CN109496706A - A kind of sterile brachyplast of Mao Xu Cao quickly breed before explant preprocess method - Google Patents
A kind of sterile brachyplast of Mao Xu Cao quickly breed before explant preprocess method Download PDFInfo
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- CN109496706A CN109496706A CN201811487048.4A CN201811487048A CN109496706A CN 109496706 A CN109496706 A CN 109496706A CN 201811487048 A CN201811487048 A CN 201811487048A CN 109496706 A CN109496706 A CN 109496706A
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- transplanted seedling
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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
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
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Abstract
Explant preprocess method before quickly being bred the present invention relates to a kind of sterile brachyplast of Mao Xu Cao, comprising the following steps: (1) carry out disinfection processing to culture medium；(2) to sealable vinyl house pre-sterilization；(3) transplanted seedling selection and trimming disinfection；(4) transplanted seedling is colonized；(5) nursery stock periodically sterilizes；(6) clip shoot.Operation of the present invention is simple, and Disinfection Effect is good, and production cost is low, can quickly breed for batch production and provide enough explants；And this method pollution rate is low, minimum 2.0%.
Description
Technical field
Before quickly being bred the present invention relates to field of plant tissue culture technique more particularly to a kind of sterile brachyplast of Mao Xu Cao
Explant preprocess method.
Background technique
Mao Xu Cao (Clerodendranthus spicatus), Labiatae perennial herb, stem is upright, 1-1.5 meters high,
Four prismatics, corolla grey violet or white, flower, the fruiting period 5-11 month；It is mainly distributed on Guangdong And Hainan, south Guangxi, south of Yunnan, Taiwan
And Fujian；It is often born at hayashishita humidity, is also seen sometimes without on shady level land.Mao Xu Cao aerial part is used as medicine, and sweet light slight bitter has
Stronger antibacterial, antiinflammation can be such that renal tubule destroys and mitigate, and messangial cell structural lesions are reduced, and complete glomerulus number increases
Add, have protection and improvement result to the damage of nephridial tissue, can increase glomerular filtration and kidney circulation blood volume, toxic metabolic is promoted to produce
The exclusion of object reduces internal blood uric acid.Since it is with important pharmacological action, the market demand is growing.Using plant group
Knitting culture, quickly breeding, health seedling needed for providing Mao Xu Cao large-scale plantation have become urgent problem.
The type that Plant Tissue Breeding is quickly bred has: sterile Spur Type, from proliferous type of sprouting, Organogenesis and embryo shape
Body generation type, however the scope of application and advantage of different proliferation types are different, therefore in production in practice according to floristic
The proliferation type that habit and feature selection are suitble to.Sterile Spur Type quick breeding technology is that material to be bred is cut into band
1 leaf stem section of 1 leaf, is transferred in seedling medium bottle, the 1 bud stem that seedlings can be grown up to after certain time culture, are cut into 1 leaf of band again
Section, squamous subculture and Cheng great Miao.By seedlings, dissection is transferred repeatedly when squamous subculture, to obtain more tender stem rapidly.Then will
A part of tender stem is transferred on root media, can turn out complete test tube seedling.This method mainly utilizes top
Advantage, it is rapid to can be used for shoot growth, or to the higher herbaceous plant of Plant Tissue Breeding seedling quality requirement and a part of woody
Plant.The technology seedling is fast, and without the callus induction stage, stabilization characteristics of genetics, incubation is simple, the scope of application
Extensively, it moves to cut out and is easy to survive.
Mao Xu Cao wild resource is increasingly reduced under a large amount of diggings of people, and Mao Xu Cao planting material depends on skewer
It inserts and the technologies such as in vitro micropropogation provides health seedling.Sterile brachyplast quick breeding technology is an important in vitro micro- numerous skill
Art, but because its explant be usually directly derived from big Tanaka growth by a large amount of bacteriums and the stem with bud of absorbability fungal contamination
Deng, cause tissue cultures pollution rate very high so that quick reproduction technique efficiency it is very low in addition failure.Many researchers use more bacterium
The reagents such as spirit, Bravo, Sandofan, thiophanate-methyl, gentamicin, prochloraz, tetracycline are individually or admixing medical solutions impregnate
Explant is as the pretreatment before disinfection, or benefit training base fungicide is added on solid medium, these processing are in certain journey
Pollution rate is reduced on degree, but cannot be tackled the problem at its root.Therefore, new explant high-effective disinfecting method is explored, it is right
Pollution problem has important meaning during solving the sterile brachyplast fast breeding of Mao Xu Cao seedling.
Summary of the invention
According to the above-mentioned deficiencies of the prior art, the present invention provides the explant before a kind of sterile brachyplast of Mao Xu Cao quickly breeds
Preprocess method carries out matrix, transplanted seedling and the growing space of cultivating Mao Xu Cao explant using multiple spectrum fungicide
Comprehensive disinfection treatment, pollution rate is low, to improve efficiency and yield that the sterile brachyplast of Mao Xu Cao quickly breeds.
In order to achieve the object of the present invention, the invention adopts the following technical scheme:
A kind of sterile brachyplast of Mao Xu Cao quickly breed before explant preprocess method, comprising the following steps:
Step 1 carries out disinfection processing to culture medium: with 50% 350 times of liquid of water-soluble ambam, using by every square metre of culture substrate
3 kilograms of dilution sprinklings uniformly, are used to plant to the slightly dry flowerpot that can be packed into of soil；
Step 2, to sealable vinyl house pre-sterilization: it is moved into first 15 days in Mao Xu Cao transplanted seedling, it is primary every 5 days, first
The secondary prochloraz for being 0.4% with concentration, second is 0.2% Mancozeb with concentration, and using concentration for the third time is 0.2% Dai Sen
MnZn medicine liquid spray vinyl house ground and surrounding, disinfection treatment time are the morning, closed plastic greenhouse after spray；
Step 3, transplanted seedling selection and trimming disinfection: before transplanted seedling field planting, select the full blade no disease and pests harm of sprout, growth strong
Strong Mao Xu Cao is carefully dug, and is shaked off the soil of root and is rinsed with clear water, then at branch from the ground 3-4cm trimming and
It cuts off old leaf and retains petiole, the Mancozeb medicine that the prochloraz or concentration for being 0.4% with concentration by the branch trimmed are 0.2%
Liquid soaking disinfection 1-2h；
Transplanted seedling field planting: transplanted seedling described in step 3 is colonized in the flower equipped with culture substrate described in step 1 by step 4
In basin, and move into vinyl house culture described in step 2；
Step 5, nursery stock periodically sterilize: after the field planting of transplanted seedling described in step 4, it is 0.4% that concentration was used alternatingly every 7 days
Prochloraz and concentration be 0.2% Mancozeb or to be applied alone prochloraz that concentration is 0.4% also or concentration is applied alone be 0.2%
Mancozeb medicine liquid spray seedling plant, ground and vinyl house surrounding it is primary, the disinfection treatment time be the morning；
Step 6, clip shoot: it is new when extracting young tender shoots section out at the top of the nursery stock described in the step 5, it cuts, takes out from its base portion
Explant material of the bud section as tissue cultures.
It is cut in the step six to newly extract young tender shoots section growth 8cm-10cm Shi Congqi base portion out.
Compared with prior art, the beneficial effects of the present invention are: (1) easy to operate, Disinfection Effect is good；(2) this method
Production cost is low, can quickly breed for batch production and provide enough explants；(3) pollution rate is low, and minimum 2.0%.
Specific embodiment
Embodiment 1
A kind of sterile brachyplast of Mao Xu Cao quickly breed before the explant processing method, comprising the following steps:
Step 1 carries out disinfection processing to culture medium: with 50% 350 times of liquid of water-soluble ambam, using by every square metre of culture substrate
3 kilograms of dilution sprinklings uniformly, are used to plant to the slightly dry flowerpot that can be packed into of soil；
Step 2, to sealable vinyl house pre-sterilization: it is moved into first 15 days in Mao Xu Cao transplanted seedling, it is primary every 5 days, first
The secondary prochloraz for being 0.4% with concentration, second is 0.2% Mancozeb with concentration, and using concentration for the third time is 0.2% Dai Sen
MnZn medicine liquid spray vinyl house ground and surrounding, disinfection treatment time are the morning, closed plastic greenhouse after spray；
Step 3, transplanted seedling selection and trimming disinfection: before transplanted seedling field planting, select the full blade no disease and pests harm of sprout, growth strong
Strong Mao Xu Cao is carefully dug, and is shaked off the soil of root and is rinsed with clear water, is then trimmed and is cut at branch from the ground 3cm
Old leaf is gone to retain petiole, the Mancozeb medical fluid that the prochloraz or concentration for being 0.4% with concentration by the branch trimmed are 0.2%
Soaking disinfection 1h；
Transplanted seedling field planting: transplanted seedling described in step 3 is colonized in the flower equipped with culture substrate described in step 1 by step 4
In basin, and move into vinyl house culture described in step 2；
Step 5, nursery stock periodically sterilize: after the field planting of transplanted seedling described in step 4, it is 0.4% that concentration was used alternatingly every 7 days
Prochloraz and concentration be 0.2% Mancozeb medicine liquid spray seedling plant, ground and vinyl house surrounding it is primary, at disinfection
The reason time is the morning；
Step 6, clip shoot: it is new when extracting young tender shoots section out at the top of the nursery stock described in the step 5, to newly extract young tender shoots Duan Sheng out
Long 8cm Shi Congqi base portion is cut, and as the explant material of tissue cultures, paper slip is taken back laboratory with sterilizing brown paper.
Embodiment 2
A kind of sterile brachyplast of Mao Xu Cao quickly breed before the explant processing method, comprising the following steps:
Step 1 carries out disinfection processing to culture medium: with 50% 350 times of liquid of water-soluble ambam, using by every square metre of culture substrate
3 kilograms of dilution sprinklings uniformly, are used to plant to the slightly dry flowerpot that can be packed into of soil.
Step 2, to sealable vinyl house pre-sterilization: it is moved into first 15 days in Mao Xu Cao transplanted seedling, it is primary every 5 days,
The prochloraz for being for the first time 0.4% with concentration, it is 0.2% Mancozeb that second, which is used concentration, is for the third time 0.2% with concentration
Mancozeb medicine liquid spray vinyl house ground and surrounding, disinfection treatment time are the morning, closed plastic greenhouse after spray；
Step 3, transplanted seedling selection and trimming disinfection: before transplanted seedling field planting, select the full blade no disease and pests harm of sprout, growth strong
Strong Mao Xu Cao is carefully dug, and is shaked off the soil of root and is rinsed with clear water, then at branch from the ground 3.5cm trimming and
It cuts off old leaf and retains petiole, the Mancozeb medicine that the prochloraz or concentration for being 0.4% with concentration by the branch trimmed are 0.2%
Liquid soaking disinfection 1.5h；
Transplanted seedling field planting: transplanted seedling described in step 3 is colonized in the flower equipped with culture substrate described in step 1 by step 4
In basin, and move into vinyl house culture described in step 2；
Step 5, nursery stock periodically sterilize: after the field planting of transplanted seedling described in step 4, it is 0.4% miaow that concentration was applied alone every 7 days
Amide medicine liquid spray seedling plant, ground and vinyl house surrounding are primary, and the disinfection treatment time is the morning；
Step 6, clip shoot: it is new when extracting young tender shoots section out at the top of the nursery stock described in the step 5, to newly extract young tender shoots Duan Sheng out
Long 9cm Shi Congqi base portion is cut, and as the explant material of tissue cultures, paper slip is taken back laboratory with sterilizing brown paper.
Embodiment 3
A kind of sterile brachyplast of Mao Xu Cao quickly breed before the explant processing method, comprising the following steps:
Step 1 carries out disinfection processing to culture medium: with 50% 350 times of liquid of water-soluble ambam, using by every square metre of culture substrate
3 kilograms of dilution sprinklings uniformly, are used to plant to the slightly dry flowerpot that can be packed into of soil；
Step 2, to sealable vinyl house pre-sterilization: it is moved into first 15 days in Mao Xu Cao transplanted seedling, it is primary every 5 days, first
The secondary prochloraz for being 0.4% with concentration, second is 0.2% Mancozeb with concentration, and using concentration for the third time is 0.2% Dai Sen
MnZn medicine liquid spray vinyl house ground and surrounding, disinfection treatment time are the morning, closed plastic greenhouse after spray；
Step 3, transplanted seedling selection and trimming disinfection: the Mao Xu Cao of the full blade no disease and pests harm of selection sprout, robust growth is small
The heart is dug, and is shaked off the soil of root and is rinsed with clear water, is then trimmed and is cut off old leaf at branch from the ground 4cm and retain leaf
Handle, the Mancozeb medical fluid soaking disinfection 2h that the prochloraz or concentration for being 0.4% with concentration by the branch trimmed are 0.2%；
Transplanted seedling field planting: transplanted seedling described in step 3 is colonized in the flower equipped with culture substrate described in step 1 by step 4
In basin, and move into vinyl house culture described in step 2；
Step 5, nursery stock periodically sterilize: after the field planting of transplanted seedling described in step 4, it is 0.2% generation that concentration was applied alone every 7 days
Gloomy MnZn medicine liquid spray seedling plant, ground and vinyl house surrounding are primary, and the disinfection treatment time is the morning；
Step 6, clip shoot: at the top of the nursery stock described in the step 5 it is new extract young tender shoots section out after, to newly extract young tender shoots Duan Sheng out
Long 10cm Shi Congqi base portion is cut, and as the explant material of tissue cultures, paper slip is taken back laboratory with sterilizing brown paper.
Later, the young tender shoots section that embodiment 1,2 and 3 obtains is cut into 1 bud stem of 1 leaf on the superclean bench in laboratory
Section simultaneously immerses concentration to sterilize 30s in 75% alcohol, then flushing 2 times is constantly shaken with sterile water, and being then immersed in concentration is
8min is impregnated in 0.1% mercuric chloride solution, is constantly shaken therebetween, after then constantly shaking flushing 7 times with sterile water again, with nothing
Bacterium filter paper blots surface moisture, and bud section is inoculated on MS culture medium and is cultivated, and condition of culture is arranged in artificial climate incubator
Are as follows: the day night time are as follows: 15h/9h；Intensity of illumination 2500Lx；Day night cultivation temperature is 23 DEG C/18 DEG C；Relative humidity are as follows: 85%.
Experimental result
Statistical result shows to use the pollution rate of not pretreated crop field explant for 100% after inoculation 30 days；Periodically to close
The pollution rate for closing the prochloraz explant that plant spray concentration in space is 0.4% is 5.1%；Periodically to plant in confined space
The pollution rate for the Mancozeb explant that spray concentration is 0.2% is 6.5%；Periodically plant in confined space is alternately sprayed every secondary
The pollution rate for spilling the prochloraz that concentration is 0.4% and the Mancozeb explant that concentration is 0.2% is 2.0%.Wherein with periodically right
It is to take the generation for being 0.2% every the prochloraz and concentration that spray concentration is 0.4% secondary alternately that plant, which carries out disinfection, in confined space
The effect of gloomy MnZn is best, followed by independent spray concentration be 0.4% prochloraz, be again independent spray concentration be 0.2%
Mancozeb.
Claims (2)
1. a kind of sterile brachyplast of Mao Xu Cao quickly breed before explant preprocess method, which comprises the following steps:
Step 1 carries out disinfection processing to culture medium: with 50% 350 times of liquid of water-soluble ambam, using by every square metre of culture substrate
3 kilograms of dilution sprinklings uniformly, are used to plant to the slightly dry flowerpot that can be packed into of soil；
Step 2, to sealable vinyl house pre-sterilization: it is moved into first 15 days in Mao Xu Cao transplanted seedling, it is primary every 5 days, first
The secondary prochloraz for being 0.4% with concentration, second is 0.2% Mancozeb with concentration, and using concentration for the third time is 0.2% Dai Sen
MnZn medicine liquid spray vinyl house ground and surrounding, disinfection treatment time are the morning, closed plastic greenhouse after spray；
Step 3, transplanted seedling selection and trimming disinfection: before transplanted seedling field planting, select the full blade no disease and pests harm of sprout, growth strong
Strong Mao Xu Cao is carefully dug, and is shaked off the soil of root and is rinsed with clear water, then at branch from the ground 3-4cm trimming and
It cuts off old leaf and retains petiole, the Mancozeb medicine that the prochloraz or concentration for being 0.4% with concentration by the branch trimmed are 0.2%
Liquid soaking disinfection 1-2h；
Transplanted seedling field planting: transplanted seedling described in step 3 is colonized in the flower equipped with culture substrate described in step 1 by step 4
In basin, and move into vinyl house culture described in step 2；
Step 5, nursery stock periodically sterilize: after the field planting of transplanted seedling described in step 4, it is 0.4% that concentration was used alternatingly every 7 days
Prochloraz and concentration be 0.2% Mancozeb or to be applied alone prochloraz that concentration is 0.4% also or concentration is applied alone be 0.2%
Mancozeb medicine liquid spray seedling plant, ground and vinyl house surrounding it is primary, the disinfection treatment time be the morning；
Step 6, clip shoot: it is new when extracting young tender shoots section out at the top of the nursery stock described in the step 5, it cuts, takes out from its base portion
Explant material of the bud section as tissue cultures.
2. the sterile brachyplast of a kind of Mao Xu Cao according to claim 1 quickly breed before explant preprocess method, it is special
Sign is, cuts in the step 6 to newly extract young tender shoots section growth 8cm-10cm Shi Congqi base portion out.
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| CN201811487048.4A CN109496706B (en) | 2018-12-06 | 2018-12-06 | Pretreatment method for explant before aseptic short branch rapid propagation of Orthosiphon aristatus |
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| CN201811487048.4A CN109496706B (en) | 2018-12-06 | 2018-12-06 | Pretreatment method for explant before aseptic short branch rapid propagation of Orthosiphon aristatus |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112655553A (en) * | 2020-12-08 | 2021-04-16 | 广西壮族自治区亚热带作物研究所（广西亚热带农产品加工研究所） | Rapid sterile short-shoot propagation method for Orthosiphon aristatus |
| CN115024225A (en) * | 2022-07-06 | 2022-09-09 | 天津博奥聚能生物科技有限公司 | Culture method of cold-resistant orthosiphon aristatus |
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| CN104871752A (en) * | 2014-02-28 | 2015-09-02 | 中国热带农业科学院橡胶研究所 | Explant pretreatment method before inducing and culturing multiple shoot of rubber tree nursery stock |
| CN106258290A (en) * | 2015-06-12 | 2017-01-04 | 宋维成 | A kind of cultural method of kidney tea |
| CN106717788A (en) * | 2016-11-11 | 2017-05-31 | 西双版纳丰能现代农业科技开发有限公司 | A kind of method for culturing seedlings of Mao Xu Cao |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112655553A (en) * | 2020-12-08 | 2021-04-16 | 广西壮族自治区亚热带作物研究所（广西亚热带农产品加工研究所） | Rapid sterile short-shoot propagation method for Orthosiphon aristatus |
| CN115024225A (en) * | 2022-07-06 | 2022-09-09 | 天津博奥聚能生物科技有限公司 | Culture method of cold-resistant orthosiphon aristatus |
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