CN101904301B - Method for inducing embryoids of sugarcane by using lingfasu - Google Patents
Method for inducing embryoids of sugarcane by using lingfasu Download PDFInfo
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- CN101904301B CN101904301B CN2010102161546A CN201010216154A CN101904301B CN 101904301 B CN101904301 B CN 101904301B CN 2010102161546 A CN2010102161546 A CN 2010102161546A CN 201010216154 A CN201010216154 A CN 201010216154A CN 101904301 B CN101904301 B CN 101904301B
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Abstract
The invention discloses a method for inducing embryoids of sugarcane by using lingfasu, which comprises the following steps of: preparing an improved solid medium by adding 0.6 to 1.0 mg.L<-1> lingfasu into MS serving as a minimal medium; culturing sugarcane spire slices serving as explants in a dark box for 8 to 12 days; and transferring the sugarcane spire slices into a condition where light and dark are alternate every day, and continuing to culture the sugarcane spire slices, so that the cultured materials produce the embryoids of various shapes one by one, wherein the production rate after 20 days is more than or equal to 85 percent.
Description
Technical field
The present invention relates to a kind of method of inducing the sugarcane embryoid with spirit hair element.
Technical background
" embryoid " also claims " somatic embryo ", be called for short " body embryo ", it is the important materials that obtains regeneration plant and manufacture of intraocular seed, also be from anatomy and inherent Mechanism of Physiological and Biochemical aspect research cell differentiation, therefore organ builds up and ontogenetic important materials, the research of embryoid has been become a popular domain of plant biological educational circles in modern age.Research to the sugarcane embryoid both at home and abroad also has many reports.But the minimal medium that uses is confined to MS and N substantially at present
6Two kinds, the growth regulator that is used for the inducing embryoid body generation all is a synthetic bar none then, has certain toxicity and residual benzene analog derivative 2,4-D (2,4 dichlorophenoxyacetic acid).And single with 2, during 4-D, the embryoid incidence only has tens percent.The people is also arranged with 2, the composite again BA (N of 4-D
6-BA, N
6-benzyladenine) or lactoalbumin hydrolysate, in the hope of improving the embryoid incidence, but also can only reach 30 to 50 percent.Spirit hair element (Lingfasu, LFS, code name PGR-08) is as a kind of novel, biogenic plant growth regulator, nonhazardous, noresidue.In the research that is acknowledged as the traditional Chinese medicine material pseudo-ginseng embryoid that is difficult to organize training, successfully induce Different Organs such as stem segment, petiole and blade all to obtain a large amount of embryoids (Xu Hongyuan etc., traditional Chinese medicine, 2004,27 (10): 711-712 with LFS; CHINA JOURNAL OF CHINESE MATERIA MEDICA, 2007,32 (6): 481-483).But, do not retrieve the report that LFS induces the sugarcane embryoid as yet.
Summary of the invention
The technical problem to be solved in the present invention provides a kind ofly induces the method for sugarcane embryoid with spirit hair element, and it can induce the sugarcane embryoid.
The present invention solves the problems of the technologies described above by the following technical programs: a kind ofly induce the method for sugarcane embryoid with spirit hair element, it is characterized in that this method comprises the steps:
1, with sugarcane strain taper with 70% alcohol disinfecting appearance, will be cut to the 0.3-0.5cm square from growing point 0.01-4.0cm young leaflet tablet,
2, above-mentioned square is connected on the dark cultivation of carrying out 24 hours unglazed photographs of whole day on MS+LFS0.6～1.0mg/L solid culture medium, obtains callus after 8-12 days;
3, above-mentioned callus is carried out light application time 10～12 hours every day following every day not changing culture medium condition, the light of light intensity 2000～2500LX, secretly alternately cultivate, the embryoid that obtains coming in every shape after 7-10 days,
4, above-mentioned embryoid is forwarded on MS+LFS0.6～1.0mg/L fresh culture cultivate, changed a fresh culture in every 20-30 days, obtain yellow embryoid.
Beneficial effect of the present invention:
Use the present invention, can induce that generation is loosely organized in a large number, graininess, color be the high-quality sugarcane embryoid of ecru.
Embodiment
Below in conjunction with embodiment technical scheme of the present invention is described in further detail.
Embodiment 1 is for adopting MS+2,4-D 2.0mgL in the table 1
-1Induce the Comparative Examples of sugarcane embryoid.
Embodiment 2 a kind ofly induces the method for sugarcane embryoid with spirit hair element for of the present invention, comprises the steps:
1. select the stem footpath thicker and do not have the sugarcane strain taper of obvious sick worm damage symptom from sugarcane field,, on superclean bench, remove taper Lao Ye with 70% alcohol disinfecting appearance, will be from growing point 0.01-4.0cm young leaflet tablet, be cut into size is the 0.3-0.5cm square;
2. above-mentioned square is connected on MS+LFS0.6～1.0mg/L solid culture medium and secretly cultivates, 24 hours unglazed photographs of whole day obtained callus after 8-12 days;
3. above-mentioned 2 material is not more being renewed under the culture medium condition, carrying out light every day, secretly alternately cultivate, every day, light application time was 10～12 hours, light intensity 2000～2500LX, and the embryoid that comes in every shape after 7-10 days forms, incidence 〉=85%.Spire number of slices * 100% of the spire number of slices/inoculation of incidence=generation embryoid.
4. above-mentioned 3 material is forwarded on MS+LFS0.6～1.0mg/L fresh culture and cultivate, changed a fresh culture in every 20-30 days, that large tracts of land forms is loosely organized, graininess, color rice are yellow embryoid.
The experimental result of two kinds of different medium of table 1
Annotate: sugar cane breed is No. 22, new platform sugar.
From table 1 experimental data as can be seen, under identical inoculation number of slices situation, the embryoid number of slices that goes out that adopts the present invention to obtain is Duoed 47 than the embryoid number of slices that goes out that traditional medium obtains, the embryoid incidence is up to 93.3%.
Claims (1)
1. induce the method for sugarcane embryoid with spirit hair element for one kind, it is characterized in that this method comprises the steps:
1.1 sugarcane strain taper with 70% alcohol disinfecting appearance, will be cut to the 0.3-0.5cm square from growing point 0.01-4.0cm young leaflet tablet,
1.2 above-mentioned square is connected on the dark cultivation of carrying out 24 hours unglazed photographs of whole day on MS+LFS0.6～1.0mg/L solid culture medium, obtains callus after 8-12 days;
1.3 above-mentioned callus is carried out light application time 10～12 hours every day following every day not changing culture medium condition, the light of light intensity 2000～2500LX, secretly alternately cultivate, the embryoid that obtains coming in every shape after 7-10 days,
1.4 above-mentioned embryoid forwarded on MS+LFS0.6～1.0mg/L fresh culture cultivates, changed a fresh culture in every 20-30 days, obtain yellow embryoid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010102161546A CN101904301B (en) | 2010-07-02 | 2010-07-02 | Method for inducing embryoids of sugarcane by using lingfasu |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010102161546A CN101904301B (en) | 2010-07-02 | 2010-07-02 | Method for inducing embryoids of sugarcane by using lingfasu |
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| Publication Number | Publication Date |
|---|---|
| CN101904301A CN101904301A (en) | 2010-12-08 |
| CN101904301B true CN101904301B (en) | 2011-07-20 |
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| CN2010102161546A Expired - Fee Related CN101904301B (en) | 2010-07-02 | 2010-07-02 | Method for inducing embryoids of sugarcane by using lingfasu |
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Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104351058A (en) * | 2014-11-24 | 2015-02-18 | 广西大学 | Method for inducing sugarcane embryoids by spermine |
| CN104396753B (en) * | 2014-11-24 | 2016-06-29 | 广西大学 | A kind of method of spermine breeding sugar-cane tissue culture seedlings |
| CN106472311B (en) * | 2016-10-14 | 2018-10-19 | 广西大学 | With the method for spirit hair element evoking tobacco callus |
| CN106613945B (en) * | 2016-10-14 | 2018-11-02 | 广西大学 | With the method for spirit hair element evoking tobacco embryoid |
| CN116491421A (en) * | 2023-05-26 | 2023-07-28 | 广西大学 | Gene gun-mediated sugarcane embryo genetic transformation method |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101731145A (en) * | 2009-11-30 | 2010-06-16 | 广西大学 | Method for enhancing root inducing of Angustmycin in production of Grosvener Siraitia tissue culture plantlet |
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2010
- 2010-07-02 CN CN2010102161546A patent/CN101904301B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101731145A (en) * | 2009-11-30 | 2010-06-16 | 广西大学 | Method for enhancing root inducing of Angustmycin in production of Grosvener Siraitia tissue culture plantlet |
Non-Patent Citations (2)
| Title |
|---|
| 宋磊，等.用灵发素(LFS)建立翅子罗汉果组织培养技术.《种子》.2008,第27卷(第12期), * |
| 许鸿源,等.灵发素对三七胚状体发生及其成苗的影响.《中药材》.2004,第27卷(第10期), * |
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