CN108401911A - Jade dew tissue culture medium (TCM) and preparation method thereof - Google Patents
Jade dew tissue culture medium (TCM) and preparation method thereof Download PDFInfo
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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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Abstract
The invention discloses a kind of beautiful dew tissue culture medium (TCM)s and preparation method thereof, it includes inducing culture, proliferated culture medium, strong seedling culture base and root media and preparation method thereof, wherein, inducing culture is to increase citric acid, 6 benayl aminopurines, methyl α-naphthyl acetate, 6 chaff adenine phosphates, sugar and carragheen on the basis of MS culture mediums；Proliferated culture medium is to increase citric acid, sugar and carragheen on the basis of 1/2MS culture mediums；Strong seedling culture base is to increase citric acid, mashed potatoes, apple butter, banana puree, sugar and carragheen on the basis of 1/2MS culture mediums；Root media is to increase citric acid, mashed potatoes, apple butter, banana puree, indolebutyric acid, activated carbon, sugar and carragheen on the basis of 1/2MS culture mediums.It has many advantages, such as that hormone dosage is few, breeding is fast, quantity is big, goes out full stand, variety pure, at low cost, wide adaptability for beautiful dew tissue culture.
Description
Technical field
The invention belongs to field of plant tissue culture technique, especially a kind of beautiful dew tissue culture medium (TCM) and preparation method thereof.
Background technology
Jade dew is the soft leaf system kind in A Fu flowers section category succulents of volume 12, and plant is small and exquisite, and plant type is compact, layer
Secondary clearly demarcated, blade plumpness is full, and leaf color is glittering and translucent, and leaf " window " is transparent and has green texture, and resistance is strong, has certain sky
Gas purification function has catered to modern people and has advocated health and pursue beautiful theory, had very high ornamental value and economic valence
Value.
Succulent fertility is poor, mostly uses sowing greatly, leaf is inserted and the propagation methods such as plant division, but these traditional breedings
Means have that breeding coefficient is relatively low and breeding cycle length, it is difficult to meet production needs, therefore famous-object valence in the market
Lattice are high.Compared with traditional division propagation mode, jade dew is quickly bred using tissue culture technique, it can
Breeding coefficient and speed are greatly improved, and is not influenced by outside environmental elements such as season and weathers, whole year can carry out numerous
It grows.Therefore, carrying out industrialization production to succulent by tissue cultures has preferable economic benefit and market prospects.
Invention content
It is an object of the invention to overcome above-mentioned problems of the prior art, provide a kind of beautiful dew tissue culture medium (TCM) and
Preparation method carries out tissue cultures using the culture medium and beautiful dew explant blade, can greatly increase breeding coefficient, solve
Beautiful dew low reproduction rate, the problem of production cycle length under natural conditions.
The technical proposal of the invention is realized in this way：A kind of beautiful dew tissue culture medium (TCM), it includes inducing culture, proliferation
Culture medium, strong seedling culture base and root media, it is characterised in that：
The inducing culture includes following components:MS basal mediums 4736mg/L, 25~35mg/L of citric acid, 6- benzyl ammonia
2~4mg/L of base purine, 0.05~0.15 mg/L of methyl α-naphthyl acetate, 0.5~1.5mg/L of 6-Furfurylaminopurine, sugar 29000~
32000mg/L, 7000~9000mg/L of carragheen；
The proliferated culture medium includes following components:1/2MS basal mediums 2471mg/L, 25~35mg/L of citric acid, sugar
29000~32000mg/L, 7000~9000mg/L of carragheen；
The strong seedling culture base includes following components:1/2MS basal mediums 2471mg/L, 25~35mg/L of citric acid, potato
16000~20000mg/L of mud, 16000~20000mg/L of apple butter, 16000~20000mg/L of banana puree, sugar 19000~
22000mg/L, 7000~9000mg/L of carragheen；
The root media includes following components:1/2MS culture medium pulvis 2471mg/L, 25~35mg/L of citric acid, potato
16000~20000mg/L of mud, 16000~20000mg/L of apple butter, 16000~20000mg/L of banana puree, indolebutyric acid
0.2-0.4 mg/L, 400~700mg/L of activated carbon, 19000~22000mg/L of sugar, 7000~9000mg/L of carragheen.
The present invention additionally provides a kind of system of beautiful dew tissue culture medium (TCM) on the basis of above-mentioned beautiful dew tissue cultures based formulas
Preparation Method, it includes the preparation method of inducing culture, proliferated culture medium, strong seedling culture base and root media, and feature exists
In：
The preparation method of the inducing culture weighs MS culture mediums pulvis, citric acid, sugar, carragheen in appearance by formula rate
In device, add distilled water, dissolves by heating；Formula rate addition, 6-benzyl aminopurine, methyl α-naphthyl acetate, 6-Furfurylaminopurine are pressed again；Add steaming
Distilled water is settled to 1L；Then pH value 6 is adjusted；It is finally divided in 20 small triangular flasks, bottleneck is tamping with sealed membrane, in high temperature 121
~126 DEG C, 0.1~0.14 megapascal of high pressure sterilizes 15~20 minutes, and taking-up shakes up cooling obtained inducing culture while hot；
The preparation method of the proliferated culture medium, by formula rate weigh 1/2MS culture mediums pulvis, citric acid, sugar, carragheen in
In container, add distilled water, dissolves by heating；Distilled water is added to be settled to 1L；Then pH value 6 is adjusted；Finally it is divided in 12 tissue cultures
In bottle, bottleneck is stoppered with rubber plug, then bottleneck is tamping with sealed membrane, in 121~126 DEG C of high temperature, 0.1~0.14 megapascal of high pressure is gone out
Bacterium 15~20 minutes, taking-up shake up cooling obtained proliferated culture medium while hot；
The preparation method of the strong seedling culture base, by formula rate weigh 1/2MS culture mediums pulvis, citric acid, sugar, carragheen in
In container, add distilled water, dissolves by heating；Mashed potatoes, apple butter, banana puree is added by formula rate again to stir evenly；Add distillation
Water is settled to 1L；Then pH value 6 is adjusted；It is finally divided in 12 tissue culture flasks, stoppers bottleneck with rubber plug, then use sealed membrane
It is tamping bottleneck, in 121~126 DEG C of high temperature, 0.1~0.14 megapascal of high pressure sterilizes 15~20 minutes, and taking-up shakes up cooling system while hot
Obtain strong seedling culture base；
The preparation method of the root media, by formula rate weigh 1/2MS culture mediums pulvis, citric acid, activated carbon, sugar,
Carragheen adds distilled water in container, dissolves by heating；Mashed potatoes, apple butter, banana puree, indolebutyric acid is added by formula again to stir
It mixes uniformly；Distilled water is added to be settled to 1L；Then pH value 6 is adjusted；It is finally divided in 12 tissue culture flasks, bottle is stoppered with rubber plug
Mouthful, then it is tamping bottleneck with sealed membrane, in 121~126 DEG C of high temperature, 0.1~0.14 megapascal of high pressure sterilizes 15~20 minutes, takes out
Cooling obtained root media is shaken up while hot.
The mashed potatoes is that potato is cleaned peeling is cooked, using smashing alms bowl to pieces or tissue mashing machine smashs to obtain potato to pieces
Mud.
The apple butter is to be enucleated apple peel, using smashing alms bowl to pieces or tissue mashing machine smashs to pieces to obtain apple butter.
The banana puree is to remove the peel banana, using smashing alms bowl to pieces or tissue mashing machine smashs to pieces to obtain banana puree.
The present invention compared to the prior art the advantages of be：
（1）The culture medium of the present invention includes inducing culture, proliferated culture medium, strong seedling culture base and root media, using outer
Implant is directly inoculated into the induction that Multiple Buds are carried out in inducing culture, Multiple Buds are cut to be inoculated into proliferated culture medium carries out
Shoot proliferation culture, is then transplanted to nurturing staff on strong seedling culture base again, is finally transplanted on root media and continues to train
It supports and root growth is promoted to cultivate the tissue-culture container seedling for meeting transplanting standard.The culture of different formulations is used in different cultivation stages
The characteristics of base, the nutritional need of adaptation tissue cultures different phase, seedling directly is grown from Multiple Buds, simplifies incubation step,
Reduce production cost.
（2）Organic matter mashed potatoes, apple butter and the banana puree added in the strong seedling culture base and root media of the present invention,
The seedling grown can be promoted neat, healthy and strong.
（3）Beautiful dew tissue cultures are carried out using the culture medium of the present invention, survival rate is 94~98%, and pollution rate is 2~3%,
Breeding coefficient is 8~10 times.
（4）The present invention is few with hormone dosage for beautiful dew tissue culture, breeding is fast, quantity is big, goes out full stand, variety pure, cost
The market competitiveness can be improved in the advantages that low, wide adaptability, promotes beautiful dew industry development.Beautiful dew conditions of tissue culture after optimization,
Reveal large-scale industrialized production suitable for jade.
Specific implementation mode
To keep technical scheme of the present invention clearer, embodiment of the present invention is made into one below by specific embodiment
Step ground detailed description.
Embodiment 1：A kind of beautiful dew tissue culture medium (TCM), it includes inducing culture, proliferated culture medium, strong seedling culture base and life
Root culture medium, it is characterised in that：
The inducing culture includes following components:MS basal mediums 4736mg/L, citric acid 30mg/L, 6- benzyl amino are fast
Purine 3mg/L, methyl α-naphthyl acetate 0.1mg/L, 6-Furfurylaminopurine 1mg/L, sugar 30500mg/L, carragheen 8000mg/L；
The proliferated culture medium includes following components:1/2MS basal mediums 2471mg/L, citric acid 30mg/L, sugar
30500mg/L, carragheen 8000mg/L；
The strong seedling culture base includes following components:1/2MS basal mediums 2471mg/L, citric acid 30mg/L, mashed potatoes
18000mg/L, apple butter 18000mg/L, banana puree 18000mg/L, sugar 20500mg/L, carragheen 8000mg/L；
The root media includes following components:1/2MS culture medium pulvis 2471mg/L, citric acid 30mg/L, mashed potatoes
18000mg/L, apple butter 18000mg/L, banana puree 18000mg/L, 0.3 mg/L of indolebutyric acid, activated carbon 550mg/L, sugar
20500mg/L, carragheen 8000mg/L.
Embodiment 2：A kind of beautiful dew tissue culture medium (TCM), it includes inducing culture, proliferated culture medium, strong seedling culture base and life
Root culture medium, it is characterised in that：
The inducing culture includes following components:MS basal mediums 4736mg/L, citric acid 25mg/L, 6- benzyl amino are fast
Purine 4mg/L, methyl α-naphthyl acetate 0.05mg/L, 6-Furfurylaminopurine 1.5mg/L, sugar 29000mg/L, carragheen 9000mg/L；
The proliferated culture medium includes following components:1/2MS basal mediums 2471mg/L, citric acid 25mg/L, sugar
32000mg/L, carragheen 7000mg/L；
The strong seedling culture base includes following components:1/2MS basal mediums 2471mg/L, citric acid 35mg/L, mashed potatoes
16000mg/L, apple butter 20000mg/L, banana puree 16000mg/L, sugar 19000mg/L, carragheen 9000mg/L；
The root media includes following components:1/2MS culture medium pulvis 2471mg/L, citric acid 35mg/L, mashed potatoes
20000mg/L, apple butter 16000mg/L, banana puree 20000mg/L, indolebutyric acid 0.2mg/L, activated carbon 700mg/L, sugar
19000mg/L, carragheen 9000mg/L.
Embodiment 3：A kind of beautiful dew tissue culture medium (TCM), it includes inducing culture, proliferated culture medium, strong seedling culture base and life
Root culture medium, it is characterised in that：
The inducing culture includes following components:MS basal mediums 4736mg/L, citric acid 35mg/L, 6- benzyl amino are fast
Purine 2mg/L, methyl α-naphthyl acetate 0.15mg/L, 6-Furfurylaminopurine 0.5mg/L, sugar 32000mg/L, carragheen 7000mg/L；
The proliferated culture medium includes following components:1/2MS basal mediums 2471mg/L, citric acid 35mg/L, sugar
29000mg/L, carragheen 9000mg/L；
The strong seedling culture base includes following components:1/2MS basal mediums 2471mg/L, citric acid 25mg/L, mashed potatoes
20000mg/L, apple butter 16000mg/L, banana puree 20000mg/L, sugar 22000mg/L, carragheen 7000mg/L；
The root media includes following components:1/2MS culture medium pulvis 2471mg/L, citric acid 25mg/L, mashed potatoes
16000mg/L, apple butter 20000mg/L, banana puree 16000mg/L, indolebutyric acid 0.4mg/L, activated carbon 400mg/L, sugar
22000mg/L, carragheen 7000mg/L.
Embodiment 4：A kind of preparation method of beautiful dew tissue culture medium (TCM), using the formula rate of embodiment 1, respectively by as follows
Step and method prepare inducing culture, proliferated culture medium, strong seedling culture base and root media；
The preparation method of the inducing culture weighs MS culture mediums pulvis, citric acid, sugar, carragheen in appearance by formula rate
In device, add distilled water, dissolves by heating；Formula rate addition, 6-benzyl aminopurine, methyl α-naphthyl acetate, 6-Furfurylaminopurine are pressed again；Add steaming
Distilled water is settled to 1L；Then pH value 6 is adjusted；It is finally divided in 20 small triangular flasks, bottleneck is tamping with sealed membrane, in high temperature 124
DEG C, 0.12 megapascal of high pressure sterilizes 18 minutes, and taking-up shakes up cooling obtained inducing culture while hot；
The preparation method of the proliferated culture medium, by formula rate weigh 1/2MS culture mediums pulvis, citric acid, sugar, carragheen in
In container, add distilled water, dissolves by heating；Distilled water is added to be settled to 1L；Then pH value 6 is adjusted；Finally it is divided in 12 tissue cultures
In bottle, bottleneck is stoppered with rubber plug, then bottleneck is tamping with sealed membrane, in 124 DEG C of high temperature, 0.12 megapascal of high pressure sterilizes 18 minutes, takes
Go out and shakes up cooling obtained proliferated culture medium while hot；
The preparation method of the strong seedling culture base, by formula rate weigh 1/2MS culture mediums pulvis, citric acid, sugar, carragheen in
In container, add distilled water, dissolves by heating；Mashed potatoes, apple butter, banana puree is added by formula rate again to stir evenly；Add distillation
Water is settled to 1L；Then pH value 6 is adjusted；It is finally divided in 12 tissue culture flasks, stoppers bottleneck with rubber plug, then use sealed membrane
It is tamping bottleneck, in 121~126 DEG C of high temperature, 0.12 megapascal of high pressure sterilizes 18 minutes, and taking-up shakes up cooling obtained strong sprout training while hot
Support base；
The preparation method of the root media, by formula rate weigh 1/2MS culture mediums pulvis, citric acid, activated carbon, sugar,
Carragheen adds distilled water in container, dissolves by heating；Mashed potatoes, apple butter, banana puree, indolebutyric acid is added by formula again to stir
It mixes uniformly；Distilled water is added to be settled to 1L；Then pH value 6 is adjusted；It is finally divided in 12 tissue culture flasks, bottle is stoppered with rubber plug
Mouthful, then it is tamping bottleneck with sealed membrane, in 124 DEG C of high temperature, 0.12 megapascal of high pressure sterilizes 18 minutes, and taking-up shakes up cooling system while hot
Obtain root media.
Claims (2)
1. a kind of beautiful dew tissue culture medium (TCM), it includes inducing culture, proliferated culture medium, strong seedling culture base and root media,
It is characterized in that：
The inducing culture includes following components:MS basal mediums 4736mg/L, 25~35mg/L of citric acid, 6- benzyl ammonia
2~4mg/L of base purine, 0.05~0.15 mg/L of methyl α-naphthyl acetate, 0.5~1.5mg/L of 6-Furfurylaminopurine, sugar 29000~
32000mg/L, 7000~9000mg/L of carragheen；
The proliferated culture medium includes following components:1/2MS basal mediums 2471mg/L, 25~35mg/L of citric acid, sugar
29000~32000mg/L, 7000~9000mg/L of carragheen；
The strong seedling culture base includes following components:1/2MS basal mediums 2471mg/L, 25~35mg/L of citric acid, potato
16000~20000mg/L of mud, 16000~20000mg/L of apple butter, 16000~20000mg/L of banana puree, sugar 19000~
22000mg/L, 7000~9000mg/L of carragheen；
The root media includes following components:1/2MS culture medium pulvis 2471mg/L, 25~35mg/L of citric acid, potato
16000~20000mg/L of mud, 16000~20000mg/L of apple butter, 16000~20000mg/L of banana puree, indolebutyric acid
0.2-0.4 mg/L, 400~700mg/L of activated carbon, 19000~22000mg/L of sugar, 7000~9000mg/L of carragheen.
2. the Preparation Method of beautiful dew tissue culture medium (TCM) described in claim 1, it includes inducing culture, proliferated culture medium, strong sprout
The preparation method of culture medium and root media, it is characterised in that：
The preparation method of the inducing culture weighs MS culture mediums pulvis, citric acid, sugar, carragheen in appearance by formula rate
In device, add distilled water, dissolves by heating；Formula rate addition, 6-benzyl aminopurine, methyl α-naphthyl acetate, 6-Furfurylaminopurine are pressed again；Add steaming
Distilled water is settled to 1L；Then pH value 6 is adjusted；It is finally divided in 20 small triangular flasks, bottleneck is tamping with sealed membrane, in high temperature 121
~126 DEG C, 0.1~0.14 megapascal of high pressure sterilizes 15~20 minutes, and taking-up shakes up cooling obtained inducing culture while hot；
The preparation method of the proliferated culture medium, by formula rate weigh 1/2MS culture mediums pulvis, citric acid, sugar, carragheen in
In container, add distilled water, dissolves by heating；Distilled water is added to be settled to 1L；Then pH value 6 is adjusted；Finally it is divided in 12 tissue cultures
In bottle, bottleneck is stoppered with rubber plug, then bottleneck is tamping with sealed membrane, in 121~126 DEG C of high temperature, 0.1~0.14 megapascal of high pressure is gone out
Bacterium 15~20 minutes, taking-up shake up cooling obtained proliferated culture medium while hot；
The preparation method of the strong seedling culture base, by formula rate weigh 1/2MS culture mediums pulvis, citric acid, sugar, carragheen in
In container, add distilled water, dissolves by heating；Mashed potatoes, apple butter, banana puree is added by formula rate again to stir evenly；Add distillation
Water is settled to 1L；Then pH value 6 is adjusted；It is finally divided in 12 tissue culture flasks, stoppers bottleneck with rubber plug, then use sealed membrane
It is tamping bottleneck, in 121~126 DEG C of high temperature, 0.1~0.14 megapascal of high pressure sterilizes 15~20 minutes, and taking-up shakes up cooling system while hot
Obtain strong seedling culture base；
The preparation method of the root media, by formula rate weigh 1/2MS culture mediums pulvis, citric acid, activated carbon, sugar,
Carragheen adds distilled water in container, dissolves by heating；Mashed potatoes, apple butter, banana puree, indolebutyric acid is added by formula again to stir
It mixes uniformly；Distilled water is added to be settled to 1L；Then pH value 6 is adjusted；It is finally divided in 12 tissue culture flasks, bottle is stoppered with rubber plug
Mouthful, then it is tamping bottleneck with sealed membrane, in 121~126 DEG C of high temperature, 0.1~0.14 megapascal of high pressure sterilizes 15~20 minutes, takes out
Cooling obtained root media is shaken up while hot.
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Application publication date: 20180817 |