CN111528090A - Strawberry tissue culture seedling hardening matrix for improving hardening quality of strawberry tissue culture seedlings and preparation and use methods and application thereof - Google Patents
Strawberry tissue culture seedling hardening matrix for improving hardening quality of strawberry tissue culture seedlings and preparation and use methods and application thereof Download PDFInfo
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- CN111528090A CN111528090A CN202010370368.2A CN202010370368A CN111528090A CN 111528090 A CN111528090 A CN 111528090A CN 202010370368 A CN202010370368 A CN 202010370368A CN 111528090 A CN111528090 A CN 111528090A
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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
- 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
- A01G22/05—Fruit crops, e.g. strawberries, tomatoes or cucumbers
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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
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
-
- 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
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
- A01G24/12—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material containing soil minerals
- A01G24/15—Calcined rock, e.g. perlite, vermiculite or clay aggregates
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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
Abstract
The invention relates to a strawberry tissue culture seedling hardening matrix for improving the hardening quality of strawberry tissue culture seedlings, wherein each 10L of the matrix comprises the following components in percentage by weight: grass carbon: 2.5-4L; vermiculite: 2.5-3.5L; perlite: 2-3L; sodium hypochlorite or clear water: 1000-; trace fertilizer: the element ratio of N to P to K is 3:2: 2. The invention relates to a hardening and transforming substrate for strawberry tissue culture seedlings, which is prepared by uniformly mixing turf, vermiculite, perlite and other trace element fertilizer auxiliary materials according to a specific proportion and prewetting by sodium hypochlorite and has certain water holding capacity and excellent fertilizer holding effect.
Description
Technical Field
The invention belongs to the technical field of molecular breeding, and particularly relates to a strawberry tissue culture seedling hardening matrix for improving the hardening quality of strawberry tissue culture seedlings, and a preparation method and application thereof.
Background
Strawberry is a perennial herb of the genus strawberry of the family rosaceae. Modern large fruit type cultivated strawberries are mainly pineapple strawberries (Fragaria × ananassa Duch). The seedling gardening market demand of the strawberry is high, the agricultural production technology is high, the virus-free seedling production difficulty is high, and the virus-free seedling technical level in China is low. The breeding of early strawberry seedlings in China is mainly carried out by utilizing stolons for asexual propagation, but the method has lower propagation coefficient and poorer seedling quality, and the plant diseases and insect pests which harm strawberries are easy to spread in the propagation process, thereby limiting the development of the strawberry industry in China. The plant tissue culture technology is a mode of in vitro regeneration of plant tissue organs, and achieves the purpose of high-efficiency and quick propagation through less plant tissues, and in recent years, the purity of strawberry varieties is effectively guaranteed and the harm of diseases and pests is weakened along with the introduction of the plant tissue culture technology to the propagation of strawberries.
The tissue culture seedling in the plant tissue culture technology does not adapt to the environment outside the test tube, in order to enable the tissue culture seedling to adapt to the external growth environment, the acclimatization is an important ring in the breeding process of the strawberry tissue culture seedling, and the acclimatization survival rate of the strawberry tissue culture seedling is related to the popularization and application of the plant tissue culture and virus-free seedling production technology in industrial seedling culture.
In the prior art, coconut chaff, river sand and the like are mainly used as culture hardening matrix of strawberry tissue culture seedlings, and then clear water is poured.
The coconut husk and river sand substrates have the following disadvantages:
1. poor stability of water and fertilizer holding, low organic matter content and few microbial flora.
2. The substrate is easy to contain salt, the root growth of the seedling is greatly influenced, and the seedling growth is weak and the survival rate is low.
Through a search, no published patent literature relevant to the present patent application has been found.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provides a strawberry tissue culture seedling hardening matrix for improving the hardening quality of strawberry tissue culture seedlings, and a preparation method and application thereof.
The technical scheme adopted by the invention for solving the technical problems is as follows:
a hardening matrix for improving the hardening quality of strawberry tissue culture seedlings comprises the following components in percentage by weight in 10L of matrix:
grass carbon: 2.5-4L;
vermiculite: 2.5-3.5L;
perlite: 2-3L;
sodium hypochlorite or clear water: 1000-;
trace fertilizer: the element ratio of N to P to K is 3:2: 2.
Moreover, the fiber length of the turf is 1-10 mm; the particle size of the vermiculite is 3-5 mm; the particle size of the perlite is 3-6 mm; the available chlorine of the sodium hypochlorite is 0.01 percent.
The preparation method of the strawberry tissue culture seedling acclimatization substrate for improving the acclimatization quality of the strawberry tissue culture seedling comprises the following steps:
the grass carbon, the vermiculite and the perlite are uniformly mixed, then the grass carbon, the vermiculite and the perlite which are uniformly mixed are pre-wetted by adding sodium hypochlorite or clear water, then the trace fertilizer is added, and the mixture is uniformly mixed, so that the strawberry tissue culture seedling hardening matrix for improving the hardening quality of the strawberry tissue culture seedlings is obtained, wherein the water content of the matrix is 50-60%.
The use method of the strawberry tissue culture seedling acclimatization substrate for improving the acclimatization quality of the strawberry tissue culture seedling comprises the following steps:
the strawberry tissue culture seedlings are processed by chlorothalonil and then cultured by utilizing a strawberry tissue culture seedling hardening matrix.
Moreover, the chlorothalonil is a 2g/L aqueous solution of the chlorothalonil with the content of the effective component of 75 percent.
And, the strawberry tissue culture seedling is soaked in 2g/L chlorothalonil water solution for 30min for disinfection treatment.
The application of the strawberry tissue culture seedling hardening matrix for improving the hardening quality of the strawberry tissue culture seedlings is to improve the hardening quality of the strawberry tissue culture seedlings.
The invention has the advantages and positive effects that:
1. the invention relates to a hardening and transforming substrate for strawberry tissue culture seedlings, which is prepared by uniformly mixing turf, vermiculite, perlite and other trace element fertilizer auxiliary materials according to a specific proportion and prewetting by sodium hypochlorite and has certain water holding capacity and excellent fertilizer holding effect.
2. The mixed matrix prepared by the invention is free from exogenous pollution by processing, is beneficial to the mass propagation of virus-free strawberry seedlings, and avoids the infection of exogenous viruses and germs in the refining link.
3. The prepared mixed matrix ensures the water retention of the matrix, ensures the permeability of the matrix to be excellent, and comprises but not limited to promoting the growth of the root system and the growth of new leaves of strawberry tissue culture seedlings, promoting the generation of microbial floras which are mutually beneficial to symbiosis with the root system of strawberry plants in the matrix, and being beneficial to the plants to finish heterotrophic culture from the culture medium and alternating to autotrophic culture of the plants under the photosynthesis.
4. The mixed matrix prepared by the invention obviously improves the survival rate of the strawberry tissue culture seedlings and promotes the accumulation of nutrients of the strawberry seedlings in the acclimatization growth of the strawberry tissue culture seedlings, and the indexes include but are not limited to the improvement of the height of strawberry plants, the length of root systems, the number of new leaves, the fresh weight of plants, the dry weight and the like.
5. Compared with the traditional river sand and other matrixes, the mixed matrix prepared by the invention has the advantages of lower unit volume weight of the matrix and lower preparation cost of the matrix, and is an excellent culture matrix for hardening the tissue culture seedlings of strawberries.
Detailed Description
The following detailed description of the embodiments of the present invention is provided for the purpose of illustration and not limitation, and should not be construed as limiting the scope of the invention.
The raw materials used in the invention are conventional commercial products unless otherwise specified; the methods used in the present invention are conventional in the art unless otherwise specified.
A hardening matrix for improving the hardening quality of strawberry tissue culture seedlings comprises the following components in percentage by weight in 10L of matrix:
grass carbon: 2.5-4L;
vermiculite: 2.5-3.5L;
perlite: 2-3L;
sodium hypochlorite or clear water: 1000-;
trace fertilizer: the element ratio of N to P to K is 3:2: 2.
Preferably, the length of the fiber of the turf is 1-10 mm; the particle size of the vermiculite is 3-5 mm; the particle size of the perlite is 3-6 mm; the available chlorine of the sodium hypochlorite is 0.01 percent.
The preparation method of the strawberry tissue culture seedling acclimatization substrate for improving the acclimatization quality of the strawberry tissue culture seedling comprises the following steps:
the grass carbon, the vermiculite and the perlite are uniformly mixed, then the grass carbon, the vermiculite and the perlite which are uniformly mixed are pre-wetted by adding sodium hypochlorite or clear water, then the trace fertilizer is added, and the mixture is uniformly mixed, so that the strawberry tissue culture seedling hardening matrix for improving the hardening quality of the strawberry tissue culture seedlings is obtained, wherein the water content of the matrix is 50-60%.
The use method of the strawberry tissue culture seedling acclimatization substrate for improving the acclimatization quality of the strawberry tissue culture seedling comprises the following steps:
the strawberry tissue culture seedlings are processed by chlorothalonil and then cultured by utilizing a strawberry tissue culture seedling hardening matrix.
Preferably, the chlorothalonil is a 2g/L aqueous solution of chlorothalonil with the content of effective components of 75%.
Preferably, the strawberry tissue culture seedling is soaked in 2g/L chlorothalonil water solution for 30min for disinfection treatment.
The application of the strawberry tissue culture seedling hardening matrix for improving the hardening quality of the strawberry tissue culture seedlings is to improve the hardening quality of the strawberry tissue culture seedlings.
Specifically, the preparation and detection are as follows:
moving the strawberries from the illumination culture room to a greenhouse facility for early-stage hardening off for about 3d, enabling the environmental temperature in the tissue culture bottle to be approximately 22-24 ℃ with the external environmental temperature, and appropriately shading.
After the early-stage seedling hardening is finished, the rooting strawberry tissue culture seedlings with uniform and strong plants and complete root systems are selected and used for seedling hardening tests.
Before transplanting, the tissue culture seedling of the strawberry is soaked in 2g/L of chlorothalonil water solution for 30min for disinfection treatment.
Grass carbon, vermiculite, perlite and other trace fertilizer auxiliary materials are uniformly mixed according to a specific proportion, and are pre-wetted by sodium hypochlorite to form a mixture with a certain degree of moisture. The grass carbon, the vermiculite, the perlite and other trace element fertilizer auxiliary materials are uniformly mixed according to a specific proportion, and then the mixed matrix with specific water holding capacity and certain fertilizer holding capacity can be prepared.
Selecting 3 matrixes of grass carbon, vermiculite and perlite for combination, and designing 7 matrixes in volume ratio for treatment: treatment 1: vermiculite, treatment 2: peat, treatment 3: perlite, treatment 4: the volume ratio of the grass carbon to the vermiculite to the perlite is 3:2: 1, and the treatment is carried out by the following steps: the volume ratio of the grass carbon to the vermiculite to the perlite is 1: 1, and the treatment is carried out according to the following steps: the volume ratio of the grass carbon to the vermiculite to the perlite is 2: 1, and the treatment is carried out according to the following steps: the volume ratio of the grass carbon to the vermiculite to the perlite is 2: 1.
When transplanting the tissue culture seedlings, keeping the water content of the matrix at about 80%, covering the matrix by a small plastic shed to ensure that the environmental conditions in the shed are 70-80% of relative humidity and 22-24 ℃, and watering once at intervals of 4 days every time to ensure that the culture matrix is kept moist.
After culturing for 30 days, counting the survival rate of the strawberry tissue culture seedlings, randomly selecting 10 strawberry tissue culture seedlings from 7 matrixes with different volume ratios as measuring plants, washing the measuring plants with clear water, measuring the fresh weight of the measuring plants on an electronic balance, recording the fresh weight, and measuring the height and the number of new leaves of each strawberry tissue culture seedling by using a vernier caliper.
1. Influence of different acclimatization substrates on strawberry tissue culture seedling transplanting survival rate
As can be seen from table 1, after 30d of transplanting, the volume ratio of grass carbon to vermiculite to perlite in 5-1 is higher than that in other matrix treatment groups, the survival rate is 100%, and the plants grow robustly. The next was treatment 7, but the plants of the strawberry tissue culture seedlings were short. The survival rate of the treatment 1 is higher than that of the treatment 3 than that of the treatment 2, and the treatment 2 is pure turf treatment which may cause poor air permeability and root respiration to cause plant death.
The transplanting of the strawberry tissue culture seedling from the culture medium into the seedling hardening matrix is a process from 'heterotrophic' to 'autotrophic' with the nutrition supplied by the culture medium, and the process is a process for gradually exercising the strawberry tissue culture seedling for adaptation. In the transplanting and acclimatizing process of the strawberry tissue culture seedling, the capabilities of fertilizer preservation, ventilation, water preservation and the like in the matrix are closely related to the survival rate of the tissue culture seedling and the early growth of young plants. The survival rate and the plant state of the strawberry tissue culture seedling are visible, the survival rate of the strawberry tissue culture seedling is the highest when the volume ratio of the matrix is 1: 1, and the strawberry tissue culture seedling grows the most robust.
TABLE 1 Effect of different acclimatization media on the survival rate of strawberry tissue culture seedling transplantation
2. Influence of different acclimatization matrixes on the height of transplanted strawberry tissue culture seedlings
As can be seen from Table 2, the plant height of the treatment 5 in terms of the growth amount of the strawberry tissue culture seedlings is significantly higher than that of other matrixes by 48.13cm, the plant height of the treatment 3 is significantly lower than that of the other matrixes by 24.87cm, and the other matrixes have no obvious difference in the range of 38.85 cm-23.29 cm. The strawberry tissue culture seedlings grow robustly in the treatment 5, and can meet the nutrient elements required in the growth and development of the plants. The pure substrate of treatment 3 is easy to cause poor growth, dwarfing, yellowing and the like because of lack of nutrient elements for plant growth.
In the transplanting and hardening of the strawberry tissue culture seedling, the strawberry tissue culture seedling with strong plant, vigorous growth and good development is selected, which is beneficial to the transplanting and survival of the strawberry tissue culture seedling. The higher the plant height of the strawberry tissue culture seedling is, the better the plant height is, and the seedling may be caused by overhigh elongation or excessive growth. The growth of strawberry tissue culture seedlings can be promoted by proper matrix proportion, the space for root growth can be increased by transplanting and acclimatizing the strawberry tissue culture seedlings, the water and fertilizer retention performance is good, the strawberry tissue culture seedlings can grow more favorably, and the tissue culture seedlings can grow by transplanting and acclimatizing the strawberry tissue culture seedlings. The experiment analysis shows that the strawberry tissue culture seedling has the largest growth amount in the matrix volume ratio of turf, vermiculite and perlite which is 1: 1, so that the treatment 5 is the most suitable matrix ratio of the strawberry tissue culture seedling.
TABLE 2 influence of different acclimatization media on the height of the transplanted strawberry tissue culture seedlings
3. Influence of different acclimatization substrates on new leaf number of transplanted strawberry tissue culture seedlings
The strawberry tissue culture seedlings are planted and refined in matrixes with different volume ratios, and the number of new leaves of the strawberry tissue culture seedlings is observed and recorded, and the results are shown in table 3. The treatment 1, the treatment 2 and the treatment 5 are all 6 pieces in the aspect of extracting new leaves from the strawberry tissue culture seedling, and no obvious difference exists. The treatment 4 and the treatment 7 are 5 pieces in the aspect of extracting new leaves from the strawberry tissue culture seedlings. Treatment 3 was significantly lower than treatment 4 and treatment 7 was 5 pieces, and treatment 6 was significantly lower than the other substrates were 4 pieces. In the tissue culture seedling transplantation and acclimatization, the treatment 6 is obviously lower than other matrixes, and the bottom leaves are often wilted and withered and yellowed. And the number of the new leaves extracted by other matrixes is not obviously different.
The number of new leaves of the strawberry tissue culture seedling extracted in different matrixes has different degrees, the strawberry tissue culture seedling has wilting and slight yellow leaves with different degrees in the process of adapting to a new environment, and new leaves are gradually extracted after a period of adaptation. In conclusion, the reasonable configuration matrix can meet the requirements of growth and development of strawberry tissue culture seedlings and nutrient elements required for extracting new leaves and the like. The humus soil has better fertilizer and water retention performance than nutrient soil, and is beneficial to the extension of root systems, thereby being beneficial to the growth of new leaves on the overground part. According to experimental analysis, the influence of different refining matrixes on the number of new leaves of the strawberry tissue culture seedlings is not great.
TABLE 3 Effect of different acclimatizing bases on the number of new leaves transplanted to tissue culture seedlings of strawberry
4. Influence of different acclimatization substrates on fresh weight of transplanted strawberry tissue culture seedlings
The fresh weight change degree of the strawberry tissue culture seedlings before and after growth is compared by observing the strawberry tissue culture seedlings in matrixes with different volume ratios and recording data. From table 4, it can be seen that the fresh weight of the strawberry tissue culture seedling in treatment 5 is significantly higher than that of other substrates by 0.75g, the fresh weight of the strawberry tissue culture seedling in treatment 1 is significantly lower than that of the other substrates by 0.40g, and the other substrates have no significant difference in the range of 0.50g to 0.60 g. The fresh weight of the treated 5 is the largest and better than that of other matrixes, and the nutrient elements in the matrixes can meet the growth and development of the strawberry tissue culture seedlings. The fresh weight of treatment 1 was minimal and inferior to that of the other substrates, probably due to environmental discomfort during the growth of the strawberry tissue culture seedlings.
TABLE 4 Effect of different acclimatizing bases on the number of new leaves transplanted to tissue culture seedlings of strawberry
Although the embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that: various substitutions, changes and modifications are possible without departing from the spirit and scope of the invention and the appended claims, and therefore the scope of the invention is not limited to the embodiments disclosed.
Claims (7)
1. A strawberry tissue culture seedling acclimatization substrate for improving the acclimatization quality of strawberry tissue culture seedlings is characterized in that: each 10L of the matrix comprises the following components in percentage by weight:
grass carbon: 2.5-4L;
vermiculite: 2.5-3.5L;
perlite: 2-3L;
sodium hypochlorite or clear water: 1000-;
trace fertilizer: the element ratio of N to P to K is 3:2: 2.
2. The strawberry tissue culture seedling acclimatization substrate for improving strawberry tissue culture seedling acclimatization quality according to claim 1, characterized in that: the fiber length of the turf is 1-10 mm; the particle size of the vermiculite is 3-5 mm; the particle size of the perlite is 3-6 mm; the available chlorine of the sodium hypochlorite is 0.01 percent.
3. The method for preparing a strawberry tissue culture seedling acclimatization substrate for improving the acclimatization quality of strawberry tissue culture seedlings according to claim 1 or 2, wherein the method comprises the following steps: the method comprises the following steps:
the grass carbon, the vermiculite and the perlite are uniformly mixed, then the grass carbon, the vermiculite and the perlite which are uniformly mixed are pre-wetted by adding sodium hypochlorite or clear water, then the trace fertilizer is added, and the mixture is uniformly mixed, so that the strawberry tissue culture seedling hardening matrix for improving the hardening quality of the strawberry tissue culture seedlings is obtained, wherein the water content of the matrix is 50-60%.
4. The method for using the strawberry tissue culture seedling acclimatization substrate for improving the acclimatization quality of strawberry tissue culture seedlings according to claim 1 or 2, characterized by comprising the following steps: the method comprises the following steps:
the strawberry tissue culture seedlings are processed by chlorothalonil and then cultured by utilizing a strawberry tissue culture seedling hardening matrix.
5. Use according to claim 4, characterized in that: the chlorothalonil is 2g/L aqueous solution of chlorothalonil with the effective component content of 75%.
6. Use according to claim 4, characterized in that: and (3) soaking the strawberry tissue culture seedlings in 2g/L chlorothalonil water solution for 30min, and performing disinfection treatment.
7. The application of the strawberry tissue culture seedling acclimatization substrate for improving the acclimatization quality of strawberry tissue culture seedlings according to claim 1 or 2 in the aspect of improving the acclimatization quality of strawberry tissue culture seedlings.
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