CN114600891A - Composition and method for seed germination - Google Patents
Composition and method for seed germination Download PDFInfo
- Publication number
- CN114600891A CN114600891A CN202210228705.3A CN202210228705A CN114600891A CN 114600891 A CN114600891 A CN 114600891A CN 202210228705 A CN202210228705 A CN 202210228705A CN 114600891 A CN114600891 A CN 114600891A
- Authority
- CN
- China
- Prior art keywords
- agar
- gibberellin
- seeds
- component
- acer truncatum
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
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Images
Classifications
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- A—HUMAN NECESSITIES
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- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
- A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
- A01N43/06—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom five-membered rings
- A01N43/12—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom five-membered rings condensed with a carbocyclic ring
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
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- A—HUMAN NECESSITIES
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- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/02—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing liquids as carriers, diluents or solvents
- A01N25/04—Dispersions, emulsions, suspoemulsions, suspension concentrates or gels
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/44—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a nitrogen atom attached to the same carbon skeleton by a single or double bond, this nitrogen atom not being a member of a derivative or of a thio analogue of a carboxylic group, e.g. amino-carboxylic acids
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- A—HUMAN NECESSITIES
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- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
- A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
- A01N43/14—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings
- A01N43/16—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings with oxygen as the ring hetero atom
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N45/00—Biocides, pest repellants or attractants, or plant growth regulators, containing compounds having three or more carbocyclic rings condensed among themselves, at least one ring not being a six-membered ring
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Environmental Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Engineering & Computer Science (AREA)
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- Pest Control & Pesticides (AREA)
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- Agronomy & Crop Science (AREA)
- Chemical & Material Sciences (AREA)
- Dispersion Chemistry (AREA)
- Toxicology (AREA)
- Soil Sciences (AREA)
- Pretreatment Of Seeds And Plants (AREA)
Abstract
The present invention relates to a composition for seed germination comprising: a first component that promotes plant growth; a second component that provides soluble sugars for initial plant growth; storing a third component of the first and second components, wherein the first and second components are mixed in the third component in a gel form, and allowing the plants to simultaneously contact air and the first and second components mixed in the third component at the initial growth. The present invention relates to a composition capable of increasing the germination rate of seeds by increasing the efficiency of uptake of diterpene compounds by the seeds.
Description
Technical Field
The invention relates to the technical field of agricultural planting, in particular to a composition and a method for seed germination.
Background
Acer Truncatum (Acer Truncatum), also known as Acer Truncatum, is a tree of Sapindales, Aceraceae, Acer genus, deciduous tree. The height of the Acer truncatum adults is 8-10 m, and the diameter at breast height is 80-180 cm. The acer truncatum is originally produced in the north of China and is distributed scattered in secondary forests. The crown of the acer truncatum is wide and round, the bark is gray yellow to gray, the acer truncatum has longitudinal split stripes, the small branches are bright and unhaired, and the branches grow for 1 year. The leaves of Acer Truncatum Bunge are pale reddish brown or green with crimson, and then gray. The acer truncatum bunge has the advantages of opposite single leaves, 5-shaped palms, 5-10 cm long, 6-15 cm wide, full edges, gradually tapered tip at the tip and winged fruit. The acer truncatum crown is shade, the tree appearance is beautiful, the leaf shape is beautiful, and the leaf color is gorgeous. The leaves in autumn are mostly bright red or golden yellow, and are famous autumn leaves and tree species in northern China. The acer truncatum has a special status in landscaping due to the characteristics of barren resistance, drought resistance, low temperature resistance, strong adaptability and the like.
Because the special physiological dormancy type of the acer truncatum, the germination rate of acer truncatum seeds is only 4%, and in the prior art, a new acer truncatum single plant individual is obtained mostly through a tissue transplanting or grafting mode. Chinese patent application publication No. CN107173139A discloses a seedling growing method of acer truncatum, which comprises the following steps: 1) selecting land and preparing land; 2) processing acer truncatum slips; 3) transplanting acer truncatum; 4) managing after the transplanting; 5) seedling stage management: and (3) applying the mixed fertilizer for 1 time in 8-9 months in the first year, wherein the mixed fertilizer comprises a vinasse, bean cake powder and a compound fertilizer, digging a fertilizing pit in each acer truncatum seedling pit, loading the compound fertilizer into the fertilizing pit, and then returning soil, wherein the fertilizing amount in each pit is 40-60 g, the fertilizing period is 6-10 months, and the acer truncatum trees can be listed in the fifth year after planting. However, the transplanting or grafting means requires a great deal of energy for maintenance, resulting in a waste of energy during the cultivation process.
The Chinese patent application with publication number CN107223512A discloses a method for breeding and strengthening ornamental Acer truncatum seedlings, which comprises the following steps: selecting proper land and conditionsThe test has the advantages that the test has good irrigation conditions, flat terrain, loose soil texture, good section drainage and sandy loam texture, the test has convenient irrigation conditions, loam texture and moderate fertility; preparing soil and making bed, deeply turning over garden before winter and applying base fertilizer every hm for soil curing2Applying wet manure 45t, diamine and calcium superphosphate of 750kg respectively, and performing shallow ploughing and land preparation to form furrows before sowing in spring, wherein the width of each furrow is 1.2-1.5 m; the method comprises the steps of treating seeds before sowing, wherein germination accelerating and direct sowing of dry seeds are carried out, wherein the germination accelerating comprises seed soaking for 1 day, 2 days or 3 days by clear water, seed soaking and germination accelerating of a bamboo basket are carried out after seed soaking, the germination accelerating method of the bamboo basket comprises the steps of soaking full seeds for one day by warm water (35-41 ℃) one week before sowing, taking out the seeds, putting the seeds into the bamboo basket, covering a straw curtain, placing the bamboo basket at the room temperature of 20-30 ℃, watering and turning for 2-3 times every day until the 'split-tip white-exposed' seeds reach 20%, and then sowing. In actual operation, the operation of peeling off the seed coat is complicated and easy to damage the seedling, and the pollution of the seed embryo is easy to cause in the peeling process, so that the seed is dead.
The invention designs a composition and a method capable of increasing the germination rate of acer truncatum seeds, and enables acer truncatum to germinate through simple culture medium batch induction based on the composition and the method, thereby reducing the cultivation cost of acer truncatum.
Furthermore, on the one hand, due to the differences in understanding to the person skilled in the art; on the other hand, since the applicant has studied a great deal of literature and patents when making the present invention, but the disclosure is not limited thereto and the details and contents thereof are not listed in detail, it is by no means the present invention has these prior art features, but the present invention has all the features of the prior art, and the applicant reserves the right to increase the related prior art in the background.
Disclosure of Invention
In view of the deficiencies of the prior art, the present invention provides a composition and method for seed germination.
A composition for seed germination comprising: a first component consisting of a plant growth hormone; a second component that provides soluble sugars for initial plant growth; a third component that provides both air and moisture for the initial growth of the plant. The first and second components are mixed in a third component in the form of a gel, and the plant is allowed to simultaneously contact air and the first and second components mixed in the third component at the time of initial growth. The plant growth comprises four processes of seed imbibition, germination and seedling growth, wherein the composition mainly acts on the germination and germination processes of seeds, on one hand, the success rate of the germination and the germination of the seeds can be improved, and on the other hand, the germination and the germination rate of the seeds can be improved.
The technical scheme has the advantages that: in the prior art, the commercial cultivation of seeds mostly adopts a soaking method of clear water or a germinator mixed with clear water to break the dormancy condition of the acer truncatum seeds. However, water and oxygen are indispensable in the germination process of acer truncatum seeds. In contrast to the anoxic germination process of acer truncatum seeds soaked in water, the third component can provide a support carrier that exposes a portion of the surface of the acer truncatum seeds to air and absorbs oxygen. The surface of the acer truncatum bunge attached carrier can absorb soluble sugar and diterpene compounds mixed in the carrier to grow normally. Diterpene compounds and soluble sugars are both small molecules. The diterpene compound and the soluble sugar can quickly enter the acer truncatum seeds, so that the influence of the high-thickness seed coats of the acer truncatum on the efficiency of the acer truncatum seeds for absorbing the diterpene compound is avoided.
According to a preferred embodiment, the first component can comprise diterpene compounds which are involved in mediating and inhibiting soluble starch synthesis in the primary growth of plants. During the germination process of the seeds, the generation of soluble sugar can inhibit the germination process. The invention adopts diterpene compounds which can inhibit the synthesis of soluble starch, thereby regulating the germination process of acer truncatum seeds at the physiological mechanism level. Preferably, the content of the first component can be in the range of 0.1mg/L to 1 mg/L.
According to a preferred embodiment, the second component further comprises a small molecule peptide consisting of hydrolysed casein. The hydrolyzed casein is a mixture of various amino acids, and has good promoting effect on embryoid and adventitious bud differentiation. Preferably, the concentration of hydrolyzed casein in the composition ranges from 0.1mg/L to 0.5 g/L.
According to a preferred embodiment, the second component comprises 0.5g/L hydrolysed casein and 30g/L sucrose.
According to a preferred embodiment, the third component comprises water and a coagulating agent in a form capable of forming a solid which supports and grows plants or plant tissue on its surface. The third component forms a gel having a pore size such that the first component and the second component are stored in the pore size of the gel. Preferably, the carrier comprises agar powder.
According to a preferred embodiment, the diterpene compound comprises gibberellin 3. Gibberellin 3 can be present in the composition as a primary component that promotes germination of acer truncatum seeds. The component gibberellin 3, also known as GA, in the compositions referred to in this application3。
According to a preferred embodiment, the first component comprises 0.1mg/L to 1mg/L of GA3。
According to a preferred embodiment, the third component comprises water and agar.
A composition for seed germination of Acer truncatum Bunge comprises agar and gibberellin 3. The mixture ratio of the agar and the gibberellin 3 is any one of the following a1) -a 5): a1) the method comprises the following steps 0.05mg/L gibberellin 3: 7g/L agar; a2) the method comprises the following steps 0.1mg/L gibberellin 3: 7g/L agar; a3) the method comprises the following steps 0.25mg/L gibberellin 3: 7g/L agar; a4) the method comprises the following steps 0.5mg/L gibberellin 3: 7g/L agar; a5) the method comprises the following steps 1.0mg/L gibberellin 3: 7g/L agar.
A method for seed germination, contacting a plant or plant tissue with a composition according to any one of the following a1) to a 5): a1) the method comprises the following steps 0.05mg/L gibberellin 3: 7g/L agar; a2) the method comprises the following steps 0.1mg/L gibberellin 3: 7g/L agar; a3) the method comprises the following steps 0.25mg/L gibberellin 3: 7g/L agar; a4) the method comprises the following steps 0.5mg/L gibberellin 3: 7g/L agar; a5) the method comprises the following steps 1.0mg/L gibberellin 3: 7g/L agar.
A method of preparing a composition for seed germination comprising the steps of: dissolving sucrose and agar in water, and heating until solid components are completely dissolved; adding 0.05-1.0 mg/L gibberellin 3, uniformly stirring, and adjusting the pH value to be weakly acidic to obtain the composition for seed germination of acer truncatum bunge. Preferably, the sucrose content of the composition is 3%. The content of agar in the composition was 7 g/L.
According to a preferred embodiment, the composition comprises from 20 to 30g/L of sucrose, from 6 to 0.7g/L of agar, from 0.1 to 1g/L of hydrolysed casein and from 0.1 to 1mg/L of GA3The pH of the solution is 5.6-5.8.
According to a preferred embodiment, the plant or plant tissue is soaked in water before being contacted with the composition in such a way that dormancy of said plant or plant tissue is broken. The germination speed of the acer truncatum seeds is increased by soaking and subsequently adding gibberellin, so that the germination success rate of the acer truncatum seeds is improved.
According to a preferred embodiment, the plant tissue is a plant seed or callus involved in the initial growth of the plant. In the prior art, callus of acer truncatum can also realize efficient germination and growth by using the composition based on directional induction of a culture medium.
Drawings
FIG. 1 is a line graph of germination percentage of Acer truncatum seeds provided by the present invention;
FIG. 2 is a line graph of the germination rate of the seeds of the Acer truncatum Bunge of the invention;
FIG. 3 is a line graph showing the germination percentage of "brilliant red" acer truncatum seeds provided by the present invention.
Detailed Description
The following detailed description is made with reference to the accompanying drawings.
The acer truncatum has ornamental value, industrial value and practical value. In the aspect of ornamental value, the acer truncatum bunge is a multi-effect beneficial tree species integrating edible oil, tanning materials, proteins, medicinal, chemical, water and soil conservation, special materials and landscaping and appreciation. The acer truncatum is a tree species for greening roadways, courtyards and scenic spots, and has beautiful tree shape and dense branches and leaves. The autumn leaves of Acer truncatum have early color change and long duration, and are changed into yellow, orange and red. Acer truncatum is an excellent foliage tree species. In urban greening, the acer truncatum is suitable for scattered planting near buildings, in courtyards and in green lands; the method can also achieve better effect by planting the plants in the suburb park by utilizing the sloping fields. The leaves of Acer Truncatum Bunge are rich in color, the spring leaves are red and bright, the autumn leaves are golden, the leaves can be picked for several times, and the new leaves are small and red after picking, so the Acer Truncatum Bunge is a very special pile scene material. The acer truncatum has strong adaptability and extensive management, and is a main tree species of stump bonsai in Beijing areas.
In the aspect of industrial value, the Acer Truncatum Bunge wood is tough and delicate and can be used as an appliance. The seeds can be used for oil extraction, and can be used for food and industrial purposes. The bark fiber can be used for making paper and cotton substitute.
In terms of edible value, the oil of acer truncatum has a similar fatty acid composition to that of sesame oil and peanut oil which are generally edible, and the content of essential fatty acid is high. The oil of Acer Truncatum Bunge can be used for frying. The hot-pressed acer truncatum buge oil is used as oral health-care oil, has better taste than seabuckthorn oil, and has no peculiar smell. The acer truncatum buge oil has rich fat-soluble vitamin content, high vitamin E content and good oxidation resistance, and is more storage-resistant than sea buckthorn oil and walnut oil. The oil of the acer truncatum bunge kernel is rich in various essential fatty acids and fat-soluble vitamins of a human body, wherein the content of unsaturated fatty acids is up to 92 percent, particularly the content of nervonic acid is 5.52 percent, and the acer truncatum bunge kernel has extremely high health care effect.
Acer truncatum, as a plant with high economic value, has important effects on the cost due to the growth rate and the cultivation environment. The dormancy type of the acer truncatum seeds is physiological dormancy, and the germination rate of newly harvested seeds is only 4 percent. Thus, acer truncatum, which is cultured independently on an individual plant depending on seed germination, needs a culture composition and method capable of enhancing the seed germination rate thereof.
The seed dormancy refers to the characteristic that the seeds with germination vigor still cannot germinate under the appropriate germination conditions. Generally, the degree of dormancy of seeds is related to the water content of the seeds, and when the water content in the seeds decreases to a certain degree, the seeds enter a dormant state. In order to release dormancy of seeds and promote germination of seeds, dry after-ripening, stratification treatment, light irradiation, and treatment with hormones such as gibberellin are often used.
In the prior art, the seeds are germinated by simple soaking in clear water or adding a germinant for soaking. The seeds are deprived of oxygen during germination by the seeds, thereby causing a problem of bad seeds. The acer truncatum seeds are too thick in skin and have endogenous hormones or substances for inhibiting seed germinationThe seed embryo is blocked in the germination process, so that the problem that how to break the dormancy of acer truncatum seeds, rapidly produce a large number of aseptic seedlings and ensure the material source established by the acer truncatum genetic system is urgently needed to be solved at present. The experiment innovating the traditional germination accelerating method and utilizing the tissue culture technology, provides a culture method for promoting the germination of acer truncatum seeds. By adjusting temperature and GA3Working concentration and changing the germination environment of the acer truncatum seeds to improve the germination rate of the acer truncatum seeds. The application carries out contrast research through sprouting different varieties acer truncatum seeds to improve the germination rate of acer truncatum.
Example 1
Three varieties of acer truncatum seeds are adopted in the test, namely common acer truncatum seeds, acer truncatum seeds with the variety of beautiful red and acer truncatum seeds with the variety of brilliant red. The MS medium used in this experiment contained: 1/2MS + 3% sucrose +0.5g/L hydrolysed casein +7g/L agar, pH 5.8.
In particular, the testing step comprises pretreatment, inoculation and cultivation of the seeds.
The pretreatment of the seeds comprises the following steps:
selecting three plump acer truncatum seeds, removing fruit wings, cleaning with clear water dropwise added with Tween 20, and transferring the seeds to an aseptic beaker;
adding sterile water, rinsing for 3 minutes, sterilizing with 75% alcohol for 1 minute, and rinsing with sterile water for 3 minutes for 2 times;
cleaning with sterile water, disinfecting with a sodium hypochlorite solution for the first time, adding a 20% sodium hypochlorite solution (containing 1-2 drops of Tween 20) into a test tube, rinsing for 10 minutes, rinsing with sterile water for 3 times, 3 minutes each time, and continuously shaking to fully wash seeds;
repeating the sodium hypochlorite solution sterilization process once;
the seeds were soaked in sterile water for 10 minutes.
The inoculation comprises the following steps:
sterilizing the seeds, and then sucking water by using filter paper;
dividing three varieties of acer truncatum seeds into a control group and a first treatment group;
soaking three varieties of acer truncatum seeds of a control group in clear water for 4 hours, sterilizing, sucking water, and inoculating on a blank MS culture medium;
three varieties of acer truncatum seeds of the first treatment group are soaked in clear water for 4 hours, sterilized and dried by suction, and then inoculated in additional GA3In the MS medium of (1), wherein, GA3The concentrations were 0(CK), 0.05mg/L, 0.1mg/L, 0.25mg/L, 0.5mg/L and 1.0mg/L, respectively.
The cultivation comprises the following steps:
the inoculated seeds are placed in a refrigerator at 15 ℃ and cultured in the dark. After about 1 week, when seeds germinate in each culture dish, taking out and transferring to a culture room for illumination culture under the conditions of 16 hours of illumination/8 hours of darkness and the temperature (24 +/-2) DEG C.
The germination rates were counted on days 7, 14, and 21, respectively, and the growth state of the seedlings was observed. The germination rate is (number of germinated seeds/total number of inoculated seeds) ×%. The embryo root elongation of 2mm is taken as the standard of seed germination. The test results were processed and analyzed by Excel, and plotted by Origin software.
As shown in FIGS. 1-3, the seeds of Acer truncatum were inoculated with GA at different concentrations3The germination rate of the MS medium (2) was determined in 21 days, GA3The treatment has slightly different pregermination effects on the acer truncatum seeds of 3 varieties. The content of common Acer Truncatum Bunge seed in GA of 0, 0.1, 0.25, 0.5 and 1.0mg/L3The germination rates under catalysis are 41%, 78%, 61%, 36% and 31% in sequence; the germination rates of the 'Lihong' acer truncatum seeds are 74%, 92%, 86%, 68% and 60% in sequence.
To verify 0-0.1 mg/L GA3Whether the peak value still exists in the middle, and GA is adjusted3The working concentration of (1) is 0, 0.05, 0.1, 0.25 and 0.5mg/L, and the germination rate of the "brilliant red" acer truncatum is 73%, 72%, 85%, 75% and 70% in sequence according to statistical analysis for 21 days.
According to GA at different concentrations in this experiment3The cultivation of (1) shows that gibberellin 3 with a concentration of 0.25mg/L in MS medium has an obvious effect on seed germination of Acer truncatum BungeThe application is as follows. Inoculating Acer Truncatum Bunge seed, processing in dark at 15 deg.C, and adding 0.1mg/L GA3Is suitable concentration for breaking seed dormancy and promoting seed germination. But GA3Has different promoting effects on acer truncatum of different varieties, has the most obvious pregermination effect in the seeds of the common acer truncatum, and has the GA content of 0.1mg/L3And 0.25mg/L GA3The germination rate is obviously improved under the treatment; because the seeds of the Lihong acer truncatum have plump kernels, the germination rate of a control group can reach 74 percent and is higher than 0.5mg/L GA3And 1.0mg/L GA 368% and 60% of (B), but less than 0.1mg/L of GA3And 0.25mg/L GA 392% and 86%; in "brilliant red" acer truncatum, 0.1mg/L GA3The treated seeds still have the highest germination rate, but the germination accelerating effect is not obvious from the control group, and the germination of the seeds is inhibited by other concentration treatments.
According to a preferred embodiment, the concentration of diterpenoids in the present invention can be in the range of 0.1-0.5 mg/L, wherein 0.1mg/L GA3Is the optimum concentration for promoting the germination of the acer truncatum seeds and breaking the dormancy of the acer truncatum seeds.
According to a preferred embodiment, the GA3When the concentration is increased to 0.5mg/L or more, particularly 1mg/L, the method not only can inhibit the germination of seeds, but also seedlings germinated from the seeds are easy to have deformity, short and thick stem nodes and abnormal expansion of roots in the later growth and development process. Therefore, the concentration of the diterpenoid compound applicable to the seeds can be in the range of 0.1-0.25 mg/L. Addition of GA based on the growth status of seedlings at each inoculation concentration3The acer truncatum seeded on the culture medium is easier to have lateral buds than acer truncatum of a control group, the high growth is not obviously limited, and other phenotypic characteristics have no difference with seedlings of the control group, so that GA in the concentration range is not added3The addition of the compound has no negative influence on the production standard of the aseptic seedlings, and the composition and the cultivation method related in the invention can meet the material guarantee established by a subsequent regeneration system and even a genetic transformation system.
According to a preferred embodiment, the method is used for promoting germination of acer truncatum seeds3Mixed in the second groupThe soluble sugar can be sucrose, glucose, lactose or maltose.
Example 2
This example was carried out in substantially the same manner as in example 1 except for the following.
The test adopts three varieties of acer truncatum seeds, namely common acer truncatum seeds, beautiful red acer truncatum seeds and gorgeous red acer truncatum seeds.
In particular, the test steps comprise pretreatment, inoculation and cultivation of the seeds.
The pretreatment of the seeds comprises the following steps:
selecting three plump acer truncatum seeds, removing fruit wings, cleaning with clear water dropwise added with Tween 20, and transferring the seeds to an aseptic beaker;
adding sterile water for rinsing for 3 minutes, then sterilizing for 1 minute by using 75% alcohol, and then rinsing for 3 minutes for 2 times by using sterile water;
cleaning with sterile water, disinfecting with a sodium hypochlorite solution for the first time, adding a 20% sodium hypochlorite solution (containing 1-2 drops of Tween 20) into a test tube, rinsing for 10 minutes, rinsing with sterile water for 3 times, 3 minutes each time, and continuously shaking to fully wash seeds;
repeating the sodium hypochlorite solution sterilization process once;
the seeds were soaked in sterile water for 10 minutes.
The inoculation comprises the following steps:
sterilizing the seeds, and then sucking water by using filter paper;
dividing three varieties of acer truncatum seeds into a control group, a first treatment group and a second treatment group;
soaking three varieties of acer truncatum seeds of a control group in clear water for 4 hours, sterilizing, sucking water, and inoculating on a blank MS culture medium;
selecting GA with moderate concentration from three varieties of acer truncatum seeds of the first treatment group3(0.25mg/L) the seed soaking treatment was carried out for 4 hours, sterilized and dried by suction, and then inoculated on a blank MS medium;
three varieties of acer truncatum seeds of the second treatment group are soaked in clear water for 4 hours, sterilized and inoculated in additional GA after moisture is absorbed3In the MS medium of (1), wherein, GA3The concentrations were 0.25mg/L, respectively.
The test results are shown in the following table 1, wherein the germination rates of the acer truncatum seeds under different inoculation matrixes are respectively as follows:
TABLE 1
As shown in Table 1, the seeds were inoculated with 0.25mg/L GA after soaking in clean water for 4 hours3The germination effect of the seeds on the MS culture medium is the best, and the germination rate can reach 61% in 21 days; 0.25mg/L GA3The germination accelerating effect of the seed after soaking for 4 hours is next to that of the seed after inoculating on a blank MS culture medium, and the germination rate in 21 days is 47%; the 21-day germination rate of the control group is only 41 percent. Additional GA3The MS culture medium proves to have a certain positive effect on the germination of the acer truncatum seeds, and is a proper treatment mode.
The seed germination rate of acer truncatum seeds is low, the seed embryos have no dormancy phenomenon, the seed coats of acer truncatum are considered by Linshijie and the like to be main factors influencing the germination of the acer truncatum seeds, firstly, the seed coats are too thick to block the germination, and secondly, the seed coats contain endogenous hormones or substances for inhibiting the seed germination. However, the germination rate of the in vitro embryo can reach 100%, but the peeling process is complex and easy to cause secondary pollution, and the method cannot be used as a means for mass production of aseptic seedlings. On one hand, the invention adopts the germinator capable of promoting the seed germination aiming at the seed germination of the acer truncatum bunge, and on the other hand, the germinator can avoid seed coats and enter the seed bodies based on a special application method, so that the promoting efficiency of the germinator on the seed germination is increased.
In the prior art, diterpenoid compounds (including GA) are used1、GA2、GA3Etc.), break seed dormancy, weaken seed coat resistance in the later stage of seed germination, and help radicle break through seed coat to successfully germinate. However, in the prior art, the diterpenoid compounds mostly soak the seeds in the form of aqueous solution to influence the seed hairAnd (4) bud.
In the present invention, because the seed skin of acer truncatum is thicker, GA is used3In the seed soaking process, the hormone absorption degree of the seeds is not uniform, the effect is not obvious, so that the GA is applied3Is mixed with nutrients required for germination and stored in a gel-like solid carrier to allow GA to grow3Can form a synergistic passive transportation along the concentration gradient difference with the nutrient substances in the seed absorption process. Different hormone application modes are adopted to influence the germination rate of acer truncatum seeds, and GA is obtained under the same concentration3The germination rate of the soaked mode is lower than that of the GA3Addition to MS medium. Hormone is added into the culture medium, the culture medium continuously provides nutrition for the seeds in the germination process, the seeds can contact the hormone for a longer time, and the hormone is absorbed more sufficiently, so that when the acer truncatum aseptic seedlings are produced, the acer truncatum seeds are added to GA3The culture medium ensures that the acer truncatum seeds have high germination rate and form a more convenient operation process.
According to a preferred embodiment, the seeds of acer truncatum have more endophytes, the ordinary sterilization mode cannot completely remove bacteria and fungi, and the seeds are easy to be polluted greatly in the tissue culture process. Most of the prior art uses mercuric chloride to disinfect seeds of acer truncatum. Although highly toxic disinfectants such as mercuric chloride have a certain sterilization effect, the harm is large, and the promotion and the use are not suitable. Based on GA3Under the condition of supplying the same concentration, the hormone for nutrition and germination is provided for the acer truncatum seeds in a mode of soaking in clear water or supplying culture medium. The Tween 20 can enable the disinfectant to deeply contact with the seed coat, and effectively adsorb endophytes of seeds. In the experiment, the sodium hypochlorite solution dropwise added with tween 20 is used for disinfecting the acer truncatum seeds, so that the acer truncatum bunge is non-toxic and environment-friendly, and the pollution rate of the seeds can be effectively reduced.
Based on the test, the invention considers that the low-temperature condition has positive effect on the germination of the seeds of three varieties of acer truncatum bunge. The seed germination temperature of the acer truncatum bunge of the invention can be 15 ℃.
According to a preferred embodiment, the GA concentration is in the range of 0.1-0.25 mg/L3Mixing with sucrose, adding agar to form flat culture medium, and inoculating Acer Truncatum Bunge seedInoculating on the surface of culture medium, and processing in dark at 15 deg.C until Acer Truncatum Bunge seed germinates.
According to a preferred embodiment, the carrier present in the third component of the present invention can comprise one or more of agar-agar, gelatin, foamed cotton, pectin, silica gel, carrageenan, xanthan gum or cellulose. The carrier can be a MS medium comprising water and agar.
It should be noted that the above-mentioned embodiments are exemplary, and that those skilled in the art, having benefit of the present disclosure, may devise various arrangements that are within the scope of the present disclosure and that fall within the scope of the invention. It should be understood by those skilled in the art that the present specification and figures are illustrative only and are not limiting upon the claims. The scope of the invention is defined by the claims and their equivalents. The present description contains several inventive concepts, such as "preferably", "according to a preferred embodiment" or "optionally", each indicating that the respective paragraph discloses a separate concept, the applicant reserves the right to submit divisional applications according to each inventive concept. Throughout this document, the features referred to as "preferably" are only an optional feature and should not be understood as necessarily requiring that such applicant reserves the right to disclaim or delete the associated preferred feature at any time.
Claims (10)
1. A composition for germination of seeds, comprising:
a first component consisting of a plant growth hormone;
a second component that provides soluble sugars for initial plant growth;
storing a third component of the first component and the second component,
wherein the first and second components are mixed in the third component in a gel form and allow the plants to simultaneously contact air and the first and second components mixed in the third component at the time of initial growth.
2. The composition of claim 1, wherein the first component can comprise a diterpene compound that is involved in mediating and inhibiting soluble starch synthesis in the primary growth of plants.
3. The composition of claim 1 or 2, wherein the soluble sugar is a monosaccharide or disaccharide.
4. The composition according to any one of claims 1 to 3, wherein the second component further comprises a polypeptide consisting of hydrolysed casein.
5. The composition of any one of claims 1 to 4, wherein the disaccharide can be sucrose.
6. The composition of any one of claims 1 to 5, wherein the third component comprises agar.
7. A composition according to any one of claims 1 to 6, wherein the diterpene compound comprises gibberellin 3.
8. A composition for promoting seed germination, comprising agar and gibberellin 3, wherein the ratio of the agar to the gibberellin 3 is any one of the following a1) -a 5):
a1) the method comprises the following steps 0.05mg/L gibberellin 3: 7g/L agar;
a2) the method comprises the following steps 0.1mg/L gibberellin 3: 7g/L agar;
a3) the method comprises the following steps 0.25mg/L gibberellin 3: 7g/L agar;
a4) the method comprises the following steps 0.5mg/L gibberellin 3: 7g/L agar;
a5) the method comprises the following steps 1.0mg/L gibberellin 3: 7g/L agar.
9. A method for seed germination, characterized in that a plant or plant tissue is contacted with a composition according to any of the following a1) -a 5):
a1) the method comprises the following steps 0.05mg/L gibberellin 3: 7g/L agar;
a2) the method comprises the following steps 0.1mg/L gibberellin 3: 7g/L agar;
a3) the method comprises the following steps 0.25mg/L gibberellin 3: 7g/L agar;
a4) the method comprises the following steps 0.5mg/L gibberellin 3: 7g/L agar;
a5) the method comprises the following steps 1.0mg/L gibberellin 3: 7g/L agar.
10. A method of preparing a composition for seed germination comprising the steps of:
dissolving sucrose and agar in water, and heating until solid components are completely dissolved;
adding 0.05-1.0 mg/L gibberellin 3, uniformly stirring, and adjusting the pH value to be weakly acidic to obtain the composition for seed germination of acer truncatum.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN116671437A (en) * | 2022-07-27 | 2023-09-01 | 北京市园林绿化科学研究院 | Acer truncatum culture medium and application thereof |
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| CN108077070A (en) * | 2016-11-21 | 2018-05-29 | 四川七彩林业开发有限公司 | A kind of maple tissue cultures culture medium and cultural method |
| CN110604057A (en) * | 2019-10-17 | 2019-12-24 | 中国林业科学研究院亚热带林业研究所 | Regeneration method of camellia oleifera regeneration system |
| CN112806264A (en) * | 2021-01-26 | 2021-05-18 | 中国科学院华南植物园 | Tissue culture and rapid propagation method for Acer catalpa |
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| CN103404438A (en) * | 2013-08-06 | 2013-11-27 | 巴中市光雾山植物研究所 | Acer paimatum seed tissue culture method |
| CN108077070A (en) * | 2016-11-21 | 2018-05-29 | 四川七彩林业开发有限公司 | A kind of maple tissue cultures culture medium and cultural method |
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| CN116671437A (en) * | 2022-07-27 | 2023-09-01 | 北京市园林绿化科学研究院 | Acer truncatum culture medium and application thereof |
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