CN110063260B - Method for rapidly propagating longstalck peaches - Google Patents
Method for rapidly propagating longstalck peaches Download PDFInfo
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- CN110063260B CN110063260B CN201910466294.XA CN201910466294A CN110063260B CN 110063260 B CN110063260 B CN 110063260B CN 201910466294 A CN201910466294 A CN 201910466294A CN 110063260 B CN110063260 B CN 110063260B
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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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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
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Abstract
The invention discloses a method for rapidly propagating longstalck peaches, which comprises the steps of inoculating embryos of treated longstalck peaches seeds into a first culture medium for induction culture to obtain cluster buds, wherein the first culture medium comprises a culture substrate, 6-benzylaminopurine and 2, 4-dichlorophenoxyacetic acid; inoculating the cluster buds into a second culture medium for multiplication culture, wherein the second culture medium comprises a culture substrate, 6-benzylaminopurine and 2, 4-dichlorophenoxyacetic acid; inoculating the cluster buds subjected to multiplication culture into a third culture medium for induction culture to obtain rooted seedlings, wherein the third culture medium comprises a culture substrate, naphthylacetic acid and indolebutyric acid; transplanting the rooted seedlings into a substrate for cultivation. The method has the advantages that the amygdalus pedunculata pall can be rapidly and efficiently cultured, the cluster bud induction rate, the multiplication coefficient and the rooting rate in the culture process are all high levels, the culture process is not influenced by seasons and environments, and the method can be popularized and applied in a large scale.
Description
Technical Field
The invention belongs to the technical field of plant cultivation, and particularly relates to a method for quickly propagating longstalck peaches.
Background
The longstalck peaches (amygdalus pedunculate pall) is an important ecological greening tree species and economic tree species and has wide application and practical value. Due to desert burying and artificial damage, the quantity of longstalck peaches population is continuously reduced, natural distribution is gradually withered, and the longstalck peaches population is listed as second-level endangered plants. The longstalck peaches have hard endocarp, so the longstalck peaches are difficult to germinate in natural environment and have low survival rate, and the longstalck peaches are difficult to survive after being transplanted due to developed main root systems, so a large number of transplanted seedlings with developed root systems need to be propagated by using a tissue culture method.
In the prior art, the Sunzhizhuangye researches the influence of different exogenous plant growth regulators, culture medium types, and other factors for culturing test-tube plantlets on the proliferation efficiency and the rooting of the test-tube plantlets by taking the stem tips of the longstalck peaches as explants; the influence of the combination of lanthanum nitrate and auxin with different concentrations on the rooting of the test-tube plantlets of the amygdalus pedunculata is researched by extreme difference analysis. However, no report is found on the research of inducing cluster buds by using longstalck amygdalus seed embryos in the prior art.
Disclosure of Invention
The invention aims to provide a method for quickly propagating amygdalus pedunculata pall, which can quickly and efficiently culture amygdalus pedunculata pall by adopting a specific culture medium to culture amygdalus pedunculata pall seeds.
The purpose of the invention is realized by the following technical scheme:
a method for rapidly propagating longstalck peaches comprises the following cultivation steps: pretreating mature seeds of longstalck peaches to obtain embryos; inoculating the embryo into a first culture medium for induction culture to obtain cluster buds, wherein the first culture medium comprises a culture substrate, 0.4-0.6mg/L of 6-benzylaminopurine and 0.05-0.15mg/L of 2, 4-dichlorophenoxyacetic acid; inoculating the cluster buds into a second culture medium for multiplication culture, wherein the second culture medium comprises the culture substrate, 0.05-0.15mg/L of 6-benzylaminopurine and 0.25-0.35mg/L of 2, 4-dichlorophenoxyacetic acid; inoculating the cluster buds after propagation culture into a third culture medium for induction culture to obtain rooted seedlings, wherein the third culture medium comprises the culture substrate, 0.05-0.15mg/L of naphthylacetic acid and 0.25-0.35mg/L of indolebutyric acid; transplanting the rooted seedlings into a matrix for cultivation, wherein the matrix comprises grass carbon, perlite and vermiculite.
The first culture medium comprises 0.5mg/L of 6-benzylaminopurine and 0.1mg/L of 2, 4-dichlorophenoxyacetic acid, the second culture medium comprises 0.1mg/L of 6-benzylaminopurine and 0.3mg/L of 2, 4-dichlorophenoxyacetic acid, and the third culture medium comprises 0.1mg/L of naphthylacetic acid and 0.3mg/L of indolebutyric acid.
The pH values of the first culture medium, the second culture medium and the third culture medium are 5.8-6.0, and the culture substrate further comprises an MS culture medium, 28-32g/L of sucrose and 5-7g/L of agar.
The culture substrate comprises 30g/L of sucrose and 6g/L of agar.
The incubation time in the first medium and the second medium was 21 days, and the incubation time in the third medium was 35 days.
The ratio of the grass carbon, the perlite and the vermiculite in the matrix is 6:3: 1.
In each cultivation step, the temperature is 21-25 ℃, the illumination intensity is 1700-1900Lx, and the illumination time is 14-16 h/d.
In each cultivation step, the temperature is 23 ℃, the illumination intensity is 1800Lx, and the illumination time is 15 h/d.
The pretreatment comprises the following treatment steps: removing pulp and endocarp, soaking in 2% NaCl solution for 30 min, cleaning, standing, soaking in 75% ethanol for 30 s, soaking in 0.1% mercuric chloride for 10 min, and removing embryo with part of cotyledon; and (3) washing with sterile water before and after each treatment step.
The invention has the advantages that: can quickly and efficiently cultivate the longstalck peaches, the induction rate, the multiplication coefficient and the rooting rate of the cluster buds in the cultivation process are all high, the cultivation process is not influenced by seasons and environment, and the method can be popularized and applied in a large scale.
Detailed Description
The features of the present invention and other related features are described in further detail below by way of examples to facilitate understanding by those skilled in the art:
example (b): the embodiment specifically relates to a method for rapidly propagating amygdalus pedunculata, which can rapidly and efficiently culture amygdalus pedunculata seeds by adopting a specific culture medium.
The method for rapidly propagating the longstalck peaches in the embodiment comprises the following cultivation steps: pretreating mature seeds of longstalck peaches to obtain embryos; inoculating the embryo into a first culture medium for induction culture to obtain cluster buds, inoculating the cluster buds into a second culture medium for propagation culture, inoculating the cluster buds subjected to propagation culture into a third culture medium for induction culture to obtain rooted seedlings, transplanting the rooted seedlings into a substrate for culture, and growing into the longstalck peaches with strong roots in the rooted seedling substrate.
In this embodiment, the pretreatment includes the following steps: picking up mature seeds of longstalck peaches, removing pulp, drying, mechanically breaking hard endocarp, washing the kernels with the peeled pericarps for 2 times with water, soaking the kernels in a NaCl solution with the concentration of 2% for 30 minutes in a clean environment, then washing with sterile water for 4-5 times, standing for 24 hours, soaking the kernels in 75% alcohol for 30 seconds with sterile water, washing with sterile water for 3-5 times, soaking with 0.1% mercuric chloride for 10 minutes, then washing with sterile water for 7-8 times, finally removing the seed coats with a scalpel and leaving embryos with parts of cotyledons, and leaving the embryos for cultivation. The process can ensure that the subsequent embryos can be effectively developed by using tissues influencing the development of the embryos except the embryos, and the inventory rate of the seeds is improved.
In this embodiment, the first medium comprises a culture substrate, 0.4-0.6 mg/L6-benzylaminopurine, and 0.05-0.15 mg/L2, 4-dichlorophenoxyacetic acid. The second culture medium comprises culture substrate, 0.05-0.15 mg/L6-benzylaminopurine, and 0.25-0.35 mg/L2, 4-dichlorophenoxyacetic acid. The third culture medium comprises culture substrate, 0.05-0.15mg/L naphthylacetic acid, 0.25-0.35mg/L indolebutyric acid. The culture substrate comprises MS culture medium, 28g/L-32g/L sucrose and 5g/L-7g/L agar. The pH of the first culture medium, the second culture medium and the third culture medium is 5.8-6.0. The matrix comprises grass carbon, perlite and vermiculite, and the ratio of the grass carbon to the perlite to the vermiculite is 6:3: 1.
In this example, the incubation times in the first and second media were 21 days, and the incubation time in the third medium was 35 days. In each cultivation step, the temperature is 21-25 ℃, the illumination intensity is 1700-1900Lx, and the illumination time is 14-16 h/d.
In this example, the first medium, the second medium, and the third medium were prepared in different amounts based on the experimental data in tables 1 to 3. The experimental groups A1-A5, the experimental groups B1-B5 and the experimental groups C1-C5 are mutually contrasted, and comparison is carried out according to the induction rate, the multiplication coefficient and the rooting rate of the cluster buds obtained through experiments, the content proportion in the embodiment can ensure that the induction rate of the cluster buds is higher than 80%, the multiplication coefficient is higher than 4.2 and the rooting rate is higher than 70%, a good cultivation effect can be obtained, and the cultivation success rate is stable.
In this example, according to the experimental data in tables 1 to 3, the first medium contains 0.5mg/L of 6-benzylaminopurine and 0.1mg/L of 2, 4-dichlorophenoxyacetic acid, the second medium contains 0.1mg/L of 6-benzylaminopurine and 0.3mg/L of 2, 4-dichlorophenoxyacetic acid, and the third medium contains 0.1mg/L of naphthylacetic acid and 0.3mg/L of indolebutyric acid. The culture substrate comprises MS culture medium, 30g/L sucrose and 6g/L agar. In each cultivation step, the temperature is 23 ℃, the illumination intensity is 1800Lx, and the illumination time is 15 h/d. The factor proportion can ensure that the cluster bud induction rate is 85 percent, the multiplication coefficient is 4.5 and the rooting rate is 75 percent in the propagation process of the longstalck peaches, and the culture effect is better.
Table 1: induced cultivation of cluster buds under different contents of first culture medium
| A1 | A2 | A3 | A4 | A5 | |
| 6-benzylaminopurine (mg/L) | 0.3 | 0.4 | 0.5 | 0.6 | 0.8 |
| 2, 4-Dichlorophenoxyacetic acid (mg/L) | 0 | 0.05 | 0.1 | 0.15 | 0.3 |
| Sucrose (g/L) | 20 | 28 | 30 | 32 | 40 |
| Agar (g/L) | 2 | 5 | 6 | 7 | 10 |
| Induction rate of cluster buds | 50% | 80% | 85% | 80% | 60% |
Table 2: proliferation status of cluster buds in different contents of second medium
| B1 | B2 | B3 | B4 | B5 | |
| 6-benzylaminopurine (mg/L) | 0 | 0.05 | 0.1 | 0.15 | 0.3 |
| 2, 4-Dichlorophenoxyacetic acid (mg/L) | 0 | 0.25 | 0.3 | 0.35 | 0.6 |
| Sucrose (g/L) | 20 | 28 | 30 | 32 | 40 |
| Agar (g/L) | 2 | 5 | 6 | 7 | 10 |
| Coefficient of proliferation | 2 | 4.2 | 4.5 | 4.2 | 3 |
Table 3: rooting condition of cluster buds with different contents in second culture medium
| C1 | C2 | C3 | C4 | C5 | |
| Naphthylacetic acid (mg/L) | 0 | 0.05 | 0.1 | 0.15 | 0.3 |
| Indolebutyric acid (mg/L) | 0 | 0.25 | 0.3 | 0.35 | 0.6 |
| Sucrose (g/L) | 20 | 28 | 30 | 32 | 40 |
| Agar (g/L) | 2 | 5 | 6 | 7 | 10 |
| Rooting rate | 30% | 70% | 75% | 70% | 50% |
The embodiment has the following advantages: can quickly and efficiently cultivate the longstalck peaches, the induction rate, the multiplication coefficient and the rooting rate of the cluster buds in the cultivation process are all high, the cultivation process is not influenced by seasons and environment, and the method can be popularized and applied in a large scale.
Claims (9)
1. A method for rapidly propagating longstalck peaches is characterized by comprising the following culturing steps: pretreating mature seeds of longstalck peaches to obtain embryos;
inoculating the embryo into a first culture medium for induction culture to obtain cluster buds, wherein the first culture medium consists of a culture substrate, 6-benzylaminopurine and 2, 4-dichlorophenoxyacetic acid, and each liter of culture medium is added with 0.4-0.6mg of 6-benzylaminopurine and 0.05-0.15mg of 2, 4-dichlorophenoxyacetic acid;
inoculating the cluster buds into a second culture medium for multiplication culture, wherein the second culture medium consists of a culture substrate, 6-benzylaminopurine and 2, 4-dichlorophenoxyacetic acid, and each liter of culture medium is added with 0.05-0.15mg of 6-benzylaminopurine and 0.25-0.35mg of 2, 4-dichlorophenoxyacetic acid;
inoculating the cluster buds after propagation culture into a third culture medium for induction culture to obtain rooted seedlings, wherein the third culture medium consists of a culture substrate, naphthylacetic acid and indolebutyric acid, and each liter of culture medium is added with 0.05-0.15mg of naphthylacetic acid and 0.25-0.35mg of indolebutyric acid;
transplanting the rooted seedlings into a matrix for cultivation, wherein the matrix comprises grass carbon, perlite and vermiculite;
the culture substrate consists of an MS culture medium, sucrose and agar, wherein each liter of MS culture medium is added with 28-32g of sucrose and 5-7g of agar.
2. The method of claim 1, wherein the first culture medium is supplemented with 0.5mg of 6-benzylaminopurine and 0.1mg of 2, 4-dichlorophenoxyacetic acid per liter of the culture medium, the second culture medium is supplemented with 0.1mg of 6-benzylaminopurine and 0.3mg of 2, 4-dichlorophenoxyacetic acid per liter of the culture medium, and the third culture medium is supplemented with 0.1mg of naphthylacetic acid and 0.3mg of indolebutyric acid per liter of the culture medium.
3. The method for rapid propagation of Amygdalus pedunculata as claimed in claim 1, wherein the pH of the first culture medium, the second culture medium and the third culture medium is 5.8-6.0.
4. The method for rapidly propagating longstalck peaches according to claim 1, wherein 30g of sucrose and 6g of agar are added to each liter of MS culture medium in the culture substrate.
5. The method for rapid propagation of Amygdalus pedunculata as claimed in claim 1, wherein the incubation time in the first culture medium and the second culture medium is 21 days, and the incubation time in the third culture medium is 35 days.
6. The method for rapidly propagating amygdalus pedunculata pall as claimed in claim 1, wherein the ratio of the peat, the perlite and the vermiculite in the matrix is 6:3: 1.
7. The method for rapidly propagating longstalck peaches as claimed in claim 1, wherein in each of the cultivating steps, the temperature is 21-25 ℃, the illumination intensity is 1700-1900Lx, and the illumination time is 14-16 h/d.
8. The method of claim 7, wherein in each of the culturing steps, the temperature is 23 ℃, the illumination intensity is 1800Lx, and the illumination time is 15 h/d.
9. The method for rapidly propagating longstalck peaches as claimed in claim 1, wherein the pretreatment comprises the following steps: removing pulp and endocarp, soaking in 2% NaCl solution for 30 min, cleaning, standing, soaking in 75% ethanol for 30 s, soaking in 0.1% mercuric chloride for 10 min, and removing embryo with part of cotyledon; and (3) washing with sterile water before and after each treatment step.
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