CN107581073A - A kind of Morinda officinalis group culturation rapid propagating technology - Google Patents
A kind of Morinda officinalis group culturation rapid propagating technology Download PDFInfo
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- CN107581073A CN107581073A CN201711125336.0A CN201711125336A CN107581073A CN 107581073 A CN107581073 A CN 107581073A CN 201711125336 A CN201711125336 A CN 201711125336A CN 107581073 A CN107581073 A CN 107581073A
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Abstract
The invention discloses a kind of Morinda officinalis group culturation rapid propagating technology, Morinda officinalis is madder wort, and whole year can harvest, and is preferred with autumn, winter harvesting.Take 5 ~ 7 years raw roots, remove lateral root, clean, shine to 6 seventy percent dry, gently flattened with mallet, then dried again, be cut into 6 ~ 12cm length, get product.Scalded if steaming half an hour or being steeped with boiling water, color is more purple, and matter is more preferably.Root is in oblate cylindricality or net post shape, slightly curved song, has vertical wrinkle and the band got deeply stuck in, surface dark gray or sepia, some micro-strip purples.Matter is tough and tensile, and plane of rupture is uneven.Gas is without sweet and slightly puckery.It is preferred with dry, sturdy, the aobvious beaded of body, meat thickness, color purple person.The present invention is using Morinda officinalis seed as explant, by callus induction, propagation, break up, take root, the process such as acclimatization and transplantses have successfully been obtained the in vitro plant again of Morinda officinalis, establish Morinda officinalis tissue culture rapid propagation technique system, to save the sowing quantity in cultivation production, the yield of its medicinal material is improved.
Description
Technical field
The present invention relates to Plant Tissue Breeding class, specifically, is related to a kind of Morinda officinalis group culturation rapid propagating technology.
Background technology
Morinda officinalis is madder wort, originates in Guangdong, Guangxi, and whole year can harvest, and is preferred with autumn, winter harvesting.Take
5 ~ 7 years raw roots, remove lateral root, clean, and shine to 6 seventy percent dry, are gently flattened with mallet, then dry again, be cut into 6 ~ 12cm
It is long, get product.Scalded if steaming half an hour or being steeped with boiling water, color is more purple, and matter is more preferably.Morinda offcinalis How Tiangeng is in oblate cylindricality or net post
Shape, slightly curved song have vertical wrinkle and the band got deeply stuck in, and some hangs contracting or the dialysis of crust transverse direction and exposes watercore, forms beaded, long
It is short, 1 ~ 2cm of diameter；Surface dark gray or sepia, some micro-strip purples.Matter is tough and tensile, and plane of rupture is uneven, and skin zone's thickness 5 ~
7mm, lavender, easily peel off, 2 ~ 4mm of woody part diameter.Gas is without sweet and slightly puckery.With dry, sturdy, the aobvious beaded of body, meat thickness, color
Purple person is preferred.Morinda officinalis contains phytosterol, anthraquinone, carbohydrate and vitamin C.With kidney-replenishing, strengthening the bones and muscles, wind-damp dispelling.For waist
Knees soreness, joint pain, cold and pain in the lower abdomen, impotence, seminal emission, arthralgia pain due to rheumatism, muscles and bones is withered soft.At present, Morinda officinalis is mainly with seed
Breeding, but its medicinal effects is also seed, and this reduces the yield of its medicinal material to a certain extent.Therefore, the present invention is with Morinda offcinalis How
Its seed is explant, by callus induction, propagation, break up, take root, the process such as acclimatization and transplantses have successfully been obtained Morinda officinalis
In vitro plant again, establishes Morinda officinalis tissue culture rapid propagation technique system, to save the sowing quantity in cultivation production, improves it
The yield of medicinal material and further carry out Study on Genetic Transformation for Morinda officinalis technical support is provided.
The content of the invention
It is an object of the invention to provide a kind of Morinda officinalis group culturation rapid propagating technology is gone out, the present invention is using Morinda officinalis seed as explant
Body, by callus induction, propagation, break up, take root, the process such as acclimatization and transplantses have successfully been obtained the in vitro plant again of Morinda officinalis,
Morinda officinalis tissue culture rapid propagation technique system is established, it is achieved thereby that the purpose of the present invention.
A kind of Morinda officinalis tissue-culturing rapid propagation technology of the present invention, it is characterised in that comprise the following steps：
（1）The acquisition of aseptic seedling：Full Morinda officinalis seed is chosen, kind of a skin is peelled off with surgical scissors in superclean bench, is used
75% alcohol disinfecting 62s, then 16min is sterilized with 0.1% mercuric chloride solution, finally with rinsed with sterile water 6 times, table is blotted with aseptic filter paper
It is inoculated into after the moisture of face in seed germination medium, daily illumination 26 hours, intensity of illumination 2200lx, cultivation temperature after inoculation
For 30 DEG C, relative air humidity counts germination rate after being cultivated 22 days under conditions of being 75%, and described germination medium is：1/2MS
+ 0.06mg/LNAA+32g/L sucrose+6.2g/L agar, pH 5.9；
（2）Callus induces：When the cotyledon of aseptic seedling just deploys, the stem apex of aseptic seedling is taken to be cut into as explant with scalpel
0.6cm or so length, it is inoculated in the culture medium for callus induction and cultivates, daily illumination 16 hours, intensity of illumination after inoculation
For 2200lx, cultivation temperature is 30 DEG C, and relative air humidity counts inductivity after being cultivated 32 days under conditions of being 75%, described
Calli induction media is：MS+3.2mg/L6-BA+0.6mg/LNAA+1.2mg/L KT+32g/L sucrose+6.2g/L agar, pH
For 5.9；
（3）Multiplying culture：By step（2）Obtained callus is induced to cut into 0.6cm3The fritter of size is simultaneously transferred to propagation training
Foster base carries out expanding numerous, daily illumination 15 hours, intensity of illumination 3200lx after inoculation, and cultivation temperature is 30 DEG C, and air is relatively wet
Spend to count proliferative conditions after cultivating 32 days under conditions of 75%, described proliferated culture medium is：MS+2.2mg/L6-BA+
0.6mg/L NAA+32g/L sucrose+6.2g/L agar, pH 5.9；
（4）Differentiation culture：By step（3）Breed obtained callus and cut into 0.6cm3The fritter of size is simultaneously transferred to differentiation training
The generation of base evoking adventive bud, daily illumination 15 hours, intensity of illumination 3200lx after inoculation are supported, cultivation temperature is 30 DEG C, empty
Gas relative humidity counts differentiation situation after being cultivated 32 days under conditions of being 75%, and described differential medium is：MS+1.2mg/LKT
+ 2.2mg/LNAA+32g/L sucrose+6.2g/L agar, pH 5.9；
（5）Culture of rootage：By step（4）Differentiation obtains adventitious bud and cuts and be seeded in root media from base portion to be induced
Take root, daily illumination 15 hours, intensity of illumination 3200lx after inoculation, cultivation temperature is 30 DEG C, and relative air humidity is 75%
Under the conditions of cultivate 32 days after statistics take root situation, described root media is：1/2MS+1.2mg/LIBA+0.6mg/L NAA+
1.2% activated carbon+32g/L sucrose+6.2g/L agar, pH 5.9；
（6）Acclimatization and transplantses：After taking root one month, its bottleneck is opened, and pours into sterilized water, to ensure its moisture supply, is taken after 8 days
Go out rooted seedling and clean institute's band culture medium on root, be transplanted to matrix by peat soil:Perlite:Vermiculite=4:2:In the matrix of 1 composition,
Covered with the film for leaving air-vent, be placed on shady and cool ventilative interior, throw off film within 16 days, then outside scenery, every other day
A water is poured, transplanting counts survival rate after 32 days.
Compared with existing skill it is an advantage of the invention that：Using Morinda officinalis seed as explant, by callus induction, increase
Grow, break up, taking root, the process such as acclimatization and transplantses have successfully been obtained the in vitro plant again of Morinda officinalis, it is quick to establish Morinda officinalis tissue cultures
Reproduction technique system, to save the sowing quantity in cultivation production, improve the yield of its medicinal material and further carry out something lost for Morinda officinalis
Pass Study on Transformation and provide powerful technique support.
Embodiment
Following examples are the further explanations to the present invention, are not limitations of the present invention.
Embodiment 1
（1）The acquisition of aseptic seedling：Full Morinda officinalis seed is chosen, kind of a skin is peelled off with surgical scissors in superclean bench, is used
75% alcohol disinfecting 38s, then 0.1% mercuric chloride solution sterilization 6min, finally with rinsed with sterile water 2 times, surface is blotted with aseptic filter paper
It is inoculated into after moisture in seed germination medium.Daily illumination 22 hours is placed in after inoculation, intensity of illumination 800lx, is placed in training
It is 24 DEG C to support temperature, and germination rate reaches 100% after relative air humidity is cultivated 18 days under conditions of being 75%.Described sprouting culture
Base is：1/2MS+0.01mg/L NAA+12g/L sucrose+3.2g/L agar, pH 5.4.
（2）Callus induces：When the cotyledon of aseptic seedling just deploys, the stem apex of aseptic seedling is taken to use scalpel as explant
0.3cm or so length is cut into, is inoculated in the culture medium for callus induction and cultivates.Daily illumination 10 hours is placed in after inoculation,
Intensity of illumination is 800lx, is placed in cultivation temperature as 24 DEG C, inductivity after relative air humidity is cultivated 28 days under conditions of being 75%
Reach 92%.Described calli induction media is：MS+1.3mg/L 6-BA+0.2mg/LNAA+0.6mg/L KT+23g/L sugarcanes
Sugar+4.3g/L agar, pH 5.3.
（3）Multiplying culture：By step（2）Obtained callus is induced to cut into 0.3cm3The fritter of size is simultaneously transferred to increasing
Grow culture medium expand it is numerous.Daily illumination is placed in after inoculation 10 hours, intensity of illumination 1800lx, is placed in cultivation temperature as 24
DEG C, proliferation times reach 4.32 times after relative air humidity is cultivated 28 days under conditions of being 75%.Described proliferated culture medium is：
MS+1.4mg/L 6-BA+0.2mg/L NAA+18g/L sucrose+3.6g/L agar, pH 5.3.
（4）Differentiation culture：By step（3）Breed obtained callus and cut into 0.3cm3The fritter of size is simultaneously transferred to point
Change the generation of culture medium evoking adventive bud.Daily illumination 10 hours, intensity of illumination 1800lx after inoculation, being placed in cultivation temperature is
24 DEG C, differentiation rate reaches 92% after relative air humidity is cultivated 28 days under conditions of being 75%, and average adventitious bud number is 4.2.Institute
The differential medium stated is：MS+0.5mg/LKT+1.3mg/L NAA+28g/L sucrose+4.3g/L agar, pH 5.3.
（5）Culture of rootage：By step（4）Differentiation obtains adventitious bud and cuts and be seeded in root media from base portion to carry out
Root induction.Daily illumination 10 hours, intensity of illumination 2200lx after inoculation, cultivation temperature is placed in as 24 DEG C, air is relatively wet
Spend for rooting rate 92% after cultivating 28 days under conditions of 75%.Described root media is：1/2MS+0.5mg/L IBA+
0.1mg/L NAA+0.3% activated carbon+13g/L sucrose+3.3g/L agar, pH 5.3.
（6）Acclimatization and transplantses：After taking root one month, its bottleneck is opened, and pours into sterilized water, to ensure its moisture supply, 5 days
Rooted seedling is taken out afterwards and cleans institute's band culture medium on root, is transplanted to matrix by peat soil:Perlite:Vermiculite=2:1:The base of 1 composition
In matter.Covered with the film for leaving air-vent, be placed on shady and cool ventilative interior, film thrown off in 10 days, be subsequently placed in outdoor
Culture, pours a water every other day, and survival rate is to more than 91% after transplanting 28 days.
Embodiment 2
（1）The acquisition of aseptic seedling：Full Morinda officinalis seed is chosen, kind of a skin is peelled off with surgical scissors in superclean bench, is used
75% alcohol disinfecting 46s, then 0.1% mercuric chloride solution sterilization 9min, finally with rinsed with sterile water 6 times, surface is blotted with aseptic filter paper
It is inoculated into after moisture in seed germination medium.Daily illumination 25 hours is placed in after inoculation, intensity of illumination 1600lx, is placed in training
It is 26 DEG C to support temperature, and germination rate reaches 100% after relative air humidity is cultivated 21 days under conditions of being 75%.Described sprouting culture
Base is：1/2MS+0.06mg/L NAA+19g/L sucrose+3.9g/L agar, pH 5.8.
（2）Callus induces：When the cotyledon of aseptic seedling just deploys, the stem apex of aseptic seedling is taken to use scalpel as explant
0.6cm or so length is cut into, is inoculated in the culture medium for callus induction and cultivates.Daily illumination 15 hours is placed in after inoculation,
Intensity of illumination is 1600lx, is placed in cultivation temperature as 28 DEG C, inductivity after relative air humidity is cultivated 32 days under conditions of being 75%
Reach 91%.Described calli induction media is：MS+2.1mg/L 6-BA+0.6mg/LNAA+1.2mg/L KT+29g/L sugarcanes
Sugar+5.6g/L agar, pH 5.8.
（3）Multiplying culture：By step（2）Obtained callus is induced to cut into 0.6cm3The fritter of size is simultaneously transferred to increasing
Grow culture medium expand it is numerous.Daily illumination is placed in after inoculation 14 hours, intensity of illumination 2600lx, is placed in cultivation temperature as 28
DEG C, proliferation times reach 5.08 times after relative air humidity is cultivated 31 days under conditions of being 75%.Described proliferated culture medium is：
MS+2.1mg/L 6-BA+0.6mg/L NAA+26g/L sucrose+4.6g/L agar, pH 5.8.
（4）Differentiation culture：By step（3）Breed obtained callus and cut into 0.6cm3The fritter of size is simultaneously transferred to point
Change the generation of culture medium evoking adventive bud.Daily illumination is placed in after inoculation 14 hours, intensity of illumination 2600lx, is placed in culture temperature
Spend for 28 DEG C, differentiation rate reaches 94% after relative air humidity is cultivated 31 days under conditions of being 75%, and average adventitious bud number is 4.98
It is individual.Described differential medium is：MS+1.2mg/LKT+2.2mg/L NAA+31g/L sucrose+5.6g/L agar, pH 5.6.
（5）Culture of rootage：By step（4）Differentiation obtains adventitious bud and cuts and be seeded in root media from base portion to carry out
Root induction.Daily illumination is placed in after inoculation 15 hours, intensity of illumination 3100lx, it is 28 DEG C to be placed in cultivation temperature, air phase
Rooting rate 96% after being cultivated 31 days under conditions of being 75% to humidity.Described root media is：1/2MS+1.2mg/L IBA+
0.6mg/L NAA+0.9% activated carbon+21g/L sucrose+4.6g/L agar, pH 5.6.
（6）Acclimatization and transplantses：After taking root one month, its bottleneck is opened, and pours into sterilized water, to ensure its moisture supply, 6 days
Rooted seedling is taken out afterwards and cleans institute's band culture medium on root, is transplanted to matrix by peat soil:Perlite:Vermiculite=2:1:The base of 1 composition
In matter.Covered with the film for leaving air-vent, be placed on shady and cool ventilative interior, film thrown off in 15 days, be subsequently placed in outdoor
Culture, pours a water every other day, and survival rate is to 95% after transplanting 31 days.
Claims (1)
1. a kind of Morinda officinalis group culturation rapid propagating technology, it is characterised in that comprise the following steps：
（1）The acquisition of aseptic seedling：Full Morinda officinalis seed is chosen, kind of a skin is peelled off with surgical scissors in superclean bench, is used
75% alcohol disinfecting 62s, then 16min is sterilized with 0.1% mercuric chloride solution, finally with rinsed with sterile water 6 times, table is blotted with aseptic filter paper
It is inoculated into after the moisture of face in seed germination medium, daily illumination 26 hours, intensity of illumination 2200lx, cultivation temperature after inoculation
For 30 DEG C, relative air humidity counts germination rate after being cultivated 22 days under conditions of being 75%, and described germination medium is：1/2MS
+ 0.06mg/LNAA+32g/L sucrose+6.2g/L agar, pH 5.9；
（2）Callus induces：When the cotyledon of aseptic seedling just deploys, the stem apex of aseptic seedling is taken to be cut into as explant with scalpel
0.6cm or so length, it is inoculated in the culture medium for callus induction and cultivates, daily illumination 16 hours, intensity of illumination after inoculation
For 2200lx, cultivation temperature is 30 DEG C, and relative air humidity counts inductivity after being cultivated 32 days under conditions of being 75%, described
Calli induction media is：MS+3.2mg/L6-BA+0.6mg/LNAA+1.2mg/L KT+32g/L sucrose+6.2g/L agar, pH
For 5.9；
（3）Multiplying culture：By step（2）Obtained callus is induced to cut into 0.6cm3The fritter of size is simultaneously transferred to propagation training
Foster base carries out expanding numerous, daily illumination 15 hours, intensity of illumination 3200lx after inoculation, and cultivation temperature is 30 DEG C, and air is relatively wet
Spend to count proliferative conditions after cultivating 32 days under conditions of 75%, described proliferated culture medium is：MS+2.2mg/L6-BA+
0.6mg/L NAA+32g/L sucrose+6.2g/L agar, pH 5.9；
（4）Differentiation culture：By step（3）Breed obtained callus and cut into 0.6cm3The fritter of size is simultaneously transferred to differentiation training
The generation of base evoking adventive bud, daily illumination 15 hours, intensity of illumination 3200lx after inoculation are supported, cultivation temperature is 30 DEG C, empty
Gas relative humidity counts differentiation situation after being cultivated 32 days under conditions of being 75%, and described differential medium is：MS+1.2mg/LKT
+ 2.2mg/LNAA+32g/L sucrose+6.2g/L agar, pH 5.9；
（5）Culture of rootage：By step（4）Differentiation obtains adventitious bud and cuts and be seeded in root media from base portion to be induced
Take root, daily illumination 15 hours, intensity of illumination 3200lx after inoculation, cultivation temperature is 30 DEG C, and relative air humidity is 75%
Under the conditions of cultivate 32 days after statistics take root situation, described root media is：1/2MS+1.2mg/LIBA+0.6mg/L NAA+
1.2% activated carbon+32g/L sucrose+6.2g/L agar, pH 5.9；
（6）Acclimatization and transplantses：After taking root one month, its bottleneck is opened, and pours into sterilized water, to ensure its moisture supply, is taken after 8 days
Go out rooted seedling and clean institute's band culture medium on root, be transplanted to matrix by peat soil:Perlite:Vermiculite=4:2:In the matrix of 1 composition,
Covered with the film for leaving air-vent, be placed on shady and cool ventilative interior, throw off film within 16 days, then outside scenery, every other day
A water is poured, transplanting counts survival rate after 32 days.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108575743A (en) * | 2018-03-29 | 2018-09-28 | 中国长江三峡集团有限公司 | A kind of regenerated tissue culture and rapid propagation method of induction Radix Mussaendae plumule |
| CN109662028A (en) * | 2018-11-28 | 2019-04-23 | 西南林业大学 | A method of passing through the beautiful blade tea of test tube seedling continuous production promise |
| CN112931224A (en) * | 2021-04-16 | 2021-06-11 | 广州中医药大学(广州中医药研究院) | Tissue culture method of morinda officinalis |
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| CN1379972A (en) * | 2002-05-16 | 2002-11-20 | 李旭群 | Morinda tissue culture method |
| CN102960251A (en) * | 2012-12-03 | 2013-03-13 | 无限极（中国）有限公司 | Method and culture medium for obtaining morinda officinalis body cell regeneration plant |
| CN104041417A (en) * | 2014-07-10 | 2014-09-17 | 黄振忠 | Morinda officinalis tissue culture breeding method |
| CN104782495A (en) * | 2015-05-02 | 2015-07-22 | 冯文杰 | Tissue culture and rapid propagation method for euphorbia lathyris |
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- 2017-11-14 CN CN201711125336.0A patent/CN107581073A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1379972A (en) * | 2002-05-16 | 2002-11-20 | 李旭群 | Morinda tissue culture method |
| CN102960251A (en) * | 2012-12-03 | 2013-03-13 | 无限极（中国）有限公司 | Method and culture medium for obtaining morinda officinalis body cell regeneration plant |
| CN104041417A (en) * | 2014-07-10 | 2014-09-17 | 黄振忠 | Morinda officinalis tissue culture breeding method |
| CN104782495A (en) * | 2015-05-02 | 2015-07-22 | 冯文杰 | Tissue culture and rapid propagation method for euphorbia lathyris |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108575743A (en) * | 2018-03-29 | 2018-09-28 | 中国长江三峡集团有限公司 | A kind of regenerated tissue culture and rapid propagation method of induction Radix Mussaendae plumule |
| CN108575743B (en) * | 2018-03-29 | 2020-05-29 | 中国长江三峡集团有限公司 | Tissue culture rapid propagation method for inducing regeneration of mussaenda pubescens embryo |
| CN109662028A (en) * | 2018-11-28 | 2019-04-23 | 西南林业大学 | A method of passing through the beautiful blade tea of test tube seedling continuous production promise |
| CN112931224A (en) * | 2021-04-16 | 2021-06-11 | 广州中医药大学(广州中医药研究院) | Tissue culture method of morinda officinalis |
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Application publication date: 20180116 |