CN110521595B - Method for promoting stem elongation in vitro culture of oriental cherry - Google Patents
Method for promoting stem elongation in vitro culture of oriental cherry Download PDFInfo
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- CN110521595B CN110521595B CN201910828691.7A CN201910828691A CN110521595B CN 110521595 B CN110521595 B CN 110521595B CN 201910828691 A CN201910828691 A CN 201910828691A CN 110521595 B CN110521595 B CN 110521595B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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Abstract
The invention discloses a method for promoting stem elongation in-vitro culture of oriental cherry, and belongs to the technical field of plant tissue culture. The invention discloses a method for promoting stem elongation in vitro culture of oriental cherry, which comprises the steps of sterilizing an explant, performing cold treatment and culturing under red light; the problems of short and small plant growth and blocked stem extension in the tissue culture process of the oriental cherry seedlings are solved; the dormancy of the buds is broken by adopting low temperature, the growth of the stems is promoted by adopting hormone and red light irradiation, and the length of the stem section reaches 4.23 cm.
Description
Technical Field
The invention relates to the technical field of plant tissue culture, in particular to a method for promoting stem elongation in-vitro culture of oriental cherry.
Background
The oriental cherry (cerasus sp.) belongs to Rosaceae, Prunoideae, and Oriental cherry, and has a height of 15-25 m. The bark is chestnut brown and smooth; oval to oval, caudate, sharp or heavy sawtooth at the edge, short thorn and no hair on both sides. White or light pink, single petal, 3-5 flowers forming short umbrella house, no fragrance, bell-shaped calyx, and no hair. The stone fruit is spherical, red first and purple brown later. In the flowering period of 4 months, flowers and leaves are placed together. Cherry blossom is a sunny one, deep and fertile one with good drainage, and has strong adsorption capacity to harmful gases such as sodium hydroxide and carbon dioxide, so it can be used to purify air. The main variants are: the cherry blossom with heavy petals, the cherry blossom with weeping branches, the rose blossom with magnificent petals and the like are world famous ornamental woody flowers, have beautiful tree shapes, are competitive in flowers in spring, are light and delicate, are suitable for being planted in groups in a sheet form, are used for landscape architecture or street trees, can also be used in the fields of food, wine, pharmacy, health products, daily chemical industry and the like, and have higher ornamental value and economic value.
The oriental cherry varieties are more than 150 in total in the world, and are mainly distributed in europe in the northern hemisphere, warm regions in north america, china in asia, japan, and korea. Wherein, about 50 oriental cherry blossoms exist in China, and are mainly distributed in Yangtze river basin, west and southwest areas. The traditional oriental cherry propagation mainly comprises seed sowing, grafting, cuttage and the like. Although the seed propagation is fast and convenient, the seed is not easy to germinate under natural conditions, and simultaneously the seed propagation is influenced by multiple factors such as collection, storage, modulation time, germination rate and the like, the grafting is the most common propagation method for oriental cherry at present, but the industrialized production is difficult to carry out due to the fact that a proper stock cannot be found, and oriental cherry is difficult to root by cutting. The tissue culture technology not only has the advantages of large propagation coefficient, short seedling period and good genetic character stability, but also can obtain a large amount of high-quality seedlings in a short period. As tissue culture techniques mature, more and more researchers and researchers are working on them. Mainly comprises embryo culture, bud differentiation culture, leaf culture and other organs, but the cherry tissue culture seedling is shorter and the stem section of the cherry is difficult to elongate and grow. Therefore, it is a problem to be solved by those skilled in the art to provide a method for promoting stem elongation in vitro culture of oriental cherries.
Disclosure of Invention
In view of the above, the present invention provides a method for promoting stem elongation in vitro culture of oriental cherry.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for promoting stem elongation in vitro culture of oriental cherry comprises the following specific steps:
(1) collecting explants in 4-6 months;
(2) placing the explant in a sterilized beaker, sealing the opening with gauze, and washing for 1.5-2h under tap water; soaking in 5% sodium hypochlorite for 15min, and washing with sterile water for 2-3 times; soaking 0.2% benlate and 0.2% captan for 10min, and washing with sterile water;
(3) inoculating the sterilized explants in WPM +0.2 mg-L-16-BA+0.1mg·L-1Culturing in NAA culture medium at 5.5 deg.C for 42 d; the illumination intensity is 20 mu mol/(m) by adopting red light for illumination2S) the illumination time is 12 h/d;
(4) after the cold treatment is finished, transferring the tissue culture seedlings to the condition of 21-25 ℃ for continuous culture for 28 days, and irradiating by adopting red light with the illumination intensity of 20 mu mol/(m)2S) and the illumination time is 12 h/d.
Further, the explant is selected from robust branches without diseases and insect pests, full axillary buds and good growth condition.
Further, the explant is a 2-3cm stem section with 1-2 axillary buds.
Further, the pH of the culture medium in the step (3) is 5.8-6.0.
According to the technical scheme, compared with the prior art, the invention discloses a method for promoting stem elongation in-vitro culture of oriental cherry, and solves the problems of short plant growth and stem elongation resistance of oriental cherry seedlings in the tissue culture process; the dormancy of the buds is broken by adopting low temperature, the growth of the stems is promoted by adopting hormone and red light irradiation, and the length of the stem section reaches 4.23 cm.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.
FIG. 1 is a photograph of a sterile seedling obtained by the cultivation in example 1;
FIG. 2 is a photograph of a sterile seedling obtained by the comparative example culture.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The Oriental cherry plant is selected from scientific research nursery of Jiangsu institute of agriculture and forestry and occupational technology.
Example 1
A method for promoting stem elongation in vitro culture of oriental cherry comprises the following specific steps:
(1) selecting a strong annual branch with no plant diseases and insect pests, full axillary buds and good growth condition from a mother plant in a sunny day of 4 months at about ten am, cutting off leaves, and cutting into 2-3cm stem segments with 1-2 axillary buds to serve as explants;
(2) placing the explant in a sterilized beaker, sealing the opening with gauze, and washing for 1.5-2h under tap water; soaking in 5% sodium hypochlorite for 15min, and washing with sterile water for 2-3 times; soaking 0.2% benlate and 0.2% captan for 10min, and washing with sterile water;
(3) inoculating the sterilized explants in WPM +0.2 mg-L-16-BA+0.1mg·L-1Culturing in NAA medium (pH 5.8-6.0) at 5.5 deg.C for 42 d; the illumination intensity is 20 mu mol/(m) by adopting red light for illumination2S) the illumination time is 12 h/d;
(4) after the cold treatment is finished, transferring the tissue culture seedlings to the condition of 21-25 ℃ for continuous culture for 28 days, and irradiating by adopting red light with the illumination intensity of 20 mu mol/(m)2S) and the illumination time is 12 h/d.
Comparative example
An in vitro culture method of oriental cherry comprises the following specific steps:
(1) selecting a strong annual branch with no plant diseases and insect pests, full axillary buds and good growth condition from a mother plant in a sunny day of 4 months at about ten am, cutting off leaves, and cutting into 2-3cm stem segments with 1-2 axillary buds to serve as explants;
(2) placing the explant in a sterilized beaker, sealing the opening with gauze, and washing for 1.5-2h under tap water; soaking in washing powder for 10-15min, washing with sterile water, soaking in mixed solution of 2000ppm carbendazim and 600ppm sodium penicillin for 10min, and washing with sterile water for 2-3 times; placing on a clean bench for inoculation; connecting 1 material per bottle, connecting 40 bottles, and repeating for 2 times;
(3) inoculating the sterilized explants in WPM +0.5 mg-L-16-BA+0.1mg·L-1Culturing in NAA medium (pH 5.8-6.0) at 21-25 deg.C for 70 days under illumination of 12h/d and 2000 lx.
After the culture is finished, photographing is carried out on the sterile seedlings obtained in the example 1 and the comparative example, and the results are shown in the figure 1-2; and the stem length of the aseptic seedlings was measured, and the stem length of the aseptic seedlings of example 1 was 4.23cm on average, and the stem length of the aseptic seedlings of comparative example was 2.47cm on average.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (1)
1. A method for promoting stem elongation in vitro culture of oriental cherry is characterized by comprising the following specific steps:
(1) collecting explants in 4-6 months; the explant is selected from robust branches without diseases and insect pests, full axillary buds and good growth condition; the explant is a 2-3cm stem section with 1-2 axillary buds;
(2) washing the explant for 1.5-2 h; soaking in 5% sodium hypochlorite for 15min, and washing with sterile water for 2-3 times; soaking 0.2% benlate and 0.2% captan for 10min, and washing with sterile water;
(3) inoculating the sterilized explants in WPM +0.2 mg-L-16-BA+0.1mg·L-1Culturing in NAA culture medium at 5.5 deg.C for 42 d; the illumination intensity is 20 mu mol/(m) by adopting red light for illumination2S) the illumination time is 12 h/d; the pH of the culture medium is 5.8-6.0;
(4) after the cold treatment is finished, transferring the tissue culture seedlings to the condition of 21-25 ℃ for continuous culture for 28 days, and irradiating by adopting red light with the illumination intensity of 20 mu mol/(m)2S) and the illumination time is 12 h/d.
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