CN105580733A - Culture medium formula for rapid propagation of morning glory - Google Patents
Culture medium formula for rapid propagation of morning glory Download PDFInfo
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- CN105580733A CN105580733A CN201510990256.6A CN201510990256A CN105580733A CN 105580733 A CN105580733 A CN 105580733A CN 201510990256 A CN201510990256 A CN 201510990256A CN 105580733 A CN105580733 A CN 105580733A
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
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Abstract
The invention relates to a culture medium formula for rapid propagation of morning glory. The formula comprises a callus induction culture medium, a callus differentiation culture medium and a rooting culture medium. The culture medium formula is specially applicable to rapid propagation of the morning glory, the callus induction culture medium can induce tissue to form callus rapidly, the callus differentiation culture medium is used for promoting callus differentiation and increasing the multiplication coefficient, and the rooting culture medium can greatly promote the rooting rate of the tissue so as to increase the survival rate of the morning glory. The formula comprising the three culture media has a synergistic effect, facilitates rapid propagation of the tissue of the morning glory and lays a foundation for research on the morning glory color gene engineering.
Description
Technical field
The invention belongs to Plant Tissue Breeding field, relate to a kind of morning glory fast propagating culture medium formula.
Background technology
Morning glory is common climing property showy flowers of herbaceous plants summer and autumn, can do to shelter from heat or light before garth and room window, small-sized frame, hedge wallBeautify, also can do to be planted; Except cultivation is for viewing and admiring, seed is conventional Chinese medicine, ugly ox of name (Yunnan), blackUgly, semen pharbitidis, two ugly (the sub-mixing of black, white race), be used as medicine multiplex black ugly, the less use of semen pharbitidis. Morning glory has water pouring diuresis,By phlegm, effect of desinsection. Pattern is one of most important characteristics of plants ' aesthetics, be also flowers improvement highest priority itOne. Conventional hybridization, mutation breeding and directed selection technology have very large drawback, make traditional breeding technology create new flowerOn look, be difficult to have breakthrough. Engineered rise provides brand-new thinking for improveing and modify pattern, can be directedModify pattern and retain other original proterties, change the method for plant pattern by importing foreign gene, multipleOn plant, succeed. As the classical material of flower color gene engineering research, the molecular biology research of petunia is carried outEarly, more deep, be one of the separation of flower color gene, Main Patterns plant of conversion, but morning glory is in this respectResearch or space state, tissue cultivate be genetically modified basis.
Cultured in vitro is again in Plant Tissue Breeding (broad sense), refers to isolate from plant the tissue, the organ that suit the requirementsOr cell, protoplast etc., by sterile working, under manual control condition, cultivate to obtain the complete of regenerationPlant or production have the technology of other products of economic worth. Plant Tissue Breeding (narrow sense) refers to use plant each several partTissue, cultivates acquisition regeneration plant as formed layer, parenchymal tissue, mesophyll tissue, endosperm etc., also refers to cultivatingIn process, produce the cultivation of callus from each organ, callus is again through being differentiated to form aftergrowth again.
In Plant Tissue Breeding, to use various culture mediums. Culture medium is raw for microorganism, plant tissue and animal tissueLong and maintain the nutriment of the artificial preparation of use, generally all contain carbohydrate, nitrogen substance, inorganic salts and (comprise micro-Secondary element) and vitamin and water etc. Different culture media can be according to actual needs, adds that some self cannot be synthesizedCompound, i.e. growth factor.
But at present, be not also suitable for specially the culture medium of morning glory tissue cultivation, had a strong impact on to morning glory thisPlant the research of classical material.
Summary of the invention
The first object of the present invention is to provide a kind of morning glory fast propagating culture medium formula, is suitable for specially morning glory groupKnit cultivation, can carry out on the one hand the Fast-propagation of famous and precious kind, on the other hand grinding for morning glory flower color gene engineeringStudy carefully and lay the foundation.
The second object of the present invention is to provide the purposes of above-mentioned morning glory fast propagating culture medium formula.
The present invention is achieved through the following technical solutions:
One, a kind of morning glory fast propagating culture medium formula, comprises callus inducing medium, Calli DifferentiationThree kinds of culture medium and root medias, wherein,
Callus inducing medium formula: 1 liter of MS culture medium, 6-benzyl aminopurine 0.8-1.4 mg/litre, naphthalene secondAcid 1.5-2.5 mg/litre, sucrose 30 grams per liters, plant agar 7-8 grams per liter;
Calli Differentiation culture medium prescription: 1 liter of MS culture medium, 6-benzyl aminopurine 0.5-2.0 mg/litre, naphthalene secondAcid 0.1-0.5 mg/litre, sucrose 30 grams per liters, plant agar 7-8 grams per liter;
Prescription of rooting medium: 500 milliliters of MS culture mediums, heteroauxin 0.5-1.5 mg/litre, sucrose 20 grams per liters,Plant agar 7-8 grams per liter, 500 milliliters, water.
Two, according to above-mentioned a kind of morning glory fast propagating culture medium formula, comprise callus inducing medium, healThree kinds of injured tissue differential medium and root medias, wherein,
Callus inducing medium formula: 1 liter of MS culture medium, 6-benzyl aminopurine 1.0-1.2 mg/litre, naphthalene secondAcid 1.5-2.0 mg/litre, sucrose 30 grams per liters, plant agar 7-8 grams per liter;
Calli Differentiation culture medium prescription: 1 liter of MS culture medium, 6-benzyl aminopurine 1.0-1.5 mg/litre, naphthalene secondAcid 0.1-0.3 mg/litre, sucrose 30 grams per liters, plant agar 7-8 grams per liter;
Prescription of rooting medium: 20 grams of 500 milliliters of MS culture mediums, heteroauxin 0.75-1.25 mg/litre, sucrose/Liter, plant agar 7-8 grams per liter, 500 milliliters, water.
MS culture medium is as the minimal medium of the technical program, for those skilled in the art's common practise, containedComponent and content as table 1:
The formula of table 1MS culture medium
Three, the application of above-mentioned culture medium prescription in morning glory Fast-propagation is cultivated.
Adopt the good effect of technique scheme: culture medium prescription of the present invention be suitable for specially to morning glory fastBreeding is used, and callus inducing medium can form callus by induced tissue fast, and Calli Differentiation is cultivatedBase is used for promoting Calli Differentiation, improves growth coefficient, and root media can promote the rooting rate of tissue greatly,And then the survival rate of raising morning glory; Three culture medium prescriptions synergy of the present invention, facilitates the fast of morning glory tissueSpeed breeding, for the research of morning glory flower color gene engineering lays the foundation.
Detailed description of the invention
Below by embodiment and test example, technical scheme of the present invention is described further, but should not be construed as rightRestriction of the present invention:
Embodiment 1
A morning glory fast propagating culture medium formula, comprises callus inducing medium, Calli Differentiation cultivationThree kinds of base and root medias, wherein,
Callus inducing medium formula: 1 liter of MS culture medium, 6-benzyl aminopurine 0.8 mg/litre, methyl α-naphthyl acetate 1.0Mg/litre, sucrose 30 grams per liters, plant agar 7 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Calli Differentiation culture medium prescription: 1 liter of MS culture medium, 6-benzyl aminopurine 0.5 mg/litre, methyl α-naphthyl acetate 0.1Mg/litre, sucrose 30 grams per liters, plant agar 7 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Prescription of rooting medium: 500 milliliters of MS culture mediums, heteroauxin 0.5 mg/litre, sucrose 20 grams per liters, plantThing agar 7 grams per liters, 500 milliliters, water. When preparation, first measure MS culture medium, add heteroauxin, add sucrose,Add water, after sucrose dissolved, adjust pH to 5.8, add plant agar, heating is melted plant agar, and high temperature is highPress sterilizing (121 DEG C, 25 minutes), be cooled to solid for subsequent use.
Embodiment 2
A morning glory fast propagating culture medium formula, comprises callus inducing medium, Calli Differentiation cultivationThree kinds of base and root medias, wherein,
Callus inducing medium formula: 1 liter of MS culture medium, 6-benzyl aminopurine 0.8 mg/litre, methyl α-naphthyl acetate 1.5Mg/litre, sucrose 30 grams per liters, plant agar 8 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Calli Differentiation culture medium prescription: 1 liter of MS culture medium, 6-benzyl aminopurine 0.5 mg/litre, methyl α-naphthyl acetate 0.3Mg/litre, sucrose 30 grams per liters, plant agar 8 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Prescription of rooting medium: 500 milliliters of MS culture mediums, heteroauxin 0.75 mg/litre, sucrose 20 grams per liters,Plant agar 7 grams per liters, 500 milliliters, water. When preparation, first measure MS culture medium, add heteroauxin, add sugarcaneSugar, adds water, and after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating is melted plant agar, heightTemperature autoclaving (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Embodiment 3
A morning glory fast propagating culture medium formula, comprises callus inducing medium, Calli Differentiation cultivationThree kinds of base and root medias, wherein,
Callus inducing medium formula: 1 liter of MS culture medium, 6-benzyl aminopurine 0.8 mg/litre, methyl α-naphthyl acetate 2.0Mg/litre, sucrose 30 grams per liters, plant agar 7 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Calli Differentiation culture medium prescription: 1 liter of MS culture medium, 6-benzyl aminopurine 0.5 mg/litre, methyl α-naphthyl acetate 0.5Mg/litre, sucrose 30 grams per liters, plant agar 7 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Prescription of rooting medium: 500 milliliters of MS culture mediums, heteroauxin 1.0 mg/litre, sucrose 20 grams per liters, plantThing agar 7 grams per liters, 500 milliliters, water. When preparation, first measure MS culture medium, add heteroauxin, add sucrose,Add water, after sucrose dissolved, adjust pH to 5.8, add plant agar, heating is melted plant agar, and high temperature is highPress sterilizing (121 DEG C, 25 minutes), be cooled to solid for subsequent use.
Embodiment 4
A morning glory fast propagating culture medium formula, comprises callus inducing medium, Calli Differentiation cultivationThree kinds of base and root medias, wherein,
Callus inducing medium formula: 1 liter of MS culture medium, 6-benzyl aminopurine 0.8 mg/litre, methyl α-naphthyl acetate 2.5Mg/litre, sucrose 30 grams per liters, plant agar 8 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Calli Differentiation culture medium prescription: 1 liter of MS culture medium, 6-benzyl aminopurine 0.1 mg/litre, methyl α-naphthyl acetate 0.1Mg/litre, sucrose 30 grams per liters, plant agar 8 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Prescription of rooting medium: 500 milliliters of MS culture mediums, heteroauxin 1.25 mg/litre, sucrose 20 grams per liters,Plant agar 8 grams per liters, 500 milliliters, water. When preparation, first measure MS culture medium, add heteroauxin, add sugarcaneSugar, adds water, and after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating is melted plant agar, heightTemperature autoclaving (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Embodiment 5
A morning glory fast propagating culture medium formula, comprises callus inducing medium, Calli Differentiation cultivationThree kinds of base and root medias, wherein,
Callus inducing medium formula: 1 liter of MS culture medium, 6-benzyl aminopurine 1.0 mg/litre, methyl α-naphthyl acetate 1.0Mg/litre, sucrose 30 grams per liters, plant agar 7 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Calli Differentiation culture medium prescription: 1 liter of MS culture medium, 6-benzyl aminopurine 1.0 mg/litre, methyl α-naphthyl acetate 0.3Mg/litre, sucrose 30 grams per liters, plant agar 7 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Prescription of rooting medium: 500 milliliters of MS culture mediums, heteroauxin 1.5 mg/litre, sucrose 20 grams per liters, plantThing agar 7 grams per liters, 500 milliliters, water. When preparation, first measure MS culture medium, add heteroauxin, add sucrose,Add water, after sucrose dissolved, adjust pH to 5.8, add plant agar, heating is melted plant agar, and high temperature is highPress sterilizing (121 DEG C, 25 minutes), be cooled to solid for subsequent use.
Embodiment 6
A morning glory fast propagating culture medium formula, comprises callus inducing medium, Calli Differentiation cultivationThree kinds of base and root medias, wherein,
Callus inducing medium formula: 1 liter of MS culture medium, 6-benzyl aminopurine 1.0 mg/litre, methyl α-naphthyl acetate 1.5Mg/litre, sucrose 30 grams per liters, plant agar 8 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Calli Differentiation culture medium prescription: 1 liter of MS culture medium, 6-benzyl aminopurine 1.0 mg/litre, methyl α-naphthyl acetate 0.5Mg/litre, sucrose 30 grams per liters, plant agar 8 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Prescription of rooting medium: 500 milliliters of MS culture mediums, heteroauxin 0.5 mg/litre, sucrose 20 grams per liters, plantThing agar 8 grams per liters, 500 milliliters, water. When preparation, first measure MS culture medium, add heteroauxin, add sucrose,Add water, after sucrose dissolved, adjust pH to 5.8, add plant agar, heating is melted plant agar, and high temperature is highPress sterilizing (121 DEG C, 25 minutes), be cooled to solid for subsequent use.
Embodiment 7
A morning glory fast propagating culture medium formula, comprises callus inducing medium, Calli Differentiation cultivationThree kinds of base and root medias, wherein,
Callus inducing medium formula: 1 liter of MS culture medium, 6-benzyl aminopurine 1.0 mg/litre, methyl α-naphthyl acetate 2.0Mg/litre, sucrose 30 grams per liters, plant agar 7 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Calli Differentiation culture medium prescription: 1 liter of MS culture medium, 6-benzyl aminopurine 1.5 mg/litre, methyl α-naphthyl acetate 0.1Mg/litre, sucrose 30 grams per liters, plant agar 7 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Prescription of rooting medium: 500 milliliters of MS culture mediums, heteroauxin 0.75 mg/litre, sucrose 20 grams per liters,Plant agar 7 grams per liters, 500 milliliters, water. When preparation, first measure MS culture medium, add heteroauxin, add sugarcaneSugar, adds water, and after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating is melted plant agar, heightTemperature autoclaving (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Embodiment 8
A morning glory fast propagating culture medium formula, comprises callus inducing medium, Calli Differentiation cultivationThree kinds of base and root medias, wherein,
Callus inducing medium formula: 1 liter of MS culture medium, 6-benzyl aminopurine 1.0 mg/litre, methyl α-naphthyl acetate 2.5Mg/litre, sucrose 30 grams per liters, plant agar 8 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Calli Differentiation culture medium prescription: 1 liter of MS culture medium, 6-benzyl aminopurine 1.5 mg/litre, methyl α-naphthyl acetate 0.3Mg/litre, sucrose 30 grams per liters, plant agar 8 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Prescription of rooting medium: 500 milliliters of MS culture mediums, heteroauxin 1.0 mg/litre, sucrose 20 grams per liters, plantThing agar 8 grams per liters, 500 milliliters, water. When preparation, first measure MS culture medium, add heteroauxin, add sucrose,Add water, after sucrose dissolved, adjust pH to 5.8, add plant agar, heating is melted plant agar, and high temperature is highPress sterilizing (121 DEG C, 25 minutes), be cooled to solid for subsequent use.
Embodiment 9
A morning glory fast propagating culture medium formula, comprises callus inducing medium, Calli Differentiation cultivationThree kinds of base and root medias, wherein,
Callus inducing medium formula: 1 liter of MS culture medium, 6-benzyl aminopurine 1.2 mg/litre, methyl α-naphthyl acetate 1.0Mg/litre, sucrose 30 grams per liters, plant agar 7 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Calli Differentiation culture medium prescription: 1 liter of MS culture medium, 6-benzyl aminopurine 1.5 mg/litre, methyl α-naphthyl acetate 0.5Mg/litre, sucrose 30 grams per liters, plant agar 7 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Prescription of rooting medium: 500 milliliters of MS culture mediums, heteroauxin 1.25 mg/litre, sucrose 20 grams per liters,Plant agar 7 grams per liters, 500 milliliters, water. When preparation, first measure MS culture medium, add heteroauxin, add sugarcaneSugar, adds water, and after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating is melted plant agar, heightTemperature autoclaving (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Embodiment 10
A morning glory fast propagating culture medium formula, comprises callus inducing medium, Calli Differentiation cultivationThree kinds of base and root medias, wherein,
Callus inducing medium formula: 1 liter of MS culture medium, 6-benzyl aminopurine 1.2 mg/litre, methyl α-naphthyl acetate 1.5Mg/litre, sucrose 30 grams per liters, plant agar 8 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Calli Differentiation culture medium prescription: 1 liter of MS culture medium, 6-benzyl aminopurine 2.0 mg/litre, methyl α-naphthyl acetate 0.1Mg/litre, sucrose 30 grams per liters, plant agar 8 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Prescription of rooting medium: 500 milliliters of MS culture mediums, heteroauxin 1.5 mg/litre, sucrose 20 grams per liters, plantThing agar 8 grams per liters, 500 milliliters, water. When preparation, first measure MS culture medium, add heteroauxin, add sucrose,Add water, after sucrose dissolved, adjust pH to 5.8, add plant agar, heating is melted plant agar, and high temperature is highPress sterilizing (121 DEG C, 25 minutes), be cooled to solid for subsequent use.
Embodiment 11
A morning glory fast propagating culture medium formula, comprises callus inducing medium, Calli Differentiation cultivationThree kinds of base and root medias, wherein,
Callus inducing medium formula: 1 liter of MS culture medium, 6-benzyl aminopurine 1.2 mg/litre, methyl α-naphthyl acetate 2.0Mg/litre, sucrose 30 grams per liters, plant agar 7 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Calli Differentiation culture medium prescription: 1 liter of MS culture medium, 6-benzyl aminopurine 2.0 mg/litre, methyl α-naphthyl acetate 0.3Mg/litre, sucrose 30 grams per liters, plant agar 7 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Prescription of rooting medium: 500 milliliters of MS culture mediums, heteroauxin 0.5 mg/litre, sucrose 20 grams per liters, plantThing agar 7 grams per liters, 500 milliliters, water. When preparation, first measure MS culture medium, add heteroauxin, add sucrose,Add water, after sucrose dissolved, adjust pH to 5.8, add plant agar, heating is melted plant agar, and high temperature is highPress sterilizing (121 DEG C, 25 minutes), be cooled to solid for subsequent use.
Embodiment 12
A morning glory fast propagating culture medium formula, comprises callus inducing medium, Calli Differentiation cultivationThree kinds of base and root medias, wherein,
Callus inducing medium formula: 1 liter of MS culture medium, 6-benzyl aminopurine 1.2 mg/litre, methyl α-naphthyl acetate 2.5Mg/litre, sucrose 30 grams per liters, plant agar 8 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Calli Differentiation culture medium prescription: 1 liter of MS culture medium, 6-benzyl aminopurine 2.0 mg/litre, methyl α-naphthyl acetate 0.5Mg/litre, sucrose 30 grams per liters, plant agar 8 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Prescription of rooting medium: 500 milliliters of MS culture mediums, heteroauxin 0.75 mg/litre, sucrose 20 grams per liters,Plant agar 8 grams per liters, 500 milliliters, water. When preparation, first measure MS culture medium, add heteroauxin, add sugarcaneSugar, adds water, and after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating is melted plant agar, heightTemperature autoclaving (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Embodiment 13
A morning glory fast propagating culture medium formula, comprises callus inducing medium, Calli Differentiation cultivationThree kinds of base and root medias, wherein,
Callus inducing medium formula: 1 liter of MS culture medium, 6-benzyl aminopurine 1.4 mg/litre, methyl α-naphthyl acetate 1.0Mg/litre, sucrose 30 grams per liters, plant agar 7 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Calli Differentiation culture medium prescription: 1 liter of MS culture medium, 6-benzyl aminopurine 1.0 mg/litre, methyl α-naphthyl acetate 0.1Mg/litre, sucrose 30 grams per liters, plant agar 7 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Prescription of rooting medium: 500 milliliters of MS culture mediums, heteroauxin 1.0 mg/litre, sucrose 20 grams per liters, plantThing agar 7 grams per liters, 500 milliliters, water. When preparation, first measure MS culture medium, add heteroauxin, add sucrose,Add water, after sucrose dissolved, adjust pH to 5.8, add plant agar, heating is melted plant agar, and high temperature is highPress sterilizing (121 DEG C, 25 minutes), be cooled to solid for subsequent use.
Embodiment 14
A morning glory fast propagating culture medium formula, comprises callus inducing medium, Calli Differentiation cultivationThree kinds of base and root medias, wherein,
Callus inducing medium formula: 1 liter of MS culture medium, 6-benzyl aminopurine 1.4 mg/litre, methyl α-naphthyl acetate 1.5Mg/litre, sucrose 30 grams per liters, plant agar 8 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Calli Differentiation culture medium prescription: 1 liter of MS culture medium, 6-benzyl aminopurine 1.0 mg/litre, methyl α-naphthyl acetate 0.3Mg/litre, sucrose 30 grams per liters, plant agar 8 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Prescription of rooting medium: 500 milliliters of MS culture mediums, heteroauxin 1.25 mg/litre, sucrose 20 grams per liters,Plant agar 8 grams per liters, 500 milliliters, water. When preparation, first measure MS culture medium, add heteroauxin, add sugarcaneSugar, adds water, and after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating is melted plant agar, heightTemperature autoclaving (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Embodiment 15
A morning glory fast propagating culture medium formula, comprises callus inducing medium, Calli Differentiation cultivationThree kinds of base and root medias, wherein,
Callus inducing medium formula: 1 liter of MS culture medium, 6-benzyl aminopurine 1.4 mg/litre, methyl α-naphthyl acetate 2.0Mg/litre, sucrose 30 grams per liters, plant agar 7 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Calli Differentiation culture medium prescription: 1 liter of MS culture medium, 6-benzyl aminopurine 1.5 mg/litre, methyl α-naphthyl acetate 0.1Mg/litre, sucrose 30 grams per liters, plant agar 7 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Prescription of rooting medium: 500 milliliters of MS culture mediums, heteroauxin 1.5 mg/litre, sucrose 20 grams per liters, plantThing agar 7 grams per liters, 500 milliliters, water. When preparation, first measure MS culture medium, add heteroauxin, add sucrose,Add water, after sucrose dissolved, adjust pH to 5.8, add plant agar, heating is melted plant agar, and high temperature is highPress sterilizing (121 DEG C, 25 minutes), be cooled to solid for subsequent use.
Embodiment 16
A morning glory fast propagating culture medium formula, comprises callus inducing medium, Calli Differentiation cultivationThree kinds of base and root medias, wherein,
Callus inducing medium formula: 1 liter of MS culture medium, 6-benzyl aminopurine 1.4 mg/litre, methyl α-naphthyl acetate 2.5Mg/litre, sucrose 30 grams per liters, plant agar 8 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Calli Differentiation culture medium prescription: 1 liter of MS culture medium, 6-benzyl aminopurine 1.5 mg/litre, methyl α-naphthyl acetate 0.3Mg/litre, sucrose 30 grams per liters, plant agar 8 grams per liters. When preparation, first measure MS culture medium, add 6-benzyl ammoniaBase purine, methyl α-naphthyl acetate, add sucrose, after sucrose dissolved, adjusts pH to 5.8, adds plant agar, and heating makesPlant agar melts, and autoclave sterilization (121 DEG C, 25 minutes), is cooled to solid for subsequent use.
Prescription of rooting medium: 500 milliliters of MS culture mediums, heteroauxin 1.0 mg/litre, sucrose 20 grams per liters, plantThing agar 8 grams per liters, 500 milliliters, water. When preparation, first measure MS culture medium, add heteroauxin, add sucrose,Add water, after sucrose dissolved, adjust pH to 5.8, add plant agar, heating is melted plant agar, and high temperature is highPress sterilizing (121 DEG C, 25 minutes), be cooled to solid for subsequent use.
Test example 1
This test example explanation Fast-propagation process to morning glory and the optimization of various parameters with culture medium.
1, seed disinfection
Get healthy full morning glory seed, first rinse 10min with running water, then with sterile distilled water flushing 5 times, incite somebody to actionIt is put in sterilized triangular flask, and then the Ethanol Treatment 5min that is first 75% by percent by volume uses quality percentageProcess 10min than the mercury chloride that is 0.1%, finally rinse 5 times with sterile distilled water, for subsequent use.
2, the acquisition of morning glory aseptic seedling
Seed after sterilization is inoculated in MS culture medium, and (25 ± 2) DEG C dark cultivation 5 days then cultivated under light,Cultivate and within 14 days, obtain the high aseptic seedling of 4-5cm.
3, the induction of callus
The hypocotyl of aseptic seedling is cut into the long segment of 0.4-0.6cm and puts into and cultivate callus inducing medium and cultivate,Callus inducing medium refers to the MS that is added with 0.5-1.5mg/L6-benayl aminopurine, 1.0-2.5mg/L methyl α-naphthyl acetateSolid medium (embodiment 1-16). While cultivating 5d, hypocotyl starts to expand, and while cultivating 15d, forms straightThe callus of footpath 1.0-1.2cm. Experimental result is as table 2:
Table 2 callus inducing medium experimental result
It is as shown in the table, and callus induction rate reaches as high as 99%.
4. Calli Differentiation is cultivated
4-5 rear stable callus of subculture is inoculated in Calli Differentiation culture medium and cultivated, Calli DifferentiationCulture medium refers to the MS solid medium (reality that is added with 0.5-2.0mg/L6-benayl aminopurine, 0.1-0.5mg/L methyl α-naphthyl acetateExecute routine 1-12). Experimental result is as table 3:
Table 3 Calli Differentiation culture medium experimental result
| 6-benzyl aminopurine | Methyl α-naphthyl acetate | Phenylacetic acid % | Growth coefficient | |
| Embodiment 1 | 0.5 | 0.1 | 33 | 1.2 |
| Embodiment 2 | 0.5 | 0.3 | 41 | 1.5 |
| Embodiment 3 | 0.5 | 0.5 | 40 | 1.6 |
| Embodiment 4 | 1.0 | 0.1 | 88 | 4.0 |
| Embodiment 5 | 1.0 | 0.3 | 89 | 5.8 |
| Embodiment 6 | 1.0 | 0.5 | 68 | 2.4 |
| Embodiment 7 | 1.5 | 0.1 | 89 | 6.0 |
| Embodiment 8 | 1.5 | 0.3 | 88 | 5.5 |
| Embodiment 9 | 1.5 | 0.5 | 70 | 3.0 |
| Embodiment 10 | 2.0 | 0.1 | 61 | 2.4 |
| Embodiment 11 | 2.0 | 0.3 | 65 | 2.5 |
| Embodiment 12 | 2.0 | 0.5 | 52 | 1.8 |
It is as shown in the table, and phenylacetic acid is up to 89%, and value-added coefficient is 4-6.
5, culture of rootage
The group training seedling that Calli Differentiation is gone out is cultivated 4-5cm when high, and be inoculated in root media and cultivated,Root media refers to the 1/2MS solid medium (embodiment 1-5) of the heteroauxin that is added with 0.5-1.5mg/L.Experimental result is as table 4:
Table 4 root media experimental result
It is as shown in the table, and rooting rate reaches 100%, and the number of taking root is for 8-10 bar.
6, group training transplantation of seedlings
In the time that plant root length is grown to 1.5-2cm, open sealed membrane and carry out hardening 2-3 days, be transplanted into the weight of Sha ︰ vermiculiteThan being 1 ︰ 1, being that 121 DEG C and pressure are in the matrix of 100kPa sterilizing through excess temperature, front 5-6 days builds sealed membrane,Maintenance humidity is 80%-90%, the 15-18 days that grows under 23-27 DEG C of condition, within every 3 days during this time, water MS culture medium withoutMachine salting liquid, moves to afterwards in outdoor soil and grows, and has greatly improved the survival rate of morning glory.
Culture medium prescription of the present invention is suitable for the Fast-propagation of morning glory to use specially, callus inducing mediumCan form fast callus by induced tissue, Calli Differentiation culture medium is used for promoting Calli Differentiation, improvesGrowth coefficient, root media can promote the rooting rate of tissue greatly, and then improves the survival rate of morning glory; ThisThree culture medium prescriptions of invention act synergistically, and facilitate the Fast-propagation of morning glory tissue, are morning glory flower color gene workThe research of journey lays the foundation.
Claims (3)
1. a morning glory fast propagating culture medium formula, is characterized in that: comprise callus inducing medium, healThree kinds of injured tissue differential medium and root medias, wherein,
Callus inducing medium formula: 1 liter of MS culture medium, 6-benzyl aminopurine 0.8-1.4 mg/litre, naphthalene secondAcid 1.5-2.5 mg/litre, sucrose 30 grams per liters, plant agar 7-8 grams per liter;
Calli Differentiation culture medium prescription: 1 liter of MS culture medium, 6-benzyl aminopurine 0.5-2.0 mg/litre, naphthalene secondAcid 0.1-0.5 mg/litre, sucrose 30 grams per liters, plant agar 7-8 grams per liter;
Prescription of rooting medium: 500 milliliters of MS culture mediums, heteroauxin 0.5-1.5 mg/litre, sucrose 20 grams per liters,Plant agar 7-8 grams per liter, 500 milliliters, water.
2. a kind of morning glory fast propagating culture medium formula according to claim 1, is characterized in that: comprise moreThree kinds of injured tissue inducing culture, Calli Differentiation culture medium and root medias, wherein,
Callus inducing medium formula: 1 liter of MS culture medium, 6-benzyl aminopurine 1.0-1.2 mg/litre, naphthalene secondAcid 1.5-2.0 mg/litre, sucrose 30 grams per liters, plant agar 7-8 grams per liter;
Calli Differentiation culture medium prescription: 1 liter of MS culture medium, 6-benzyl aminopurine 1.0-1.5 mg/litre, naphthalene secondAcid 0.1-0.3 mg/litre, sucrose 30 grams per liters, plant agar 7-8 grams per liter;
Prescription of rooting medium: 20 grams of 500 milliliters of MS culture mediums, heteroauxin 0.75-1.25 mg/litre, sucrose/Liter, plant agar 7-8 grams per liter, 500 milliliters, water.
3. the application of the culture medium prescription described in claim 1 or 2 in morning glory Fast-propagation is cultivated.
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| JP2004305101A (en) * | 2003-04-08 | 2004-11-04 | Keiichi Shimizu | Method for efficiently proliferating morning glory |
| CN103053429A (en) * | 2013-02-06 | 2013-04-24 | 郑州师范学院 | Method for regenerating semen pharbitidis in vitro embryonic axis plant |
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