CN104542274A - Culture medium of induced haplobiont for culturing eggplant anther and method of culture medium - Google Patents
Culture medium of induced haplobiont for culturing eggplant anther and method of culture medium Download PDFInfo
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Abstract
The invention relates to a culture medium of an induced haplobiont for culturing eggplant anther and a method of the culture medium, and belongs to the technical field of plant biology. The method for cultivating the eggplant anther provided by the invention comprises the following steps: with the eggplant anther as a material, carrying out anther culture to induce callus; carrying out differentiation culture on the callus; carrying out rooting culture on differentiated seedlings; carrying out acclimatization and transplanting; and carrying out haplobiont doubling germination, so as to finish regeneration of an eggplant plant. The culture medium provided by the invention is low in cost and good in culture effect; the method for culturing the eggplant anther by virtue of the culture medium provided by the invention has the advantages of high callus induction rate, simple culture method and short culture period; the culture medium and the eggplant anther culture technology provided by the invention are applied to eggplant genetic breeding; the excellent genotype individual bodies of the eggplant are greatly enriched; the breeding period is obviously shortened; and a reliable technical support is provided for variety improvement of the eggplant.
Description
Technical field
The present invention relates to medium and the method thereof of a kind of eggplant anther culture induction haplobiont of plant biotechnology field.
Background technology
Plant anther culture technique has broad application prospects in Crop Genetic Breeding theory and practice research, homozygous doubled haploid can be obtained fast and effectively by anther culture, avoid and need inbreeding of more generation just can obtain stable strain of isozygotying by traditional breeding method means, thus substantially reduce breeding cycle, improve efficiency of selection, reduce costs.In addition, the double haploid obtained by anther culture can be directly used in genetic map structure, cultivate new varieties or as Parents for breed improvement.Utilize anther culture to induce monoploid or double haploid on a lot of plant, to have successful report, utilize the New idioplasm resource of acquisition to carry out the selection and popularization of new varieties simultaneously, and achieve good Social and economic benef@.
Along with the development of plant anther culture technique, the seed selection for New Eggplant Varieties opens a new way.Utilize eggplant flower pesticide can obtain pure lines and mutant fast, Innovation Germplasm resource, has great importance to the breed improvement of eggplant.The deficiency of the application of current eggplant anther culture in actual breeding has callus induction rate lower, and regeneration plant differentiation rate is low, haplobiont to add multiplying power low, cultivation cycle is long.Therefore, the multiplying power that adds how improving the inductivity of callus in eggplant anther culture and the differentiation rate of Calli Differentiation plant and haplobiont is the difficult problem being badly in need of in the research of eggplant anther culture technique solving.
Summary of the invention
The object of the invention is to overcome the deficiency that in existing anther culture technique, callus induction rate is low, the deficiency that Calli Differentiation Plantlet Differentiation rate is low and haplobiont add the low deficiency of multiplying power, provide medium and the method thereof of a kind of eggplant anther culture induction haplobiont, callus can be formed by rapid induction eggplant flower pesticide by this method, that improves the differentiation-inducing rate of callus and haplobiont adds multiplying power, thus abundant suitable eggplant anther culture condition, lay the foundation for setting up efficient eggplant anther plant regenerating system.
To achieve these goals, the present invention is by the following technical solutions:
A kind of medium of eggplant anther culture induction haplobiont, it is characterized in that: described medium comprises eggplant Induction of Anther Culture Callus inducing culture and Calli Differentiation medium, described eggplant Induction of Anther Culture Callus inducing culture is: MS minimal medium, 0.2-0.5mg/L 2,4-D, 0.5-1mg/L KT, 5-10mg/L Vc, 3% sucrose, 5g/L agar, pH value 5.8; Described Calli Differentiation medium is: MS minimal medium, 0.01-0.05mg/L IAA, 1.0-2.0mg/L 6-BA, 0.1-0.5g/L active carbon, 2% sucrose, 5.5g/L agar, pH value 5.8.
Described eggplant Induction of Anther Culture Callus inducing culture is: MS minimal medium, 0.2mg/L 2,4-D, 0.9mg/L KT, 10mg/L Vc, 3% sucrose, 5g/L agar, pH value 5.8; Described Calli Differentiation medium is: MS minimal medium, 0.02mg/L IAA, 2.0mg/L 6-BA, 0.5g/L active carbon, 2% sucrose, 5.5g/L agar, pH value 5.8.
The cultivation of described eggplant flower pesticide comprises the steps:
1) grow from eggplant plant and choose bud to the position of eggplant or four door socket eggplants, in bud, anther cell is in monokaryon mid-term to mid-late uninucleate stage, bud diameter 7-10mm, and flower pesticide is that yellowish green colour cast is green;
2) pretreatment: the bud waterproof bag of return collection or blake bottle are closed, is placed in 4-5 DEG C of refrigerating chamber and preserves 12-24 h;
3) sterilizing: first carry out bud surface sterilization 30s with the alcohol of 70% or 75%, again with the volumetric concentration of clorox stoste be 7% solution soak 15-20min, wherein in clorox stoste, the mass concentration of clorox is 10%, afterwards with sterile water washing 3-4 time, each 3min, then blots bud with aseptic paper for subsequent use;
4) inoculate: on aseptic working platform, from the bud of bacterium of having gone out, strip flower pesticide gently, be inoculated in and be equipped with in the Φ 60mm culture dish of eggplant Induction of Anther Culture Callus inducing culture, each culture dish is equipped with 10mL medium, and each culture dish connects the flower pesticide of 1-2 bud;
5) Heat thermostability: the culture dish having inoculated flower pesticide is put in incubator and carries out Heat thermostability, dark culturing, heat-shock temperature is 35 ~ 40 DEG C, process 4 ~ 9d;
6) anther culture: the flower pesticide after high temperature Heat thermostability, is placed on 25-28 DEG C, cultivates under intensity of illumination 1500-2000Lx, illumination 12-16h/d, and transfer every 15-20 d secondary, callus appears in 30-40d;
7) callus plantlet: in the middle part of flower pesticide or top callus lines grow to diameter and be about 10mm, callus is transferred on Calli Differentiation medium, subculture is cut once every 15-20 d, 30-35 d callus starts to differentiate green bud point, squamous subculture condition: intensity of illumination 1500-2000Lx, illumination 12-16h/d, temperature is 25-28 DEG C, through squamous subculture, the Calli Differentiation of 45%-55% is that ripe embryoid directly generates the full complete plantlets of root, stem and leaf tool;
8) callus of regeneration plant root induction: 8%-12% directly differentiates the regeneration bud of unrooted, when bud seedling grows to 2-3cm, cut the seedling of band portion callus, be transferred in MS medium, at intensity of illumination 1500-2000Lx, illumination 12-16h/d, temperature is under 25-28 DEG C of culture of rootage condition, grows adventive root after 10-15d, generate whole plant, rooting rate is 90%;
9) Ploidy detection: adopt tip of a root growing point chromosome G banding detection method, in aseptic working platform, from blake bottle, the take root tender tip of a root of children of regrowth of clip detects;
10) haplobiont doubles: step 9 is detected gained haplobiont and adopt growing point infusion method, the outside spire of growing point of haplobiont is peelled off between the lights after transplanting, vegetative cone is leaked outside, 0.3-1.0% colchicin+1.5%DMSO solution parcel vegetative cone is dipped with absorbent cotton, outside is again with black plastic film parcel, process 1 ~ 3h, the normal eggplant male and female flowering of dliploid can be obtained, add multiplying power and reach 60%, the method can be carried out at the different times of eggplant growth, has broken colchicine and has doubled to process the limitation can only carried out in seedling stage.
In described step (5), be put in incubator by the culture dish having inoculated flower pesticide and carry out Heat thermostability, dark culturing, heat-shock temperature is 36 DEG C, processes 5 d.
In described step (10), dip 0.5 % colchicin+1.5%DMSO solution parcel vegetative cone with absorbent cotton, outside is again with black plastic film parcel.
Compared with prior art, the present invention adds appropriate VC and active carbon and uses IAA in minimal medium, the inductivity with callus is high, Calli Differentiation efficiency is high, culture effect is good, not easily the advantage such as brownization, and the callus tissue culture time is long, can squamous subculture 2 years, continuous regeneration induction plant from the callus cultivated.The inductivity of callus reaches as high as 90%, and shoot regeneration frequency is up to 47%, and monoploid rate can reach 60%, and the colchicine process of suitable concentration and method makes haploid multiplying power that adds up to 60%; Anther culture method of the present invention is applied in eggplant genetic breeding, greatly can improve the inductivity of eggplant anther culture regeneration plant, enrich eggplant superior genotypes individual, improve eggplant haploid breeding approach, shorten breeding cycle, for quickening eggplant genetic breeding and the research of plant of Solanaceae vitro anther culture provide technology and theoretical foundation.
Accompanying drawing explanation
Fig. 1 is the flower pesticide schematic diagram that one embodiment of the present of invention have just been inoculated
Fig. 2 is the flower pesticide schematic diagram that one embodiment of the present of invention are expanded
Fig. 3 is the flower pesticide schematic diagram that one embodiment of the present of invention produce callus
Fig. 4 is the schematic diagram that one embodiment of the present of invention Calli Differentiation goes out eggplant bud seedling
Fig. 5 is the various regeneration plant schematic diagram of one embodiment of the present of invention root leaf bud
Fig. 6 is that one embodiment of the present of invention regrowth transplants schematic diagram
Fig. 7 is one embodiment of the present of invention colchicine-induced schematic diagram.
Embodiment
Embodiment 1: a kind of medium of eggplant anther culture induction haplobiont and method thereof, its step is as follows:
1) grow from eggplant plant and choose bud to the position of eggplant or four door socket eggplants, in bud, anther cell is in monokaryon mid-term to mid-late uninucleate stage, bud diameter 10mm, and flower pesticide is that yellowish green colour cast is green;
2) pretreatment: the bud waterproof bag of return collection or blake bottle are closed, and are placed in 4-5 DEG C of refrigerating chamber and preserve 24 h;
3) sterilizing: first carry out bud surface sterilization 30s with the alcohol of 75%, again with mass concentration be 7% clorox Stock solutions soak 20min, wherein in clorox stoste, the mass concentration of clorox is 10%, 4 times are washed afterwards with sterile water, each 3min, then blots bud with aseptic paper for subsequent use;
4) inoculate: on aseptic working platform, from the bud of bacterium of having gone out, strip flower pesticide gently, be inoculated in the Φ 60mm culture dish of the inducing culture that eggplant Induction of Anther Culture Callus is housed, each culture dish is equipped with 10mL medium, and each culture dish connects the flower pesticide of 1-2 bud;
5) Heat thermostability: the culture dish having inoculated flower pesticide is put in incubator and carries out Heat thermostability, dark culturing, heat-shock temperature is 36 DEG C, process 5d;
6) anther culture: the flower pesticide after high temperature Heat thermostability, is placed on 25-28 DEG C, cultivates under intensity of illumination 1500-2000Lx, illumination 12-16h/d, and depending on medium situation, transfer medium once every 20d, about 35d there will be callus;
7) callus plantlet: in the middle part of flower pesticide or top callus lines grow to diameter and be about 10mm, by Transformation of Callus on Calli Differentiation medium, subculture is cut once at interval of 15-20 d, 30-40 d callus starts to differentiate green bud point, squamous subculture condition: intensity of illumination 1500-2000Lx, illumination 12-16h/d, temperature is 25-28 DEG C, through squamous subculture, the Calli Differentiation of about 50% is that ripe embryoid directly generates the full complete plantlets of root, stem and leaf tool;
8) regeneration plant root induction: the callus of about 10% directly differentiates the regeneration bud of unrooted, when bud seedling grows to 3cm, cut the seedling of band portion callus, be transferred in MS medium, at intensity of illumination 1500-2000Lx, illumination 12-16h/d, temperature is under 25-28 DEG C of culture of rootage condition, after 15d, has adventive root to occur;
9) Ploidy detection: adopt tip of a root growing point chromosome G banding detection method, in aseptic working platform, from blake bottle, the take root tender tip of a root of children of regrowth of clip detects;
10) haplobiont doubles: step 9 is detected gained haplobiont and adopt growing point infusion method to double, the outside spire of growing point of haplobiont is peelled off between the lights after transplanting, vegetative cone is leaked outside, 0.5% colchicin+1.5%DMSO solution is dipped with absorbent cotton, parcel vegetative cone, outside with black plastic film parcel, processes 2h again, the normal eggplant male and female flowering of dliploid can be obtained, add multiplying power and reach 60%.
Used medium:
The main component of callus inducing medium is MS minimal medium, 2,4-D, KT, Vc, sucrose and agar, wherein the concentration of 2,4-D is the concentration of 0.2mg/L, KT is 0.9mg/L, the concentration of Vc is 10mg/L, and the concentration of sucrose is 3%, and the concentration of agar is 5g/L, and pH value is 5.8;
The main component of Calli Differentiation medium is: MS minimal medium, IAA, 6-BA, active carbon, sucrose, agar, wherein the concentration of IAA is the concentration of 0.02mg/L, 6-BA is 2.0mg/L, the concentration of active carbon is 0.5g/L, the concentration of sucrose is 2%, and the concentration of agar is 5.5g/L, and pH value is 5.8.
Embodiment 2: a kind of medium of eggplant anther culture induction haplobiont and method thereof, its step is as follows:
1) grow from eggplant plant and choose bud to the position of eggplant or four door socket eggplants, in bud, anther cell is in monokaryon mid-term to mid-late uninucleate stage, bud diameter 10mm, and flower pesticide is that yellowish green colour cast is green;
2) pretreatment: the bud waterproof bag of return collection or blake bottle are closed, and are placed in 4-5 DEG C of refrigerating chamber and preserve 24 h;
3) sterilizing: first carry out bud surface sterilization 30s with the alcohol of 75%, again with mass concentration be 7% clorox Stock solutions soak 20min, wherein in clorox stoste, the mass concentration of clorox is 10%, 4 times are washed afterwards with sterile water, each 3min, then blots bud with aseptic paper for subsequent use;
4) inoculate: on aseptic working platform, from the bud of bacterium of having gone out, strip flower pesticide gently, be inoculated in the Φ 60mm culture dish of the inducing culture that eggplant Induction of Anther Culture Callus is housed, each culture dish is equipped with 10mL medium, and each culture dish connects the flower pesticide of 1-2 bud;
5) Heat thermostability: the culture dish having inoculated flower pesticide is put in incubator and carries out Heat thermostability, dark culturing, heat-shock temperature is 40 DEG C, process 4d;
6) anther culture: the flower pesticide after high temperature Heat thermostability, is placed on 25-28 DEG C, cultivates under intensity of illumination 1500-2000Lx, illumination 12-16h/d, and depending on medium situation, transfer medium once every 20d, about 35d there will be callus;
7) callus plantlet: in the middle part of flower pesticide or top callus lines grow to diameter and be about 8mm, by Transformation of Callus on Calli Differentiation medium, subculture is cut once at interval of 15-20 d, 30-40 d callus starts to differentiate green bud point, squamous subculture condition: intensity of illumination 1500-2000Lx, illumination 12-16h/d, temperature is 25-28 DEG C, through squamous subculture, the Calli Differentiation of about 50% is that ripe embryoid directly generates the full complete plantlets of root, stem and leaf tool;
8) regeneration plant root induction: the callus of about 10% directly differentiates the regeneration bud of unrooted, when bud seedling grows to 3cm, cut the seedling of band portion callus, be transferred in MS medium, at intensity of illumination 1500-2000Lx, illumination 12-16h/d, temperature is under 25-28 DEG C of culture of rootage condition, after 15d, has adventive root to occur;
9) Ploidy detection: adopt tip of a root growing point chromosome G banding detection method, in superclean bench, from blake bottle, the take root tender tip of a root of children of regrowth of clip detects;
10) haplobiont doubles: step 9 is detected gained haplobiont and adopt growing point infusion method to double, haplobiont peels off the outside spire of growing point of haplobiont between the lights after transplanting, vegetative cone is leaked outside, 0.3% colchicin+1.5%DMSO solution is dipped with absorbent cotton, parcel vegetative cone, outside with black plastic film parcel, processes 2h again, the normal eggplant male and female flowering of dliploid can be obtained, add multiplying power and reach 60%.
Used medium:
In callus inducing medium, the concentration of the concentration of 2,4-D to be the concentration of 0.4mg/L, KT be 0.5mg/L, Vc is 5mg/L, and the concentration of sucrose is 3%, and the concentration of agar is 5g/L, and pH value is 5.8;
In Calli Differentiation medium, the concentration of IAA is the concentration of 0.05mg/L, 6-BA is 1.0mg/L, and the concentration of active carbon is 0.2g/L, and the concentration of sucrose is 2%, and the concentration of agar is 5.5g/L, and pH value is 5.8.
Embodiment 3: a kind of medium of eggplant anther culture induction haplobiont and method thereof, its step is as follows:
1) grow from eggplant plant and choose bud to the position of eggplant or four door socket eggplants, in bud, anther cell is in monokaryon mid-term to mid-late uninucleate stage, bud diameter 10mm, and flower pesticide is that yellowish green colour cast is green;
2) pretreatment: the bud waterproof bag of return collection or blake bottle are closed, and are placed in 4-5 DEG C of refrigerating chamber and preserve 24 h;
3) sterilizing: first carry out bud surface sterilization 30s with the alcohol of 70%, again with mass concentration be 7% clorox Stock solutions soak 20min, wherein in clorox stoste, the mass concentration of clorox is 10%, 4 times are washed afterwards with sterile water, each 3min, then blots bud with aseptic paper for subsequent use;
4) inoculate: on aseptic working platform, from the bud of bacterium of having gone out, strip flower pesticide gently, be inoculated in the Φ 60mm culture dish of the inducing culture that eggplant Induction of Anther Culture Callus is housed, each culture dish is equipped with 10mL medium, and each culture dish connects the flower pesticide of 1-2 bud;
5) Heat thermostability: the culture dish having inoculated flower pesticide is put in incubator and carries out Heat thermostability, dark culturing, heat-shock temperature is 36 DEG C, process 9d;
6) anther culture: the flower pesticide after high temperature Heat thermostability, is placed on 25-28 DEG C, cultivates under intensity of illumination 1500-2000Lx, illumination 12-16h/d, and depending on medium situation, transfer medium once every 20d, about 35d there will be callus;
7) callus plantlet: in the middle part of flower pesticide or top callus lines grow to diameter and be about 9mm, by Transformation of Callus on Calli Differentiation medium, subculture is cut once at interval of 15-20 d, 30-40 d callus starts to differentiate green bud point, squamous subculture condition: intensity of illumination 1500-2000Lx, illumination 12-16h/d, temperature is 25-28 DEG C, through squamous subculture, the Calli Differentiation of about 50% is that ripe embryoid directly generates the full complete plantlets of root, stem and leaf tool;
8) regeneration plant root induction: the callus of about 10% directly differentiates the regeneration bud of unrooted, when bud seedling grows to 3cm, cut the seedling of band portion callus, be transferred in MS medium, at intensity of illumination 1500-2000Lx, illumination 12-16h/d, temperature is under 25-28 DEG C of culture of rootage condition, after 15d, has adventive root to occur;
9) Ploidy detection: adopt tip of a root growing point chromosome G banding detection method, in superclean bench, from blake bottle, the take root tender tip of a root of children of regrowth of clip detects;
10) haplobiont doubles: step 9 is detected gained haplobiont and adopt growing point infusion method to double, haplobiont peels off the outside spire of growing point of haplobiont between the lights after transplanting, vegetative cone is leaked outside, 0.8% colchicin+1.5%DMSO solution is dipped with absorbent cotton, parcel vegetative cone, outside with black plastic film parcel, processes 2h again, the normal eggplant male and female flowering of dliploid can be obtained, add multiplying power and reach 60%.
Used medium:
In callus inducing medium, the concentration of the concentration of 2,4-D to be the concentration of 0.3mg/L, KT be 0.7 mg/L, Vc is 8mg/L, and the concentration of sucrose is 3%, and the concentration of agar is 5g/L, and pH value is 5.8;
In Calli Differentiation medium, the concentration of IAA is the concentration of 0.03mg/L, 6-BA is 1.5mg/L, and the concentration of active carbon is 0.4g/L, and the concentration of sucrose is 2%, and the concentration of agar is 5.5g/L, and pH value is 5.8.
The inductivity of different cultivars material callus is 35%-90%, and higher is purple long eggplant, and minimum is green eggplant.Shoot regeneration frequency is 11%-47%, and monoploid rate is 40%-60%.It is better that 0.5% colchicin+1.5% DMSO doubles effect, adds multiplying power and reach 60%.
Use in the present invention 2,4-D, KT, IAA, 6-BA belong to the plant hormone needed for Plant Tissue Breeding.
Claims (6)
1. the medium of an eggplant anther culture induction haplobiont, it is characterized in that: described medium comprises eggplant Induction of Anther Culture Callus inducing culture and Calli Differentiation medium, described eggplant Induction of Anther Culture Callus inducing culture is: MS minimal medium, 0.2-0.5mg/L 2,4-D, 0.5-1mg/L KT, 5-10mg/L Vc, 3% sucrose, 5g/L agar, pH value 5.8; Described Calli Differentiation medium is: MS minimal medium, 0.01-0.05mg/L IAA, 1.0-2.0mg/L 6-BA, 0.1-0.5g/L active carbon, 2% sucrose, 5.5g/L agar, pH value 5.8.
2. the medium of a kind of eggplant anther culture haplobiont according to claim 1, is characterized in that: described eggplant Induction of Anther Culture Callus inducing culture is: MS minimal medium, 0.2mg/L 2,4-D, 0.9mg/L KT, 10mg/L Vc, 3% sucrose, 5g/L agar, pH value 5.8; Described Calli Differentiation medium is: MS minimal medium, 0.02mg/L IAA, 2.0mg/L 6-BA, 0.5g/L active carbon, 2% sucrose, 5.5g/L agar, pH value 5.8.
3. the method for a kind of eggplant anther culture induction haplobiont according to claim 1, is characterized in that: the cultivation of described flower pesticide comprises the steps:
1) grow from eggplant plant and choose bud to the position of eggplant or four door socket eggplants, in bud, anther cell is in monokaryon mid-term to mid-late uninucleate stage, bud diameter 7-10mm, and flower pesticide is that yellowish green colour cast is green;
2) pretreatment: the bud waterproof bag of return collection or blake bottle are closed, is placed in 4-5 DEG C of refrigerating chamber and preserves 12-24h;
3) sterilizing: the alcohol with 70% or 75% carries out bud surface sterilization 30s; Soak 15-20min with the solution that the volumetric concentration of clorox stoste is 7% again, wherein in clorox stoste, the mass concentration of clorox is 10%; With sterile water washing 3-4 time, each 3min, then blots bud with aseptic paper for subsequent use;
4) inoculate: on aseptic working platform, from the bud of bacterium of having gone out, strip flower pesticide gently, be inoculated in the culture dish of Φ 60mm of the inducing culture that eggplant Induction of Anther Culture Callus is housed, each culture dish is equipped with 10mL medium, and each culture dish connects the flower pesticide of 1-2 bud;
5) Heat thermostability: the culture dish having inoculated flower pesticide is put in incubator and carries out Heat thermostability, dark culturing, heat-shock temperature is 35 ~ 40 DEG C, process 4 ~ 9d;
6) anther culture: the flower pesticide after high temperature Heat thermostability, is placed on 25-28 DEG C, cultivates under intensity of illumination 1500-2000Lx, illumination 12-16h/d, and depending on medium situation, transfer medium once every 15-20d, 30 ~ 40d there will be callus;
7) callus plantlet: in the middle part of flower pesticide or top callus lines grow to diameter about 7 ~ 10mm, by Transformation of Callus on Calli Differentiation medium, subculture is cut once at interval of 15-20 d, 30-45 d callus starts to differentiate green bud point, squamous subculture condition: intensity of illumination 1500-2000Lx, illumination 12-16h/d, temperature is 25-28 DEG C, through squamous subculture, the Calli Differentiation of 45%-55% is that ripe embryoid directly generates the full complete plantlets of root, stem and leaf tool;
8) callus of regeneration plant root induction: 8%-12% directly differentiates the regeneration bud of unrooted, when bud seedling grows to 3cm, cut the seedling of band portion callus, be transferred in MS medium, at intensity of illumination 1500-2000Lx, illumination 12-16h/d, temperature is under 25-28 DEG C of culture of rootage condition, grows adventive root after 10-15d, generate whole plant, rooting rate is 90%;
9) Ploidy detection: adopt tip of a root growing point chromosome G banding detection method, in aseptic working platform, from blake bottle, the take root tender tip of a root of children of regrowth of clip detects;
10) haplobiont doubles: step 9 is detected gained haplobiont and adopt growing point infusion method to double, the outside spire of growing point of haplobiont is peelled off between the lights after transplanting, vegetative cone is leaked outside, 0.3-1.0% colchicin+1.5%DMSO solution is dipped with absorbent cotton, parcel vegetative cone, outside with black plastic film parcel, processes 1 ~ 3h again, the normal eggplant male and female flowering of dliploid can be obtained, add multiplying power and reach 60%.
4. the method for a kind of eggplant anther culture induction haplobiont according to claim 3, it is characterized in that: in described step (5), the culture dish having inoculated flower pesticide is put in incubator and carries out Heat thermostability, dark culturing, heat-shock temperature is 36 DEG C, process 5d.
5. the method for a kind of eggplant anther culture induction haplobiont according to claim 3, it is characterized in that: in described step (7), in the middle part of flower pesticide or top callus lines grow to diameter 8mm, by Transformation of Callus on Calli Differentiation medium.
6. the method for a kind of eggplant anther culture induction haplobiont according to claim 3, it is characterized in that: in described step (10), dip 0.5% colchicin+1.5%DMSO solution parcel vegetative cone with absorbent cotton, outside with black plastic film parcel, processes 2h again.
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| CN115644067A (en) * | 2022-12-09 | 2023-01-31 | 云南省农业科学院药用植物研究所 | Method for producing polygonatum kingianum dihaploid |
| CN116806694A (en) * | 2023-08-16 | 2023-09-29 | 金陵科技学院 | Eggplant doubled haploid anther culture method |
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| CN105165620A (en) * | 2015-10-09 | 2015-12-23 | 金陵科技学院 | Eggplant flower bud surface disinfection technology |
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| CN106770250A (en) * | 2016-12-21 | 2017-05-31 | 农业部环境保护科研监测所 | A kind of method of the resistance to cadmium performance of quick detection paddy rice |
| CN106937594A (en) * | 2017-03-07 | 2017-07-11 | 重庆市农业科学院 | A kind of method for promoting eggplant pollen embryo to breed |
| CN106962187A (en) * | 2017-03-24 | 2017-07-21 | 金陵科技学院 | A kind of eggplant haplobiont method for doubling |
| CN111374048A (en) * | 2020-04-28 | 2020-07-07 | 北京市海淀区植物组织培养技术实验室 | Chromosome doubling method of pepper or eggplant haploid plant |
| CN111374048B (en) * | 2020-04-28 | 2022-03-08 | 北京市海淀区植物组织培养技术实验室 | Chromosome doubling method of pepper or eggplant haploid plant |
| CN113728923A (en) * | 2021-09-30 | 2021-12-03 | 云南省农业科学院热区生态农业研究所 | Method for inducing tomato anther callus to rapidly proliferate |
| CN115644067A (en) * | 2022-12-09 | 2023-01-31 | 云南省农业科学院药用植物研究所 | Method for producing polygonatum kingianum dihaploid |
| CN116806694A (en) * | 2023-08-16 | 2023-09-29 | 金陵科技学院 | Eggplant doubled haploid anther culture method |
| CN116849128A (en) * | 2023-08-23 | 2023-10-10 | 金陵科技学院 | Method for culturing anther of Kazakii autotetraploid plant |
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