CN103651114A - Tissue culture and rapid propagation method for cardamine hupingshanesis - Google Patents
Tissue culture and rapid propagation method for cardamine hupingshanesis Download PDFInfo
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- CN103651114A CN103651114A CN201210539976.7A CN201210539976A CN103651114A CN 103651114 A CN103651114 A CN 103651114A CN 201210539976 A CN201210539976 A CN 201210539976A CN 103651114 A CN103651114 A CN 103651114A
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- hairy bittercress
- tall bottle
- spout hairy
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Abstract
The invention relates to the field of cardamine hupingshanesis culture and in particular relates to a tissue culture and rapid propagation method for cardamine hupingshanesis. The tissue culture and rapid propagation method comprises the following steps of material taking and treating, callus induction, induction of adventitious buds, rooting culture and transplanting. The culture method disclosed by the invention is simple and convenient; conventional culture mediums are used; the cost is low, and the survival rate is high; and therefore, the tissue culture and rapid propagation method is a rapid culture method for the cardamine hupingshanesis, and is deserved to be popularized.
Description
Technical field
The present invention relates to tall bottle with spout Hairy Bittercress and cultivate field, the tissue that relates in particular to a kind of tall bottle with spout Hairy Bittercress is cultivated and method for quickly breeding.
Background technology
Tall bottle with spout Hairy Bittercress, is China's distinctive Cruciferae cardamine novel species, and the local common people do wild vegetable often searches for food, and individual plant weighs 400 grams, has superpower selenium rich ability, but it is lower to be limited to its output, is badly in need of a kind of easy culture technique.
Summary of the invention
The tissue that the invention provides a kind of tall bottle with spout Hairy Bittercress is cultivated and method for quickly breeding, and its method is easy, survival rate is high.
The embodiment of the present invention provides a kind of tissue of tall bottle with spout Hairy Bittercress to cultivate and method for quickly breeding, and it comprises the steps:
Draw materials and process: get stem, spire or the petiole of tall bottle with spout Hairy Bittercress, clear water is disinfected after rinsing;
Callus of induce: tall bottle with spout Hairy Bittercress is seeded on inductive differentiation medium, under 20 ~ 25 degrees Celsius of temperature, 20 ~ 27 moles of/square of meter per seconds of intensity of illumination, the illumination environment of 10 ~ 17 hours/day, cultivate 13 ~ 15 days, tall bottle with spout Hairy Bittercress explant has green callus to form as seen;
The induction of indefinite bud: the continuous growth of callus of formation 25 ~ 35 days, to callus, form green bud point, continue to cultivate 12 ~ 17 days, a large amount of Multiple Buds generate;
Culture of rootage: when Multiple Buds grows to 2-3 centimetre, be inoculated on root media after Multiple Buds is cut, 25 ~ 30 days root growths, become regrowth;
Transplant: select the regrowth of well developed root system, 4 ~ 8 days afterwash medium of hardening, are transplanted in vermiculite, peat soil and perlitic mixed-matrix, and covered with plastic film moves under natural conditions and grows after growing under low light environment 8 ~ 12 days.
To being further improved to of technique scheme: described in draw materials and treatment step in tall bottle with spout Hairy Bittercress material is cut into 0.5 centimetre of blockage of 0.5 cm x after disinfecting.
Preferably, the described step of disinfecting is specially: 2 ~ 6 degrees Celsius of low temperature treatment after 6 ~ 15 hours, with 50% ~ 90% alcohol-pickled 2 ~ 10 seconds, with aseptic water washing for several times, mercuric chloride solution soaking disinfection number minute with 0.05% ~ 0.2%, then with aseptic water washing for several times.
Wherein, described inductive differentiation medium is: MS+6-BA 2.0 mg/litre+NAA 0.1, and add 30 grams per liter sucrose and 7 grams per liter agar, pH was 5.8,110 ~ 130 degrees Celsius of lower moist heat sterilizations 10 ~ 30 minutes.
Wherein, described root media was: 1/2MS+NAA 0.3, and adds 30 grams per liter sucrose and 7 grams per liter agar, and pH is 5.8,110 ~ 130 degrees Celsius of lower moist heat sterilizations 10 ~ 30 minutes.
In described transplant step, vermiculite, peat soil and perlitic ratio are 1:1:1.
Describedly draw materials and process, clear water washing time is 1 ~ 2 hour.
Preferably, in the induction of described callus of induce, indefinite bud and culture of rootage step, culture environment is 20 ~ 25 degrees Celsius of temperature, 20 ~ 27 moles of/square of meter per seconds of intensity of illumination, illumination 10 ~ 17 hours/day.
The tissue of the present embodiment tall bottle with spout Hairy Bittercress is cultivated and method for quickly breeding, and beneficial effect is:
By on medium after the induction of callus of induce, indefinite bud and culture of rootage transplanting breeding, cultural method is easy, all uses conventional medium, cost is low, survival rate is high, is a kind of fast culture process of the tall bottle with spout Hairy Bittercress being worthy to be popularized.
Embodiment
For making the object, technical solutions and advantages of the present invention clearer, below the present invention is described in further detail.
Embodiment 1:
The present embodiment provides a kind of tissue of tall bottle with spout Hairy Bittercress to cultivate and method for quickly breeding, and it comprises the steps:
Make medium:
Inductive differentiation medium: with MS+6-BA 2.0 mg/litre+NAA 0.1, and add 30 grams per liter sucrose and 7 grams per liter agar, regulate pH to 5.8,121 degrees Celsius of lower moist heat sterilizations 20 minutes.
Root media: with 1/2MS+NAA 0.3, and add 30 grams per liter sucrose and 7 grams per liter agar, regulate pH to 5.8,121 degrees Celsius of lower moist heat sterilizations 20 minutes.
Draw materials and process: stem, spire or the petiole of getting tall bottle with spout Hairy Bittercress, clear water rinses 2 hours, 4 degrees Celsius of low temperature treatment after 12 hours, with 75% alcohol-pickled 7 seconds, with aseptic water washing 3 times, mercuric chloride solution soaking disinfection with 0.1% 10 minutes, then use aseptic water washing 3 times, tall bottle with spout Hairy Bittercress material is cut into the blockage of 0.5 centimetre of 0.5 cm x.
Callus of induce: tall bottle with spout Hairy Bittercress is seeded on inductive differentiation medium, under 23 degrees Celsius of temperature, 24 moles of/square of meter per seconds of intensity of illumination, the illumination environment of 14 hours/day, cultivate, after 4 days there is callus in medium, present light green, after 14 days, 80% above petiole explant has green callus to form as seen.
The induction of indefinite bud: the continued growth 30 days under identical medium and culture environment of the callus of formation, to callus, form green bud point, continue to cultivate 15 days, a large amount of Multiple Buds generate.
Culture of rootage: when Multiple Buds grows to 2-3 centimetre, be inoculated on root media after Multiple Buds is cut, cultivate root growth after 30 days under identical culture environment, rooting rate is 100%, becomes regrowth.
Transplant: select the regrowth of well developed root system, 7 days afterwash medium of hardening, are transplanted in the mixed-matrix of vermiculite, peat soil and perlite ratio 1:1:1, covered with plastic film, moves to growth under natural conditions under low light environment after growing 10 days.
After testing, the tall bottle with spout Hairy Bittercress regrowth survival rate of above-mentioned transplanting reaches more than 90%.
Embodiment 2:
The present embodiment provides a kind of tissue of tall bottle with spout Hairy Bittercress to cultivate and method for quickly breeding, and it comprises the steps:
Make medium:
Inductive differentiation medium: with MS+6-BA 2.0 mg/litre+NAA 0.1, and add 30 grams per liter sucrose and 7 grams per liter agar, regulate pH to 5.8,110 degrees Celsius of lower moist heat sterilizations 10 minutes.
Root media: with 1/2MS+NAA 0.3, and add 30 grams per liter sucrose and 7 grams per liter agar, regulate pH to 5.8,110 degrees Celsius of lower moist heat sterilizations 10 minutes.
Draw materials and process: stem, spire or the petiole of getting tall bottle with spout Hairy Bittercress, clear water rinses 1 hour, 2 degrees Celsius of low temperature treatment after 6 hours, with 50% alcohol-pickled 2 seconds, with aseptic water washing 3 times, mercuric chloride solution soaking disinfection with 0.05% 10 minutes, then use aseptic water washing 3 times, tall bottle with spout Hairy Bittercress material is cut into the blockage of 0.5 centimetre of 0.5 cm x.
Callus of induce: tall bottle with spout Hairy Bittercress is seeded on inductive differentiation medium, under 20 degrees Celsius of temperature, 20 moles of/square of meter per seconds of intensity of illumination, the illumination environment of 10 hours/day, cultivate, after 3 days there is callus in medium, present light green, after 13 days, 80% above petiole explant has green callus to form as seen.
The induction of indefinite bud: the continued growth 25 days under identical medium and culture environment of the callus of formation, to callus, form green bud point, continue to cultivate 12 days, a large amount of Multiple Buds generate.
Culture of rootage: when Multiple Buds grows to 2-3 centimetre, be inoculated on root media after Multiple Buds is cut, cultivate root growth after 25 days under identical culture environment, rooting rate is 100%, becomes regrowth.
Transplant: select the regrowth of well developed root system, 4 days afterwash medium of hardening, are transplanted in the mixed-matrix of vermiculite, peat soil and perlite ratio 1:1:1, covered with plastic film, moves to growth under natural conditions under low light environment after growing 8 days.
Embodiment 3:
The present embodiment provides a kind of tissue of tall bottle with spout Hairy Bittercress to cultivate and method for quickly breeding, and it comprises the steps:
Make medium:
Inductive differentiation medium: with MS+6-BA 2.0 mg/litre+NAA 0.1, and add 30 grams per liter sucrose and 7 grams per liter agar, regulate pH to 5.8,130 degrees Celsius of lower moist heat sterilizations 30 minutes.
Root media: with 1/2MS+NAA 0.3, and add 30 grams per liter sucrose and 7 grams per liter agar, regulate pH to 5.8,130 degrees Celsius of lower moist heat sterilizations 30 minutes.
Draw materials and process: stem, spire or the petiole of getting tall bottle with spout Hairy Bittercress, clear water rinses 2 hours, 6 degrees Celsius of low temperature treatment after 15 hours, with 90% alcohol-pickled 10 seconds, with aseptic water washing 3 times, mercuric chloride solution soaking disinfection with 0.2% 10 minutes, then use aseptic water washing 3 times, tall bottle with spout Hairy Bittercress material is cut into the blockage of 0.5 centimetre of 0.5 cm x.
Callus of induce: tall bottle with spout Hairy Bittercress is seeded on inductive differentiation medium, under 25 degrees Celsius of temperature, 27 moles of/square of meter per seconds of intensity of illumination, the illumination environment of 17 hours/day, cultivate, after 4 days there is callus in medium, present light green, after 15 days, 80% above petiole explant has green callus to form as seen.
The induction of indefinite bud: the continued growth 35 days under identical medium and culture environment of the callus of formation, to callus, form green bud point, continue to cultivate 17 days, a large amount of Multiple Buds generate.
Culture of rootage: when Multiple Buds grows to 2-3 centimetre, be inoculated on root media after Multiple Buds is cut, cultivate root growth after 30 days under identical culture environment, rooting rate is 100%, becomes regrowth.
Transplant: select the regrowth of well developed root system, 8 days afterwash medium of hardening, are transplanted in the mixed-matrix of vermiculite, peat soil and perlite ratio 1:1:1, covered with plastic film, moves to growth under natural conditions under low light environment after growing 12 days.
After testing, the tall bottle with spout Hairy Bittercress regrowth survival rate of above-mentioned transplanting reaches more than 90%.
Be more than the preferred embodiment of the present invention, it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention, can also make some improvements and modifications, these improvements and modifications are also considered as protection scope of the present invention." substitute and also can.
Claims (8)
1. the tissue of tall bottle with spout Hairy Bittercress is cultivated and a method for quickly breeding, it is characterized in that, comprises the following steps of carrying out successively:
Draw materials and process: get stem, spire or the petiole of tall bottle with spout Hairy Bittercress, clear water is disinfected after rinsing;
Callus of induce: tall bottle with spout Hairy Bittercress is seeded on inductive differentiation medium and is cultivated 13 ~ 15 days, and tall bottle with spout Hairy Bittercress explant has green callus to form as seen;
The induction of indefinite bud: the continuous growth of callus of formation 25 ~ 35 days, to callus, form green bud point, continue to cultivate 12 ~ 17 days, a large amount of Multiple Buds generate;
Culture of rootage: when Multiple Buds grows to 2-3 centimetre, be inoculated on root media after Multiple Buds is cut, 25 ~ 30 days root growths, become regrowth;
Transplant: select the regrowth of well developed root system, 4 ~ 8 days afterwash medium of hardening, are transplanted in vermiculite, peat soil and perlitic mixed-matrix, and covered with plastic film moves under natural conditions and grows after growing under low light environment 8 ~ 12 days.
2. the tissue of tall bottle with spout Hairy Bittercress is cultivated and method for quickly breeding as claimed in claim 1, it is characterized in that,
Described draw materials and treatment step in tall bottle with spout Hairy Bittercress material is cut into 0.5 centimetre of blockage of 0.5 cm x after disinfecting.
3. the tissue of tall bottle with spout Hairy Bittercress is cultivated and method for quickly breeding as claimed in claim 1, it is characterized in that:
The described step of disinfecting is specially: 2 ~ 6 degrees Celsius of low temperature treatment after 6 ~ 15 hours, and with 50% ~ 90% alcohol-pickled 2 ~ 10 seconds, with aseptic water washing for several times, the mercuric chloride solution soaking disinfection number minute with 0.05% ~ 0.2%, then with aseptic water washing several.
4. the tissue of tall bottle with spout Hairy Bittercress is cultivated and method for quickly breeding as claimed in claim 1, it is characterized in that:
Described inductive differentiation medium is: MS+6-BA 2.0 mg/litre+NAA 0.1, and add 30 grams per liter sucrose and 7 grams per liter agar, pH was 5.8,110 ~ 130 degrees Celsius of lower moist heat sterilizations 10 ~ 30 minutes.
5. the tissue of tall bottle with spout Hairy Bittercress is cultivated and method for quickly breeding as claimed in claim 1, it is characterized in that:
Described root media was: 1/2MS+NAA 0.3, and adds 30 grams per liter sucrose and 7 grams per liter agar, and pH is 5.8,110 ~ 130 degrees Celsius of lower moist heat sterilizations 10 ~ 30 minutes.
6. the tissue of tall bottle with spout Hairy Bittercress is cultivated and method for quickly breeding as claimed in claim 1, it is characterized in that:
In described transplant step, vermiculite, peat soil and perlitic ratio are 1:1:1.
7. the tissue of tall bottle with spout Hairy Bittercress is cultivated and method for quickly breeding as claimed in claim 1, it is characterized in that:
Describedly draw materials and process, clear water washing time is 1 ~ 2 hour.
8. the tissue of tall bottle with spout Hairy Bittercress is cultivated and method for quickly breeding as claimed in claim 1, it is characterized in that:
In the induction of described callus of induce, indefinite bud and culture of rootage step, culture environment is 20 ~ 25 degrees Celsius of temperature, 20 ~ 27 moles of/square of meter per seconds of intensity of illumination, illumination 10 ~ 17 hours/day.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210539976.7A CN103651114A (en) | 2012-12-14 | 2012-12-14 | Tissue culture and rapid propagation method for cardamine hupingshanesis |
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| CN201210539976.7A CN103651114A (en) | 2012-12-14 | 2012-12-14 | Tissue culture and rapid propagation method for cardamine hupingshanesis |
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| CN103651114A true CN103651114A (en) | 2014-03-26 |
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| CN201210539976.7A Pending CN103651114A (en) | 2012-12-14 | 2012-12-14 | Tissue culture and rapid propagation method for cardamine hupingshanesis |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108450339A (en) * | 2018-05-14 | 2018-08-28 | 安徽过湾农业科技有限公司 | A kind of tissue culture and rapid proliferation method of Hairy Bittercress |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007028979A1 (en) * | 2005-09-06 | 2007-03-15 | Isis Innovation Limited | Plant transformation |
| CN102301871A (en) * | 2010-11-26 | 2012-01-04 | 恩施清江生物工程有限公司 | Selenium-enriching cultivation method for cardamine hupingshanensis |
| CN102511283A (en) * | 2011-12-19 | 2012-06-27 | 苏州硒谷科技有限公司 | Method of artificially rearing seedlings of Cardamine hupingshanensis |
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2012
- 2012-12-14 CN CN201210539976.7A patent/CN103651114A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007028979A1 (en) * | 2005-09-06 | 2007-03-15 | Isis Innovation Limited | Plant transformation |
| CN102301871A (en) * | 2010-11-26 | 2012-01-04 | 恩施清江生物工程有限公司 | Selenium-enriching cultivation method for cardamine hupingshanensis |
| CN102511283A (en) * | 2011-12-19 | 2012-06-27 | 苏州硒谷科技有限公司 | Method of artificially rearing seedlings of Cardamine hupingshanensis |
Non-Patent Citations (3)
| Title |
|---|
| TOMOO MAEDA ET.AL.,: "Development of Asexual Propagation System via in Vitro Culture in Cardamine yezoensis Maxim. and its Application to Hydroponic Cultivation", 《J. JAPAN. SOC. HORT. SCI.》, vol. 77, no. 3, 31 December 2008 (2008-12-31), pages 270 - 276 * |
| 张宇等: "大叶碎米荠的组织培养和快速繁殖", 《植物生理学通讯》, vol. 42, no. 2, 30 April 2006 (2006-04-30), pages 4 - 1 * |
| 徐全乐等: "碎米荠的组织培养与快速繁殖", 《植物生理学通讯》, vol. 45, no. 7, 31 July 2009 (2009-07-31), pages 2 - 4 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108450339A (en) * | 2018-05-14 | 2018-08-28 | 安徽过湾农业科技有限公司 | A kind of tissue culture and rapid proliferation method of Hairy Bittercress |
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Owner name: ENSHI SUNSE BIOLOGICAL ENGINEERING CO., LTD. Free format text: FORMER OWNER: HUBEI SUNSE BIOTECHNOLOGY CO., LTD. Effective date: 20150625 |
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Effective date of registration: 20150625 Address after: Enshi City, Hubei Province, 445000 National Road No. 1203 building B Champs ruidu Applicant after: ENSHI SHENGXI BIOLOGICAL ENGINEERING CO., LTD. Address before: 430000 A-7 building, small and Medium Enterprise Park Service Center, hi tech Zone, East Lake hi tech Zone, Wuhan, Hubei Applicant before: Hubei Shengxi Biotechnology Co., Ltd. |
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Application publication date: 20140326 |