wt bananas juice+6.8-7.5g/L fine jade
Fat, Ph5.6-5.8.
6. the method for fast culture Pseudobulbus Bletillae (Rhizoma Bletillae) seedling according to claim 1 is it is characterised in that in step (4), described squama
The group of stem inducing culture is divided into：1/2MS culture medium+1.0mg/L NAA+7.5%wt bananas juice+6.8～7.5g/L agar,
PH5.6～5.8.
7. the method for fast culture Pseudobulbus Bletillae (Rhizoma Bletillae) seedling according to claim 1 is it is characterised in that in step (5), described tames and dociles
Change cultivation matrix compares 1: 1: 0.5 by rural area soil, peat soil and Vermiculitum with weight and forms.
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| CN109511549A (en) * | 2018-11-25 | 2019-03-26 | 福建省亚热带植物研究所 | One kind inhibiting protocorm differentiation and reversal method in bletilla striata seeds incubation |
| CN112741003A (en) * | 2021-03-10 | 2021-05-04 | 广西壮族自治区中国科学院广西植物研究所 | Method for promoting rapid seedling formation of bletilla striata seeds |
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Application publication date: 20170222 |