CN109362564A - A kind of coriander tissue culture and rapid propagation method - Google Patents
A kind of coriander tissue culture and rapid propagation method Download PDFInfo
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- CN109362564A CN109362564A CN201811412316.6A CN201811412316A CN109362564A CN 109362564 A CN109362564 A CN 109362564A CN 201811412316 A CN201811412316 A CN 201811412316A CN 109362564 A CN109362564 A CN 109362564A
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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
- A01H4/001—Culture apparatus for tissue culture
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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
- A01G22/15—Leaf crops, e.g. lettuce or spinach
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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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
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- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Developmental Biology & Embryology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a kind of coriander tissue culture and rapid propagation method, the problems such as variety deterioration caused by artificial cultivation variet complexity and long-term vegetative propagation, serious, the yield decline of disease.Plant Tissue Breeding has the characteristics that breeding is fast, spends less, growth coefficient is high; the present invention is using stem with bud as explant; by induction, be proliferated, take root and acclimatization and transplants and etc. establish coriander tissue culture quick breeding system; sapling multiplication speed and scale can not only be expanded; the new varieties of high yield and high quality can also be selected on this basis; the economic benefit for increasing plantation coriander, for alleviating the coriander realistic problem that supply falls short of demand, protection wild resource has great importance.
Description
Technical field
The present invention relates to the methods of Plant Tissue Breeding in agricultural biotechnologies, specifically, being related to a kind of coriander tissue culture
Quick-breeding method.
Background technique
Coriander alias: caraway, coriandrum, circle Sui.Annual book on Chinese herbal medicine, it is hairless, it is aromatic.There are production in each provinces and regions in the whole nation.Acrid flavour, property
Temperature.With sweating promoting eruption, help digestion lower gas, stomach invigorating, anti-inflammatory.That there are breeding coefficients is not high for the Traditional breeding processes of coriander, growth week
The problems such as phase length, variety deterioration, limits the quickly work such as breeding and breeding of new variety.And in vitro culture can make up traditional breeding
The deficiency of method, the expanding propagation excellent variety in short period of time, and new product can be cultivated in a short time using genetic transfoumation
Kind.Therefore, the foundation of coriander vitro Regeneration System, for alleviating the realistic problem that supply falls short of demand, protection wild resource has weight
The meaning wanted.
Summary of the invention
The purpose of the present invention is to provide corianders using stem with bud as explant, by induction, is proliferated, takes root and hardening
Transplanting and other steps establish coriander tissue culture quick breeding system, to realize the purpose of the present invention.
A kind of coriander tissue culture and rapid propagation method of the invention, includes the following steps:
Step 1, Fiber differentiation: after the stem section for acquiring back washes away surface attachments with clear water, liquid detergent impregnates 16min, then
Flowing water rinses 3h, in superclean bench with 30s is sterilized in 75% ethanol solution, is washed 7 times with sterile afterwards, then with 0.4% mercuric chloride
Solution disinfection 26min is carried out bud and is lured with being inoculated into induced medium after sucking moisture with aseptic filter paper after aseptic water washing 8 times
Culture is led, it is first dark culture 11 days full under the conditions of 30 DEG C after inoculation, it is subsequently placed in daily illumination 15 hours, intensity of illumination is
3200lx is placed in culture under conditions of cultivation temperature is 30 DEG C until formation adventitious bud;Induced medium are as follows: MS+3.8mg/L
6-BA+1.5mg/L NAA+31g/L sucrose+7.8g/L agar, pH 5.3；
Step 2, Multiplying culture: cultivating obtained sprout for step 1, cut bud from base portion and be transferred on proliferated culture medium carry out after
It is commissioned to train feeding, it is first dark culture 4 days full under the conditions of 30 DEG C after inoculation, it is subsequently placed in daily illumination 17 hours, intensity of illumination is
3200lx is placed under conditions of cultivation temperature is 30 DEG C and cultivates, and subculture is primary within 31 days;Proliferated culture medium are as follows: MS+3.0mg/L
6-BA+40g/L sucrose+8.0g/L agar, pH 5.3；
Step 3, culture of rootage: by the sprout of step 1 or step 2 it is long to 5cm high when, be cut into simple bud and be inoculated into culture of rootage
Culture of rootage is carried out on base, it is first dark culture 4 days full under the conditions of 30 DEG C after inoculation, it is subsequently placed in daily illumination 15 hours, illumination
Intensity is 5100lx, and cultivation temperature is cultivated under conditions of being 30 DEG C to taking root;Root media are as follows: 1/2MS+2.6mg/L GGR
+ 0.6mg/L IBA+32g/L sucrose+7.8g/L agar, pH 5.3；
Step 4, acclimatization and transplants: by the half-open placement hardening 2 under field conditions (factors) of the culture bottle cap of the rooting tube plantlet of high about 7cm
It, then standard-sized sheet bottle cap is added suitable tap water and is placed in natural lighting lower refining seedling 8 days, and test tube seedling is taken out from culture bottle,
Root culture medium is washed off, is planted by peat soil: land: small in daily illumination 13 in the matrix that yellow sand mud=3:2:1 is mixed into
When, intensity of illumination 5100lx, cultivation temperature is 30 DEG C, is cultivated in the incubator that air humidity is 91% 8 days, then transplanting is fixed
It plants in big Tanaka.
It is an advantage of the invention that as demand increases year by year, but inadequate resource, artificial cultivation variet complexity and long-term nothing
The problems such as serious, the yield decline of variety deterioration, disease caused by sexual reproduction.Plant Tissue Breeding, which has, breeds fast, cost
Less, the features such as growth coefficient is high, the present invention, by induction, are proliferated, take root and acclimatization and transplants etc. using stem with bud as explant
Step establishes coriander tissue culture quick breeding system, can not only expand sapling multiplication speed and scale, can also be on this basis
The new varieties for selecting high yield and high quality increase the economic benefit of plantation, wild for alleviating the realistic problem, protection that supply falls short of demand
Resource has great importance.
Specific embodiment
It the following examples are further illustrations of the invention, is not limitation of the present invention.
Embodiment 1:
(1) Fiber differentiation: acquire back stem section Yi Cheongju washing remove surface attachments after, laundry take liquid impregnate 10min, then flow
Water rinses 6h, in superclean bench with 15s is sterilized in 75% ethanol solution, is washed 8 times with sterile afterwards, then molten with 0.1% mercuric chloride
Liquid disinfectant 15min carries out bud induction with being inoculated into induced medium after sucking moisture with aseptic filter paper after aseptic water washing 8 times
Culture.It is first dark culture 12 days full under the conditions of 26 DEG C after inoculation, it is subsequently placed in daily illumination 16 hours, intensity of illumination is
2600lx, adventitious bud can be differentiated by being placed under conditions of cultivation temperature is 26 DEG C to cultivate 30 days, bear adventitious bud after 50 days,
Inductivity is 85%.The induced medium are as follows: MS+1.5mg/L 6-BA+0.8mg/L NAA+25g/L sucrose+4.5g/L fine jade
Rouge, pH 5.6.
(2) Multiplying culture: cultivating obtained sprout for step (1), cuts bud from base portion and to be transferred to proliferated culture medium enterprising
Row squamous subculture, it is first dark culture 6 days full under the conditions of 27 DEG C after inoculation, it is subsequently placed in daily illumination 16 hours, intensity of illumination is
2500lx, sprouting can be differentiated for 16 days by being placed under conditions of cultivation temperature is 26 DEG C to cultivate, and subculture is primary within 35 days, growth coefficient
It is 5.8.The proliferated culture medium are as follows: MS+0.6mg/L 6-BA+23g/L sucrose+4.8g/L agar, pH 5.6.
(3) culture of rootage: by the sprout of step (1) or step (2) it is long to 9cm high when, be cut into simple bud and be inoculated into life
Culture of rootage is carried out on root culture medium, it is first dark culture 7 days full under the conditions of 30 DEG C after inoculation, it is small to be subsequently placed in daily illumination 19
When, intensity of illumination 4000lx, cultivation temperature is cultivated 37 days and can be taken root under conditions of being 30 DEG C, rooting rate 96%, average raw
Root item number is 8.5.The root media are as follows: 1/2MS+1.3mg/L GGR+0.5mg/L IBA+33g/L sucrose+
5.8g/L agar, pH 5.6.
(4) acclimatization and transplants: by the half-open placement hardening 8 under field conditions (factors) of the culture bottle cap of the rooting tube plantlet of high about 7cm
It, then standard-sized sheet bottle cap is added suitable tap water and is placed in natural lighting lower refining seedling 10 days, and test tube seedling is taken out from culture bottle,
Root culture medium is washed off, is planted by peat soil: land: small in daily illumination 15 in the matrix that yellow sand mud=3:2:2 is mixed into
When, intensity of illumination 3500lx, cultivation temperature is 26 DEG C, is cultivated in the incubator that air humidity is 81% 10 days, then transplanting is fixed
It plants in big Tanaka, transplanting survival rate 94%.
Embodiment 2:
(1) Fiber differentiation: acquire back stem section Yi Cheongju washing remove surface attachments after, laundry take liquid impregnate 18min, then flow
Water rinses 9h, in superclean bench with 35s is sterilized in 75% ethanol solution, is washed 10 times with sterile afterwards, then molten with 0.1% mercuric chloride
Liquid disinfectant 20min is carried out bud and is lured with being inoculated into induced medium after sucking moisture with aseptic filter paper after aseptic water washing 10 times
Lead culture.It is first dark culture 15 days full under the conditions of 30 DEG C after inoculation, it is subsequently placed in daily illumination 10 hours, intensity of illumination is
3100lx, adventitious bud can be differentiated by being placed under conditions of cultivation temperature is 30 DEG C to cultivate 22 days, bear adventitious bud after 43 days,
Inductivity is 88%.The induced medium are as follows: MS+6.0mg/L 6-BA+1.0mg/L NAA+30g/L sucrose+5.5g/L fine jade
Rouge, pH 5.3.
(2) Multiplying culture: cultivating obtained sprout for step (1), cuts bud from base portion and to be transferred to proliferated culture medium enterprising
Row squamous subculture, it is first dark culture 9 days full under the conditions of 30 DEG C after inoculation, it is subsequently placed in daily illumination 19 hours, intensity of illumination is
3000lx, sprouting can be differentiated for 19 days by being placed under conditions of cultivation temperature is 30 DEG C to cultivate, and subculture is primary within 35 days, growth coefficient
It is 6.5.The proliferated culture medium are as follows: MS+1.5mg/L 6-BA+25g/L sucrose+5.8g/L agar, pH 5.3.
(3) culture of rootage: by the sprout of step (1) or step (2) it is long to 8cm high when, be cut into simple bud and be inoculated into life
Culture of rootage is carried out on root culture medium, it is first dark culture 8 days full under the conditions of 26 DEG C after inoculation, it is small to be subsequently placed in daily illumination 19
When, intensity of illumination 3500lx, cultivation temperature is cultivated 35 days and can be taken root under conditions of being 26 DEG C, rooting rate 96%, average raw
Root item number is 8.6.The root media are as follows: 1/2MS+1.5mg/L GGR+0.8mg/L IBA+32g/L sucrose+
6.5g/L agar, pH 5.3.
(4) acclimatization and transplants: by the half-open placement hardening 5 under field conditions (factors) of the culture bottle cap of the rooting tube plantlet of high about 10cm
It, then standard-sized sheet bottle cap is added suitable tap water and is placed in natural lighting lower refining seedling 7 days, and test tube seedling is taken out from culture bottle,
Root culture medium is washed off, is planted by peat soil: land: small in daily illumination 16 in the matrix that yellow sand mud=3:2:2 is mixed into
When, intensity of illumination 3600lx, cultivation temperature is 30 DEG C, is cultivated in the incubator that air humidity is 91% 8 days, then transplanting is fixed
It plants in big Tanaka, transplanting survival rate 95%.
Claims (1)
1. a kind of coriander tissue culture and rapid propagation method, it is characterised in that the following steps are included:
Step 1, Fiber differentiation: after the stem section for acquiring back washes away surface attachments with clear water, liquid detergent impregnates 16min, then
Flowing water rinses 3h, in superclean bench with 30s is sterilized in 75% ethanol solution, is washed 7 times with sterile afterwards, then with 0.4% mercuric chloride
Solution disinfection 26min is carried out bud and is lured with being inoculated into induced medium after sucking moisture with aseptic filter paper after aseptic water washing 8 times
Culture is led, it is first dark culture 11 days full under the conditions of 30 DEG C after inoculation, it is subsequently placed in daily illumination 15 hours, intensity of illumination is
3200lx is placed in culture under conditions of cultivation temperature is 30 DEG C until formation adventitious bud;Induced medium are as follows: MS+3.8mg/L
6-BA+1.5mg/L NAA+31g/L sucrose+7.8g/L agar, pH 5.3；
Step 2, Multiplying culture: cultivating obtained sprout for step 1, cut bud from base portion and be transferred on proliferated culture medium carry out after
It is commissioned to train feeding, it is first dark culture 4 days full under the conditions of 30 DEG C after inoculation, it is subsequently placed in daily illumination 17 hours, intensity of illumination is
3200lx is placed under conditions of cultivation temperature is 30 DEG C and cultivates, and subculture is primary within 31 days;Proliferated culture medium are as follows: MS+3.0mg/L
6-BA+40g/L sucrose+8.0g/L agar, pH 5.3；
Step 3, culture of rootage: by the sprout of step 1 or step 2 it is long to 5cm high when, be cut into simple bud and be inoculated into culture of rootage
Culture of rootage is carried out on base, it is first dark culture 4 days full under the conditions of 30 DEG C after inoculation, it is subsequently placed in daily illumination 15 hours, illumination
Intensity is 5100lx, and cultivation temperature is cultivated under conditions of being 30 DEG C to taking root;Root media are as follows: 1/2MS+2.6mg/L GGR
+ 0.6mg/L IBA+32g/L sucrose+7.8g/L agar, pH 5.3；
Step 4, acclimatization and transplants: by the half-open placement hardening 2 under field conditions (factors) of the culture bottle cap of the rooting tube plantlet of high about 7cm
It, then standard-sized sheet bottle cap is added suitable tap water and is placed in natural lighting lower refining seedling 8 days, and test tube seedling is taken out from culture bottle,
Root culture medium is washed off, is planted by peat soil: land: small in daily illumination 13 in the matrix that yellow sand mud=3:2:1 is mixed into
When, intensity of illumination 5100lx, cultivation temperature is 30 DEG C, is cultivated in the incubator that air humidity is 91% 8 days, then transplanting is fixed
It plants in big Tanaka.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104719154A (en) * | 2015-03-05 | 2015-06-24 | 罗焕荣 | Swordlike atractylodes rhizome tissue culture and rapid propagation method |
| CN104719155A (en) * | 2015-03-05 | 2015-06-24 | 罗焕荣 | Phoebe bournei tissue culture and rapid propagation method |
| CN104982330A (en) * | 2015-06-11 | 2015-10-21 | 刘祖英 | Method for tissue culture and rapid propagation of strobilanthes cusia |
| CN107494277A (en) * | 2017-10-15 | 2017-12-22 | 陈金水 | A kind of gingko tissue culture and rapid propagation method |
-
2018
- 2018-11-25 CN CN201811412316.6A patent/CN109362564A/en not_active Withdrawn
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104719154A (en) * | 2015-03-05 | 2015-06-24 | 罗焕荣 | Swordlike atractylodes rhizome tissue culture and rapid propagation method |
| CN104719155A (en) * | 2015-03-05 | 2015-06-24 | 罗焕荣 | Phoebe bournei tissue culture and rapid propagation method |
| CN104982330A (en) * | 2015-06-11 | 2015-10-21 | 刘祖英 | Method for tissue culture and rapid propagation of strobilanthes cusia |
| CN107494277A (en) * | 2017-10-15 | 2017-12-22 | 陈金水 | A kind of gingko tissue culture and rapid propagation method |
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Application publication date: 20190222 |