CN111149465A - A method for promoting morphological post-ripening of Epimedium koreanum seed embryos - Google Patents
A method for promoting morphological post-ripening of Epimedium koreanum seed embryos Download PDFInfo
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Classifications
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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
-
- 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
- A01C1/06—Coating or dressing seed
-
- 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
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Soil Sciences (AREA)
- Botany (AREA)
- Pretreatment Of Seeds And Plants (AREA)
Abstract
本发明属于植物的繁殖技术领域，具体涉及一种促进朝鲜淫羊藿种胚形态后熟的方法。本申请的促进朝鲜淫羊藿种胚形态后熟的方法，通过选种，并将选取的种子用油菜花提取液浸泡10‑20小时后，倾去水，再将种子与河砂拌匀后装入保温板槽中，覆盖黑色带孔塑料薄膜置于适宜的温度和湿度环境中培养，获得具有子叶胚的种子，该种子的处理时间仅为45天，可操作性强，子叶胚发生率达到95％以上；最后将具有子叶胚的种子连同河砂直接散播于土畦上，并覆盖细土，浇水后覆盖落叶松松针，获得朝鲜淫羊藿育苗，该育苗的发芽率可达90％以上，大幅提高了子叶胚发生率、缩短了处理时间和提高了发芽率，可广泛应用于朝鲜淫羊藿的工厂化育苗和规模化栽培。
The invention belongs to the technical field of plant propagation, in particular to a method for promoting the morphological after-ripening of Epimedium korea chinensis seed embryos. The method for promoting the after-ripening of the embryonic shape of Epimedium korea korea of the present application is to select seeds, soak the selected seeds in rape flower extract for 10-20 hours, pour off the water, and then mix the seeds and river sand evenly. Put it into a heat preservation board tank, cover it with black perforated plastic film and place it in a suitable temperature and humidity environment to cultivate, to obtain seeds with cotyledon embryos. The treatment time of the seeds is only 45 days, and the operability is strong. Finally, the seeds with cotyledon embryos and river sand are directly scattered on the soil border, covered with fine soil, and covered with larch pine needles after watering to obtain Epimedium korea seedlings, and the germination rate of the seedlings can reach 90% The above can greatly improve the cotyledon embryo occurrence rate, shorten the treatment time and improve the germination rate, and can be widely used in the factory seedling breeding and large-scale cultivation of Epimedium korea.
Description
Technical Field
The invention belongs to the technical field of plant propagation, and particularly relates to a method for promoting morphological after-ripening of Korean epimedium seed embryos.
Background
Epimedium koreanum Nakai is a perennial root herbaceous plant of Epimedium of berberidaceae, and is centrally distributed in Changbai mountain area of northeast China. In the multi-growing forests or shrubs, the elevation of 400 + 1500 m is favored to grow in the places with loose soil, rich soil and high humus content. Korean epimedium is a traditional Chinese tonifying traditional Chinese medicinal material, is collected from pharmacopoeia of the people's republic of China, is called to have the effects of warming and invigorating kidney yang, strengthening muscles and bones, dispelling wind-damp and the like, belongs to Changbai mountain genuine Chinese medicinal materials, and can be used for treating symptoms such as impotence, spermatorrhea, premature ejaculation, urinary incontinence, cold joint pain, irregular menstruation and the like. The medicinal components of the epimedium are as follows: icariin, emodin, liquiritigenin, hyperoside, quercetin, undecane, sterol, palmitic acid, ginkgol, magnoflorine, anthocyanidin, caffeic acid, etc.; the functions are as follows: dilating coronary artery blood vessel, lowering blood pressure, relieving cough, eliminating phlegm, relieving asthma, resisting bacteria and viruses, and relieving inflammation, and can be used for treating sexual impotence, premature ejaculation, rheumatic arthritis and gynecological diseases. Modern medical research indicates that the epimedium koreanum has a plurality of new pharmacological actions and purposes, such as anti-aging, bone growth promotion, hematopoietic system disorder improvement and the like, besides the traditional medical value and application range, so that the product is one of the most popular products in the fields of modern medicine and food, and is a plant medicinal material with great development value.
The Korean epimedium is a rare Chinese herbal medicine, has large market demand and is in a state of short supply throughout the year. Due to the rush harvesting of the drug traders, the drug farmers start the rush harvesting before and after the flowering, seeds are difficult to obtain, natural updating fails, wild resources are reduced year by year, and meanwhile, the feasibility of artificial cultivation is hindered due to the absence of a large number of seedlings. The reason for the failure of artificial cultivation is that after the pod is mature, the seed embryo in the seed is in the stage of the globular embryo, and the seed embryo is likely to germinate only in the stage of the cotyledon embryo due to the seed embryo, namely, the globular embryo can grow into the germinated cotyledon embryo after the development of the heart-shaped embryo. Therefore, the research on the method for promoting the morphological after-ripening of the embryo has an important role in the industrial seedling culture and large-scale artificial cultivation of the facing endangered epimedium koreanum.
Disclosure of Invention
In order to solve the problems that species are endangered, seeds are difficult to obtain, artificial cultivation cannot be carried out and the like due to the fact that the demand of the Korean epimedium is large, the collection and consumption amount exceeds the regeneration capacity of natural resources, and the like, the invention provides a method for promoting the morphological after-ripening of the Korean epimedium.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows: a method for promoting morphological after-ripening of Epimedium koreanum seed embryo comprises the following steps:
s1, seed selection: collecting mature fruit pods of epimedium koreanum in late ten (6) months, taking out seeds, putting the seeds into clear water, and pouring off the seeds floating on the water surface and reserving the seeds sinking to the water bottom for later use;
s2, seed embryo culture: soaking the seeds in the step S1 in the rape flower extracting solution for 10-20 hours, pouring water, uniformly stirring the seeds and river sand, putting the mixture into a heat preservation plate groove, covering a black plastic film with holes, and culturing in an environment with proper temperature and humidity to obtain seeds with cotyledon embryos;
s3, multiplication culture: and (5) directly and uniformly spreading the seeds with the cotyledon embryos and the river sand in the step S2 on soil furrows, covering fine soil, watering, and covering larch pine needles to obtain the seedlings of the Korean epimedium herb.
Further, in step S2, the mass concentration of the rape flower is 50-150 g.L-1。
Further, the mass concentration of the rape flower in the step S2 is 100--1。
Further, the ambient temperature of the hotplate slot in step S2 is 15-35 ℃.
Further, the ambient temperature of the hotplate slot in step S2 is 25-35 ℃.
Further, the environmental humidity of the heat-preserving plate groove in the step S2 is 65-78%.
Further, the mass ratio of the seeds to the river sand in the step S2 is 1: 2-5.
Further, the soaking time with the rape flower extracting solution in the step S2 is 12-15 hours.
Furthermore, the hole diameter of the black plastic film with holes is 0.2-0.5 cm.
Further, in step S3, the thickness of the covering fine soil is 3-4cm, and the thickness of the covering larch pine needle is 3-4 cm.
The invention provides a method for promoting the morphological after-ripening of Korean epimedium seed embryo, which comprises the steps of selecting seeds, soaking the selected seeds in rape flower extract for 10-20 hours, pouring off water, uniformly stirring the seeds and river sand, putting the mixture into a heat preservation plate groove, covering a black plastic film with holes, and culturing in a proper temperature and humidity environment to obtain seeds with cotyledon embryo, wherein the Korean epimedium seed has short treatment time of only 45 days and strong operability, and the cotyledon embryo occurrence rate reaches more than 95%; finally, directly and uniformly spreading the seeds with the cotyledon embryos and river sand on soil beds, covering fine soil, watering, and covering pine needles of larch to obtain seedlings of Korean epimedium, wherein the germination rate of the seedlings can reach over 90%; the method not only solves the key technology of morphological after-ripening of the seed embryo of the Korean epimedium, but also greatly improves the occurrence rate of the cotyledon embryo, shortens the processing time and improves the germination rate, and can be popularized and applied to industrial seedling culture and large-scale cultivation of the Korean epimedium.
Drawings
In FIG. 1, a is the harvested seed embryo (globular embryo); b is the seed embryo (heart shaped embryo) 25 days after treatment; c is the seed embryo (cotyledon embryo) 45 days after treatment.
Detailed Description
The technical solutions in the present invention will be described clearly and completely with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments.
As shown in FIG. 1, a method for promoting morphological after-ripening of Epimedium koreanum seed embryo comprises the following steps:
s1, seed selection: collecting mature fruit pods of epimedium koreanum in late ten (6) months, taking out seeds, putting the seeds into clear water, and pouring off the seeds floating on the water surface and reserving the seeds sinking to the water bottom for later use;
collecting mature fruit pods (yellow, with slight cracks) of Epimedium koreanum in late 6 months, taking out the seeds, putting the seeds into clear water, pouring off the seeds floating on the water surface, and reserving the seeds sinking to the water bottom for later use;
s2, seed embryo culture: soaking the seeds in the step S1 in the rape flower extracting solution for 10-20 hours, pouring water, uniformly stirring the seeds and river sand, putting the mixture into a heat preservation plate groove, covering a black plastic film with holes, and culturing in an environment with proper temperature and humidity to obtain seeds with cotyledon embryos;
wherein the mass concentration of the rape flower is 50-150 g.L-1Preferably 100-150 g.L-1(ii) a Soaking in rape flower extractive solution for 12-15 hr;
the environmental temperature of the heat preservation plate groove is 15-35 ℃, preferably 25-35 ℃; the environmental humidity of the heat-preservation plate groove is 65-78%, preferably 75%;
the mass ratio of the seeds to the river sand is 1:2-5, preferably 1: 3; the hole diameter of the black plastic film with holes is 0.2-0.5cm, preferably 0.3cm, so that the culture environment of the seeds has good air permeability, and the germination of the seeds is effectively promoted;
dividing seeds sinking to water bottom into multiple parts (each part contains 100 seeds), and adding rape flower extractive solution (50-100 g.L)-1) Soaking for 12-15 hr, draining, mixing with river sand (1:3), placing into a thermal insulation plate groove, covering with black plastic film with holes (hole diameter of about 0.3 cm), treating at different temperatures (15-25 deg.C), and controlling relative humidity at about 75%;
mixing the seeds with sand, treating the mixture in different temperature (15-25 ℃) environments, taking seeds every 15 days to observe the development condition of the seed embryos, counting the occurrence rate of the cotyledon embryos, taking the percentage of every 100 seed embryos developed into the cotyledon embryos as a subject to be inspected, namely the occurrence rate of the cotyledon embryos is equal to the number of the cotyledon embryos/100 multiplied by 100%, designing an experiment by a uniform design method, and selecting U for improving the speed and the occurrence rate of the cotyledon embryos of the Korean epimedium seeds6(62) Uniformly treating 100 seeds, repeating the treatment for 3 times, taking an average value, and determining the mass concentration of the rape flower extract and the optimal amount of the treatment temperature to the cotyledon embryo of the Korean epimedium seed embryo;
s3, multiplication culture: directly and uniformly spreading the seeds with the cotyledon embryos and the river sand in the step S2 on soil furrows, covering fine soil, watering, and covering larch and pine needles to obtain Korean epimedium seedlings;
the processed seeds of the epimedium koreanum with cotyledon embryos can be directly and uniformly spread on a prepared soil bed together with river sand, fine soil with the thickness of about 3-4cm is covered, and larch and pine needles with the thickness of about 3-4cm are covered after water is poured. Seeds in the upper ten days in 5 months after winter can directly germinate, grow and extend out of the soil;
s4: results and analysis
The applicant successfully establishes a 1 hectare (15 mu) germplasm garden through a wild resource root division method in recent 7 years, obtains a large amount of seeds in the germplasm garden, and finds that the germination rate of the seeds after sowing is less than 0.2%, and the seeds in the seeds are in a spherical embryo stage after the pods are mature, and the seeds are likely to germinate only in a cotyledon embryo stage, namely the spherical embryos develop into germinable cotyledon embryos through heart-shaped embryos. Therefore, the applicant develops a series of researches on the morphological after-ripening of the seed embryo, obtains the method for rapidly and simply promoting the morphological after-ripening of the seed embryo through the 3-year attack, and finds that the mass concentration of the rape flower extracting solution and the culture environment temperature have important influence on the cotyledon embryogenesis of the Korean epimedium seed embryo. The following is a main analysis of the influence of the mass concentration and temperature of the rape flower extract on the cotyledon embryogenesis of the Korean epimedium seed embryo.
Influence of rape flower extract quality concentration and temperature on seed embryo cotyledon embryogenesis of Korean herba Epimedii
TABLE 1U of major factors affecting embryogenesis of seed embryo of Epimedium koreanum6(62) Experimental design and results
After 45 days of Korean epimedium seed treatment, the regression equation of Y-66.1 +0.101X can be obtained after the obtained data (table 1) are analyzed and treated by uniform design software1+0.437X2Significance level α ═ 0.05, complex correlation coefficient R ═ 0.9365, residual standard deviation S ═ 1.08, test value FtMore than 10.70 > more than critical value F(0.05,2,3)9.552, the regression equation has significance, and the influence of rape flower extract and temperature on the embryogenesis of Korean epimedium seed embryos has significance. The U can be known by calculating the contribution value and the contribution rate of the rape flower extract and the temperature to the cotyledon embryogenesis of the Korean epimedium seed embryo1＝17.2，U1/U＝68.6％；U2＝12.8，U251.1 percent of the total amount of the extracted rape flower, and the contribution value and the contribution rate of the temperature to the generation of the seed cotyledon embryo of the epimedium koreanum due to the fact that the temperature is lower than the contribution value and the contribution rate of the extracted rape flower to the generation of the seed cotyledon embryo of the epimedium koreanum, the significance of the temperature to the generation of the seed cotyledon embryo of the epimedium koreanum is further tested2More than 10.94 > than the critical value F(0.05,1,3)10.13, the temperature proved to be significant for the development of the korea epimedium seed embryo cotyledon embryo. The rape flower extract and the temperature are the samePositively correlated with the occurrence rate of cotyledon embryo, and guessing that the mass concentration and the temperature of the rape flower extracting solution are respectively higher than 100 g.L-1And at 25 ℃, the incidence rate of the seed embryo cotyledon embryo of the Korean epimedium herb can be higher. To verify the guessing possibility, the mass concentrations of the rape flower extract were set at 100, 110, 120, 130, 140 and 150 g.L-1Validation tests supplemented with 6 treatments at 25, 27, 29, 31, 33 and 35 ℃ were repeated 3 times. The operation method is the same as 1.1 and 1.2, and after the rape flower extract is treated for 45 days, the statistical result shows that the mass concentration of the rape flower extract is 130 g.L-1And the occurrence rate of the cotyledon embryos is the highest when the temperature is 31-33 ℃, the average occurrence rate is more than 95 percent and is higher than that of 6 treated cotyledon embryos listed in Table 1.
According to the above experimental data processing and analysis, the optimal conditions for morphological after-ripening of the seed embryo of the epimedium koreanum are as follows: the mass concentration of the seed is 130 g.L-1Soaking the rape flower extract for 12-15 hours, pouring off water, uniformly stirring seeds and river sand (1:3), putting into a heat preservation plate groove, covering a black plastic film with holes (the diameter of the holes is about 0.3 cm), and treating for 45 days at the temperature of 31-33 ℃, wherein the relative humidity is kept at about 75%, the occurrence rate of cotyledon embryos is highest, and the average occurrence rate is more than 95%. The mass concentration of the rape flower extract exceeds 130 g.L-1The incidence rate of cotyledon embryo is reduced on the contrary, and the seed rot phenomenon is serious when the temperature exceeds 33 ℃.
After the seeds are sowed, seeds of the Korean epimedium can germinate, grow and extend out of the soil surface in the first ten days in 5 months in spring of the next year, and the germination rate reaches more than 90 percent. The method has the advantages of short time for treating the seeds of the epimedium koreanum, strong operability, extremely high cotyledon embryogenesis rate and germination rate and the like, and can be widely applied to industrial seedling culture and large-scale cultivation of the epimedium koreanum.
The method for promoting the morphological after-ripening of the Korean epimedium seed embryo has the advantages of short time for treating the Korean epimedium seed, strong operability, extremely high cotyledon embryo occurrence rate and the like, the treatment time is only 45 days, and the cotyledon embryo occurrence rate reaches more than 95%; has the advantages of extremely high germination rate of Korean epimedium seeds and the like, and the germination rate can reach more than 90 percent; not only solves the key technology of morphological after-ripening of the seed embryo of the Korean epimedium, but also greatly improves the occurrence rate of the cotyledon embryo, shortens the processing time and improves the germination rate, and can be popularized and applied to the industrial seedling culture and the large-scale cultivation of the Korean epimedium.
The above description is only a preferred embodiment of the present invention, but the design concept of the present invention is not limited thereto, and any insubstantial modifications made by using the design concept should fall within the scope of infringing on the protection scope of the present invention.
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