CN103749305A - Tissue culture method of baptisia australis - Google Patents
Tissue culture method of baptisia australis Download PDFInfo
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- CN103749305A CN103749305A CN201410024961.6A CN201410024961A CN103749305A CN 103749305 A CN103749305 A CN 103749305A CN 201410024961 A CN201410024961 A CN 201410024961A CN 103749305 A CN103749305 A CN 103749305A
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Abstract
The invention relates to a tissue culture method of baptisia australis. The tissue culture method comprises the steps of sterile treatment, differentiation and proliferation, strong seedling of adventitious buds, rooting culture, acclimatization and transplanting, and the like. The step-by-step tissue culture of the baptisia australis is carried out by using a bud induction medium, a proliferation medium, a strong seedling medium and a rooting medium. Compared with the prior art, the tissue culture method has the advantages that the baptisia australis is subjected to step-by-step tissue culture by selecting the appropriate mediums, so that the reproduction speed and the evenness degree of nursery stocks are increased, the original female parent characters are maintained well, and in addition, the survival rate of transplanting can reach 85%.
Description
Technical field
The present invention relates to a kind of method for plant tissue culture, especially relate to a kind of method of cultivating the blue beans in Australia of organizing.
Background technology
The blue beans in Australia are that the counterfeit indigo of pulse family belongs to perennial root flower plant.Originate in Australia, be often born in border.Stem is upright, high about 50-100cm left and right, Cheng Congzhuan.Winglike compound leaf, alternate, raceme are born in stem top,
Corolla butterfly, blueness.The florescence 4-5 month.Fruit ellipse or Long Circle; Seed brown color, kidney shape.Can survive the winter by outdoor perennial root.Sprout strong shoot from base portion the beginning of spring.Happiness cools, and draining is good, ventilates, and sun-drenched place, avoids sultry wet environment.The blue beans in Australia are relatively afraid of dark hygrothermal environment, and the environment of cultivation must ventilate, on the sunny side.Can large area sheet plant do quilt, also can group planting or be disposed at Hua Jingzhong, be also the excellent material that garth beautifies.From the new varieties of external introduction, introduce a fine variety negligible amounts, its plant division is slow, the huge market demand, seedling supply is restricted.Therefore need to develop tissue culture technology to improve the regularity of reproduction speed and nursery stock, keep better original maternal character, but how the condition of How to choose tissue culture medium (TCM), group training is to need problem demanding prompt solution.
Summary of the invention
Object of the present invention is exactly in order to overcome the defect that above-mentioned prior art exists, to provide a kind of substep tissue that carries out to cultivate, improve survival rate, guarantee that the tissue of maternal character cultivates the method for the blue beans in Australia.
Object of the present invention can be achieved through the following technical solutions:
Organize a method of cultivating the blue beans in Australia, adopt following steps:
(1) aseptic process
Win the terminal bud that the blue beans plant in Australia has just sprouted, water rinses 2h, with the alcohol immersion 30s of 75v/v%, 1v/v%0 mercuric chloride soaks 15min, then uses aseptic water washing 5-6 time, aseptic filter paper to blot surperficial moisture content, bud section is cut into 1cm and is inoculated on bud inducing culture, control cultivation temperature is 24-26 ℃, and illumination is 70-90 μ mol/ms, carries out bud induction and cultivates;
(2) differentiation and proliferation
Be inoculated in bud section on bud inducing culture and cultivating after 4 weeks callus as seen, then cultivate after 1 month band bud callus is inoculated on proliferated culture medium, control cultivation temperature is 24-26 ℃, and illumination is 70-90 μ mol/ms, carries out differentiation and proliferation cultivation;
(3) indefinite bud strong sprout
On proliferated culture medium, induce the indefinite bud of growing thickly, indefinite bud is divided into little Cong or simple bud is placed in strong seedling culture base, controlling cultivation temperature is 24-26 ℃, and illumination is 70-90 μ mol/ms, carries out strong seedling culture, within 30 days, grows to 2-3cm;
(4) culture of rootage
By the root induction in root media of transferring of the indefinite bud in strong seedling culture base, control cultivation temperature is 24-26 ℃, and illumination is 70-90 μ mol/ms, and after 10 days, seedling base section dissolves the former base of root of many whites, after 30 days, grows to 2-3em;
(5) acclimatization and transplants
40 days root systems of culture of rootage grow to 1-2cm, and the aseptic seedling of selecting well developed root system robust growth, indoor uncork hardening 7 days, is taken out afterwash root agar, tame after 40 days, to transplant outdoorly in greenhouse, give rich water quality management.
The nutrient component of bud inducing culture is MS+6-BA1.0mg/L+NAA0.1mg/L, MS+6-BA3.0mg/L+NAA0.3mg/L or MS+6-BA5.0mg/L+NAA0.5mg/L.
As preferred embodiment, the nutrient component of bud inducing culture is MS+6-BA3.0mg/L+NAA0.3mg/L.
The nutrient component of proliferated culture medium is MS+6-BA1mg/L-+NAA0.1mg/L, MS+6-BA2mg/L+NAA0.2mg/L or MS+6-BA3mg/L+NAA0.3mg/L.
As preferred embodiment, the nutrient component of proliferated culture medium is MS+6BA2mg/L+NAA0.2mg/L.
The nutrient component of strong seedling culture base is MS+6-BA0.5mg/L+NAA0.1mg/L, MS+6-BA1.0mg/L+NAA0.1mg/L or MS+6-BA2mg/L+NAA0.2mg/L.
As preferred embodiment, the nutrient component of strong seedling culture base is MS+6-BA0.5mg/L+NAA0.1mg/L.
The nutrient component of root media is MS+NAA0.05mg/L, MS+NAA0.1mg/L or MS+NAA0.2mg/L.
As preferred embodiment, the nutrient component of root media is MS+NAA0.1mg/L.
Described bud inducing culture, proliferated culture medium, strong seedling culture base, root media also comprises the basis of sucrose 30g/L, agar 6g/L composition, and the pH value of above-mentioned medium is controlled at 5.8.
Compared with prior art, the present invention cultivates by selecting applicable medium to carry out substep tissue to the blue beans in Australia, thereby improves the regularity of reproduction speed and nursery stock, keeps better original maternal character, and transplanting survival rate can reach 85% in addition.
Embodiment
Below in conjunction with specific embodiment, the present invention is described in detail.
Embodiment 1
Organize a method of cultivating the blue beans in Australia, adopt following steps:
(1) aseptic process
Win the terminal bud that the blue beans plant in Australia has just sprouted, water rinses 2h, with the alcohol immersion 30s of 75v/v%, 1v/v%0 mercuric chloride soaks 15min, use again aseptic water washing 5-6 time, aseptic filter paper blots surperficial moisture content, bud section is cut into 1em and is inoculated on bud inducing culture, the nutrient component of bud inducing culture is MS+6-BA1.0mg/L+NAA0.1mg/L, the basis that also comprises sucrose 30g/L, agar 6g/L composition, the pH value of medium is 5.8, controlling cultivation temperature is 24 ℃, illumination is 70 μ mol/ms, carries out bud induction and cultivates;
(2) differentiation and proliferation
The bud section being inoculated on bud inducing culture is being cultivated visible callus after 4 weeks, cultivate again after 1 month band bud callus is inoculated on proliferated culture medium, the nutrient component of proliferated culture medium is MS+6-BA1mg/L-+NAA0.1mg/L, also comprise the basis of sucrose 30g/L, agar 6g/L composition, the pH value of medium is 5.8, controlling cultivation temperature is 24 ℃, and illumination is 70 μ mol/ms, carries out differentiation and proliferation cultivation;
(3) indefinite bud strong sprout
On proliferated culture medium, induce the indefinite bud of growing thickly, indefinite bud is divided into little Cong or simple bud is placed in strong seedling culture base, the nutrient component of strong seedling culture base is MS+6-BA0.5mg/L+NAA0.1mg/L, also comprise the basis of sucrose 30g/L, agar 6g/L composition, the pH value of medium is 5.8, and controlling cultivation temperature is 24 ℃, and illumination is 70 μ mol/ms, carry out strong seedling culture, within 30 days, grow to 2-3cm;
(4) culture of rootage
By the root induction in root media of transferring of the indefinite bud in strong seedling culture base, the nutrient component of root media is MS+NAA0.05mg/L, also comprise the basis of sucrose 30g/L, agar 6g/L composition, the pH value of medium is 5.8, controlling cultivation temperature is 24 ℃, illumination is 70 μ mol/ms, and after l0 days, seedling base section dissolves the former base of root of many whites, after 30 days, grows to 2-3cm;
(5) acclimatization and transplants
40 days root systems of culture of rootage grow to 1-2cm, and the aseptic seedling of selecting well developed root system robust growth, indoor uncork hardening 7 days, is taken out afterwash root agar, tame after 40 days, to transplant outdoorly in greenhouse, give rich water quality management.
Embodiment 2
Organize a method of cultivating the blue beans in Australia, adopt following steps:
(1) aseptic process
Win the terminal bud that the blue beans plant in Australia has just sprouted, water rinses 2h, with the alcohol immersion 30s of 75v/v%, 1v/v%0 mercuric chloride soaks 15min, use again aseptic water washing 5 times, aseptic filter paper blots surperficial moisture content, bud section is cut into lcm and is inoculated on bud inducing culture, the nutrient component of bud inducing culture is MS+6-BA3.0mg/L+NAA0.3mg/L, the basis that also comprises sucrose 30g/L, agar 6g/L composition, the pH value of medium is 5.8, controlling cultivation temperature is 25 ℃, illumination is 80 μ mol/ms, carries out bud induction and cultivates;
(2) differentiation and proliferation
The bud section being inoculated on bud inducing culture is being cultivated visible callus after 4 weeks, cultivate again after 1 month band bud callus is inoculated on proliferated culture medium, the nutrient component of proliferated culture medium is MS+6BA2mg/L+NAA0.2mg/L, also comprise the basis of sucrose 30g/L, agar 6g/L composition, the pH value of medium is 5.8, controlling cultivation temperature is 25 ℃, and illumination is 80 μ mol/ms, carries out differentiation and proliferation cultivation;
(3) indefinite bud strong sprout
On proliferated culture medium, induce the indefinite bud of growing thickly, indefinite bud is divided into little Cong or simple bud is placed in strong seedling culture base, the nutrient component of strong seedling culture base is MS+6-BA0.5mg/L+NAA0.1mg/L, also comprise the basis of sucrose 30g/L, agar 6g/L composition, the pH value of medium is 5.8, and controlling cultivation temperature is 25 ℃, and illumination is 80 μ mol/ms, carry out strong seedling culture, within 30 days, grow to 2-3cm;
(4) culture of rootage
By the root induction in root media of transferring of the indefinite bud in strong seedling culture base, the nutrient component of root media is MS+NAA0.1mg/L, also comprise the basis of sucrose 30g/L, agar 6g/L composition, the pH value of medium is 5.8, controlling cultivation temperature is 25 ℃, illumination is 80 μ mol/ms, and after 10 days, seedling base section dissolves the former base of root of many whites, after 30 days, grows to 2-3cm;
(5) acclimatization and transplants
40 days root systems of culture of rootage grow to 1-2cm, and the aseptic seedling of selecting well developed root system robust growth, indoor uncork hardening 7 days, is taken out afterwash root agar, tame after 40 days, to transplant outdoorly in greenhouse, give rich water quality management.
Embodiment 3
Organize a method of cultivating the blue beans in Australia, adopt following steps:
(1) aseptic process
Win the terminal bud that the blue beans plant in Australia has just sprouted, water rinses 2h, with the alcohol immersion 30s of 75v/v%, 1v/v%0 mercuric chloride soaks 15min, use again aseptic water washing 6 times, aseptic filter paper blots surperficial moisture content, bud section is cut into 1cm and is inoculated on bud inducing culture, the nutrient component of bud inducing culture is MS+6-BA5.0mg/L+NAA0.5mg/L, the basis that also comprises sucrose 30g/L, agar 6g/L composition, the pH value of medium is 5.8, controlling cultivation temperature is 26 ℃, illumination is 90 μ mol/ms, carries out bud induction and cultivates;
(2) differentiation and proliferation
The bud section being inoculated on bud inducing culture is being cultivated visible callus after 4 weeks, cultivate again after 1 month band bud callus is inoculated on proliferated culture medium, the nutrient component of proliferated culture medium is MS+6-BA3mg/L+NAA0.3mg/L, also comprise the basis of sucrose 30g/L, agar 6g/L composition, the pH value of medium is 5.8, controlling cultivation temperature is 26 ℃, and illumination is 90 μ mol/ms, carries out differentiation and proliferation cultivation;
(3) indefinite bud strong sprout
On proliferated culture medium, induce the indefinite bud of growing thickly, indefinite bud is divided into little Cong or simple bud is placed in strong seedling culture base, the nutrient component of strong seedling culture base is MS+6-BA2mg/L+NAA0.2mg/L, also comprise the basis of sucrose 30g/L, agar 6g/L composition, the pH value of medium is 5.8, and controlling cultivation temperature is 26 ℃, and illumination is 90 μ mol/ms, carry out strong seedling culture, within 30 days, grow to 2-3cm;
(4) culture of rootage
By the root induction in root media of transferring of the indefinite bud in strong seedling culture base, the nutrient component of root media is MS+NAA0.2mg/L, also comprise the basis of sucrose 30g/L, agar 6g/L composition, the pH value of medium is 5.8, controlling cultivation temperature is 26 ℃, illumination is 90 μ mol/ms, and after 10 days, seedling base section dissolves the former base of root of many whites, grows to 2-3cm after 30 days:
(5) acclimatization and transplants
40 days root systems of culture of rootage grow to 1-2cm, and the aseptic seedling of selecting well developed root system robust growth, indoor uncork hardening 7 days, is taken out afterwash root agar, tame after 40 days, to transplant outdoorly in greenhouse, give rich water quality management.
Claims (10)
1. organize a method of cultivating the blue beans in Australia, it is characterized in that, the method adopts following steps:
(1) aseptic process
Win the terminal bud that the blue beans plant in Australia has just sprouted, water rinses 2h, with the alcohol immersion 30s of 75v/v%, 1v/v%0 mercuric chloride soaks 15min, then uses aseptic water washing 5-6 time, aseptic filter paper to blot surperficial moisture content, bud section is cut into 1cm and is inoculated on bud inducing culture, control cultivation temperature is 24-26 ℃, and illumination is 70-90 μ mol/ms, carries out bud induction and cultivates;
(2) differentiation and proliferation
Be inoculated in bud section on bud inducing culture and cultivating after 4 weeks callus as seen, then cultivate after 1 month band bud callus is inoculated on proliferated culture medium, control cultivation temperature is 24-26 ℃, and illumination is 70-90 μ mol/ms, carries out differentiation and proliferation cultivation;
(3) indefinite bud strong sprout
On proliferated culture medium, induce the indefinite bud of growing thickly, indefinite bud is divided into little Cong or simple bud is placed in strong seedling culture base, controlling cultivation temperature is 24-26 ℃, and illumination is 70-90 μ mol/ms, carries out strong seedling culture, within 30 days, grows to 2-3cm;
(4) culture of rootage
By the root induction in root media of transferring of the indefinite bud in strong seedling culture base, control cultivation temperature is 24-26 ℃, and illumination is 70-90 μ mol/ms, and after 10 days, seedling base section dissolves the former base of root of many whites, after 30 days, grows to 2-3cm;
(5) acclimatization and transplants
40 days root systems of culture of rootage grow to 1-2cm, and the aseptic seedling of selecting well developed root system robust growth, indoor uncork hardening 7 days, is taken out afterwash root agar, tame after 40 days, to transplant outdoorly in greenhouse, give rich water quality management.
2. a kind of method of cultivating the blue beans in Australia of organizing according to claim 1, it is characterized in that, the nutrient component of described bud inducing culture is MS+6-BA1.0mg/L+NAA0.1mg/L, MS+6-BA3.0mg/L+NAA0.3mg/L or MS+6-BA5.0mg/L+NAA0.5mg/L.
3. a kind of method of cultivating the blue beans in Australia of organizing according to claim 2, is characterized in that, the nutrient component of described bud inducing culture is MS+6-BA3.0mg/L+NAA0.3mg/L.
4. a kind of method of cultivating the blue beans in Australia of organizing according to claim 1, it is characterized in that, the nutrient component of described proliferated culture medium is MS+6-BA1mg/L-+NAA0.1mg/L, MS+6-BA2mg/L+NAA0.2mg/L or MS+6-BA3mg/L+NAA0.3mg/L.
5. a kind of method of cultivating the blue beans in Australia of organizing according to claim 4, is characterized in that, the nutrient component of described proliferated culture medium is MS+6BA2mg/L+NAA0.2mg/L.
6. a kind of method of cultivating the blue beans in Australia of organizing according to claim 1, it is characterized in that, the nutrient component of described strong seedling culture base is MS+6-BA0.5mg/L+NAA0.1mg/L, MS+6-BA1.0mg/L+NAA0.1mg/L or MS+6-BA2mg/L+NAA0.2mg/L.
7. a kind of method of cultivating the blue beans in Australia of organizing according to claim 6, is characterized in that, the nutrient component of described strong seedling culture base is MS+6-BA0.5mg/L+NAA0.1mg/L.
8. a kind of method of cultivating the blue beans in Australia of organizing according to claim 1, is characterized in that, the nutrient component of described root media is MS+NAA0.05mg/L, MS+NAA0.1mg/L or MS+NAA0.2mg/L.
9. a kind of method of cultivating the blue beans in Australia of organizing according to claim 8, is characterized in that, the nutrient component of described root media is MS+NAA0.1mg/L.
10. a kind of method of cultivating the blue beans in Australia of organizing according to claim 1, it is characterized in that, described bud inducing culture, proliferated culture medium, strong seedling culture base, root media also comprises the basis of sucrose 30g/L, agar 6g/L composition, and the pH value of above-mentioned medium is controlled at 5.8.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101869065A (en) * | 2009-04-24 | 2010-10-27 | 上海上房园林植物研究所 | Tissue culture method for purple prairie clover |
| CN101869072A (en) * | 2009-04-24 | 2010-10-27 | 上海上房园林植物研究所 | Tissue culture method of golden-heart Iris pseudacorus |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101869065A (en) * | 2009-04-24 | 2010-10-27 | 上海上房园林植物研究所 | Tissue culture method for purple prairie clover |
| CN101869072A (en) * | 2009-04-24 | 2010-10-27 | 上海上房园林植物研究所 | Tissue culture method of golden-heart Iris pseudacorus |
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Inventor after: Chen Jianhua Inventor after: Huang Jianrong Inventor after: Cui Xinhong Inventor after: Chen Di Inventor after: Zhang Chendi Inventor after: Shen Qin Inventor before: Chen Jianhua Inventor before: Huang Jianrong Inventor before: Shen Qin |
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Application publication date: 20140430 |