CN109618930A - The method for cultivating giantreed using flower training monoploid - Google Patents
The method for cultivating giantreed using flower training monoploid Download PDFInfo
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- CN109618930A CN109618930A CN201811619500.8A CN201811619500A CN109618930A CN 109618930 A CN109618930 A CN 109618930A CN 201811619500 A CN201811619500 A CN 201811619500A CN 109618930 A CN109618930 A CN 109618930A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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- Developmental Biology & Embryology (AREA)
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Abstract
The invention discloses the methods for cultivating giantreed using flower training monoploid, method and step is as follows: S1: the bud that mid-late uninucleate stage is on the big giantreed of good stress resistance, plant is removed, then carry out disinfection processing, and the bud after disinfection treatment is taken out anther under anatomical lens；S2: anther obtained by S1 is seeded on callus tissue culture base and carries out induction of callus；S3: the giantreed callus that S2 culture obtains is inoculated on adventitious bud differentiation and culture base and carries out differentiation cultivation, obtains giantreed adventitious bud；S4: the resulting giantreed adventitious bud of S3 is inoculated on root media and carries out culture of rootage；S5: passing through Ploidy Identification, obtains giantreed haplobiont.The present invention improves the resistance of giantreed, shortens the cultivation period of giantreed.
Description
Technical field
The present invention relates to giantreed Cultivating techniques fields, more particularly to the method for cultivating giantreed using flower training monoploid.
Background technique
Giantreed (Arundodonax) grass family, giantreed category perennial plant, have flourishing rhizomes, and stalk is coarse upright, hard
It is tough, have most sections, Chang Shengfen, can be used for a variety of industries such as feed, ethyl alcohol, papermaking, power generation and environment remediation.
Existing giantreed breeding has the methods of sowing, plant division, cuttage, generally based on plant division method；Usually early spring when use
It seizes fastly and has been cut into 4-5 bud one clump along plant surrounding, then transplant；This method reproduction speed is slow, seedling easily band is malicious, and works
Intensity is big, transport is inconvenient, and cost of forestation is high, it is difficult to solve the problems, such as seedling supply shortage in a short time, be unfavorable for giantreed
Industrialization production.Relative to traditional method, giantreed is bred using tissue culture technology, can be obtained in the short time a large amount of whole
Neat consistent virus-free seedling, seedling early growth is with the obvious advantage, is a kind of effective way of giantreed Industrialization of seeds and seedlings breeding.
Polyploid breeding is the important means that new variety of plant is cultivated, and polyploid breeding originates from early 20th century, now
It is widely used in the cultivation of new variety of plant.Polyploid is often shown modal huge due to the increase of genome ploidy
The features such as big property and resistance enhance.Wherein triploid also has the characteristics that sterile, can be used as permanent hybrid, is also beneficial to business and protects
Shield；By genome tripling, the triploid with polyploid and cenospecies double dominant can be obtained, Heterosis is provided
There are huge property and irreplaceability.
Summary of the invention
Technical problems based on background technology, the invention proposes utilization flowers to train the method that monoploid cultivates giantreed,
The resistance for improving giantreed shortens the cultivation period of giantreed.
The method proposed by the present invention for cultivating giantreed using flower training monoploid, method and step are as follows:
S1: the bud that mid-late uninucleate stage is on the big giantreed of good stress resistance, plant is removed, then carry out disinfection place
Reason, takes out anther under anatomical lens for the bud after disinfection treatment；
S2: anther obtained by S1 being seeded on callus tissue culture base and carries out induction of callus, the callus group
The formula for knitting culture medium is 1/2MS+NAA 0.1-0.15mg/L+ sucrose 6-8g/L+ coconut palm cream 5-10g/L+ methyl α-naphthyl acetate 0.05-
0.1mg/L+6-BA(2-4mg/L)；
S3: the giantreed callus that S2 culture obtains is inoculated on adventitious bud differentiation and culture base and carries out differentiation cultivation, is obtained
Giantreed adventitious bud, the formula of the differential medium be 1/2MS+NAA 0.1-0.15mg/L+CPPU (0.8-1.2mg/L)+
Coconut palm cream 5-10g/L+6-BA (0.1-0.15mg/L)+methyl α-naphthyl acetate 0.05-0.1mg/L+2,4-D (1-3mg/L)+sucrose 6-8g/L；
S4: the resulting giantreed adventitious bud of S3 being inoculated on root media and carries out culture of rootage, the root media
Formula are as follows: MS+NAA 0.1-0.15mg/L+ sucrose 6-8g/L+ coconut palm cream 5-10g/L+ methyl α-naphthyl acetate 0.05-0.1mg/L+ sepiolite
0.5-1.5g/L+ active carbon 0.8-2.0g/L；
S5: passing through Ploidy Identification, obtains giantreed haplobiont.
Preferably, the disinfection treatment method in the S1 are as follows: first to the bud aseptic water washing removed, then use again
Alcohol impregnates 20-40min, then carries out surface sterilizing to bud with sodium hypochlorite again, finally uses aseptic water washing again, and like to feel
The moisture on bud surface.
Preferably, in the S2 callus condition of culture are as follows: dark condition, 25 ± 2 DEG C of temperature.
Preferably, the condition of culture is broken up in the S3 are as follows: 25 ± 2 DEG C of cultivation temperature, light application time 10-14h/d, light
According to intensity 2000-3000Lux, colour temperature 4500-5500k.
Preferably, in the S4 culture of rootage condition are as follows: cultivation temperature is within the scope of 25-18 DEG C of day and night temperature, light
It is 10-14h/d, intensity of illumination 2000-3000Lux, colour temperature 4500-5500k according to the time.
Compared with prior art, the beneficial effects of the present invention are: the present invention obtains monoploid children by the Hua Peilai of giantreed
Seedling improves the resistance of giantreed, shortens the period of giantreed cultivation；Culture medium used in callus is 1/2MS+NAA
0.1-0.15mg/L+ sucrose 6-8g/L+ coconut palm cream 5-10g/L+ methyl α-naphthyl acetate 0.05-0.1mg/L+6-BA (2-4mg/L), differentiation culture
Base be 1/2MS+NAA 0.1-0.15mg/L+CPPU (0.8-1.2mg/L)+coconut palm cream 5-10g/L+6-BA (0.1-0.15mg/L)+
Methyl α-naphthyl acetate 0.05-0.1mg/L+2,4-D (1-3mg/L)+sucrose 6-8g/L, root media are MS+NAA 0.1-0.15mg/L+
Sucrose 6-8g/L+ coconut palm cream 5-10g/L+ methyl α-naphthyl acetate 0.05-0.1mg/L+ sepiolite 0.5-1.5g/L+ active carbon 0.8-2.0g/L,
The performance of the giantreed seedling obtained with above-mentioned culture medium culture is greatly improved.
Specific embodiment
Combined with specific embodiments below the present invention is made further to explain.
Embodiment 1
The method proposed by the present invention for cultivating giantreed using flower training monoploid, method and step are as follows:
S1: the bud that mid-late uninucleate stage is on the big giantreed of good stress resistance, plant is removed, then carry out disinfection place
Reason, takes out anther, disinfection treatment method for the bud after disinfection treatment under anatomical lens are as follows: first to the bud removed with sterile
Water rinses, and then impregnates 20min with alcohol again, then surface sterilizing is carried out to bud with sodium hypochlorite again, finally again with sterile
Water rinses, and likes the moisture on sense bud surface；
S2: anther obtained by S1 being seeded on callus tissue culture base and carries out induction of callus, the callus group
The formula for knitting culture medium is 1/2MS+NAA 0.1mg/L+ sucrose 6g/L+ coconut palm cream 5g/L+ methyl α-naphthyl acetate 0.05mg/L+6-BA (2mg/
L), condition of culture are as follows: dark condition, 25 ± 2 DEG C of temperature；
S3: the giantreed callus that S2 culture obtains is inoculated on adventitious bud differentiation and culture base and carries out differentiation cultivation, is obtained
Giantreed adventitious bud is obtained, the formula of the differential medium is 1/2MS+NAA 0.1mg/L+CPPU (0.8mg/L)+coconut palm cream 5g/L+
6-BA (0.1mg/L)+methyl α-naphthyl acetate 0.05mg/L+2,4-D (1mg/L)+sucrose 6g/L, breaks up the condition of culture are as follows: cultivation temperature
25 ± 2 DEG C, light application time 10h/d, intensity of illumination 2000Lux, colour temperature 4500k；
S4: the resulting giantreed adventitious bud of S3 being inoculated on root media and carries out culture of rootage, the root media
Formula are as follows: MS+NAA 0.1mg/L+ sucrose 6g/L+ coconut palm cream 5g/L+ methyl α-naphthyl acetate 0.05mg/L+ sepiolite 0.5g/L+ active carbon
0.8g/L, the condition of culture of rootage are as follows: cultivation temperature is within the scope of 25-18 DEG C of day and night temperature, light application time 10h/d, light
According to intensity 2000Lux, colour temperature 4500k；
S5: passing through Ploidy Identification, obtains giantreed haplobiont.
Embodiment 2
The method proposed by the present invention for cultivating giantreed using flower training monoploid, method and step are as follows:
S1: the bud that mid-late uninucleate stage is on the big giantreed of good stress resistance, plant is removed, then carry out disinfection place
Reason, takes out anther, disinfection treatment method for the bud after disinfection treatment under anatomical lens are as follows: first to the bud removed with sterile
Water rinses, and then impregnates 40min with alcohol again, then surface sterilizing is carried out to bud with sodium hypochlorite again, finally again with sterile
Water rinses, and likes the moisture on sense bud surface；
S2: anther obtained by S1 being seeded on callus tissue culture base and carries out induction of callus, the callus group
The formula for knitting culture medium is 1/2MS+NAA 0.15mg/L+ sucrose 8g/L+ coconut palm cream 10g/L+ methyl α-naphthyl acetate 0.1mg/L+6-BA (4mg/
L), condition of culture are as follows: dark condition, 25 ± 2 DEG C of temperature；
S3: the giantreed callus that S2 culture obtains is inoculated on adventitious bud differentiation and culture base and carries out differentiation cultivation, is obtained
Giantreed adventitious bud is obtained, the formula of the differential medium is 1/2MS+NAA 0.15mg/L+CPPU (1.2mg/L)+coconut palm cream 10g/L
+ 6-BA (0.15mg/L)+methyl α-naphthyl acetate 0.1mg/L+2,4-D (3mg/L)+sucrose 8g/L, breaks up the condition of culture are as follows: cultivation temperature
25 ± 2 DEG C, light application time 14h/d, intensity of illumination 3000Lux, colour temperature 5500k；
S4: the resulting giantreed adventitious bud of S3 being inoculated on root media and carries out culture of rootage, the root media
Formula are as follows: MS+NAA 0.15mg/L+ sucrose 8g/L+ coconut palm cream 10g/L+ methyl α-naphthyl acetate 0.1mg/L+ sepiolite 1.5g/L+ active carbon
2.0g/L, the condition of culture of rootage are as follows: cultivation temperature is within the scope of 25-18 DEG C of day and night temperature, light application time 14h/d, light
According to intensity 3000Lux, colour temperature 5500k；
S5: passing through Ploidy Identification, obtains giantreed haplobiont.
Embodiment 3
The method proposed by the present invention for cultivating giantreed using flower training monoploid, method and step are as follows:
S1: the bud that mid-late uninucleate stage is on the big giantreed of good stress resistance, plant is removed, then carry out disinfection place
Reason, takes out anther, disinfection treatment method for the bud after disinfection treatment under anatomical lens are as follows: first to the bud removed with sterile
Water rinses, and then impregnates 30min with alcohol again, then surface sterilizing is carried out to bud with sodium hypochlorite again, finally again with sterile
Water rinses, and likes the moisture on sense bud surface；
S2: anther obtained by S1 being seeded on callus tissue culture base and carries out induction of callus, the callus group
The formula for knitting culture medium is 1/2MS+NAA 0.12mg/L+ sucrose 7g/L+ coconut palm cream 8g/L+ methyl α-naphthyl acetate 0.08mg/L+6-BA (3mg/
L), condition of culture are as follows: dark condition, 25 ± 2 DEG C of temperature；
S3: the giantreed callus that S2 culture obtains is inoculated on adventitious bud differentiation and culture base and carries out differentiation cultivation, is obtained
Giantreed adventitious bud is obtained, the formula of the differential medium is 1/2MS+NAA 0.12mg/L+CPPU (1.0mg/L)+coconut palm cream 8g/L+
6-BA (0.12mg/L)+methyl α-naphthyl acetate 0.08mg/L+2,4-D (2mg/L)+sucrose 7g/L, breaks up the condition of culture are as follows: cultivation temperature
25 ± 2 DEG C, light application time 12h/d, intensity of illumination 2500Lux, colour temperature 5000k；
S4: the resulting giantreed adventitious bud of S3 being inoculated on root media and carries out culture of rootage, the root media
Formula are as follows: MS+NAA 0.12mg/L+ sucrose 7g/L+ coconut palm cream 8g/L+ methyl α-naphthyl acetate 0.08mg/L+ sepiolite 1.0g/L+ active carbon
1.4g/L, the condition of culture of rootage are as follows: cultivation temperature is within the scope of 25-18 DEG C of day and night temperature, light application time 12h/d, light
According to intensity 2500Lux, colour temperature 5000k；
S5: passing through Ploidy Identification, obtains giantreed haplobiont.
Embodiment 4
The method proposed by the present invention for cultivating giantreed using flower training monoploid, method and step are as follows:
S1: the bud that mid-late uninucleate stage is on the big giantreed of good stress resistance, plant is removed, then carry out disinfection place
Reason, takes out anther, disinfection treatment method for the bud after disinfection treatment under anatomical lens are as follows: first to the bud removed with sterile
Water rinses, and then impregnates 20min with alcohol again, then surface sterilizing is carried out to bud with sodium hypochlorite again, finally again with sterile
Water rinses, and likes the moisture on sense bud surface；
S2: anther obtained by S1 being seeded on callus tissue culture base and carries out induction of callus, the callus group
The formula for knitting culture medium is 1/2MS+NAA 0.1mg/L+ sucrose 8g/L+ coconut palm cream 5g/L+ methyl α-naphthyl acetate 0.05mg/L+6-BA (4mg/
L), condition of culture are as follows: dark condition, 25 ± 2 DEG C of temperature；
S3: the giantreed callus that S2 culture obtains is inoculated on adventitious bud differentiation and culture base and carries out differentiation cultivation, is obtained
Giantreed adventitious bud is obtained, the formula of the differential medium is 1/2MS+NAA 0.1mg/L+CPPU (1.2mg/L)+coconut palm cream 10g/L+
6-BA (0.12mg/L)+methyl α-naphthyl acetate 0.08mg/L+2,4-D (2mg/L)+sucrose 6g/L, breaks up the condition of culture are as follows: cultivation temperature
25 ± 2 DEG C, light application time 12h/d, intensity of illumination 2000Lux, colour temperature 5500k；
S4: the resulting giantreed adventitious bud of S3 being inoculated on root media and carries out culture of rootage, the root media
Formula are as follows: MS+NAA 0.1mg/L+ sucrose 6g/L+ coconut palm cream 10g/L+ methyl α-naphthyl acetate 0.1mg/L+ sepiolite 0.5g/L+ active carbon
0.8g/L, the condition of culture of rootage are as follows: cultivation temperature is within the scope of 25-18 DEG C of day and night temperature, light application time 10h/d, light
According to intensity 3000Lux, colour temperature 5500k；
S5: passing through Ploidy Identification, obtains giantreed haplobiont.
Embodiment 5
The method proposed by the present invention for cultivating giantreed using flower training monoploid, method and step are as follows:
S1: the bud that mid-late uninucleate stage is on the big giantreed of good stress resistance, plant is removed, then carry out disinfection place
Reason, takes out anther, disinfection treatment method for the bud after disinfection treatment under anatomical lens are as follows: first to the bud removed with sterile
Water rinses, and then impregnates 30min with alcohol again, then surface sterilizing is carried out to bud with sodium hypochlorite again, finally again with sterile
Water rinses, and likes the moisture on sense bud surface；
S2: anther obtained by S1 being seeded on callus tissue culture base and carries out induction of callus, the callus group
The formula for knitting culture medium is 1/2MS+NAA 0.1mg/L+ sucrose 8g/L+ coconut palm cream 8g/L+ methyl α-naphthyl acetate 0.05mg/L+6-BA (4mg/
L), condition of culture are as follows: dark condition, 25 ± 2 DEG C of temperature；
S3: the giantreed callus that S2 culture obtains is inoculated on adventitious bud differentiation and culture base and carries out differentiation cultivation, is obtained
Giantreed adventitious bud is obtained, the formula of the differential medium is 1/2MS+NAA 0.15mg/L+CPPU (1.2mg/L)+coconut palm cream 10g/L
+ 6-BA (0.1mg/L)+methyl α-naphthyl acetate 0.08mg/L+2,4-D (1mg/L)+sucrose 6g/L, breaks up the condition of culture are as follows: cultivation temperature
25 ± 2 DEG C, light application time 12h/d, intensity of illumination 2000Lux, colour temperature 5000k；
S4: the resulting giantreed adventitious bud of S3 being inoculated on root media and carries out culture of rootage, the root media
Formula are as follows: MS+NAA 0.12mg/L+ sucrose 7g/L+ coconut palm cream 8g/L+ methyl α-naphthyl acetate 0.08mg/L+ sepiolite 0.5g/L+ active carbon
1.6g/L, the condition of culture of rootage are as follows: cultivation temperature is within the scope of 25-18 DEG C of day and night temperature, light application time 12h/d, light
According to intensity 2000Lux, colour temperature 5500k；
S5: passing through Ploidy Identification, obtains giantreed haplobiont.
The foregoing is only a preferred embodiment of the present invention, but scope of protection of the present invention is not limited thereto,
Anyone skilled in the art in the technical scope disclosed by the present invention, according to the technique and scheme of the present invention and its
Inventive concept is subject to equivalent substitution or change, should be covered by the protection scope of the present invention.
Claims (5)
1. the method for cultivating giantreed using flower training monoploid, which is characterized in that method and step is as follows:
S1: the bud that mid-late uninucleate stage is on the big giantreed of good stress resistance, plant is removed, and then carry out disinfection processing, will
Bud after disinfection treatment takes out anther under anatomical lens；
S2: anther obtained by S1 being seeded on callus tissue culture base and carries out induction of callus, the callus training
The formula for supporting base is 1/2MS+NAA 0.1-0.15mg/L+ sucrose 6-8g/L+ coconut palm cream 5-10g/L+ methyl α-naphthyl acetate 0.05-0.1mg/L+
6-BA(2-4mg/L)；
S3: the giantreed callus that S2 culture obtains is inoculated on adventitious bud differentiation and culture base and carries out differentiation cultivation, obtains reed
Bamboo adventitious bud, the formula of the differential medium are 1/2MS+NAA 0.1-0.15mg/L+CPPU (0.8-1.2mg/L)+coconut palm cream
5-10g/L+6-BA (0.1-0.15mg/L)+methyl α-naphthyl acetate 0.05-0.1mg/L+2,4-D (1-3mg/L)+sucrose 6-8g/L；
S4: the resulting giantreed adventitious bud of S3 is inoculated on root media and carries out culture of rootage, the root media is matched
Side are as follows: MS+NAA 0.1-0.15mg/L+ sucrose 6-8g/L+ coconut palm cream 5-10g/L+ methyl α-naphthyl acetate 0.05-0.1mg/L+ sepiolite 0.5-
1.5g/L+ active carbon 0.8-2.0g/L；
S5: passing through Ploidy Identification, obtains giantreed haplobiont.
2. the method according to claim 1 for cultivating giantreed using flower training monoploid, which is characterized in that disappearing in the S1
Malicious processing method are as follows: first to the bud aseptic water washing removed, then 20-40min is impregnated with alcohol again, then again with secondary
Sodium chlorate carries out surface sterilizing to bud, finally uses aseptic water washing again, and likes the moisture on sense bud surface.
3. the method according to claim 1 for cultivating giantreed using flower training monoploid, which is characterized in that callus in the S2
The condition of culture of tissue are as follows: dark condition, 25 ± 2 DEG C of temperature.
4. the method according to claim 1 for cultivating giantreed using flower training monoploid, which is characterized in that break up in the S3
The condition of culture are as follows: 25 ± 2 DEG C of cultivation temperature, light application time 10-14h/d, intensity of illumination 2000-3000Lux, colour temperature
4500-5500k。
5. the method according to claim 1 for cultivating giantreed using flower training monoploid, which is characterized in that take root in the S4
The condition of culture are as follows: cultivation temperature is within the scope of 25-18 DEG C of day and night temperature, light application time 10-14h/d, intensity of illumination
2000-3000Lux, colour temperature 4500-5500k.
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811619500.8A CN109618930A (en) | 2018-12-28 | 2018-12-28 | The method for cultivating giantreed using flower training monoploid |
| EP19902458.9A EP3903577A4 (en) | 2018-12-28 | 2019-12-27 | Method for growing arundo donax plant |
| PCT/CN2019/129195 WO2020135712A1 (en) | 2018-12-28 | 2019-12-27 | Method for growing arundo donax plant |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201811619500.8A CN109618930A (en) | 2018-12-28 | 2018-12-28 | The method for cultivating giantreed using flower training monoploid |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110923264A (en) * | 2019-12-09 | 2020-03-27 | 中国科学院亚热带农业生态研究所 | Method suitable for high-efficiency tissue culture of various gramineous plants |
| WO2020135712A1 (en) * | 2018-12-28 | 2020-07-02 | 武汉兰多生物科技有限公司 | Method for growing arundo donax plant |
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| CN102550405A (en) * | 2011-12-26 | 2012-07-11 | 北京林业大学 | Breeding method of poplar haploid |
| CN103749302A (en) * | 2014-01-15 | 2014-04-30 | 江苏沿海地区农业科学研究所 | Induced acclimation and cultivation method for salt-tolerant bamboo reed seedlings |
| CN104429939A (en) * | 2014-07-18 | 2015-03-25 | 皖西学院 | Method for cultivating triploid dendrobium huoshanense by fusion breeding method of diploid cells and pollen cells |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020135712A1 (en) * | 2018-12-28 | 2020-07-02 | 武汉兰多生物科技有限公司 | Method for growing arundo donax plant |
| CN110923264A (en) * | 2019-12-09 | 2020-03-27 | 中国科学院亚热带农业生态研究所 | Method suitable for high-efficiency tissue culture of various gramineous plants |
| CN110923264B (en) * | 2019-12-09 | 2021-09-07 | 中国科学院亚热带农业生态研究所 | Method suitable for high-efficiency tissue culture of various gramineous plants |
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Application publication date: 20190416 |