CN101773039B - Transgenic cotton regenerated plant transplantation method - Google Patents
Transgenic cotton regenerated plant transplantation method Download PDFInfo
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- CN101773039B CN101773039B CN 201010103941 CN201010103941A CN101773039B CN 101773039 B CN101773039 B CN 101773039B CN 201010103941 CN201010103941 CN 201010103941 CN 201010103941 A CN201010103941 A CN 201010103941A CN 101773039 B CN101773039 B CN 101773039B
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Abstract
The invention discloses a transgenic cotton regenerated plant transplantation method, which solves the problems that the survival rate of the transgenic cotton regenerated plants transplanted by the traditional method is low and the steps are trivial. The method comprises the following steps that: 1) the cover of a bottle containing transgenic cotton regenerated plant seedlings with roots is opened for indoor seedling hardening in the bottle for 2-3 days; 2) culture medium on the roots of the transgenic cotton regenerated plants is washed, the transgenic cotton regenerated plants are planted in a container containing sterilized roseite and are watered thoroughly at one step, and the transgenic cotton regenerated plants are cultured in an incubator with humidity of 88-90 percent and temperature of 20-22 DEG C for 10-15 days; and 3) the transgenic cotton regenerated plants are transplanted into greenhouse with normal conditions and grow to be mature to open bolls. The invention adopts the low-temperature and high-humidity environments for culture, so the survival rate reaches 90-95 percent.
Description
Technical field
The present invention relates to the transgene cotton technical field, be specially a kind of transgenic cotton regenerated plant transplantation method.
Background technology
Cotton transforms the transgenosis regeneration strain that foreign gene obtains through agrobacterium-mediated transformation, through Molecular Detection, the proof foreign gene has changed in the acceptor cotton, and the regenerated plant transplantation method of conversion adopts grafting or conventional method for transplanting normally to survive, and is grown to needed transgene cotton then.
The conventional method for transplanting of transgenic cotton regenerated plant is: the seedling of earlier the triangular flask transgenic cotton regenerated plant being taken root is opened bottle cap and was placed between cultivation hardening 1-2 days; Again with transgene cotton flush away root medium, plant in the container that the sterilization vermiculite is housed, disposable water permeable; Put into humidity 70%, 27 ℃~29 ℃ incubators of temperature then, the strain of earlier whole strain lid was cultivated 8~10 days, removed the container that covers cotton plant then, cultivated about 5~7 days at incubator again; Move at last in the greenhouse of normal condition and survive the cotton boll blowing maturation.
The purpose of cotton transgenic is that the external source genes of interest is changed in the conventional cotton, in the hope of obtaining transgene cotton, has been the primary study project of country " 863 " since " 95 " always.The transgenic cotton plant that obtains costs an arm and a leg, but, because existing transgenic cotton regenerated plant transplantation method, need be at high temperature, cultivate in the incubator of low humidity, cause butt rot easily, cause the low problem of motility rate, general survival rate has only 20%~60%, and in incubation, need earlier whole strain to add a cover, and then move lid and cultivate, operating procedure is more loaded down with trivial details, so do not reach ideal results all the time.
Seek a kind of simple to operately, the transgenic cotton regenerated plant transplantation method that survival rate is high becomes the focus of present research.
Summary of the invention
The present invention adopts high temperature, low humidity in order to solve existing transgenic cotton regenerated plant transplantation method, and causes survival rate low, the problem of complex steps, and a kind of transgenic cotton regenerated plant transplantation method is provided.
Transgenic cotton regenerated plant transplantation method of the present invention may further comprise the steps:
1) transgene cotton triangular flask regeneration strain is taken root seedling bottle cap is opened and was carried out indoor in-bottle seeding 2 days~3 days;
2) with transgenic cotton regenerated plant root medium flush away, plant and be equipped with in the container of sterilization vermiculite, disposablely water permeablely, be 88%~90% in humidity, temperature is to cultivate 10~15 days in 20 ℃~22 ℃ the incubator;
3) survive the cotton boll blowing maturation in the greenhouse of immigration normal condition.
Conventional trasplanting method is to carry out under the environment of high temperature low humidity, the strain of earlier whole strain lid was cultivated 8~10 days, remove the container that covers cotton plant then, cultivate again about 5~7 days, the regeneration strain root rots easily in the environment of high temperature low humidity like this, cause whole strain death, and owing to obtain costing an arm and a leg of transgenosis regeneration strain, thereby make cost improve.The present invention is through a large amount of experiments, adopts finally that humidity is 88%~90%, temperature is 20 ℃~22 ℃ low temperature, high humidity culture environment, makes survival rate reach 90%~95%, and save the incubation step with the container cover strain in incubation.
With per 10 strains of transgenic cotton regenerated plant is one group, totally 12 groups, under different temperature and humidity environment, transplant cultivation, and observe survival rate.Concrete experimental data is as described in the following table table 1:
Table 1
As known from Table 1, under the environment of low temperature and high relative humidity, the survival rate of transgenic cotton regenerated plant reaches 90%.On March 15th, 2009, in the transplantation experiments process of transgenic cotton regenerated plant, select for use the environment of low temperature and high relative humidity to carry out, be one group with per 40 strains of regeneration strain, totally 9 groups, observe survival rate, concrete experimental data is as described in the following table table 2:
Table 2
Transgenic cotton regenerated plant transplantation method of the present invention, adopt the culture environment of low temperature and high relative humidity, improved survival rate greatly, and the whole strain of having simplified in the existing transgenic cotton regenerated plant transplantation method is added a cover this step with glass or plastic foil, is a kind of comparatively desirable transgenic cotton regenerated plant transplantation method.
Implement embodiment
Embodiment 1
The transgenic cotton regenerated plant seedling bottle cap of taking root is opened and carried out indoor in-bottle seeding 2 days; Again with the regeneration strain root medium flush away after the hardening, plant in the container that the sterilization vermiculite is housed, disposablely water permeablely, be 90% in humidity, temperature is to cultivate 15 days in 22 ℃ the incubator; Move into to cultivate in the greenhouse and survive the cotton boll blowing maturation.
Embodiment 2
The transgenic cotton regenerated plant seedling bottle cap of taking root is opened and carried out indoor in-bottle seeding 3 days; Again with the regeneration strain root medium flush away after the hardening, plant in the container that the sterilization vermiculite is housed, disposablely water permeablely, be 88% in humidity, temperature is to cultivate 10 days in 20 ℃ the incubator; Move into to cultivate in the greenhouse and survive the cotton boll blowing maturation.
Embodiment 3
The transgenic cotton regenerated plant seedling bottle cap of taking root is opened and carried out indoor in-bottle seeding 3 days; Again with the regeneration strain root medium flush away after the hardening, plant in the container that the sterilization vermiculite is housed, disposablely water permeablely, be 89% in humidity, temperature is to cultivate 13 days in 21 ℃ the incubator; Move into to cultivate in the greenhouse and survive the cotton boll blowing maturation.
Claims (1)
1. transgenic cotton regenerated plant transplantation method is characterized in that may further comprise the steps:
1) transgenic cotton regenerated plant is taken root seedling bottle cap is opened and was carried out indoor in-bottle seeding 2 days~3 days;
2) with transgenic cotton regenerated plant root medium flush away, plant and be equipped with in the container of sterilization vermiculite, disposablely water permeablely, be 88%~90% in humidity, temperature is to cultivate 10 days~15 days in 20 ℃~22 ℃ the incubator;
3) survive the cotton boll blowing maturation in the greenhouse of immigration normal condition.
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CN 201010103941 CN101773039B (en) | 2010-01-28 | 2010-01-28 | Transgenic cotton regenerated plant transplantation method |
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CN 201010103941 CN101773039B (en) | 2010-01-28 | 2010-01-28 | Transgenic cotton regenerated plant transplantation method |
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CN101773039A CN101773039A (en) | 2010-07-14 |
CN101773039B true CN101773039B (en) | 2011-05-25 |
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Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
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CN102972297B (en) * | 2012-12-05 | 2014-05-07 | 中国农业科学院生物技术研究所 | Method for cultivating regeneration plants of cotton |
CN104956930A (en) * | 2015-07-14 | 2015-10-07 | 山西省农业科学院棉花研究所 | Method for suspended transplantation of regenerated transgenic cotton plant |
CN106171430A (en) * | 2016-07-15 | 2016-12-07 | 南京农业大学 | A kind of transgenic cotton regenerated plant method for transplanting of high-survival rate |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1685795A (en) * | 2005-04-30 | 2005-10-26 | 山西省农业科学院棉花研究所 | Substrate and method of cotton regeneration test tube seedling direct transplantation |
CN101024820A (en) * | 2007-02-07 | 2007-08-29 | 三峡大学 | Method and use of setting up efficient peanut trans genosis receptor system |
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2010
- 2010-01-28 CN CN 201010103941 patent/CN101773039B/en not_active Expired - Fee Related
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1685795A (en) * | 2005-04-30 | 2005-10-26 | 山西省农业科学院棉花研究所 | Substrate and method of cotton regeneration test tube seedling direct transplantation |
CN101024820A (en) * | 2007-02-07 | 2007-08-29 | 三峡大学 | Method and use of setting up efficient peanut trans genosis receptor system |
Non-Patent Citations (2)
Title |
---|
郭余龙等.陆地棉体胚发生及再生植株移栽.《西南农业大学学报》.1999,第21卷(第6期),514-517. * |
陆瑞菊等.玉米小孢子培育再生植株的移栽及倍性评估.《中国农学通报》.2005,第21卷(第3期),84-85,221. * |
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