CN105010139A - Cultivation method for polyploid Xinjiang lavender plant - Google Patents
Cultivation method for polyploid Xinjiang lavender plant Download PDFInfo
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- CN105010139A CN105010139A CN201510393020.4A CN201510393020A CN105010139A CN 105010139 A CN105010139 A CN 105010139A CN 201510393020 A CN201510393020 A CN 201510393020A CN 105010139 A CN105010139 A CN 105010139A
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Abstract
The invention relates to a cultivation method for a polyploid Xinjiang lavender plant. The method comprises the steps: by taking sterile seedling and stem segments of a perferable superior plant of Xinjiang lavender as materials, adding colchicine with different concentrations into a culture medium for induction treatment; and after the plant is doubled, identifying the plant by applying a ploidy identification techniques to obtain the polyploid Xinjiang lavender plant. The method provides the optimum colchicine concentration, the processing time and the processing temperature for inducing the polyploid plant and a method for ploidy identification. A novel variety of the polyploid Xinjiang lavender plant can be quickly bred by virtue of the method and the breeding period is shortened, so that the cultivation rate of excellent seedlings of Xinjiang lavender is effectively increased, and the problem of germplasm degeneration easily occurred in a long-term planting process and conventional breeding of Xinjiang lavender is overcome.
Description
Technical field
The present invention relates to biological technical field, particularly relate to the breeding method of a kind of polyploid Xinjiang lavender plant.
Background technology
Lavender is cross-pollinatd plant, and due to the complex heterogeneous of this colony, offspring always occurs trait segregation, shows diversity, and merit is difficult to stably heredity and goes down, the basic reason that current China lavender kind that Here it is easily mixes, degenerates.The conventional breeding of lavender is often in order to obtain more stable isozygoty offspring and guarantee Selection effect, must under the condition of suitable controlled pollination, repeatedly select, and the not resistance to selfing of cross-pollinatd plant, vitality is caused significantly to fail from intersection, thus the comparatively large and length consuming time of breeding difficulty, germplasm character is unstable.After using polyploid breeding technology that plant chromosome is doubled, the object improving its essential oil yield is reached by the biological yield improving lavender, that Application comparison is extensive in the world at present, be applicable to the breeding method of lavender for essential oil, simultaneously in order to obtain more stable isozygoty offspring and guarantee Selection effect, at present the most directly, method uses group culturation rapid propagating technology directly to carry out vegetative propagation to the polyploid strain selected efficiently, avoid the blastation of seed and cottage propagation seedling, thus a large amount of polyploid lavender high quality seedlings can be obtained.
Summary of the invention
The object of the invention is to, the breeding method of a kind of polyploid Xinjiang lavender plant is provided, the method utilizes the aseptic seedling stem sections of the preferred excellent strain of Xinjiang lavender to be material, induction process is carried out by adding variable concentrations colchicine in the medium, after plant is doubled, use a times gonosome authenticate technology to identify again, obtain the Xinjiang lavender plant of polyploid.This method provide the best colchicine concentration of induction polyploid, processing time, treatment temperature and the method for Ploidy Identification.This method can realize Xinjiang lavender breeding of new variety fast, shorten breeding cycle, and then effectively improve the cultivation speed of Xinjiang lavender high quality seedling, overcome the trait segregation that Xinjiang lavender easily occurs in conventional breeding, merit is difficult to stably heredity and goes down, the problems such as the new varieties cultivated easily mix, degeneration.
Of the present invention is the breeding method of a kind of polyploid Xinjiang lavender plant, follows these steps to carry out:
A, select healthy and strong lavender plant stem apex, be the detergent solution agitator treating of 5%, rinse 2-4h under discharge condition by concentration, it is for subsequent use to dry rear placement;
B, alcoholic solution 20%+0.1% mercuric chloride solution 80%+ Tween 80 2-3 stem apex cleaned in step a being placed in concentration 75% drip mixed liquor and to sterilize 7min, use aseptic water washing 4-5 time again, after sterilizing filter paper suck dry moisture, be inoculated in MS+BA1.0mg/L solid culture medium and carry out aseptic seedling Fiber differentiation;
C, by after in step b, the aseptic seedling of germination and growth cuts stem section, be inoculated on the inducing solid medium MS+BA1.0mg/L containing concentration to be 2% dimethyl sulfoxide (DMSO) and concentration be 0.20% colchicine and process 1-3d, cultivation temperature 25 DEG C, intensity of illumination 2000LX, light application time 12h;
D, the stem section processed in step c is proceeded in the propagation solid culture medium of MS+BA1.0mg/L+IBA0.3mg/L in time, cultivate 25d, cultivation temperature 24 DEG C, intensity of illumination 2000LX, light application time 12h;
E, the seedling of breeding in steps d is cut into 3-4 bud one clump, is forwarded in 1/2MS+NAA0.5mg/L root media and carries out culture of rootage, cultivation temperature 24 DEG C, intensity of illumination 3000LX, light application time 16h;
F, by the plant strain growth that grows in step e to 3-4cm, when root grows 0.5-1cm, do comparative analysis with dliploid adjoining tree, according to the two in mode of appearance width of blade, thickness, stem rugosity and the size of Stomacal guard cell and the difference of density, Preliminary screening goes out polyploid plant;
The organization of root tips 4-6mm that in g, random clip step f, the plant children of Preliminary screening is tender, under temperature 4 DEG C of conditions, uses 0.002mol/L
-1oxine pretreatment 4h or the pretreatment of the saturated paracide aqueous solution, with Kano fixer 95% ethanol under temperature 4 DEG C of conditions: glacial acetic acid volume ratio=3:1 fixes 24 h, discards fixer, the tip of a root is placed in temperature 60 C water-bath 1 molL
-1hydrochloric acid hydrolysis 9 min, rinse dissociation solution well, to dye 20 min with improved phenol fuchsin, statistics root tip chromosomes observed by compressing tablet, chromosome is defined as polyploid plant more than a times of adjoining tree, or carries out ploidy analysis with flow cytometer, determines polyploid plant;
I, the polyploid plant cultivated after testing in step g washed away the remaining medium of root attachment, be colonizated in turfy soil: perlite=2: on the greenhouse seedbed of 1, cultivated water permeable, spray after 50% carbendazol wettable powder, 1000 times of liquid carry out disinfection, build Small plastic shed, be covered with film, every day ventilates 1-2 time, ventilation time is increased to whole day gradually by 10min, need not rewater before taking off film, just slightly spray some clear water on blade face when ventilating, after 20-25d, removing film gradually, carry out seedling normal management, transplanting survival rate 90%.
The breeding method of a kind of polyploid Xinjiang of the present invention lavender plant, 50% carbendazol wettable powder described in the method is commercially available prod, and wherein carbendazim is that Jin Liang Fine Chemical Co., Ltd of Henan Province produces.
By implementing the concrete summary of the invention of the present invention, following effect can be reached:
The breeding method of a kind of polyploid Xinjiang of the present invention lavender plant, the method is simple to operate, Xinjiang lavender breeding of new variety can be realized fast, shorten breeding cycle, and then effectively improve the cultivation speed of Xinjiang lavender high quality seedling, overcome the trait segregation that Xinjiang lavender easily occurs in conventional breeding, merit is difficult to stably heredity and goes down, the problems such as the new varieties cultivated easily mix, degeneration.
Accompanying drawing explanation
Fig. 1 is lavender C-197 polyploid group of the present invention training tufted seedling picture;
Fig. 2 is lavender C-197 polyploid subculture bottle seedling picture of the present invention;
Fig. 3 is the lavender 74-26 polyploid plantlet in vitro picture of field-transplanting of the present invention.
Embodiment
Method of the present invention is not limited to following embodiment; Simultaneously in following explanation, if no special instructions, % all refers to m/m mass percent; All reagent, instrument and the instrument selected in the present invention are all well known commercially available prod, but do not limit enforcement of the present invention, and other reagent more well known in the art and equipment are all applicable to the present invention.
Embodiment 1
A, select healthy and strong Xinjiang lavender main breed C-197 plant stem apex, be the detergent solution agitator treating of 5%, rinse 2-4h under discharge condition by concentration, it is for subsequent use to dry rear placement;
B, alcoholic solution 20%+0.1% mercuric chloride solution 80%+ Tween 80 2 mixed liquors stem apex cleaned in step a being placed in concentration 75% are sterilized 7min, use aseptic water washing again 4 times, after sterilizing filter paper suck dry moisture, be inoculated in MS+BA1.0mg/L solid culture medium and carry out aseptic seedling Fiber differentiation;
C, by after in step b, the aseptic seedling of germination and growth cuts stem section, be inoculated on the inducing solid medium MS+BA1.0mg/L containing concentration to be 2% dimethyl sulfoxide (DMSO) and concentration be 0.20% colchicine and process 1d, cultivation temperature 25 DEG C, intensity of illumination 2000LX, light application time 12h;
D, the stem section processed in step c is proceeded in the propagation solid culture medium of MS+BA1.0mg/L+IBA0.3mg/L in time, cultivate 25d, cultivation temperature 24 DEG C, intensity of illumination 2000LX, light application time 12h Fig. 2;
E, the seedling of breeding in steps d is cut into 3 bud one clump, is forwarded in 1/2MS+NAA0.5mg/L root media and carries out culture of rootage, cultivation temperature 24 DEG C, intensity of illumination 3000LX, light application time 16h Fig. 1;
F, by the plant strain growth that grows in step e to 3cm, when root grows 0.5cm, do comparative analysis with dliploid adjoining tree, according to the two in mode of appearance width of blade, thickness, stem rugosity and the size of Stomacal guard cell and the difference of density, Preliminary screening goes out polyploid plant;
The organization of root tips 4mm that in g, random clip step f, the plant children of Preliminary screening is tender, under temperature 4 DEG C of conditions, uses 0.002mol/L
-1oxine pretreatment 4h, with Kano fixer 95% ethanol under temperature 4 DEG C of conditions: glacial acetic acid=3:1(volume ratio) fix 24 h, discard fixer, the tip of a root is placed in temperature 60 C water-bath 1 molL
-1hydrochloric acid hydrolysis 9 min, rinse dissociation solution well, to dye 20 min with improved phenol fuchsin, statistics root tip chromosomes observed by compressing tablet, and chromosome is defined as polyploid plant more than a times of adjoining tree;
I, the polyploid plant cultivated after testing in step g washed away the remaining medium of root attachment, be colonizated in turfy soil: perlite=2: on the greenhouse seedbed of 1, cultivated water permeable, spray after 50% carbendazol wettable powder, 1000 times of liquid carry out disinfection, build Small plastic shed, be covered with film, every day ventilates 1-2 time, ventilation time is increased to whole day gradually by 10min, need not rewater before taking off film, just slightly spray some clear water on blade face when ventilating, after 20d, removing film gradually, carry out seedling normal management, transplanting survival rate 90%.
Embodiment 2
A, select healthy and strong Xinjiang lavender main breed 74-26 plant stem apex, be the detergent solution agitator treating of 5%, rinse 4h under discharge condition by concentration, it is for subsequent use to dry rear placement;
B, alcoholic solution 20%+0.1% mercuric chloride solution 80%+ Tween 80 3 mixed liquors stem apex cleaned in step a being placed in concentration 75% are sterilized 7min, use aseptic water washing again 5 times, after sterilizing filter paper suck dry moisture, be inoculated in MS+BA1.0mg/L solid culture medium and carry out aseptic seedling Fiber differentiation;
C, by after in step b, the aseptic seedling of germination and growth cuts stem section, be inoculated on the inducing solid medium MS+BA1.0mg/L containing concentration to be 2% dimethyl sulfoxide (DMSO) and concentration be 0.20% colchicine and process 3d, cultivation temperature 25 DEG C, intensity of illumination 2000LX, light application time 12h;
D, the stem section processed in step c is proceeded in the propagation solid culture medium of MS+BA1.0mg/L+IBA0.3mg/L in time, cultivate 25d, cultivation temperature 24 DEG C, intensity of illumination 2000LX, light application time 12h;
E, the seedling of breeding in steps d is cut into 4 bud one clump, is forwarded in 1/2MS+NAA0.5mg/L root media and carries out culture of rootage, cultivation temperature 24 DEG C, intensity of illumination 3000LX, light application time 16h;
F, by the plant strain growth that grows in step e to 4cm, when root grows 1cm, do comparative analysis with dliploid adjoining tree, according to the two in mode of appearance width of blade, thickness, stem rugosity and the size of Stomacal guard cell and the difference of density, Preliminary screening goes out polyploid plant;
The organization of root tips 4-6mm that in g, random clip step f, the plant children of Preliminary screening is tender, under temperature 4 DEG C of conditions, with the pretreatment of the saturated paracide aqueous solution, with Kano fixer 95% ethanol under temperature 4 DEG C of conditions: glacial acetic acid=3:1(volume ratio) fix 24 h, discard fixer, the tip of a root is placed in temperature 60 C water-bath 1 molL
-1hydrochloric acid hydrolysis 9 min, rinse dissociation solution well, to dye 20 min with improved phenol fuchsin, statistics root tip chromosomes observed by compressing tablet, carries out ploidy analysis, determine polyploid plant with flow cytometer;
I, the polyploid plant cultivated after testing in step g washed away the remaining medium of root attachment, be colonizated in turfy soil: perlite=2: on the greenhouse seedbed of 1, cultivated water permeable, spray after 50% carbendazol wettable powder, 1000 times of liquid carry out disinfection, build Small plastic shed, be covered with film, every day ventilates 1-2 time, ventilation time is increased to whole day gradually by 10min, need not rewater before taking off film, just slightly spray some clear water on blade face when ventilating, after 20-25d, removing film gradually, carry out seedling normal management, transplanting survival rate 90% Fig. 3.
Claims (1)
1. a breeding method for polyploid Xinjiang lavender plant, is characterized in that following these steps to carry out:
A, select healthy and strong lavender plant stem apex, be the detergent solution agitator treating of 5%, rinse 2-4h under discharge condition by concentration, it is for subsequent use to dry rear placement;
B, alcoholic solution 20%+0.1% mercuric chloride solution 80%+ Tween 80 2-3 stem apex cleaned in step a being placed in concentration 75% drip mixed liquor and to sterilize 7min, use aseptic water washing 4-5 time again, after sterilizing filter paper suck dry moisture, be inoculated in MS+BA1.0mg/L solid culture medium and carry out aseptic seedling Fiber differentiation;
C, by after in step b, the aseptic seedling of germination and growth cuts stem section, be inoculated on the inducing solid medium MS+BA1.0mg/L containing concentration to be 2% dimethyl sulfoxide (DMSO) and concentration be 0.20% colchicine and process 1-3d, cultivation temperature 25 DEG C, intensity of illumination 2000LX, light application time 12h;
D, the stem section processed in step c is proceeded in the propagation solid culture medium of MS+BA1.0mg/L+IBA0.3mg/L in time, cultivate 25d, cultivation temperature 24 DEG C, intensity of illumination 2000LX, light application time 12h;
E, the seedling of breeding in steps d is cut into 3-4 bud one clump, is forwarded in 1/2MS+NAA0.5mg/L root media and carries out culture of rootage, cultivation temperature 24 DEG C, intensity of illumination 3000LX, light application time 16h;
F, by the plant strain growth that grows in step e to 3-4cm, when root grows 0.5-1cm, do comparative analysis with dliploid adjoining tree, according to the two in mode of appearance width of blade, thickness, stem rugosity and the size of Stomacal guard cell and the difference of density, Preliminary screening goes out polyploid plant;
The organization of root tips 4-6mm that in g, random clip step f, the plant children of Preliminary screening is tender, under temperature 4 DEG C of conditions, uses 0.002mol/L
-1oxine pretreatment 4h or the pretreatment of the saturated paracide aqueous solution, with Kano fixer 95% ethanol under temperature 4 DEG C of conditions: glacial acetic acid volume ratio=3:1 fixes 24 h, discards fixer, the tip of a root is placed in temperature 60 C water-bath 1 molL
-1hydrochloric acid hydrolysis 9 min, rinse dissociation solution well, to dye 20 min with improved phenol fuchsin, statistics root tip chromosomes observed by compressing tablet, chromosome is defined as polyploid plant more than a times of adjoining tree, or carries out ploidy analysis with flow cytometer, determines polyploid plant;
I, the polyploid plant cultivated after testing in step g washed away the remaining medium of root attachment, be colonizated in turfy soil: perlite=2: on the greenhouse seedbed of 1, cultivated water permeable, spray after 50% carbendazol wettable powder, 1000 times of liquid carry out disinfection, build Small plastic shed, be covered with film, every day ventilates 1-2 time, ventilation time is increased to whole day gradually by 10min, need not rewater before taking off film, just slightly spray some clear water on blade face when ventilating, after 20-25d, removing film gradually, carry out seedling normal management, transplanting survival rate 90%.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1817112A (en) * | 2006-03-03 | 2006-08-16 | 中国科学院新疆理化技术研究所 | Fast lavandulol regeneration |
| CN102823492A (en) * | 2012-08-08 | 2012-12-19 | 中国科学院武汉植物园 | Method for quickly propagating lavenders |
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- 2015-07-07 CN CN201510393020.4A patent/CN105010139A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1817112A (en) * | 2006-03-03 | 2006-08-16 | 中国科学院新疆理化技术研究所 | Fast lavandulol regeneration |
| CN102823492A (en) * | 2012-08-08 | 2012-12-19 | 中国科学院武汉植物园 | Method for quickly propagating lavenders |
Non-Patent Citations (3)
| Title |
|---|
| NIGEL A.R.URWIN: ""Generation and characterisation of colchicine-induced polyploid Lavandula×intermedia"", 《EUPHYTICA》 * |
| 左利娟等: "‘优雅’薰衣草茎段再生体系的建立", 《河北林果研究》 * |
| 蒋新明等: ""薰衣草离体培养研究"", 《安徽农业科学》 * |
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Application publication date: 20151104 |