CN111512808A - Method for rapidly inducing adventitious root regeneration of brachypodium distachyon tissue culture seedlings - Google Patents
Method for rapidly inducing adventitious root regeneration of brachypodium distachyon tissue culture seedlings Download PDFInfo
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- CN111512808A CN111512808A CN202010525793.4A CN202010525793A CN111512808A CN 111512808 A CN111512808 A CN 111512808A CN 202010525793 A CN202010525793 A CN 202010525793A CN 111512808 A CN111512808 A CN 111512808A
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- tissue culture
- brachypodium distachyon
- adventitious root
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G2/00—Vegetative propagation
- A01G2/10—Vegetative propagation by means of cuttings
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G9/00—Cultivation in receptacles, forcing-frames or greenhouses; Edging for beds, lawn or the like
- A01G9/02—Receptacles, e.g. flower-pots or boxes; Glasses for cultivating flowers
- A01G9/029—Receptacles for seedlings
- A01G9/0299—Handling or transporting of soil blocks or seedlings
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Botany (AREA)
- Developmental Biology & Embryology (AREA)
- Soil Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a method for rapidly inducing adventitious root regeneration of a brachypodium distachyon tissue culture seedling. The method mainly comprises the steps of selecting rootless brachypodium distachyon tissue culture seedlings, soaking the seedlings in rooting powder solution, planting the tissue culture seedlings in a plastic basin, putting the plastic basin into a phytotron for culture, and enabling the length of an adventitious root to reach 12-15cm after two weeks; the method for rapidly rooting brachypodium distachyon integrates the traditional rooting and seedling hardening procedures, improves the survival rate of regenerated seedlings, shortens the seedling culture period, can be directly used for the optimization research of the brachypodium distachyon regeneration and genetic transformation system, and lays a foundation for accelerating the transgenic breeding of plants.
Description
Technical Field
The invention relates to the technical field of plant tissue culture, in particular to a method for rapidly inducing adventitious root regeneration of a brachypodium distachyon tissue culture seedling.
Background
An effective way for researching plant gene function by plant tissue culture technology. At present, the technology is widely applied to plant regeneration work, but the regeneration of the adventitious root is still an important rate-limiting step, particularly how to rapidly induce the formation of the adventitious root.
Brachypodium distachyon, the C3 plant which is most widely distributed in the world, is defined as a novel model plant due to the advantages of small genome, short growth cycle, small quantity, suitability for genetic transformation and the like. Traditional two ears of short stalk grass regeneration is through controlling ambient temperature and illumination, but two ears of short stalk grass plantlets that cultivate like this develop the gas pocket and cuticle incomplete development, and the chlorophyll level is on the low side, and the root is poor with the water conduction ability of stem connecting portion, causes the plant to lose water easily and dies. Therefore, the method can quickly promote the regeneration of the adventitious roots of the brachypodium distachyon and improve the physiological state of the regenerated seedlings, and is an effective way for solving the problem.
The ex vitro rooting technology can integrate rooting and domestication, improve various defects of the traditional rooting program, improve the survival rate and shorten the seedling culture period.
Disclosure of Invention
The invention aims to provide a method for rapidly inducing the adventitious root regeneration of a brachypodium distachyon tissue culture seedling, which can improve the survival rate and shorten the seedling culture period and can be used for plant regeneration and genetic transformation research.
In order to achieve the purpose, the invention adopts the following technical scheme: a method for rapidly inducing the adventitious root regeneration of a brachypodium distachyon tissue culture seedling comprises the following steps:
(1) selecting rootless brachypodium distachyon tissue culture seedlings with the height of 2-3cm, taking the brachypodium distachyon tissue culture seedlings out of a culture bottle by using tweezers, and cleaning for 3-5 times;
(2) soaking the cleaned tissue culture seedlings in 1mg/L rooting powder solution for 3-5 minutes;
(3) transplanting the tissue culture seedlings soaked with the rooting powder into a plastic planting pot filled with mixed soil, covering the mouth of the plastic planting pot with a preservative film after planting is finished, and placing the plastic planting pot in a phytotron for culturing;
(4) the preservative film is lifted and sprayed for 2-3 times every day by using 1mg/L rooting powder solution, and the length of the adventitious root is measured after two weeks and is 12-15 cm.
In the step (1), the rootless brachypodium distachyon tissue culture seedling is an adventitious bud induced by the callus of the brachypodium distachyon.
In the step (2), the preparation method of the rooting powder solution is to dissolve 1mg of rooting powder in 1L of distilled water.
In the step (3), the mixed soil is prepared by mixing nutrient soil, perlite and vermiculite in a ratio of 2:
1: 2, adding clear water until the soil is wet.
In the step (3), the culture conditions of the artificial climate chamber are as follows: the temperature is 22 +/-2 ℃, the humidity is 60 +/-10%, and the illumination is 16 h/darkness is 8 h.
The invention has the advantages that: the method changes the growth state of the tissue culture seedlings by using the strategy of ex-vitro regeneration, rapidly induces the regeneration of the adventitious roots of the brachypodium distachyon tissue culture seedlings through the synergistic effect of the rooting powder, and improves the survival rate and the lateral root number of the tissue culture seedlings.
Drawings
FIG. 1 is a view showing an initial state of transplanting brachypodium distachyon into a plastic planting pot;
FIG. 2 is a state diagram of the two-ear brachypodium distachyon transplanted in the plastic planting pot for 2 weeks.
Detailed Description
The technical solution in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings.
Example 1
A method for rapidly inducing the adventitious root regeneration of a brachypodium distachyon tissue culture seedling comprises the following steps:
(1) selecting a rootless two-spike brachypodium distachyon tissue culture seedling which is an adventitious bud induced by the two-spike brachypodium distachyon callus, taking the rootless two-spike brachypodium distachyon tissue culture seedling out of a culture bottle by using a pair of tweezers, cleaning for 3-5 times, and removing redundant culture medium and callus;
(2) dissolving 1mg of rooting powder in 1L of distilled water to obtain a 1mg/L rooting powder solution, and soaking the cleaned tissue culture seedlings in the 1mg/L rooting powder solution for 3-5 minutes;
(3) mixing nutrient soil, perlite and vermiculite in a ratio of 2: 1: 2 after the proportion mixes, add clear water to soil is moist, obtains mixed soil, and the tissue culture seedling that will soak the rooting powder solution is transplanted to the plastics that is equipped with mixed soil and is planted in the basin, and after planting, plastic planting basin mouth (as shown in figure 1) is planted to the plastic film cover, places the phytotron and cultivates, and the cultivation condition in phytotron is: the temperature is 22 +/-2 ℃, the humidity is 60 +/-10%, and the illumination is 16 h/darkness is 8 h;
(4) the preservative film is lifted and sprayed for 2-3 times with 1mg/L rooting powder solution every day, and the length of the adventitious root is measured after two weeks and is 13.5 cm.
FIG. 2 shows the state of the tissue culture seedling of brachypodium distachyon after 2 weeks of growth in a climatic chamber, and the length of the regenerated adventitious root is measured to be 13.5cm, and the plant grows robustly with 7 lateral roots. This shows that the method can promote the regeneration of adventitious roots of the brachypodium tissue culture seedling.
Example 2
Substantially the same as in example 1, except that the regenerated adventitious roots were 12cm in length and the plants grew vigorously with 4 lateral roots.
Example 3
Substantially the same as in example 1, except that the regenerated adventitious roots were 15cm in length and the plants grew vigorously with 10 lateral roots.
The above description is only a preferred embodiment of the present invention, and all equivalent changes and modifications made in accordance with the claims of the present invention should be covered by the present invention.
Claims (5)
1. A method for rapidly inducing the adventitious root regeneration of a brachypodium distachyon tissue culture seedling is characterized by comprising the following steps:
(1) selecting rootless brachypodium distachyon tissue culture seedlings with the height of 2-3cm, taking the brachypodium distachyon tissue culture seedlings out of a culture bottle by using tweezers, and cleaning for 3-5 times;
(2) soaking the cleaned tissue culture seedlings in 1mg/L rooting powder solution for 3-5 minutes;
(3) transplanting the tissue culture seedlings soaked with the rooting powder into a plastic planting pot filled with mixed soil, covering the mouth of the plastic planting pot with a preservative film after planting is finished, and placing the plastic planting pot in a phytotron for culturing;
(4) the preservative film is lifted and sprayed for 2-3 times every day by using 1mg/L rooting powder solution, and the length of the adventitious root is measured after two weeks and is 12-15 cm.
2. The method for rapidly inducing the adventitious root regeneration of a brachypodium distachyon tissue culture seedling as claimed in claim 1, wherein in the step (1), the rootless brachypodium distachyon tissue culture seedling is an adventitious bud induced by the brachypodium distachyon callus.
3. The method for rapidly inducing the adventitious root regeneration of a brachypodium distachyon tissue culture seedling of claim 1, wherein in the step (2), the rooting powder solution is prepared by dissolving 1mg of rooting powder in 1L of distilled water.
4. The method for rapidly inducing the adventitious root regeneration of a brachypodium distachyon tissue culture seedling of claim 1, wherein in the step (3), the mixed soil is nutrient soil, perlite and vermiculite, and the ratio of the nutrient soil to the perlite is 2: 1: 2, adding clear water until the soil is wet.
5. The method for rapidly inducing the adventitious root regeneration of a brachypodium distachyon tissue culture seedling of claim 1, wherein in the step (3), the culture conditions of the artificial climate chamber are as follows: the temperature is 22 +/-2 ℃, the humidity is 60 +/-10%, and the illumination is 16 h/darkness is 8 h.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2019110436734 | 2019-10-30 | ||
| CN201911043673 | 2019-10-30 |
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| CN111512808A true CN111512808A (en) | 2020-08-11 |
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| CN202010525793.4A Pending CN111512808A (en) | 2019-10-30 | 2020-06-10 | Method for rapidly inducing adventitious root regeneration of brachypodium distachyon tissue culture seedlings |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102884983A (en) * | 2012-11-02 | 2013-01-23 | 广西壮族自治区中国科学院广西植物研究所 | Method for promoting tissue culture seedling of anisetree bark to take root rapidly |
| CN103548653A (en) * | 2013-11-01 | 2014-02-05 | 重庆文理学院 | Honeysuckle tissue culture seedling non-tube rootage method |
| CN104585039A (en) * | 2015-02-09 | 2015-05-06 | 上海青浦现代农业园区发展有限公司 | Tissue culture and rapid propagation method of blueberry |
| CN109220805A (en) * | 2018-11-05 | 2019-01-18 | 贵州大学 | A kind of Ormosia hosiei tissue culture outside sprout-cultivating-bottle radication method |
| CN109566126A (en) * | 2018-12-25 | 2019-04-05 | 福建农林大学 | A kind of method of Pearl color osmanthus tissue culture outside sprout-cultivating-bottle radication |
-
2020
- 2020-06-10 CN CN202010525793.4A patent/CN111512808A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102884983A (en) * | 2012-11-02 | 2013-01-23 | 广西壮族自治区中国科学院广西植物研究所 | Method for promoting tissue culture seedling of anisetree bark to take root rapidly |
| CN103548653A (en) * | 2013-11-01 | 2014-02-05 | 重庆文理学院 | Honeysuckle tissue culture seedling non-tube rootage method |
| CN104585039A (en) * | 2015-02-09 | 2015-05-06 | 上海青浦现代农业园区发展有限公司 | Tissue culture and rapid propagation method of blueberry |
| CN109220805A (en) * | 2018-11-05 | 2019-01-18 | 贵州大学 | A kind of Ormosia hosiei tissue culture outside sprout-cultivating-bottle radication method |
| CN109566126A (en) * | 2018-12-25 | 2019-04-05 | 福建农林大学 | A kind of method of Pearl color osmanthus tissue culture outside sprout-cultivating-bottle radication |
Non-Patent Citations (1)
| Title |
|---|
| 郭书普: "《西瓜早熟秋延栽培新技术》", 30 November 1998, 北京：中国致公出版社 * |
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