CN109548652B - Aifengli callus as well as culture method and application thereof - Google Patents
Aifengli callus as well as culture method and application thereof Download PDFInfo
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- CN109548652B CN109548652B CN201811483960.2A CN201811483960A CN109548652B CN 109548652 B CN109548652 B CN 109548652B CN 201811483960 A CN201811483960 A CN 201811483960A CN 109548652 B CN109548652 B CN 109548652B
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
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Abstract
The invention discloses a culture method of callus of air pineapple and a rapid propagation method of air pineapple. The culture method of the callus of the tillandsia comprises the following steps: obtaining a root tip tissue of the tillandsia and disinfecting the root tip tissue; inoculating the sterilized root tip tissue into an induction culture medium for induction culture treatment to obtain a callus; transferring the callus to a subculture multiplication culture medium for subculture multiplication culture treatment. The explant material of the air pineapple callus culture method is easy to obtain, and basically has zero damage to the original plant; a large amount of callus can be obtained through the induction culture treatment and the subculture multiplication culture treatment of the root tip tissue, and the callus has stable differentiation capacity. The rapid propagation method of the air plants can realize rapid and mass propagation of the air plants, and the propagated air plants have stable characters.
Description
Technical Field
The invention belongs to the technical field of plant tissue culture, and particularly relates to an air pineapple callus as well as a culture method and application thereof.
Background
The air pineapple belongs to the family of Bromeliaceae (broomeliaceae), and is a monocotyledon and perennial herb. The bromeliaceae family has 3 subfamilies, 50 genera, including nearly 550 varieties and 90 varieties. The 3 subfamilies are the panicle pineapple subfamily (Pitcairnioideae), the Fehland subfamily (Tillandsioideae) and the pineapple subfamily (Bromelioideae), respectively. Among them, fox tail pineapple (tillandia Funkiana) is a common one of the pineapple, and is a typical aerial or epiphytic plant.
The main organs of the air pineapple for absorbing nutrients and moisture are sunflower-shaped scale structures covered on the surfaces of leaves. The structure can help the air pineapple to directly obtain nutrients and moisture from air with pollutants. Their root systems deteriorate and only act as a anchorage on the support, without the need for soil, so they do not have any connection to the ground.
The air pineapple is an ornamental plant which can be used for both ornamental flowers and leaves. The air pineapple has various varieties, beautiful plant shape and strong decorative property. The artificial stone can be individually stuck on ancient stumps, artificial hillstones and walls or placed in bamboo baskets and shells to decorate space, is fashionable and fresh, and is rich in natural interests; or the components can be spliced into a bonsai and decorated into a screen, so that the living room can be vigorous. In addition, the air pineapple also has the advantage of long ornamental period, and can bloom almost all the year round; the air pineapple does not have a culture medium, does not need to be provided with a flower shelf, is clean and sanitary, can be randomly placed, and is an ideal indoor ornamental plant. The air pineapple can also be used for landscape building of garden plants, landscape group of green land, or special flower exhibition of air pineapple plants in holidays. The flower is introduced to China in recent years, quickly becomes a new favorite of plant enthusiasts, is also a novel indoor ornamental flower, and gradually enters common families.
At present, the conventional plant division propagation and cutting propagation are mostly adopted for the air pineapple propagation, and the tissue culture and the sterile culture are carried out by using seeds in mature and uncracked fruit pods. The success rate of seed reproduction is high, but the time from seed growth to finished product is 5-8 years, and the seed yield is low; although the plant is fast in propagation speed, the disinfection of the explant is very difficult due to the fact that the leaves of the pineapple are fully covered with the scales; the terminal bud or the lateral bud is taken as an explant for rapid germination, so that mould pollution is often caused and the explant fails; in addition, because the differentiation degree of the tillandsia is high, and callus induction is difficult, the tissue culture of the tillandsia is still an exploration stage at present, so that flower manufacturers mostly adopt plant division propagation at present. How to effectively propagate the tillandsia is a technical problem which is constantly strived to solve by the technicians in the field.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provides an air pineapple callus as well as a culture method and application thereof, so as to solve the technical problems that the explant material for air pineapple propagation is difficult to obtain and the propagation efficiency is not ideal.
In order to achieve the above object, according to one aspect of the present invention, a method for culturing callus of tillandsia is provided. The culture method of the tillandsia callus comprises the following steps:
obtaining a root tip tissue of the tillandsia and disinfecting the root tip tissue;
inoculating the sterilized root tip tissue into an induction culture medium for induction culture treatment to obtain a callus;
transferring the callus to a subculture multiplication culture medium for subculture multiplication culture treatment.
In another aspect of the invention, an tillandsia callus is provided. The air pineapple callus is obtained by culturing the air pineapple callus by the culture method.
In another aspect of the invention, a rapid propagation method of tillandsia is provided. The rapid propagation method of the air plants comprises the step of carrying out differentiation culture on the callus of the air plants to obtain seedlings.
Compared with the prior art, the culture method of the air pineapple callus adopts the root tip tissue of the air pineapple as the explant material, is easy to obtain, basically has zero damage to the original plant, and is simple and easy to sterilize; on the other hand, by the induction culture treatment and the subculture proliferation culture treatment of the root tip tissue, a large amount of callus can be obtained and the callus has a stable differentiation ability.
The air pineapple callus is obtained by culturing the air pineapple callus by the culture method, so that the air pineapple callus can be cultured in a large scale and has stable differentiation capacity.
The rapid propagation method of the air plants directly utilizes the callus of the air plants to perform differentiation culture to form seedlings, so that the rapid propagation method of the air plants can realize rapid and mass propagation of the air plants, and the propagated air plants have stable characters.
Drawings
FIG. 1 is a process flow chart of a method for culturing callus of tillandsia provided by the embodiment of the invention.
Detailed Description
In order to make the technical problems, technical solutions and advantageous effects to be solved by the present invention more clearly apparent, the present invention is further described in detail below with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which embodiments of the invention belong. If a definition set forth in this section is contrary to or otherwise inconsistent with a definition set forth in the patents, patent applications, published patent applications, and other publications that are herein incorporated by reference, the definition set forth in this section prevails over the definition that is incorporated herein by reference.
In one aspect, the embodiment of the invention provides a culture method of callus of tillandsia. The technological process of the culture method of the callus of the tillandsia is shown in figure 1, and the culture method comprises the following steps:
s01, disinfection of explants: obtaining a root tip tissue of the tillandsia and disinfecting the root tip tissue;
s02, callus formation: inoculating the sterilized root tip tissue into an induction culture medium for induction culture treatment to obtain a callus;
s03, subculture multiplication culture of the callus: transferring the callus to a subculture multiplication culture medium for subculture multiplication culture treatment.
Wherein, in the step S01, the root tip tissue of the tillandsia is adopted as the explant material, which is easy to obtain, has almost no harm to the original plant, and the disinfection treatment is simple and easy to implement. In one embodiment, the root tip tissue is 0.5-2cm of the root tip of the air pineapple root system. Preferably 0.5-2cm of tip of the root system of the air pineapple growing vigorously.
In another embodiment, the sterilization process for the root tip tissue comprises the steps of:
cleaning the root tip tissue with sterilized water, soaking for 10-15min, and transferring into 0.1% HgCl2Sterilizing in the solution for 3-5min, and washing with sterilized water for 3-5 times.
In addition, the air pineapple can be a common fox tail air pineapple.
In step S02, the root tip tissue is inoculated into an induction medium for induction culture treatment, and the cells of the root tip tissue part are subject to cell division and proliferation to form callus. In one embodiment, the induction medium comprises the following components:
MS basic culture medium, 0.2-1.5 mg/L2, 4-dichlorophenoxyacetic acid and 0-2mg/L diethylaminoethanol caproate.
The cell division and proliferation of the root tip tissue can be effectively induced to form callus by adding a proper amount of 2, 4-dichlorophenoxyacetic acid and diethylaminoethanol caproate into the MS minimal medium.
Wherein, the root tip tissue sterilized in step S01 is placed in the induction medium for induction culture, and callus is grown on the root tip tissue. In one embodiment, the root tip tissue after the disinfection treatment is placed in the induction culture medium for induction culture for a period of preferably 2-6 weeks. The culture conditions are preferably: the temperature is 25-30 ℃, the illumination is 12-16 h/day, and the illumination intensity is 1500-2000 lx. In the above step S03, transferring the callus described in step S03 to a subculture multiplication medium to realize continuous multi-generation culture of callus. In one embodiment, the secondary multiplication medium comprises the following components:
MS basic culture medium, 6-benzylamino adenine 0.1-1.5mg/L, 2, 4-dichlorophenoxyacetic acid 0.8-1.5 mg/L.
By adding a proper amount of 6-benzylamino adenine and 2, 4-dichlorophenoxyacetic acid into the MS minimal medium, the continuous multi-generation multiplication culture of the callus can be effectively promoted to be induced, a large amount of callus can be obtained, and the stable differentiation capacity of the callus can be improved.
In a further embodiment, in the step of the secondary proliferation culture process, the callus is transferred every 30-40 days. The subculture medium used for the transfer is the subculture containing 6-benzylamino adenine and 2, 4-dichlorophenoxyacetic acid.
Wherein, the time for placing the callus cultured in step S02 into the subculture can be adjusted and controlled according to the callus demand, and in one embodiment, the subculture conditions are as follows: the temperature is 25-30 ℃, the illumination is 12-16 h/day, and the illumination intensity is 1500-2000 lx. So as to improve the subculture proliferation of the callus.
Therefore, the culture method of the callus of the air pineapple adopts the root tip tissue of the air pineapple as the explant material, so that the explant material is relatively easy to obtain, basically has no harm to the original plant and is easy to disinfect; on the other hand, by optimizing the induction medium and the subculture, the induction culture treatment and the subculture multiplication culture treatment of the root tip tissue are realized, so that a large amount of callus can be obtained and stable differentiation capability can be imparted to the callus. In addition, the culture method of the callus of the tillandsia is easy to control conditions, and the obtained callus of the tillandsia is stable in character, strong in differentiation capacity and high in efficiency.
On the other hand, on the basis of the culture method of the tillandsia calluses, the embodiment of the invention also provides the tillandsia calluses. The callus of the air plant is obtained by culturing the callus of the air plant by the culture method of the callus of the air plant. Therefore, the tillandsia calli can be cultured in a large amount, and have stable differentiation ability.
On the basis of the callus of the tillandsia and the culture method thereof, the embodiment of the invention also provides a rapid propagation method of the tillandsia. The rapid propagation method of the tillandsia comprises the step of carrying out differentiation culture on the callus of the tillandsia to form seedlings. Therefore, the rapid propagation method of the air plants can realize rapid and mass propagation of the air plants, the propagated air plants have stable characters, and the cost is reduced.
The present invention will now be described in further detail with reference to specific aerial pineapple callus and a method for culturing the same as an example.
Example 1
The embodiment provides an tillandsia callus and a culture method thereof. The culture method of the callus of the tillandsia comprises the following steps:
s11, disinfection of explants:
selecting root system of Fox tail pineapple (Tillandsia Funkiana) with vigorous growth, cutting root tip with length of 1.0cm, cleaning with sterilized water, soaking for 15min, and adding 0.1% HgCl2Sterilizing in the solution for 3min, and washing with sterilized water for 5 times;
s12, callus formation:
inoculating the root tips sterilized in the step S11 on an MS basic culture medium, inoculating one root tip in each bottle, adding 1.0 mg/L2, 4-dichlorophenoxyacetic acid and 0.5mg/L diethylaminoethanol caproate for culture treatment, and performing cell division and proliferation at the root tips for 18-22 days to form callus with the callus rate of 80%;
s13, subculture multiplication culture:
transferring the callus formed in the step S12 to an MS basal medium added with 0.5mg/L of 6-benzylamino adenine and 0.8mg/L of 2, 4-dichlorophenoxyacetic acid for subculture proliferation, and transferring every 30-40 days to obtain a large amount of callus.
Example 2
The embodiment provides an tillandsia callus and a culture method thereof. The culture method of the callus of the tillandsia comprises the following steps:
s11, disinfection of explants:
selecting root system of Fox tail pineapple (Tillandsia Funkiana) with vigorous growth, cutting root tip with length of 0.5cm, cleaning with sterilized water, and soakingSoaking for 15min, and transferring into 0.1% HgCl2Sterilizing in the solution for 4min, and washing with sterilized water for 5 times;
s12, callus formation:
inoculating the root tips sterilized in the step S11 on an MS basic culture medium, inoculating one root tip in each bottle, adding 1.2mg/L of 2, 4-dichlorophenoxyacetic acid and 0.5mg/L of diethylaminoethanol caproate for culture treatment, and performing cell division and proliferation at the root tips for 15-20 days to form callus with the callus rate of 73%;
s13, subculture multiplication culture:
transferring the callus formed in the step S12 to an MS minimal medium added with 0.8mg/L of 6-benzylamino adenine and 1.2mg/L of 2, 4-dichlorophenoxyacetic acid for subculture proliferation, and transferring every 30-40 days to obtain a large amount of callus.
Example 3
The embodiment provides an tillandsia callus and a culture method thereof. The culture method of the callus of the tillandsia comprises the following steps:
s11, disinfection of explants:
selecting root system of Fox tail pineapple (TillandsiaFunkiana) with vigorous growth, cutting root tip with length of 1.0cm, cleaning with sterilized water, soaking for 10min, and adding 0.1% HgCl2Sterilizing in the solution for 5min, and washing with sterilized water for 5 times;
s12, callus formation:
inoculating the root tips sterilized in the step S11 on an MS basic culture medium, inoculating one root tip in each bottle, adding 1.2mg/L of 2, 4-dichlorophenoxyacetic acid and 1.0mg/L of diethylaminoethanol caproate for culture treatment, and performing cell division and proliferation on the root tips to form callus in 15-18 days, wherein the callus rate is 92%;
s13, subculture multiplication culture:
transferring the callus formed in the step S12 to an MS basal medium added with 0.4mg/L of 6-benzylamino adenine and 0.8mg/L of 2, 4-dichlorophenoxyacetic acid for subculture proliferation, and transferring every 30-40 days to obtain a large amount of callus.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Claims (6)
1. A culture method of callus of tillandsia is characterized by comprising the following steps:
obtaining a root tip tissue of the tillandsia and disinfecting the root tip tissue;
inoculating the sterilized root tip tissue into an induction culture medium for induction culture treatment to obtain a callus;
transferring the callus to a subculture multiplication culture medium for subculture multiplication culture treatment;
wherein the induction medium comprises the following components:
MS basic culture medium;
0.2-1.5mg/L of 2, 4-dichlorophenoxyacetic acid;
0.5-2mg/L of diethyl aminoethyl hexanoate;
the subculture multiplication medium comprises the following components:
MS basic culture medium;
6-benzylamino adenine 0.1-1.5 mg/L;
0.8-1.5mg/L of 2, 4-dichlorophenoxyacetic acid.
2. The culture method according to claim 1, wherein the callus is transferred every 30 to 40 days in the step of the subculture proliferation treatment.
3. The culture method according to claim 1, wherein the root tip tissue is a 0.5-2cm root tip of an tillandsia root system.
4. The culture method according to any one of claims 1 to 3, wherein the Ailanthus altissima is Fox tail Ailanthus altissimaTillandsia Funckiana 。
5. The culture method according to any one of claims 1 to 3, wherein the step of sterilizing the root tip tissue comprises the steps of:
cleaning the root tip tissue with sterilized water, soaking for 10-15min, and transferring into 0.1% HgCl2Sterilizing in the solution for 3-5min, and washing with sterilized water for 3-5 times.
6. A rapid propagation method of tillandsia, comprising the step of performing differentiation culture of the callus of tillandsia obtained by the method for culturing callus of tillandsia according to any one of claims 1 to 5 to obtain a seedling.
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| CN110547196B (en) * | 2019-09-23 | 2021-09-07 | 江苏农林职业技术学院 | Method for quickly propagating air pineapple seedlings through leaf tissue culture |
| CN110547198B (en) * | 2019-09-23 | 2022-04-26 | 江苏农林职业技术学院 | Method for rapidly propagating tillandsia through seed tissue culture |
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