CN114931006B - Method for promoting germination of akebia stem seeds and method for promoting growth of akebia stem seedlings - Google Patents
Method for promoting germination of akebia stem seeds and method for promoting growth of akebia stem seedlings Download PDFInfo
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- CN114931006B CN114931006B CN202210676501.6A CN202210676501A CN114931006B CN 114931006 B CN114931006 B CN 114931006B CN 202210676501 A CN202210676501 A CN 202210676501A CN 114931006 B CN114931006 B CN 114931006B
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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
- A01C1/02—Germinating apparatus; Determining germination capacity of seeds or the like
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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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- 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
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- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
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Abstract
The invention belongs to the technical field of plant seed germination and planting, and particularly relates to a method for promoting the germination of akebia stem seeds and a method for promoting the growth of akebia stem seedlings. The invention provides a method for promoting germination of akebia stem seeds, which shows that: after the seed coat of the fresh akebia stem seeds is removed, the time for the seeds to start germination is advanced to day 4. At 20 ℃, the germination rate of the seed coat removed akebia stem seeds is 90%, the germination rate of the whole akebia stem seeds is 50.67 +/-2.07%, and the germination rate of the half akebia stem seeds is 58.33+/-2.58%. The method for promoting the growth of the seedlings of the akebia stem seeds provided by the invention has the advantages that the germination rate of the whole akebia stem seeds stacked in the whole darkness for 40 days is highest and reaches 100%, and the total length of the seedlings exceeds 2000cm. The method of the invention obviously improves the germination rate of the akebia stem seeds, accelerates the germination process of the seeds, improves the germination vigor and promotes the growth of the seedlings of the seeds.
Description
Technical Field
The invention belongs to plant seed germination and planting, and particularly relates to a method for promoting the germination of akebia stem seeds and a method for promoting the growth of akebia stem seed seedlings.
Background
Aristolochia (Akebia trifoliata subsp. Australis) is one of five major rare Chinese medicinal materials which are mainly protected and developed in China, has the effects of clearing heat and promoting urination, promoting blood circulation and dredging collaterals, resisting bacteria and diminishing inflammation, and has long medicinal history. The fruit of the akebia stem has high edible value, and the pulp contains various minerals and microelements required by human bodies and essential amino acids, and can also be used for processing jam, fruit juice and fruit wine; the flowers, leaves, fresh fruits and peel can be used for processing health-care tea; the seed oil is rich in unsaturated fatty acid, and is a high-quality health-care edible oil source. Therefore, the akebia stem is a 'precious' in whole body, is a high-quality 'green' raw material for medicine, health-care products and food industry, and has a great development prospect.
The sexual propagation mode of seed propagation is an important means and technical link for screening and cultivating the excellent variety of akebia stem. However, the germination period of the akebia stem seeds is long, the germination vigor is low and the germination rate is low due to the high degree of oil, natural gum and waxiness in the seed coats, so that the development and the utilization of resources are seriously hindered. Therefore, a method for promoting the germination of the akebia stem seeds is explored, the germination rate and the germination vigor of the seeds are improved, and important technical support is provided for sexual reproduction, cross breeding and preservation of germplasm resources of the akebia stem.
Disclosure of Invention
The invention aims to provide a method for promoting germination of akebia stem seeds, which improves the germination rate of the akebia stem seeds, accelerates the germination process of the seeds, improves the germination vigor and assists the breeding of the akebia stem seeds.
The invention provides a method for promoting germination of akebia stem seeds, which comprises the following steps:
mixing peeled akebia stem seeds with distilled water, and performing light-dark alternate culture until seed germination is finished; the peeled akebia stem seeds are seeds with the seed coats removed and the complete endosperm and embryo reserved.
Preferably, the light-dark alternate culture is light culture for 12-14 hours and dark culture for 10-12 hours, and the temperature is 10-35 ℃; the illumination intensity is 2000-3000 lux during illumination culture.
Preferably, the volume ratio of the peeled akebia stem seeds to distilled water is (1.5-2.5) g: (5-7) mL.
Preferably, the peeled akebia stem seeds are mixed with distilled water and placed in a container for light-dark alternate culture, the container comprises a culture dish, and 2 layers of filter paper or gauze are paved in the culture dish.
The invention also provides a method for promoting the growth of the akebia stem seedlings, which comprises the following steps:
mixing the whole caulis Aristolochiae with matrix particles, and placing in a container with air holes on the wall for total darkness lamination; culturing alternately in light and dark after the total darkness is laminated until the germination of seeds is finished; the matrix particles include perlite and/or wet sand.
Preferably, the temperature of the total darkness lamination is 3-5 ℃; the total dark lamination time is 35-40 d.
Preferably, the light-dark alternate culture is light culture for 12-14 hours and dark culture for 10-12 hours, and the temperature is 10-35 ℃; the illumination intensity of the illumination culture is 2000-3000 lux.
Preferably, the mass ratio of the whole seed to the perlite in the total darkness lamination is 1:4.
preferably, the whole akebia stem seeds and the matrix particles are mixed and then the mixture of the seeds and the matrix particles is put into a seed bag; placing the seed bag into the container; the seed bag comprises a nylon net bag, and the seed bag is turned once every 7 days; perlite in the container remains moist; the container comprises a plastic container.
The invention also provides application of the technical scheme in improving the germination rate and the germination vigor of the akebia stem seeds.
The invention has the beneficial effects that: the invention provides a method for promoting germination of akebia stem seeds, which comprises the following steps: mixing peeled akebia stem seeds with distilled water, and performing light-dark alternate culture until seed germination is finished; the peeled akebia stem seeds are seeds with the seed coats removed and the complete endosperm and embryo reserved. In the method provided by the invention, the peeled akebia seeds are easier to absorb water, the air permeability of the embryo is increased, the mechanical obstruction and the seed coat inhibition of the embryo are reduced, the germination of the seeds is induced by alternate light and dark culture, the germination rate of the akebia seeds is improved, the germination process of the seeds is accelerated, and the germination potential is improved. The results of the examples show that: after the seed coats of the fresh akebia stem seeds are removed, the germination starting time of the seeds is advanced from the 14 th day to the 4 th day of the whole seeds (the seeds without the seed coats removed) while the higher germination rate is maintained, so that the germination process of the seeds is quickened, and the germination vigor is improved. After the seed coats of the fresh akebia stem seeds are removed, the germination rate of the seeds is 90 percent, the germination rate of the whole akebia stem seeds is 50.67 +/-2.07 percent, and the germination rate of half akebia stem seeds is 58.33+/-2.58 percent at 20 ℃, so that the germination rate of the akebia stem seeds is obviously improved, the germination process of the seeds is accelerated, and the germination vigor is improved by applying the method disclosed by the invention.
The invention provides a method for promoting the growth of akebia stem seedlings, which comprises the following steps: mixing the whole caulis Aristolochiae with matrix particles, and placing in a container with air holes on the wall for total darkness lamination; culturing alternately in light and dark after the total darkness is laminated until the germination of seeds is finished; the matrix particles include perlite and/or wet sand. And after the total darkness lamination is finished, light-dark alternate culture is carried out, so that seedling growth is facilitated. The invention accelerates the growth of the seedlings through the total darkness lamination, improves the germination rate and the germination vigor of the akebia stem seeds through the light-dark alternate culture, promotes the germination of the akebia stem seeds under the combined action of the total darkness lamination treatment and the light-dark alternate culture, and accelerates the growth of the akebia stem seedlings. The results of the examples show that: the germination rate of the whole akebia stem seeds stacked in the whole darkness for 40 days is highest and reaches 100%, and the total length of seedlings exceeds 2000cm. Therefore, the germination rate of the akebia stem seeds is obviously improved by applying the method disclosed by the invention, the growth of the akebia stem seedlings is accelerated, and the seedling growth condition is good.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required to be used in the embodiments will be briefly described below.
FIG. 1 shows the germination process of peeled Akebia quinata seeds in the light-dark alternate culture process under different temperature conditions in example 1 and comparative examples 6-10;
FIG. 2 shows the germination lengths of peeled Akebia quinata seeds for 31 days in the light-dark alternate culture process under different temperature conditions of example 1 and comparative examples 6-10;
FIG. 3 shows the germination process of whole akebia stem seeds in the process of alternate light and dark culture under different temperature conditions in comparative examples 1-5;
FIG. 4 shows the seedling length of 31 days of germination of whole Akebia quinata seeds in the alternative light-dark culture process under different temperature conditions in comparative examples 1-5;
FIG. 5 shows the germination course of half-grain akebia stem seeds in the alternative culturing process under different temperature conditions of comparative examples 11-16;
FIG. 6 shows the seedling length of 31 days of germination of half-grain akebia stem seeds in alternative light-dark culture processes under different temperature conditions in comparative examples 11-16;
FIG. 7 is a graph showing the effect of gibberellin treatment at different concentrations on germination rate of whole Akebia quinata seeds in comparative examples 17-21;
FIG. 8 shows germination processes in the light-dark alternate culture process of whole white akebia seeds of different total darkness layering days in example 2 and comparative examples 22 to 24;
FIG. 9 shows the seedling length of 31 days for germination of whole Akebia quinata seeds on different total darkness stratification days in example 2 and comparative examples 22-24.
Detailed Description
The invention provides a method for promoting germination of akebia stem seeds, which comprises the following steps:
mixing peeled akebia stem seeds with distilled water, and performing light-dark alternate culture until seed germination is finished; the peeled akebia stem seeds are seeds with the seed coats removed and the complete endosperm and embryo reserved.
In the invention, the peeled white akebia seed is preferably a seed separated from fresh and mature black full white akebia fruits, residual pulp covered on the surfaces of the white akebia seeds is cleaned and removed, and the clean whole seeds are obtained after airing for 24 hours, the peeled white akebia seeds are obtained after the black seed coats are removed, the black seed coats of the white akebia seeds are removed to facilitate germination of the white akebia seeds, and the time for starting germination of the seeds is shortened. The invention has no special requirement on the peeling method, and the conventional peeling method is adopted to ensure that the complete endosperm and embryo are preserved after the seed coats are removed. In the embodiment of the invention, all peeled seeds, whole seeds and half seeds are treated identically, the water content is identical, and the water content of the seeds in the implementation process of the invention is not an influencing factor on the germination of Bai Mutong seeds. The judging standard of the end of the germination of the seeds in the invention is that the germination number of the seeds is not increased within 3 days of the germination of the seeds.
In the invention, the peeled akebia stem seeds are preferably placed in a container to be mixed with distilled water; before adding the akebia stem seeds, the invention preferably fills 2 layers of filter paper or gauze in a container, and then adds the akebia stem seeds and distilled water, wherein the distilled water can wet the 2 layers of filter paper or gauze filled in the container, and the distilled water can be added in a certain amount. The preferred particle number volume ratio of the peeled akebia stem seeds to distilled water in the container is (1.5-2.5) g: (5-7) mL, more preferably 2.5g:7mL of distilled water is used for promoting the peeled akebia seeds to germinate after absorbing water. The filter paper or the gauze can absorb water, and the wet filter paper or the gauze after water absorption can maintain the wet environment required by seed germination, and meanwhile, the seeds are not soaked in water, so that the contact with air is isolated.
The source of the container and distilled water in the present invention is not particularly limited, and conventional products may be used. In an embodiment of the invention, the vessel is preferably a 9cm diameter petri dish.
In the invention, the akebia stem seeds are preferably placed in a container for light-dark alternate culture, wherein the light-dark alternate culture time is preferably 12-14 h dark culture for 10-12 h, more preferably 12-13 h dark culture for 11-12 h, and even more preferably 12h dark culture for 12h. The temperature of the light-dark alternate culture is preferably 10-35 ℃, more preferably 13-20 ℃, and even more preferably 15 ℃; the illumination intensity during the illumination culture is 2000 to 3000lux, more preferably 2600 to 3000lux, and still more preferably 3000lux. In the invention, the light-dark alternate culture temperature is selected to be 10-35 ℃ close to the temperature of the white wood through distribution area; the growth state of the seedlings of the akebia stem is poor below 10 ℃, and the seeds of the akebia stem do not sprout above 40 ℃. The alternate cultivation is carried out until the germination of seeds is finished. The germination duration of the akebia stem seeds is 31 days, and the seedling length of the germinated seeds is directly measured after the germination of the seeds is finished. In the invention, the selection of the light-dark alternate culture mode, the culture temperature, the culture time and the illumination intensity is to improve the germination vigor and the germination rate of the peeled akebia seeds and accelerate the germination process of the akebia seeds.
The invention also provides a method for promoting the growth of the akebia stem seedlings, which comprises the following steps:
mixing the whole caulis Aristolochiae with matrix particles, and placing in a container with air holes on the wall for total darkness lamination; culturing alternately in light and dark after the total darkness is laminated until the germination of seeds is finished; the matrix particles include perlite and/or wet sand.
The whole-grain akebia stem seeds are subjected to no peeling treatment, namely, seeds separated from fresh and mature black full akebia stem fruits are cleaned to remove residual pulp covered on the surfaces of the akebia stem seeds, and the clean whole-grain seeds are obtained after 24 hours of airing.
The method for preparing the whole akebia stem seed comprises the steps of mixing the whole akebia stem seed with matrix particles, and placing the mixture of the seed and the matrix particles into a seed bag. In the present invention, the matrix particles comprise perlite and/or wet sand, preferably perlite or wet sand, more preferably perlite. In the embodiment of the invention, the perlite and the whole akebia stem seeds are preferably mixed and then placed into a seed bag, and the seed bag is placed into a container with ventilation holes on the wall for total darkness lamination. The perlite in the container of the invention is preferably kept moist. In the invention, before the perlite is uniformly mixed with the whole akebia stem seeds, the perlite is preferably moistened by using sterile water to obtain moistened perlite, and the moisture content of the moistened perlite is preferably 60-70%. The mass ratio of the perlite to the whole akebia stem seeds is preferably 1:4. The perlite is selected because the perlite particles are large, so that the perlite is favorable for sufficient air circulation in the whole-grain akebia stem seeds in the whole-dark lamination process, and the germination of the akebia stem seeds and the growth of seedlings are promoted. The seed bag of the present invention preferably comprises a nylon mesh bag that is preferably turned once every 7 days; in the embodiment of the invention, the seed bag is preferably a nylon net bag, and the seed bag is used for conveniently turning seeds in the layering process; meanwhile, the seed germination promoting agent has the effects of ventilation and water permeability and promotes seed germination.
The present invention preferably places the seed bag into the container; the container wall is provided with the ventilation holes, and more preferably, four side surfaces of the container are provided with the ventilation holes; the invention has no special requirements on the material of the container, and can ensure ventilation. The container of the present invention is preferably a plastic container, more preferably a plastic box. The plastic box with the ventilation holes on the periphery can keep ventilation. The invention has no special requirements on the number of the air holes, the size of the air holes or the porosity, and can ensure the air permeability.
The seed bag is placed in a plastic container with air holes on the wall for total darkness lamination, the temperature of the total darkness lamination is preferably 3-5 ℃, more preferably 4 ℃, and the total darkness lamination time is preferably 35-40 d, more preferably 38-40 d, more preferably 40d. In the present invention, the plastic box is preferably placed in a refrigerator for refrigeration throughout the total dark lamination process, with distilled water periodically to keep the perlite moist, and the seed bag is turned once a week. The stirring seed bag has the function of ventilation and can promote germination of whole akebia stem seeds. The effect of the total darkness layering in the invention is to promote the growth of seedlings of the akebia stem seeds. In the present invention, the total darkness stratification is to fully post-ripen the seed embryo and reduce substances inhibiting seed germination. In the present invention, the total darkness condition of the total darkness laminate is achieved by placing the container in a refrigerator or incubator.
After the total darkness lamination is finished, the whole akebia stem seeds after the total darkness lamination are subjected to light-dark alternate culture until the germination of the seeds is finished. The invention preferably takes out whole-grain akebia stem seeds obtained by total darkness lamination, places the whole-grain akebia stem seeds in a container, and places the container in an illumination incubator for alternate light and darkness culture to promote germination. In the invention, 2 layers of filter paper or gauze are preferably filled before the whole-grain akebia stem seeds after the total darkness lamination are added into the container, and then the whole-grain akebia stem seeds after the total darkness lamination and distilled water are added, wherein the distilled water can be used for wetting the 2 layers of filter paper filled into the container, and the distilled water can be used for adding a little surplus water amount. The preferred particle number volume ratio of the whole particle akebia stem seeds after total darkness lamination in the container to distilled water is (1.5-2.5) g: (5-7) mL, more preferably 2.5g:7mL of distilled water is used for promoting the peeled akebia seeds to germinate after absorbing water. The source of the container and distilled water in the present invention is not particularly limited, and conventional products may be used. In an embodiment of the invention, the vessel is preferably a 9cm diameter petri dish.
In the present invention, the alternate light and dark culture is preferably a light culture for 12 to 14 hours and a dark culture for 10 to 12 hours, more preferably a light culture for 12 to 13 hours and a dark culture for 11 to 12 hours, still more preferably a light culture for 12 hours and a dark culture for 12 hours. In the present invention, the temperature of the alternate light and dark culture is preferably 10 to 35 ℃, more preferably 13 to 20 ℃, and even more preferably 15 ℃; the illumination intensity during the illumination culture is 2000 to 3000lux, more preferably 2600 to 3000lux, and still more preferably 3000lux. The seedling is obtained by alternately culturing the seeds until the germination of the seeds is finished. The germination refers to the germination of the seed radicle which breaks through the seed coat by 2 mm. The growth of the seed radicle after breaking through the seed coat is the seedling growth process. And after the seeds germinate in the culture dish for 3 days, the germination process of the seeds in the culture dish is considered to be finished, namely the germination of the seeds is finished.
For further explanation of the present invention, a method for promoting germination of seeds of akebia stem and a method for promoting seedling growth of akebia stem provided by the present invention are described in detail below with reference to the accompanying drawings and examples, but they should not be construed as limiting the scope of the present invention.
EXAMPLE 1 seed germination experiment of peeled Akebia quinata
Mature and full fruits of akebia stem (Akebia trifoliata subsp. Australis) are collected, the collection area is the same county as Hunan province, and the collected fruits are transported back to the academy of chemical engineering and biology for seed cleaning and germination experiments.
Seed collection and cleaning: collecting mature fruits on plants, peeling off seeds in pulp, cleaning with clear water, placing in indoor shade, and air drying for 24h to obtain pure caulis Aristolochiae Mollissimae seeds; and then removing only the black seed coats of the pure akebia seeds, and reserving the complete endosperm and embryo seeds to obtain the peeled akebia seeds which are used for germination culture.
Seed culture: sowing peeled caulis Aristolochiae Kaempferi seeds in a culture dish filled with 2 layers of filter paper, adding 7mL distilled water, placing in an incubator with illumination intensity of 3000lux and temperature of 15 ℃ for 12h illumination and 12h dark alternate culture until seed germination culture is completed, and co-culturing for 31d.
And sowing 25 seeds in each culture dish, sowing 3 dishes in each treatment method, checking germination once a day, considering the germination of the seeds when the radicle breaks through the elongation of the seed coat to be more than 2mm, taking out the germinated seeds, and recording the germination quantity.
EXAMPLE 2 experiments on seed germination and seedling growth promotion of Whole-grain Akebia quinata
Mature and full fruits of akebia stem (Akebia trifoliata subsp. Australis) are collected, the collection area is the same county as Hunan province, and the collected fruits are transported back to the academy of chemical engineering and biology for seed cleaning and germination experiments.
Seed collection and cleaning: collecting mature fruits on plants, peeling off seeds in pulp, cleaning with clear water, placing in indoor shade, and air drying for 24h to obtain pure whole-grain akebia stem seeds.
Seed culture: mixing the whole akebia stem seeds with clean and wet perlite according to a mass ratio of 1:4, uniformly mixing and filling the mixture into a nylon Long Wangdai, and putting the nylon Long Wangdai into a plastic box filled with wet perlite, wherein ventilation holes are formed in the periphery of the plastic box so as to keep ventilation. The plastic box containing seeds is placed in a refrigerator for refrigeration at 4 ℃, distilled water is used for keeping the perlite moist at regular intervals, and the seed bags are turned once a week. After 40 days of total darkness lamination, taking out the whole akebia stem seeds, placing the seeds in a culture dish filled with 2 layers of filter paper, adding 7mL of distilled water, and placing the seeds in an incubator with the illumination intensity of 3000lux and the temperature of 15 ℃ for 12h illumination and 12h darkness alternate culture until the germination culture of the seeds is finished.
And sowing 25 seeds in each culture dish, sowing 3 dishes in each treatment method, checking germination once a day, considering the seeds to germinate when the radicle elongation exceeds 2mm, taking out the germinated seeds, and recording the germination quantity. And when the seeds are germinated for 3 days and remain unchanged, the germination is ended, and the length of the seedlings is measured by using a ruler on the day 31 days after the germination is ended.
Comparative example 1
The whole akebia stem seeds without removing the seed coats are placed in a culture dish filled with 2 layers of filter paper, 7mL of distilled water is added, and the culture is carried out in an illumination incubator with illumination intensity of 3000lux and temperature of 10 ℃ for germination, and light-dark alternate culture, namely illumination continuous culture for 12 hours and darkness continuous culture are adopted in the germination culture process, and other conditions are the same as in example 1.
Comparative example 2
The whole akebia stem seeds without seed coat removal are placed in an illumination incubator with illumination intensity of 3000lux and temperature of 20 ℃ for germination culture, and other conditions are the same as in comparative example 1.
Comparative example 3
The whole akebia stem seeds without seed coat removal are placed in an illumination incubator with illumination intensity of 3000lux and temperature of 25 ℃ for germination culture, and other conditions are the same as in comparative example 1.
Comparative example 4
The whole akebia stem seeds without seed coat removal are placed in an illumination incubator with illumination intensity of 3000lux and temperature of 30 ℃ for germination culture, and other conditions are the same as in comparative example 1.
Comparative example 5
The whole akebia stem seeds without seed coat removal are placed in an illumination incubator with illumination intensity of 3000lux and temperature of 35 ℃ for germination culture, and other conditions are the same as in comparative example 1.
Comparative example 6
The peeled akebia stem seeds were placed in an illumination incubator with an illumination intensity of 3000lux and a temperature of 10℃for germination culture, and the other conditions were the same as in example 1.
Comparative example 7
The peeled akebia stem seeds were placed in an illumination incubator with an illumination intensity of 3000lux and a temperature of 20℃for germination culture, and the other conditions were the same as in example 1.
Comparative example 8
The peeled akebia stem seeds were placed in an illumination incubator with an illumination intensity of 3000lux and a temperature of 25℃for germination culture, and the other conditions were the same as in example 1.
Comparative example 9
The peeled akebia stem seeds were placed in an illumination incubator with an illumination intensity of 3000lux and a temperature of 30℃for germination culture, and the other conditions were the same as in example 1.
Comparative example 10
The peeled akebia stem seeds were placed in an illumination incubator with an illumination intensity of 3000lux and a temperature of 35℃for germination culture, and the other conditions were the same as in example 1.
Comparative example 11
Half-grain akebia stem seeds were obtained by cutting off the whole piece of the akebia stem seeds without peeling, and the half-grain akebia stem seeds were placed in an illumination incubator having an illumination intensity of 3000lux and a temperature of 10℃for germination culture under the same conditions as in example 1.
Comparative example 12
Half-grain akebia stem seeds were obtained by cutting off the whole piece of the akebia stem seeds without peeling, and the half-grain akebia stem seeds were placed in an illumination incubator with an illumination intensity of 3000lux and a temperature of 15℃for germination culture under the same conditions as in example 1.
Comparative example 13
Half-grain akebia stem seeds were obtained by cutting off the whole piece of the akebia stem seeds without peeling, and the half-grain akebia stem seeds were placed in an illumination incubator with an illumination intensity of 3000lux and a temperature of 20℃for germination culture under the same conditions as in example 1.
Comparative example 14
Half-grain akebia stem seeds were obtained by cutting off the whole piece of the akebia stem seeds without peeling, and the half-grain akebia stem seeds were placed in an illumination incubator with an illumination intensity of 3000lux and a temperature of 25℃for germination culture under the same conditions as in example 1.
Comparative example 15
Half-grain akebia stem seeds were obtained by cutting off the whole piece of the akebia stem seeds without peeling, and the half-grain akebia stem seeds were placed in an illumination incubator with an illumination intensity of 3000lux and a temperature of 30℃for germination culture under the same conditions as in example 1.
Comparative example 16
Half-grain akebia stem seeds were obtained by cutting off the whole piece of the akebia stem seeds without peeling, and the half-grain akebia stem seeds were placed in an illumination incubator with an illumination intensity of 3000lux and a temperature of 35℃for germination culture under the same conditions as in example 1.
Comparative example 17
The whole akebia stem seeds without removing the seed coat are placed in a culture dish filled with 2 layers of filter paper, 7mL of gibberellin solution with the concentration of 0.1mmol/L is added, and the culture is carried out in an illumination incubator with the illumination intensity of 3000lux and the temperature of 20 ℃ for germination culture, and light-dark alternate culture, namely illumination continuous culture for 12 hours and dark continuous culture for 12 hours are adopted in the germination culture process, and other conditions are the same as in example 1.
Comparative example 18
The whole akebia stem seeds without removing the seed coat are placed in a culture dish filled with 2 layers of filter paper, 7mL of 1mmol/L gibberellin solution is added, and the culture is carried out in an illumination incubator with illumination intensity of 3000lux and temperature of 20 ℃ for germination culture, and light-dark alternate culture, namely illumination continuous culture for 12 hours and dark continuous culture for 12 hours are adopted in the germination culture process, and other conditions are the same as in example 1.
Comparative example 19
The whole akebia stem seeds without removing the seed coat are placed in a culture dish filled with 2 layers of filter paper, 7mL of 10mmol/L gibberellin solution is added, and the culture is carried out in an illumination incubator with illumination intensity of 3000lux and temperature of 20 ℃ for germination culture, and light-dark alternate culture, namely illumination continuous culture for 12 hours and dark continuous culture for 12 hours are adopted in the germination culture process, and other conditions are the same as in example 1.
Comparative example 20
The whole akebia stem seeds without removing the seed coat are placed in a culture dish filled with 2 layers of filter paper, 7mL of 100mmol/L gibberellin solution is added, and the culture is carried out in an illumination incubator with illumination intensity of 3000lux and temperature of 20 ℃ for germination culture, and light-dark alternate culture, namely illumination continuous culture for 12 hours and dark continuous culture for 12 hours are adopted in the germination culture process, and other conditions are the same as in example 1.
Comparative example 21
The whole akebia stem seeds without removing the seed coat are placed in a culture dish filled with 2 layers of filter paper, 7mL of 1000mmol/L gibberellin solution is added, and the culture is carried out in an illumination incubator with illumination intensity of 3000lux and temperature of 20 ℃ for germination culture, and light-dark alternate culture, namely illumination continuous culture for 12 hours and dark continuous culture for 12 hours are adopted in the germination culture process, and other conditions are the same as in example 1.
Comparative example 22
Whole-grain akebia stem seeds without seed coat removal were laminated in total darkness for 10 days under the same conditions as in example 2.
Comparative example 23
Whole-grain Akebia quinata seeds without seed coats were laminated in total darkness for 20 days under the same conditions and seed sources as in example 2.
Comparative example 24
Whole-grain Akebia quinata seeds without seed coats were laminated in total darkness for 30 days under the same conditions and seed sources as in example 2.
Application example 1 analysis of germination Rate and vigour of Akebia quinata seeds
The germination rate and vigor of the akebia stem of example 1 and comparative examples 1 to 21 were measured, germination rate (%) = number of germinated seeds/total number of seeds×100; germination vigor (%) = number of germinated seeds/total number of seeds x 100 in the first 10 days; the measurement results are shown in Table 1.
The germination process of peeled white wood through seeds in the alternate light-dark culture process in example 1 and comparative examples 6-10 is shown in fig. 1, the germination length of peeled white wood through seeds in the alternate light-dark culture process in example 1 and comparative examples 6-10 is shown in fig. 2, the germination process of whole white wood through seeds in the alternate light-dark culture process in comparative examples 1-5 is shown in fig. 3, the germination length of whole white wood through seeds in the alternate light-dark culture process in comparative examples 1-5 is shown in fig. 4, the germination process of half white wood through seeds in the alternate light-dark culture process in comparative examples 11-16 is shown in fig. 5, the germination length of half white wood through seeds in the alternate light-dark culture process in comparative examples 11-16 is shown in fig. 6, and the effect of gibberellin treatment in different concentrations in comparative examples 17-21 on germination rate of whole white wood through seeds is shown in fig. 7.
TABLE 1 germination Rate and vigour of Akebia quinata of examples 1-2 and comparative examples 1-24
| Seed of akebia stem | Treatment of | Germination temperature | Germination rate (%) | Germination potential (%) | |
| Example 1 | Peeling off | Fresh seeds | 15℃ | 96.67±2.58 | 46.67±2.58 |
| Comparative example 1 | Finishing grain | Fresh seeds | 10℃ | 97.33±2.07 | 0.00±0.00 |
| Comparative example 2 | Finishing grain | Fresh seeds | 15℃ | 96.00±3.58 | 0.00±0.00 |
| Comparative example 3 | Finishing grain | Fresh seeds | 20℃ | 50.67±2.07 | 0.00±0.00 |
| Comparative example 4 | Finishing grain | Fresh seeds | 25℃ | 4.00±3.58 | 0.00±0.00 |
| Comparative example 5 | Finishing grain | Fresh seeds | 30℃ | 0.00±0.00 | 0.00±0.00 |
| Comparative example 6 | Peeling off | Fresh seeds | 10℃ | 95.00±4.47 | 18.33±6.83 |
| Comparative example 7 | Peeling off | Fresh seeds | 20℃ | 90.00±8.30 | 50.00±11.83 |
| Comparative example 8 | Peeling off | Fresh seeds | 25℃ | 38.33±13.66 | 25.00±4.47 |
| Comparative example 9 | Peeling off | Fresh seeds | 30℃ | 20.00±11.83 | 15.00±8.94 |
| Comparative example 10 | Peeling off | Fresh seeds | 35℃ | 3.33±2.58 | 1.67±2.58 |
| Comparative example 11 | Semi-granule | Fresh seeds | 10℃ | 13.33±6.83 | 0.00±0.00 |
| Comparative example 12 | Semi-granule | Fresh seeds | 15℃ | 98.33±2.58 | 6.67±2.58 |
| Comparative example 13 | Semi-granule | Fresh seeds | 20℃ | 58.33±2.58 | 5.00±4.47 |
| Comparative example 14 | Semi-granule | Fresh seeds | 25℃ | 6.67±10.33 | 3.33±5.15 |
| Comparative example 15 | Semi-granule | Fresh seeds | 30℃ | 1.67±2.58 | 1.67±2.58 |
| Comparative example 16 | Semi-granule | Fresh seeds | 35℃ | 0.00±0.00 | 0.00±0.00 |
| Comparative example 17 | Finishing grain | GA0.1mmol/L | 20℃ | 61.33±1.53 | 0.00±0.00 |
| Comparative example 18 | Finishing grain | GA1mmol/L | 20℃ | 41.33±4.30 | 0.00±0.00 |
| Comparative example 19 | Finishing grain | GA10mmol/L | 20℃ | 34.67±2.69 | 0.00±0.00 |
| Comparative example 20 | Finishing grain | GA100mmol/L | 20℃ | 26.67±2.41 | 1.33±0.25 |
| Comparative example 21 | Finishing grain | GA1000mmol/L | 20℃ | 0.00±0.00 | 0.00±0.00 |
From fig. 1 to 6, it can be seen that the peeled white fiveleaf akebia seed germinated from day 4, the whole white fiveleaf akebia seed germinated from day 14, and the half white fiveleaf akebia seed germinated from day 5, but the germination vigor was low. Therefore, the germination rate and the germination vigor of the akebia stem seeds can be obviously improved by germinating the peeled akebia stem seeds. From a combination of the experimental results of comparative examples 17 to 21 in Table 1 and FIG. 7, it is understood that gibberellin does not promote germination of Akebia quinata seeds.
The average seedling length and the total seedling length after germination of the akebia stem of example 1 and comparative examples 1 to 16 were measured, and the results are shown in tables 2 to 4. The standard deviation was calculated using the STDEV function in Excel.
Table 2 average seedling length and total seedling length data for whole seeds at different temperatures for example 1 and comparative examples 6 to 10
| Average seedling length/mm | Standard deviation of | Total seedling length | Standard deviation of | |
| Comparative example 6 | 14.85 | 1.59 | 297.05 | 31.79 |
| Example 1 | 26.10 | 0.85 | 504.66 | 21.10 |
| Comparative example 7 | 35.18 | 4.73 | 703.65 | 94.51 |
| Comparative example 8 | 8.06 | 0.89 | 161.28 | 17.79 |
| Comparative example 9 | 0.49 | 0.37 | 9.76 | 7.47 |
| Comparative example 10 | 0.03 | 0.05 | 0.67 | 1.03 |
Note that: the average seedling length is the average seedling length of three groups of parallel experiments, and the total seedling length is the average total seedling length of three groups of parallel experiments.
TABLE 3 average and Total seed Length data for whole seeds at different temperatures for comparative examples 1 to 5
| Average seedling length/mm | Standard deviation of | Total seedling length | Standard deviation of | |
| Comparative example 1 | 10.30 | 1.61 | 257.49 | 40.26 |
| Comparative example 2 | 14.96 | 3.69 | 373.95 | 92.13 |
| Comparative example 3 | 8.93 | 0.93 | 223.28 | 23.20 |
| Comparative example 4 | 0.61 | 0.49 | 15.18 | 12.20 |
| Comparative example 5 | 0.03 | 0.06 | 0.75 | 1.39 |
Note that: the average seedling length is the average seedling length of three groups of parallel experiments, and the total seedling length is the average total seedling length of three groups of parallel experiments.
TABLE 4 average seedling length and Total seedling length data for half-sized seeds at different temperatures for comparative examples 11 to 16
Note that: the average seedling length is the average seedling length of three groups of parallel experiments, and the total seedling length is the average total seedling length of three groups of parallel experiments.
The germination rate and germination vigor of the whole white akebia seeds subjected to total darkness stratification treatment in example 2 and comparative examples 22 to 24 are shown in Table 5, the germination progress in the light-dark alternate culture process of the whole white akebia seeds in different total darkness stratification days is shown in FIG. 8, and the seedling length of the whole white akebia seeds in different total darkness stratification days is shown in FIG. 9 after germination for 31 days. As can be seen from FIG. 8, after the whole akebia stem seeds are stacked in the complete darkness for 40 days, all the seeds germinate in the 1 st day of alternate light and dark culture, and the germination rate and the germination vigor of the whole akebia stem seeds reach 100%. After the whole akebia stem seeds are fully dark laminated for 10 days, the whole akebia stem seeds start to germinate on the 7 th day. After 20d and 30d of total darkness layering of the whole akebia stem seeds, the whole akebia stem seeds start to germinate on day 1, but have low germination vigor. As can be seen from fig. 9, the total length of seedlings of 25-piece white wood seeds laminated in total darkness for 40 days exceeds 2000cm, the total length of seedlings of 25-piece white wood seeds laminated in total darkness for 10 days exceeds 700cm, the total length of seedlings of 25-piece white wood seeds laminated in total darkness for 20 days exceeds 1100cm, and the total length of seedlings of 25-piece white wood seeds laminated in total darkness for 30 days exceeds 1300cm. Therefore, the total darkness stratified 40d can obviously improve the germination rate and vigour of the whole akebia stem seeds and the total length of 25 seedlings of the akebia stem seeds.
TABLE 5 Whole particle Akebia quinata germination Rate and vigor under total darkness stratification treatment of example 2 and comparative examples 22-24
| Seed (seed) | Laminated days (heaven) | Temperature (temperature) | Germination rate | Vigour of germination | |
| Example 2 | |
40 | 15℃ | 100.00±0.00 | 100.00±0.00 |
| Comparative example 22 | Finishing |
10 | 15℃ | 97.33±2.07 | 24.00±7.16 |
| Comparative example 23 | Finishing |
20 | 15℃ | 89.33±10.59 | 58.67±16.54 |
| Comparative example 24 | Finishing grain | 30 | 15℃ | 82.67±5.47 | 74.67±2.07 |
The average seedling length and total seedling length of the akebia stem after the total darkness lamination treatment in example 2 and comparative examples 22 to 24 are shown in table 6. The standard deviation was calculated using the STDEV function in Excel.
TABLE 6 average and Total Miao Length of Akebia quinata under different total darkness stratification treatments for example 2 and comparative examples 22-24
| Average seedling length/mm | Standard deviation of | Total seedling length | Standard deviation of | |
| Comparative example 22 | 30.92 | 0.95 | 773.01 | 23.67 |
| Comparative example 23 | 41.21 | 16.64 | 1030.25 | 415.95 |
| Comparative example 24 | 51.97 | 6.64 | 1299.25 | 165.99 |
| Example 2 | 85.81 | 6.66 | 2145.33 | 166.41 |
As is clear from tables 5 to 6, the germination rate of the whole akebia stem seeds of example 2 was 100% and the germination potential was 100% after the whole-grain akebia stem seeds were subjected to total darkness stratification. The total seedling length (25 seeds) of the akebia stem seeds stacked in the total darkness for 40 days exceeds 2000cm, and the germination method provided by the invention obviously improves the germination rate and the germination vigor of the akebia stem seeds, and the average seedling length and the total seedling length of the akebia stem seeds, and is simple to operate.
Although the foregoing embodiments have been described in some, but not all, embodiments of the invention, it should be understood that other embodiments may be devised in accordance with the present embodiments without departing from the spirit and scope of the invention.
Claims (7)
1. A method for promoting germination of akebia stem seeds, which is characterized by comprising the following steps:
mixing peeled akebia stem seeds with distilled water, and placing in a container for light-dark alternate culture until the germination of the seeds is finished; the peeled akebia stem seeds are seeds with seed coats removed and complete endosperm and seed embryo reserved; the container comprises a culture dish, wherein 2 layers of filter paper or gauze are paved in the culture dish; the light-dark alternate culture is light culture for 12-14 hours and dark culture for 10-12 hours, and the temperature is 10-35 ℃; the illumination intensity is 2000-3000 lux during illumination culture.
2. The method according to claim 1, wherein the volume ratio of the peeled akebia stem seed mass to distilled water is (1.5-2.5) g: (5-7) mL.
3. A method for promoting the growth of akebia stem seedlings, which is characterized by comprising the following steps:
mixing the whole caulis Aristolochiae with matrix particles, and placing in a container with air holes on the wall for total darkness lamination; culturing alternately in light and dark after the total darkness is laminated until the germination of seeds is finished; the matrix particles comprise perlite and/or wet sand; the light-dark alternate culture is light culture for 12-14 hours and dark culture for 10-12 hours, and the temperature is 10-35 ℃; the illumination intensity of the illumination culture is 2000-3000 lux; the total darkness lamination temperature is 3-5 ℃, and the total darkness lamination time is 35-40 d.
4. The method according to claim 3, wherein the total darkness lamination temperature is 3-5 ℃.
5. The method according to claim 3, wherein the mass ratio of whole seeds to perlite in the total darkness lamination is 1:4.
6. the method of claim 3, wherein the step of mixing the whole akebia stem seeds with the base particles further comprises placing the mixture of seeds and base particles into a seed bag; placing the seed bag into the container; the seed bag comprises a nylon net bag, and the seed bag is turned once every 7 days; perlite in the container remains moist; the container comprises a plastic container.
7. The method according to any one of claims 1 to 6, for use in improving germination rate and vigor of akebia stem seeds.
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| ZA2023/03887A ZA202303887B (en) | 2022-06-15 | 2023-03-28 | Method for promoting germination of akebia trifoliata subsp.australis seeds and method for promoting growth of seedlings of akebia trifoliata subsp.australis |
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