CN107258544A - A kind of method that utilization petiole induces epipremnum aureum - Google Patents
A kind of method that utilization petiole induces epipremnum aureum Download PDFInfo
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- CN107258544A CN107258544A CN201710655041.8A CN201710655041A CN107258544A CN 107258544 A CN107258544 A CN 107258544A CN 201710655041 A CN201710655041 A CN 201710655041A CN 107258544 A CN107258544 A CN 107258544A
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- epipremnum aureum
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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
- A01H4/008—Methods for regeneration to complete plants
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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 present invention provides a kind of method that utilization petiole induces epipremnum aureum, comprises the following steps：Maternal plant is chosen, epipremnum aureum explant is taken after pretreatment；The epipremnum aureum explant is subjected to mechanical damage, and is seeded in callus inducing medium, callus is obtained；The callus is forwarded in subculture medium and cultivated, is produced.The aseptic seedling for the epipremnum aureum petiole that the present invention is cultivated by epipremnum aureum tissue culture technique, growth cycle can be shortened, avoid or reduce the loss that extensive stem rot, root-rot caused by extraneous undesirable element are brought, avoid and epipremnum aureum is bred by cuttage at present, the easily susceptible bacterium of the offspring of cutting propagation, often occur extensive stem rot, root-rot phenomenon in production, the problem of cultivation to epipremnum aureum causes certain economic loss.
Description
Technical field
The present invention relates to technical field of plant culture, specifically, it is related to a kind of method that utilization petiole induces epipremnum aureum.
Background technology
Epipremnum aureum (scientific name：Epipremnum aureum), belong to kylin leaf platymiscium, large-scale evergreen liana is grown on the torrid zone
Area, normal climbing growth is on the rock and trunk of rainforest, and its prehensile is strong, and aerial root is flourishing, can be planted with water planting.On ripe branch
Petiole is sturdy, and long 30-40 centimetres, base portion slightly expands, and upper joints are long 2.5-3 centimetres, slightly plump, and outside of belly tool sipes, leaf sheath is long,
The thin keratin of blade, emerald green, generally (particularly blade face) has most irregular gilvous patches, and full edge is inequilateral avette
Or ovate Long Circle, tip is short tapering, base portion innermost being shape, and slightly slightly, two sides is slightly swelled.
Epipremnum aureum original producton location is in the tropical rain forest of Indonesia Solomon Islands, and property likes warm, wet environment, it is desirable to soil
The loose fertile, draining of earth is good.In torrid areas, normal climbing growth is planted on rock and trunk as huge ornamental vine
There is plant thing, south China various regions.Epipremnum aureum shade tolerance is strong, is green throughout the year, there is thicker cuticula on blade face, adapts to Indoor Dry
Dry environmental condition, is excellent Ornamental Foliage House Plants.Moreover it is possible to adsorb and remove the formaldehyde in room air, benzene, trichlorine
The pollutants such as ethene, are natural " air purifiers ", with larger market prospects.
The breeding of epipremnum aureum is currently to be bred by cuttage, because epipremnum aureum growing environment is hot and humid, cutting propagation
, often there is extensive stem rot, root-rot phenomenon in the easily susceptible bacterium of offspring in production, the cultivation to epipremnum aureum causes certain economic damage
Lose.
The content of the invention
In view of this, the technical problem to be solved in the present invention be to overcome the breeding of epipremnum aureum be currently carried out by cuttage it is numerous
Grow, because epipremnum aureum growing environment is hot and humid, extensive stem often occurs in the offspring easily susceptible bacterium of cutting propagation in production
Rotten, root-rot phenomenon, the cultivation to epipremnum aureum causes the defect of certain economic loss.
To solve the above problems, the present invention provides a kind of method that utilization petiole induces epipremnum aureum, comprise the following steps：
Maternal plant is chosen, epipremnum aureum explant is taken after pretreatment；
The epipremnum aureum explant is subjected to mechanical damage, and is seeded in callus inducing medium, callus is obtained；
The callus is forwarded in subculture medium and cultivated, is produced；
The callus inducing medium 1 includes following component：
Modified MS medium 1,6- benzyls aminoadenine, 6- glycosyls adenine phosphate, methyl α-naphthyl acetate, D butyl esters, sucrose, coconut
Water and agar powder；
The callus inducing medium 2 includes following component：
Modified MS medium 2,6- benzyls aminoadenine, 6- glycosyls adenine phosphate, methyl α-naphthyl acetate, indole-3-acetic acid, sugarcane
Sugar, coconut water and agar powder.
Preferably, the selection maternal plant, takes epipremnum aureum explant after pretreatment, including：
Choose maternal plant and use carbendazim pouring root；
Take tender leaf at the maternal plant growing point；
The tender leaf is sterilized, epipremnum aureum explant is obtained.
Preferably, the selection maternal plant uses carbendazim pouring root, including：
Choose maternal plant and carry out pouring root 30 days, every 10 days 1 time using 80% carbendazim, 1000 times of solution.
Preferably, the callus inducing medium includes callus inducing medium 1 and callus induction
Culture medium 2；
It is described that the epipremnum aureum explant is subjected to mechanical damage, and be seeded in callus inducing medium, obtain callus
Tissue, including：
After the epipremnum aureum explant is trimmed, petiole section sample is obtained；
Petiole section sample is subjected to mechanical damage；
The petiole of mechanical damage section sample is seeded in callus inducing medium 1, in 25 DEG C -28 DEG C of temperature
Cultivated with conditions of humidity 50%-65%, obtain the first callus；
The callus is transferred in inducing culture 2, in 25 DEG C -28 DEG C of temperature and humidity 50%-65%
Under the conditions of cultivated, obtain the second callus.
Preferably, the callus inducing medium 1 is configured as follows：
In modified MS medium 1, according to concentration addition 3.0mg/L 6- benzyls aminoadenine, 0.1mg/L 6- sugar
Base adenine phosphate, 0.2mg/L methyl α-naphthyl acetate, 0.05mg/L D butyl esters, 30g/L sucrose, 100ml/L coconut water and 7g/L
Agar powder, regulation pH value to 5.6-5.8, produce；
The callus inducing medium 2 is configured as follows：
In modified MS medium 2, according to concentration addition 1.0mg/L 6- benzyls aminoadenine, 0.1mg/L 6- sugar
Base adenine phosphate, 0.2mg/L methyl α-naphthyl acetate, 0.1mg/L indole-3-acetic acid, 30g/L sucrose, 150ml/L coconut water and
7g/L agar powder, regulation pH value is produced to 5.6-5.8.
Preferably, the modified MS medium 1 is configured as follows：
Based on MS culture mediums, in addition to following component, and adjust content and be：
Inositol：1-5mg/L；
Thiamine hydrochloride：3-6mg/L；
Puridoxine hydrochloride：0.5-2mg/L；
Nicotinic acid：0.5-3mg/L；
Arginine：50mg/L；
Aspartic acid：100mg/L；
Glutamic acid：150mg/L；
The modified MS medium 2 is configured as follows：
Based on MS culture mediums, in addition to following component, and adjust content and be：
Inositol：1-5mg/L；
Thiamine hydrochloride：3-6mg/L；
Puridoxine hydrochloride：0.5-2mg/L；
Nicotinic acid：0.5-3mg/L；
Arginine：100mg/L；
Aspartic acid：50mg/L；
Glutamic acid：200mg/L.
Preferably, the described callus is forwarded in subculture medium is cultivated, and is produced, including：
Second callus is forwarded in urgently culture medium and cultivated under default condition of culture；
Every 40 days subcultures once, are produced.
Preferably, the default condition of culture, including：
Intensity of illumination：1500-20001x；
Light application time：12 hours/day；
Temperature：25℃-28℃；
Humidity：50%-65%.
Preferably, the subculture medium includes following component：Modified MS medium 2,6- benzyls aminoadenine, Yin
Diindyl -3- acetic acid, sucrose and agar powder.
Preferably, the subculture medium, is prepared as follows：
In modified MS medium 2, according to concentration addition 2.0mg/L 6- benzyls aminoadenine, 0.3mg/L Yin
The agar powder of diindyl -3- acetic acid, 30g/L sucrose and 7g/L, regulation pH value is produced to 5.6-5.8.
The present invention provides a kind of method that utilization petiole induces epipremnum aureum, including chooses maternal plant, takes after pretreatment outside epipremnum aureum
Implant；The epipremnum aureum explant is subjected to mechanical damage, and is seeded in callus inducing medium, callus is obtained；Will
The callus is forwarded in subculture medium and cultivated, and produces.The epipremnum aureum that the present invention is cultivated by epipremnum aureum tissue culture technique
The aseptic seedling of petiole, can shorten growth cycle, it is to avoid or extensive stem rot, root-rot are brought caused by the extraneous undesirable element of reduction
Loss, it is to avoid epipremnum aureum is bred by cuttage at present, the offspring easily susceptible bacterium of cutting propagation often goes out in production
Now extensive stem rot, root-rot phenomenon, the problem of cultivation to epipremnum aureum causes certain economic loss.
Embodiment
In order to illustrate more clearly of technical scheme, with reference to claim of the specific embodiment to the present invention
It is described in further detail, it is to be understood that illustrate only certain embodiments of the present invention below, therefore be not to be seen as
It is the restriction to scope, the still modification of anyone limited number of time made within the scope of the invention as claimed, the power in the present invention
Within sharp claimed range.
The present invention provides a kind of method that utilization petiole induces epipremnum aureum, comprises the following steps：
Maternal plant is chosen, epipremnum aureum explant is taken after pretreatment；
The epipremnum aureum explant is subjected to mechanical damage, and is seeded in callus inducing medium, callus is obtained；
The callus inducing medium 1 includes following component：
Modified MS medium 1,6- benzyls aminoadenine, 6- glycosyls adenine phosphate, methyl α-naphthyl acetate, D butyl esters, sucrose, coconut
Water and agar powder；
The callus inducing medium 2 includes following component：
Modified MS medium 2,6- benzyls aminoadenine, 6- glycosyls adenine phosphate, methyl α-naphthyl acetate, indole-3-acetic acid, sugarcane
Sugar, coconut water and agar powder.
Epipremnum aureum (scientific name：Epipremnum aureum), belong to kylin leaf platymiscium, large-scale evergreen liana is grown on the torrid zone
Area, normal climbing growth is on the rock and trunk of rainforest, and its prehensile is strong, and aerial root is flourishing, can be planted with water planting.On ripe branch
Petiole is sturdy, and long 30-40 centimetres, base portion slightly expands, and upper joints are long 2.5-3 centimetres, slightly plump, and outside of belly tool sipes, leaf sheath is long,
The thin keratin of blade, emerald green, generally (particularly blade face) has most irregular gilvous patches, and full edge is inequilateral avette
Or ovate Long Circle, tip is short tapering, base portion innermost being shape, and slightly slightly, two sides is slightly swelled.
Epipremnum aureum original producton location is in the tropical rain forest of Indonesia Solomon Islands, and property likes warm, wet environment, it is desirable to soil
The loose fertile, draining of earth is good.In torrid areas, normal climbing growth is planted on rock and trunk as huge ornamental vine
There is plant thing, south China various regions.Epipremnum aureum shade tolerance is strong, is green throughout the year, there is thicker cuticula on blade face, adapts to Indoor Dry
Dry environmental condition, is excellent Ornamental Foliage House Plants.Moreover it is possible to adsorb and remove the formaldehyde in room air, benzene, trichlorine
The pollutants such as ethene, are natural " air purifiers ", with larger market prospects.
The present invention provides a kind of method that utilization petiole induces epipremnum aureum, including chooses maternal plant, takes after pretreatment outside epipremnum aureum
Implant；The epipremnum aureum explant is subjected to mechanical damage, and is seeded in callus inducing medium, callus is obtained；Will
The callus is forwarded in subculture medium and cultivated, and produces.The epipremnum aureum that the present invention is cultivated by epipremnum aureum tissue culture technique
The aseptic seedling of petiole, can shorten growth cycle, it is to avoid or extensive stem rot, root-rot are brought caused by the extraneous undesirable element of reduction
Loss, it is to avoid epipremnum aureum is bred by cuttage at present, the offspring easily susceptible bacterium of cutting propagation often goes out in production
Now extensive stem rot, root-rot phenomenon, the problem of cultivation to epipremnum aureum causes certain economic loss.
Preferably, the selection maternal plant, takes epipremnum aureum explant after pretreatment, including：
Choose maternal plant and use carbendazim pouring root；
Take tender leaf at the maternal plant growing point；
The tender leaf is sterilized, epipremnum aureum explant is obtained.
Preferably, the selection maternal plant uses carbendazim pouring root, including：
Choose maternal plant and carry out pouring root 30 days, every 10 days 1 time using 80% carbendazim, 1000 times of solution.
It is above-mentioned, it is the step of explant is chosen, selection form is normal, eugonic healthy and strong maternal plant pretreatment (is used
80% 1000 times of carbendazol wettable powder liquid irrigating root, every 10 days are once) after 30 days, cut 2~3 childrens at Stem nematode point tender complete
Whole leaf, the explant after surface removal of impurities, cleaning is transferred to superclean bench and carried out disinfection processing, obtains sterile epipremnum aureum explant
Body.In addition, the tangent plane of epipremnum aureum maternal plant carries out protective treatment after explant collection to tangent plane, can be again after growth to be restored
Sampling.
Preferably, the callus inducing medium includes callus inducing medium 1 and callus induction
Culture medium 2；
It is described that the epipremnum aureum explant is subjected to mechanical damage, and be seeded in callus inducing medium, obtain callus
Tissue, including：
After the epipremnum aureum explant is trimmed, petiole section sample is obtained；
Petiole section sample is subjected to mechanical damage；
The petiole of mechanical damage section sample is seeded in callus inducing medium 1, in 25 DEG C -28 DEG C of temperature
Cultivated with conditions of humidity 50%-65%, obtain the first callus；
The callus is transferred in inducing culture 2, in 25 DEG C -28 DEG C of temperature and humidity 50%-65%
Under the conditions of cultivated, obtain the second callus.
It is above-mentioned, it is the Fiber differentiation process of the callus in the present invention.Specifically, retaining epipremnum aureum blade apart from phyllopodium 1
Petiole at~1.5cm, epipremnum aureum petiole is cut into chunks sample, and is carried out rip cutting in base portion and carved wound, is inoculated into callus induction training
Support in base 1, in the state of couching, half is submerged in the medium, is 25~28 DEG C in temperature, humidity is 50~65% condition
Lower carry out light culture, expands until petiole carves injury, obtains the first callus；Petiole (the first callus) is expanded by what is obtained
Transferred again after rip cutting in callus inducing medium 2, be 25~28 DEG C in temperature, humidity is 50~65% condition
Lower light culture, obtains the second callus.
Preferably, the callus inducing medium 1 is configured as follows：
In modified MS medium 1, according to concentration addition 3.0mg/L 6- benzyls aminoadenine, 0.1mg/L 6- sugar
Base adenine phosphate, 0.2mg/L methyl α-naphthyl acetate, 0.05mg/L D butyl esters, 30g/L sucrose, 100ml/L coconut water and 7g/L
Agar powder, regulation pH value to 5.6-5.8, produce；
The callus inducing medium 2 is configured as follows：
In modified MS medium 2, according to concentration addition 1.0mg/L 6- benzyls aminoadenine, 0.1mg/L 6- sugar
Base adenine phosphate, 0.2mg/L methyl α-naphthyl acetate, 0.1mg/L indole-3-acetic acid, 30g/L sucrose, 150ml/L coconut water and
7g/L agar powder, regulation pH value is produced to 5.6-5.8.
Preferably, the modified MS medium 1 is configured as follows：
Based on MS culture mediums, in addition to following component, and adjust content and be：
Inositol：1-5mg/L；
Thiamine hydrochloride：3-6mg/L；
Puridoxine hydrochloride：0.5-2mg/L；
Nicotinic acid：0.5-3mg/L；
Arginine：50mg/L；
Aspartic acid：100mg/L；
Glutamic acid：150mg/L；
The modified MS medium 2 is configured as follows：
Based on MS culture mediums, in addition to following component, and adjust content and be：
Inositol：1-5mg/L；
Thiamine hydrochloride：3-6mg/L；
Puridoxine hydrochloride：0.5-2mg/L；
Nicotinic acid：0.5-3mg/L；
Arginine：100mg/L；
Aspartic acid：50mg/L；
Glutamic acid：200mg/L.
It is above-mentioned, it is to be understood that MS culture mediums are that Murashige and Skoog were that tobacco cell culture is set in 1962
Meter, it is characterized in that inorganic salts and ion concentration are higher, is more stable ionic equilibrium solution, its nitrate content is high, its
The quantity and ratio of nutrient are suitable, can meet the nutrition and physiological requirements of plant cell, thus the scope of application is wider, Duo Shuozhi
Thing tissue-culturing quick-propagation uses it as the minimal medium of culture medium.Based on this, this culture medium is just with their name
To name.It is as shown in the table for its formula.
The MS culture medium prescription tables of table 1
Preferably, the described callus is forwarded in subculture medium is cultivated, and is produced, including：
Second callus is forwarded in urgently culture medium and cultivated under default condition of culture；
Every 40 days subcultures once, are produced.
Preferably, the default condition of culture, including：
Intensity of illumination：1500-20001x；
Light application time：12 hours/day；
Temperature：25℃-28℃；
Humidity：50%-65%.
Preferably, the subculture medium includes following component：Modified MS medium 2,6- benzyls aminoadenine, Yin
Diindyl -3- acetic acid, sucrose and agar powder.
Preferably, the subculture medium, is prepared as follows：
In modified MS medium 2, according to concentration addition 2.0mg/L 6- benzyls aminoadenine, 0.3mg/L Yin
The agar powder of diindyl -3- acetic acid, 30g/L sucrose and 7g/L, regulation pH value is produced to 5.6-5.8.
It is above-mentioned, it is the Subculture of callus.Specifically, the second callus is transferred in subculture medium
Culture, condition of culture：Intensity of illumination is 1500~20001x, and daily light application time is 12 hours, and temperature is 25~28 DEG C, humidity
For 50~65%, every 40 days subcultures are once.
Embodiment 1：
The culture of epipremnum aureum explant is carried out as follows：
Step 1, choose maternal plant and carry out pouring root 30 days, every 10 days 1 time using 80% carbendazim, 1000 times of solution；
Step 2, tender leaf at the maternal plant growing point is taken；
Step 3, the tender leaf is sterilized, obtains epipremnum aureum explant；
Step 4, after the epipremnum aureum explant is trimmed, petiole section sample is obtained；
Step 5, petiole section sample is subjected to mechanical damage；
Step 6, the callus inducing medium includes callus inducing medium 1 and callus induction is trained
Support base 2；
Step 7, the petiole section sample of the mechanical damage is seeded in callus inducing medium 1, in temperature 25
Cultivated under conditions of DEG C -28 DEG C and humidity 50%-65%, obtain the first callus；
Step 8, the callus is transferred in inducing culture 2, in 25 DEG C -28 DEG C of temperature and humidity 50%-
Cultivated under conditions of 65%, obtain the second callus；
Step 9, second callus is forwarded in urgently culture medium in intensity of illumination：1500-20001x, illumination
Time：12 hours/day, temperature：25 DEG C -28 DEG C, humidity：Cultivated under the conditions of 50%-65%；Every 40 days subcultures once, i.e.,
.
1st, modified MS medium 1 is configured as follows：
Based on MS culture mediums, adjustment wherein a great number of elements is the 1/4 of the original content of MS culture mediums, trace element
For the 1/2 of the original content of MS culture mediums；
Also include following component, and adjust content and be：
Inositol：1mg/L；
Thiamine hydrochloride：3mg/L；
Puridoxine hydrochloride：0.5mg/L；
Nicotinic acid：0.5mg/L；
Arginine：50mg/L；
Aspartic acid：100mg/L；
Glutamic acid：150mg/L；
2nd, modified MS medium 2 is configured as follows：
Based on MS culture mediums, adjustment wherein a great number of elements is the 1/4 of the original content of MS culture mediums, trace element
For the 1/2 of the original content of MS culture mediums；
Also include following component, and adjust content and be：
Inositol：1mg/L；
Thiamine hydrochloride：3mg/L；
Puridoxine hydrochloride：0.5mg/L；
Nicotinic acid：0.5mg/L；
Arginine：100mg/L；
Aspartic acid：50mg/L；
Glutamic acid：200mg/L；
3rd, callus inducing medium 1 is configured as follows：
In modified MS medium 1, according to concentration addition 3.0mg/L 6- benzyls aminoadenine, 0.1mg/L 6- sugar
Base adenine phosphate, 0.2mg/L methyl α-naphthyl acetate, 0.05mg/L D butyl esters, 30g/L sucrose, 100ml/L coconut water and 7g/L
Agar powder, regulation pH value to 5.6-5.8, produce；
4th, callus inducing medium 2 is configured as follows：
In modified MS medium 2, according to concentration addition 1.0mg/L 6- benzyls aminoadenine, 0.1mg/L 6- sugar
Base adenine phosphate, 0.2mg/L methyl α-naphthyl acetate, 0.1mg/L indole-3-acetic acid, 30g/L sucrose, 150ml/L coconut water and
7g/L agar powder, regulation pH value is produced to 5.6-5.8；
5th, subculture medium, is prepared as follows：
In modified MS medium 2, according to concentration addition 2.0mg/L 6- benzyls aminoadenine, 0.3mg/L Yin
The agar powder of diindyl -3- acetic acid, 30g/L sucrose and 7g/L, regulation pH value is produced to 5.6-5.8.
Embodiment 2：
The culture of epipremnum aureum explant is carried out as follows：
Step 1, choose maternal plant and carry out pouring root 30 days, every 10 days 1 time using 80% carbendazim, 1000 times of solution；
Step 2, tender leaf at the maternal plant growing point is taken；
Step 3, the tender leaf is sterilized, obtains epipremnum aureum explant；
Step 4, after the epipremnum aureum explant is trimmed, petiole section sample is obtained；
Step 5, petiole section sample is subjected to mechanical damage；
Step 6, the callus inducing medium includes callus inducing medium 1 and callus induction is trained
Support base 2；
Step 7, the petiole section sample of the mechanical damage is seeded in callus inducing medium 1, in temperature 25
Cultivated under conditions of DEG C -28 DEG C and humidity 50%-65%, obtain the first callus；
Step 8, the callus is transferred in inducing culture 2, in 25 DEG C -28 DEG C of temperature and humidity 50%-
Cultivated under conditions of 65%, obtain the second callus；
Step 9, second callus is forwarded in urgently culture medium in intensity of illumination：1500-20001x, illumination
Time：12 hours/day, temperature：25 DEG C -28 DEG C, humidity：Cultivated under the conditions of 50%-65%；Every 40 days subcultures once, i.e.,
.
1st, modified MS medium 1 is configured as follows：
Based on MS culture mediums, adjustment wherein a great number of elements is the 1/4 of the original content of MS culture mediums, trace element
For the 1/2 of the original content of MS culture mediums；
Also include following component, and adjust content and be：
Inositol：2mg/L；
Thiamine hydrochloride：4mg/L；
Puridoxine hydrochloride：1mg/L；
Nicotinic acid：1mg/L；
Arginine：50mg/L；
Aspartic acid：100mg/L；
Glutamic acid：150mg/L；
2nd, modified MS medium 2 is configured as follows：
Based on MS culture mediums, adjustment wherein a great number of elements is the 1/4 of the original content of MS culture mediums, trace element
For the 1/2 of the original content of MS culture mediums；
Also include following component, and adjust content and be：
Inositol：2mg/L；
Thiamine hydrochloride：4mg/L；
Puridoxine hydrochloride：1mg/L；
Nicotinic acid：1mg/L；
Arginine：100mg/L；
Aspartic acid：50mg/L；
Glutamic acid：200mg/L；
3rd, callus inducing medium 1 is configured as follows：
In modified MS medium 1, according to concentration addition 3.0mg/L 6- benzyls aminoadenine, 0.1mg/L 6- sugar
Base adenine phosphate, 0.2mg/L methyl α-naphthyl acetate, 0.05mg/L D butyl esters, 30g/L sucrose, 100ml/L coconut water and 7g/L
Agar powder, regulation pH value to 5.6-5.8, produce；
4th, callus inducing medium 2 is configured as follows：
In modified MS medium 2, according to concentration addition 1.0mg/L 6- benzyls aminoadenine, 0.1mg/L 6- sugar
Base adenine phosphate, 0.2mg/L methyl α-naphthyl acetate, 0.1mg/L indole-3-acetic acid, 30g/L sucrose, 150ml/L coconut water and
7g/L agar powder, regulation pH value is produced to 5.6-5.8；
5th, subculture medium, is prepared as follows：
In modified MS medium 2, according to concentration addition 2.0mg/L 6- benzyls aminoadenine, 0.3mg/L Yin
The agar powder of diindyl -3- acetic acid, 30g/L sucrose and 7g/L, regulation pH value is produced to 5.6-5.8.
Embodiment 3：
The culture of epipremnum aureum explant is carried out as follows：
Step 1, choose maternal plant and carry out pouring root 30 days, every 10 days 1 time using 80% carbendazim, 1000 times of solution；
Step 2, tender leaf at the maternal plant growing point is taken；
Step 3, the tender leaf is sterilized, obtains epipremnum aureum explant；
Step 4, after the epipremnum aureum explant is trimmed, petiole section sample is obtained；
Step 5, petiole section sample is subjected to mechanical damage；
Step 6, the callus inducing medium includes callus inducing medium 1 and callus induction is trained
Support base 2；
Step 7, the petiole section sample of the mechanical damage is seeded in callus inducing medium 1, in temperature 25
Cultivated under conditions of DEG C -28 DEG C and humidity 50%-65%, obtain the first callus；
Step 8, the callus is transferred in inducing culture 2, in 25 DEG C -28 DEG C of temperature and humidity 50%-
Cultivated under conditions of 65%, obtain the second callus；
Step 9, second callus is forwarded in urgently culture medium in intensity of illumination：1500-20001x, illumination
Time：12 hours/day, temperature：25 DEG C -28 DEG C, humidity：Cultivated under the conditions of 50%-65%；Every 40 days subcultures once, i.e.,
.
1st, modified MS medium 1 is configured as follows：
Based on MS culture mediums, adjustment wherein a great number of elements is the 1/4 of the original content of MS culture mediums, trace element
For the 1/2 of the original content of MS culture mediums；
Also include following component, and adjust content and be：
Inositol：5mg/L；
Thiamine hydrochloride：6mg/L；
Puridoxine hydrochloride：2mg/L；
Nicotinic acid：3mg/L；
Arginine：50mg/L；
Aspartic acid：100mg/L；
Glutamic acid：150mg/L；
2nd, modified MS medium 2 is configured as follows：
Based on MS culture mediums, adjustment wherein a great number of elements is the 1/4 of the original content of MS culture mediums, trace element
For the 1/2 of the original content of MS culture mediums；
Also include following component, and adjust content and be：
Inositol：5mg/L；
Thiamine hydrochloride：6mg/L；
Puridoxine hydrochloride：2mg/L；
Nicotinic acid：3mg/L；
Arginine：100mg/L；
Aspartic acid：50mg/L；
Glutamic acid：200mg/L；
3rd, callus inducing medium 1 is configured as follows：
In modified MS medium 1, according to concentration addition 3.0mg/L 6- benzyls aminoadenine, 0.1mg/L 6- sugar
Base adenine phosphate, 0.2mg/L methyl α-naphthyl acetate, 0.05mg/L D butyl esters, 30g/L sucrose, 100ml/L coconut water and 7g/L
Agar powder, regulation pH value to 5.6-5.8, produce；
4th, callus inducing medium 2 is configured as follows：
In modified MS medium 2, according to concentration addition 1.0mg/L 6- benzyls aminoadenine, 0.1mg/L 6- sugar
Base adenine phosphate, 0.2mg/L methyl α-naphthyl acetate, 0.1mg/L indole-3-acetic acid, 30g/L sucrose, 150ml/L coconut water and
7g/L agar powder, regulation pH value is produced to 5.6-5.8；
5th, subculture medium, is prepared as follows：
In modified MS medium 2, according to concentration addition 2.0mg/L 6- benzyls aminoadenine, 0.3mg/L Yin
The agar powder of diindyl -3- acetic acid, 30g/L sucrose and 7g/L, regulation pH value is produced to 5.6-5.8.
It should be understood that, although the present specification is described in terms of embodiments, but not each embodiment only includes one
Individual independent technical scheme, this narrating mode of specification is only that for clarity, those skilled in the art will should say
Bright book is as an entirety, and the technical scheme in each embodiment may also be suitably combined to form those skilled in the art can
With the other embodiment of understanding.
Inventor states that the present invention can only for of the invention by a series of describe in detail of those listed above
Row embodiment is illustrated, but the invention is not limited in above-mentioned detailed process equipment and technological process.And i.e. not
Mean that the present invention should rely on above-mentioned detailed process equipment and technological process and could implement.Person of ordinary skill in the field should
This understands, any improvement in the present invention, the adding of equivalence replacement and auxiliary element to each raw material of product of the present invention, specific side
Selection of formula etc., within the scope of all falling within protection scope of the present invention and being open.
Claims (10)
1. a kind of method that utilization petiole induces epipremnum aureum, it is characterised in that comprise the following steps：
Maternal plant is chosen, epipremnum aureum explant is taken after pretreatment；
The epipremnum aureum explant is subjected to mechanical damage, and is seeded in callus inducing medium, callus is obtained；
The callus is forwarded in subculture medium and cultivated, is produced；
The callus inducing medium 1 includes following component：
Modified MS medium 1,6- benzyls aminoadenine, 6- glycosyls adenine phosphate, methyl α-naphthyl acetate, D butyl esters, sucrose, coconut water and
Agar powder；
The callus inducing medium 2 includes following component：
Modified MS medium 2,6- benzyls aminoadenine, 6- glycosyls adenine phosphate, methyl α-naphthyl acetate, indole-3-acetic acid, sucrose, coconut palm
Sub- water and agar powder.
2. the method for epipremnum aureum is induced using petiole as claimed in claim 1, it is characterised in that the selection maternal plant, it is preprocessed
After take epipremnum aureum explant, including：
Choose maternal plant and use carbendazim pouring root；
Take tender leaf at the maternal plant growing point；
The tender leaf is sterilized, epipremnum aureum explant is obtained.
3. the method for epipremnum aureum is induced using petiole as claimed in claim 2, it is characterised in that the selection maternal plant uses carbendazim
Pouring root, including：
Choose maternal plant and carry out pouring root 30 days, every 10 days 1 time using 80% carbendazim, 1000 times of solution.
4. the method for epipremnum aureum is induced using petiole as claimed in claim 3, it is characterised in that
The callus inducing medium includes callus inducing medium 1 and callus inducing medium 2；
It is described that the epipremnum aureum explant is subjected to mechanical damage, and be seeded in callus inducing medium, callus is obtained,
Including：
After the epipremnum aureum explant is trimmed, petiole section sample is obtained；
Petiole section sample is subjected to mechanical damage；
The petiole of mechanical damage section sample is seeded in callus inducing medium 1, in 25 DEG C -28 DEG C of temperature and wet
Cultivated under conditions of degree 50%-65%, obtain the first callus；
The callus is transferred in inducing culture 2, in 25 DEG C -28 DEG C of temperature and humidity 50%-65% condition
It is lower to be cultivated, obtain the second callus.
5. the method for epipremnum aureum is induced using petiole as claimed in claim 4, it is characterised in that the callus inducing medium
No. 1 configures as follows：
In modified MS medium 1, according to concentration addition 3.0mg/L 6- benzyls aminoadenine, 0.1mg/L 6- glycosyl ammonia
Base purine, 0.2mg/L methyl α-naphthyl acetate, 0.05mg/L D butyl esters, 30g/L sucrose, 100ml/L coconut water and 7g/L fine jade
Cosmetics, regulation pH value is produced to 5.6-5.8；
The callus inducing medium 2 is configured as follows：
In modified MS medium 2, according to concentration addition 1.0mg/L 6- benzyls aminoadenine, 0.1mg/L 6- glycosyl ammonia
Base purine, 0.2mg/L methyl α-naphthyl acetate, 0.1mg/L indole-3-acetic acid, 30g/L sucrose, 150ml/L coconut water and 7g/L
Agar powder, regulation pH value to 5.6-5.8, produce.
6. the method for epipremnum aureum is induced using petiole as claimed in claim 5, it is characterised in that the modified MS medium 1 is pressed
Following method configuration：
Based on MS culture mediums, in addition to following component, and adjust content and be：
Inositol：1-5mg/L；
Thiamine hydrochloride：3-6mg/L；
Puridoxine hydrochloride：0.5-2mg/L；
Nicotinic acid：0.5-3mg/L；
Arginine：50mg/L；
Aspartic acid：100mg/L；
Glutamic acid：150mg/L；
The modified MS medium 2 is configured as follows：
Based on MS culture mediums, in addition to following component, and adjust content and be：
Inositol：1-5mg/L；
Thiamine hydrochloride：3-6mg/L；
Puridoxine hydrochloride：0.5-2mg/L；
Nicotinic acid：0.5-3mg/L；
Arginine：100mg/L；
Aspartic acid：50mg/L；
Glutamic acid：200mg/L.
7. the method for epipremnum aureum is induced using petiole as claimed in claim 1, it is characterised in that described that the callus is transferred
Cultivate, produce into subculture medium, including：
Second callus is forwarded in urgently culture medium and cultivated under default condition of culture；
Every 40 days subcultures once, are produced.
8. the method for epipremnum aureum is induced using petiole as claimed in claim 7, it is characterised in that the default condition of culture, including：
Intensity of illumination：1500-20001x；
Light application time：12 hours/day；
Temperature：25℃-28℃；
Humidity：50%-65%.
9. the method for epipremnum aureum is induced using petiole as claimed in claim 7, it is characterised in that the subculture medium includes as follows
Component：Modified MS medium 2,6- benzyls aminoadenine, indole-3-acetic acid, sucrose and agar powder.
10. the method for epipremnum aureum is induced using petiole as claimed in claim 7, it is characterised in that the subculture medium, by as follows
Method is prepared：
In modified MS medium 2, according to concentration addition 2.0mg/L 6- benzyls aminoadenine, 0.3mg/L indoles -3-
The agar powder of acetic acid, 30g/L sucrose and 7g/L, regulation pH value is produced to 5.6-5.8.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107624649A (en) * | 2017-11-08 | 2018-01-26 | 东方上彩现代农业有限公司 | A kind of method that epipremnum aureum callus is induced using blade |
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2017
- 2017-08-03 CN CN201710655041.8A patent/CN107258544A/en active Pending
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| 张盛胜: "绿萝离体快繁与影响叶色变异因素的研究", 《中国优秀硕士学位论文全文数据库农业科技辑》 * |
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| CN107624649A (en) * | 2017-11-08 | 2018-01-26 | 东方上彩现代农业有限公司 | A kind of method that epipremnum aureum callus is induced using blade |
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Application publication date: 20171020 |