A method for collecting and storing gordon euryale seeds for seed saving

Abstract

The application discloses a kind of collection and storage methods of gordon euryale seed for seed saving, comprising the following steps: during the period of gordon euryale mature, collect development mature gordon euryale fruit, put into plastic frame;Frame is put in water, cover, after 5-10 days, gordon euryale peel, false seed coat naturally rots in water, seed naturally sinks;Seed is washed, remove too small, deformed seed, keep seed grain full, uniform seed;Seed is put into plastic bag, add water, tie mouth, store in 5-10 DEG C refrigerator.The water in plastic bag is replaced every 10-30 days during storage.The method can store gordon euryale seed to April-May of the second year, germination is accelerated in 25-28 DEG C incubator, and germination rate is kept above 90%.After 2 years of continuous storage, seed germination rate is still above 60%.The method is simple and easy to operate, does not need expensive equipment, seed germination rate is high, suitable for large quantities of gordon euryale seed collection and long-term storage.

Description

Technical Field

[0001] This invention belongs to the food field, specifically relating to a method for collecting and storing Euryale ferox seeds for seed preservation. Background Technology

[0002] Euryale ferox ( Euryale ferox ) is a plant of the genus Euryale in the family Nymphaeaceae. Euryale ferox The mature kernel of *Euryale ferox* (Salisb.), according to the *Shennong Bencao Jing* (Shennong's Classic of Materia Medica), has the effects of tonifying the spleen, stopping diarrhea, and removing dampness. *Euryale ferox* is also a food and medicine with various health benefits, including antioxidant properties, improved immunity, and blood sugar reduction. *Euryale ferox* reproduces by seed, but its seeds are stubborn and need to be kept moist during storage to maintain a high germination rate. If *Euryale ferox* seeds are dried, their germination rate will drop significantly, and in severe cases, they may even completely lose their germination ability. In actual production, farmers often store the seeds in water based on experience, but due to the inability to guarantee environmental conditions, it is difficult to ensure the seed's germination vitality.

[0003] Recalcitrant seeds are those that are highly sensitive to water loss. These seeds require protection from both dryness and low temperatures during storage. Their germination process is influenced not only by moisture levels but also by the metabolism of stored substances and the integrity of cell structure. Recalcitrant seeds generally have a high water content at maturity, typically between 30% and 60%, and readily initiate germination after harvesting. However, dehydration inhibits germination, drastically reducing seed viability. There are two main methods for preserving these seeds: one is using a temperature- and humidity-controlled method, which avoids dehydration and low-temperature damage, extending the shelf life to several months or even over a year. The other is cryopreservation of isolated embryos (or hypocotyls) using liquid nitrogen; for example, rubber seed embryos have been successfully regenerated into plants using this method. However, the latter method is too costly and requires specialized operation, making it difficult to promote widely.

[0004] The most common method for preserving water chestnut seeds is to pack them into woven bags and then place them in an open-air pool or shallow pond. However, this method is susceptible to damage if the temperature fluctuates during storage, potentially causing premature germination and resulting in losses. Therefore, for large-scale cultivation, a more efficient seed storage method is urgently needed to ensure high germination viability and safe storage until the following spring or longer. Summary of the Invention

[0005] To address the above problems, and through extensive experimental research, this invention provides a method for collecting and storing Euryale ferox seeds for seed preservation. This method allows the seeds to germinate in a 25°C incubator during April-May of the following year, maintaining a germination rate of over 90%.

[0006] The present invention relates to a method for collecting and storing Euryale ferox seeds for seed preservation, comprising the following steps:

[0007] (1) Collection of mature water chestnut fruits: In July and August each year, when the water chestnut fruits are ripe in large quantities, select healthy water chestnut plants that are free from pests and diseases, and collect water chestnut fruits that are well-shaped, plump and mature.

[0008] (2) Pretreatment of Gorgon Fruit: Clean the outer skin of the collected gorgon fruit and place it in a plastic frame. Fill the frame halfway with tap water, put the gorgon fruit in the water, and cover it with a lid, but not too tightly, allowing air to pass through. At this time, it is summer and the temperature is high. After 5-10 days at room temperature, the gorgon fruit peel and internal tissues will naturally decompose, and the gorgon fruit seeds with aril will naturally disperse and float on the water surface. As the aril decomposes and degrades, the gorgon fruit seeds will gradually sink to the bottom of the plastic frame. This treatment process simulates the actual changes in the later stage of gorgon fruit development in nature.

[0009] (3) Collection of gorgon fruit seeds: Take out the gorgon fruit seeds that have sunk to the bottom of the frame, remove impurities, seeds that are too small, immature or deformed seeds, select gorgon fruit seeds that are plump and uniform in size, and wash them clean with tap water.

[0010] (4) Packaging of Euryale seeds: Put the cleaned Euryale seeds into thickened plastic bags at a rate of 30-50 catties per bag, add tap water, with the water level 5-10 cm above the seeds, and tie the bag tightly.

[0011] (5) Storage of Euryale ferox seeds: Place the plastic bags from (4) side by side into a large plastic frame, and then store them together in a cold storage at 5-10℃. During storage, change the water in the plastic bags every 10-30 days. Under these conditions, Euryale ferox seeds can be stored until April-May of the following year, and then germinated in a 25-28℃ incubator, with a germination rate of over 90%. After 2 years of storage, the germination rate still remains above 60%.

[0012] further:

[0013] In step (1), mature water chestnut fruits should be collected during the peak ripening period. If it is too early, the fruits will be too small, and if it is too late, the ripeness of the fruits cannot be guaranteed. Generally, it is between mid-July and late August. Select healthy water chestnut plants that are free from pests and diseases, and collect water chestnut fruits that are well-shaped, plump, and fully developed.

[0014] The beneficial effects of this invention are reflected in:

[0015] This invention introduces a method for collecting, pretreating, and storing *Euryale ferox* seeds for seed production. Following this method, *Euryale ferox* seeds can be safely stored until April or May of the following year, and germinated in an incubator at 25-28℃, maintaining a germination rate of over 90%. Even after two years of continuous storage, the germination rate remains above 60%. In this method, the pretreatment stage allows the *Euryale ferox* seeds to fully ripen naturally, ensuring complete seed development; simultaneously, constant temperature storage at 5-10℃ avoids premature germination caused by temperature fluctuations during traditional open-air storage. The method is simple and easy to implement, requires no expensive equipment, and has a high seed germination rate, making it suitable for large-scale collection and long-term storage of *Euryale ferox* seeds.

[0016] This invention relates to a pretreatment method for water chestnut seeds, referencing the natural ripening process of water chestnut fruits while also considering the effects of human intervention, such as removing the pericarp and aril after harvesting, on seed dormancy and germination rate. Four different pretreatment methods were designed: (1) whole fruit with pericarp; (2) seeds with aril removed; (3) seeds with aril removed and the hilum between the seed and aril manually cut off; (4) water chestnut seeds directly extracted from the fruit and added to water. After storage and germination, the germination rate, germination potential, and endogenous enzyme activity of water chestnut seeds obtained from different pretreatment methods were analyzed. The results showed that whole fruit with pericarp, after natural rotting in water, yielded the highest germination rate and germination potential of the water chestnut seeds. Furthermore, a significant correlation was found between the germination rate of water chestnut seeds and the activity of endogenous proteases and amylases. Related experimental results are attached. Figure 2 See Table 1. Attached Figure Description

[0017] Figure 1 This is a flowchart of the process for collecting and storing foxnut seeds.

[0018] Figure 2 The effect of pretreatment methods on the germination rate and germination potential of Gorgon fruit seeds is described in Example 1. The methods included: (1) with pericarp attached; (2) with aril attached; (3) with aril attached and hilum cut off; and (4) water added to the seeds. Figure 2 The results show that the germination rates of water chestnut seeds under different pretreatment methods are in the following order: (1) with pericarp > (2) with aril > (3) with aril and the hilum cut off > (4) water chestnut seeds with water added, which are 95.12%, 94.87%, 90.34%, and 80.77%, respectively. The germination potential and germination rate are basically consistent. The group with pericarp is slightly higher than the group with aril, but there is no significant difference between the two. This indicates that after the mature water chestnut fruit is harvested, the aril and pericarp on the outside of the water chestnut seed can still provide nutrients or transmit some kind of signal to the seed through the hilum, which can be regarded as the after-ripening process of water chestnut seed.

[0019] Figure 3This study investigated the effects of storage temperature and humidity on the germination rate and germination potential of *Euryale ferox* seeds. The results were as follows: 5℃ without water (A); 5℃ with water (B); 10℃ without water (C); 10℃ with water (D). Figure 3 As can be seen from the data, under both storage temperatures, the germination potential and germination rate of water-added seeds were significantly higher than those of unwatered seeds (P < 0.05). Furthermore, the germination rate of water-added seeds remained relatively stable during the 365-day storage period, initially increasing and then decreasing at 5℃, while the germination activity at 10℃ showed no significant change in the early stage and decreased slightly in the later stage, but remained above 90% in all cases. Figure 3 (B, D). The germination potential and germination rate of water chestnut seeds stored at 5℃ and 10℃ for 365 days without water were significantly reduced. Figure 3 (A and C). Among them, at 5℃ without water, the germination potential and germination rate showed a trend of first increasing and then decreasing with the extension of storage time. The highest value was 72.62% after 215 days of storage, and then it decreased rapidly, dropping to 11.05% after 365 days. Detailed Implementation

[0020] The following examples are provided to better explain the process of the present invention and are not intended to limit the process of the present invention.

[0021] Example 1:

[0022] The method for preparing and storing Euryale ferox seeds for seed saving in this embodiment includes the following steps:

[0023] (1) Collection of mature water chestnut fruits: In mid-July each year, when the water chestnut fruits are ripe in large quantities, select healthy water chestnut plants that are free from pests and diseases, and collect water chestnut fruits that are well-shaped, plump and mature.

[0024] (2) Pretreatment of Gorgon fruit: Clean the outer skin of the collected gorgon fruit and place it in a plastic frame. Fill the frame halfway with tap water, put the gorgon fruit in the water, and cover it with a lid, but not too tight, allowing air to pass through. At this time, it is the summer heat. After 7 days at room temperature, the gorgon fruit peel and internal tissues quickly rot, and the gorgon fruit seeds with arils are naturally released and float on the water surface. Subsequently, the arils decompose and degrade, and the gorgon fruit seeds gradually sink to the bottom of the plastic frame.

[0025] (3) Collection of gorgon fruit seeds: Take out the gorgon fruit seeds that have sunk to the bottom of the frame, remove impurities, seeds that are too small, immature or deformed seeds, select gorgon fruit seeds that are plump and uniform in size, and wash them clean with tap water.

[0026] (4) Packaging of Euryale seeds: Put the cleaned Euryale seeds into thickened plastic bags at a rate of 30 catties per bag, add tap water, and make sure the water level is about 8 cm above the seeds. Then tie the bag tightly.

[0027] (5) Storage of Euryale ferox seeds: Place the plastic bags from (4) side by side into a large plastic frame, and then store them together in a 5℃ cold storage. During storage, change the water in the plastic bags every 20 days. Under these conditions, the Euryale ferox seeds can be stored until around April 10th of the following year, and then germinated in a 25℃ incubator, with a germination rate of 92.4%. After 2 years of storage, the germination rate is 63.8%.

[0028] Figure 2 This study investigated the effects of pretreatment methods on the germination rate and germination potential of Euryale ferox seeds. The methods included: (1) placing the fruit with its pericarp in water until it completely rotted and the seeds sank to the bottom; (2) removing the seeds from the Euryale ferox fruit, keeping the aril intact, placing them in water until the aril completely rotted and the seeds sank to the bottom; (3) removing the seeds from the Euryale ferox fruit, keeping the aril intact, and manually cutting off the hilum between the seed and the aril; (4) removing the seeds from the Euryale ferox fruit, removing the aril, leaving only the seeds, and adding water. The samples obtained were placed at room temperature until all tissues except the seeds rotted. The Euryale ferox seeds were then washed out, placed in a self-sealing bag, added water, and stored at 5-10℃ for later use. Germination rate and germination potential were determined: Euryale ferox seeds were germinated at 25℃, and the germination status was regularly recorded. The germination rate and germination potential were calculated.

[0029] Table 1 below shows the effects of seed pretreatment methods on the main nutrient components and enzyme activities in *Euryale ferox* seeds. It can be seen that the key factors affecting the germination of *Euryale ferox* seeds are the activities of endogenous amylase and protease, and different pretreatment methods have a significant impact on the activity of these endogenous enzymes.

[0030]

[0031] Example 2:

[0032] The method for preparing and storing Euryale ferox seeds for seed saving in this embodiment includes the following steps:

[0033] (1) Collection of mature water chestnut fruits: In early August each year, when the water chestnut fruits are ripe in large quantities, select healthy water chestnut plants that are free from pests and diseases, and collect water chestnut fruits that are well-shaped, plump and mature.

[0034] (2) Pretreatment of Gorgon fruit: Clean the outer skin of the collected gorgon fruit and place it in a plastic frame. Fill the frame halfway with tap water, put the gorgon fruit in the water, and cover it with a lid, but not too tight, allowing air to pass through. At this time, it is the summer heat. After 10 days at room temperature, the gorgon fruit peel and internal tissues quickly rot, and the gorgon fruit seeds with arils are naturally released and float on the water surface. Subsequently, the arils decompose and degrade, and the gorgon fruit seeds gradually sink to the bottom of the plastic frame.

[0035] (3) Collection of gorgon fruit seeds: Take out the gorgon fruit seeds that have sunk to the bottom of the frame, remove impurities, seeds that are too small, immature or deformed seeds, select gorgon fruit seeds that are plump and uniform in size, and wash them clean with tap water.

[0036] (4) Packaging of Euryale seeds: Pack the cleaned Euryale seeds into thickened plastic bags at a rate of 40 jin per bag, add tap water, with the water level about 10 cm above the seeds, and tie the bag tightly.

[0037] (5) Storage of Euryale ferox seeds: Place the plastic bags from (4) side by side into a large plastic frame, and then store them together in a 10℃ cold storage. During storage, change the water in the plastic bags every 30 days. Under these conditions, the Euryale ferox seeds can be stored until around April 20th of the following year. After germination in a 28℃ incubator, the germination rate is 93.5%. After 2 years of storage, the germination rate is 64.9%.

Claims

1. A method for collecting and storing Euryale ferox seeds for seed saving, characterized in that... Includes the following steps: (1) Harvesting mature water chestnut fruits: In July and August each year, when water chestnut fruits ripen in large quantities, select healthy water chestnut plants free from pests and diseases, and harvest water chestnut fruits that are well-shaped, plump, and mature. (2) Pretreatment of water chestnut fruit: Clean the outer skin of the collected water chestnut fruit, then put the water chestnut fruit into a container filled with water, cover it and keep it breathable. After 5-10 days at room temperature, the peel and internal tissue of the water chestnut fruit will naturally rot, and the seeds of water chestnut fruit with aril will naturally be released and float on the water surface. Then the aril will rot and degrade, and the seeds of water chestnut fruit will gradually sink to the bottom of the plastic frame. (3) Collection of gorgon fruit seeds: Take out the gorgon fruit seeds that have sunk to the bottom of the frame, remove impurities, seeds that are too small, immature or deformed seeds, select gorgon fruit seeds that are plump and uniform in size, and wash them with tap water. (4) Packaging of Euryale seeds: Put the cleaned Euryale seeds into a thickened plastic bag, add tap water, and tie the bag tightly; (5) Storage of Euryale ferox seeds: Place the plastic bag in (4) into a cold storage at 5-10℃ and change the water in the plastic bag regularly during storage.

2. The method according to claim 1, characterized in that: In step (1), the harvesting time for the gorgon fruit is from mid-to-late July to late August.

3. The method according to claim 1, characterized in that: In step (4), the cleaned water chestnut seeds are packed into thickened plastic bags at a rate of 30-50 jin per bag, and tap water is added, with the water level 5-10 cm above the seeds. The bag is then tied tightly.

4. The method according to claim 1, characterized in that: In step (5), during storage, the water in the plastic bag should be replaced every 10-30 days.

5. The method according to claim 1, characterized in that: The seeds of Euryale ferox were stored until April or May of the following year and then germinated in an incubator at 25-28℃, with a germination rate of ≥90%.

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