Composition and method for long-distance transportation, storage and sowing of cyclobalanopsis glauca fruits

Abstract

The invention particularly relates to a composition and a method for long-distance transportation, storage and sowing of cyclobalanopsis glauca fruits. According to the specific technical scheme, the method for prolonging the preservation period of the cyclobalanopsis glauca fruits comprises the steps that the cyclobalanopsis glauca fruits are collected and put into an aqueous solution containing 5-aminolevulinic acid and abscisic acid, the fruits are taken out after being soaked, then the fruits are put into a sealed space, and 1-methylcyclopropene is introduced for fumigation. The invention provides a novel method for treating cyclobalanopsis glauca fruits, the method is low in cost and convenient to operate in the field, the period of validity of the treated fruits under the conventional logistics transportation condition can reach 20-25 days, and enough time is reserved for continuous collection and transportation.

Description

Technical Field

[0001] This invention belongs to the field of plant cultivation, specifically relating to a composition and method for long-distance transportation, preservation, and sowing of thin slices of oak fruit. Background Technology

[0002] *Quercus lamellosa* Sm., a species of oak in the family Fagaceae, is an important component of the semi-evergreen broad-leaved forests of the Eastern Himalayas. The fruit of *Quercus lamellosa* is round, with the seeds enclosed in a cupule approximately 10 mm thick, which neither falls off nor splits open when mature. The thick cupule stores a significant amount of nutrients and water.

[0003] Because *Quercus stenoptera* is mostly distributed in the deep forests of subtropical regions, where temperatures are high and transportation is inconvenient, the fruit is extremely prone to rotting or drying out and becoming inactive before reaching the sowing test site without suitable preservation methods. Preservation and transportation experiments have shown that under normal temperature conditions, *Quercus stenoptera* can only be preserved for 3-5 days. By the fourth day, 40-60% of the fruit begins to blacken and rot, and by the sixth day, about 90% of the fruit has turned black and rotten, with the seeds inside also blackening, rotting, and losing their viability.

[0004] Harvesting and transporting fruits in the wild is usually time-consuming, and inconvenient transportation leads to high cold chain transportation costs and is subject to transportation route restrictions.

[0005] Therefore, developing a low-cost, field-operable post-harvest processing technology for thin-sliced ​​oak fruits to enable long-distance cold chain-free transportation of the fruits has significant scientific research and application value. Summary of the Invention

[0006] The purpose of this invention is to provide a composition and method for long-distance transportation, preservation, and sowing of thin-sliced ​​oak fruits.

[0007] To achieve the above-mentioned objective, the technical solution adopted by this invention is: a composition for extending the shelf life of thin-sliced ​​oak fruit, the composition comprising: 5-aminolevulinic acid and abscisic acid. The composition also includes 1-methylcyclopropene.

[0008] Accordingly, a method for extending the shelf life of thin-sliced ​​oak fruit includes the following steps:

[0009] (1) Collect thin slices of oak fruit and soak them in an aqueous solution containing 5-aminolevulinic acid and abscisic acid for 20-30 minutes.

[0010] (2) Remove the fruit, drain the water, and place it in a dark place to dry for 12-24 hours;

[0011] (3) Place the fruit in a sealed space and fumigate with 1-methylcyclopropene for 16-24 hours;

[0012] (4) Prepare a fruit treatment agent, which includes quicklime, sodium dithionite and dried sphagnum moss. Mix the fruit treatment agent with thin slices of oak fruit and seal for storage.

[0013] Accordingly, a method for improving the germination rate of thin-layered Quercus glauca seeds and / or the survival rate of seedlings includes the following steps:

[0014] (1) Collect thin slices of oak fruit and soak them in an aqueous solution containing 5-aminolevulinic acid and abscisic acid for 20-30 minutes.

[0015] (2) Remove the fruit, drain the water, and place it in a dark place to dry for 12-24 hours;

[0016] (3) Place the fruit in a sealed space and fumigate with 1-methylcyclopropene for 16-24 hours;

[0017] (4) Prepare a fruit treatment agent, which includes: quicklime, sodium dithionite and dried sphagnum moss. Mix the fruit treatment agent with thin slices of oak fruit and seal and store.

[0018] (5) After preservation, rinse the fruit with clean water to clean the residual agent on the fruit surface before sowing.

[0019] (6) Soak the fruit in a gibberellin aqueous solution with a concentration of 40-50 ppm and a water temperature of 28-35℃ for 10-15 hours, then remove and drain.

[0020] (7) Mix the fruit with the growth promoter; the growth promoter formula includes difenoconazole, pyraclostrobin, fludioxonil and thiamethoxam.

[0021] Accordingly, a composition for improving the survival rate of *Quercus glauca* seedlings includes: difenoconazole, azoxystrobin, fludioxonil, and thiamethoxam. The composition also includes a water-retaining agent.

[0022] Accordingly, a method for improving the survival rate of *Quercus spp.* seedlings includes the following steps: mixing *Quercus spp.* seeds with a composition that can improve the survival rate of *Quercus spp.* seedlings, wherein the composition includes: difenoconazole, pyraclostrobin, fludioxonil, and thiamethoxam.

[0023] The present invention has the following beneficial effects: The present invention provides a new method for processing thin slices of oak fruit. This method is not only low in cost, but also convenient for field operation. The processed fruit can be kept for 20 to 25 days under conventional logistics and transportation conditions, which allows sufficient time for continuous collection and transportation.

[0024] The method provided in this invention utilizes a combination of agents. Specifically, treating thin-sliced ​​oak fruits with appropriate concentrations of 5-aminolevulinic acid and 1-MCP significantly inhibits fruit respiration, particularly anaerobic respiration. This prevents seed germination during transportation and avoids fruit rot and seed inactivation due to fermentation caused by anaerobic respiration. The slaked lime in the preservative is alkaline, effectively preventing mold and insects, and also appropriately absorbs carbon dioxide produced by fruit respiration and moisture from transpiration, thus preventing fruit rot and maintaining seed viability.

[0025] In addition, this invention provides an agent and method for improving seedling survival rate, which can effectively solve the problem of high seedling mortality within two months of seed emergence. Comparative experiments have shown that the germination and survival rate of fruit seedlings treated according to the method of this invention is 36-50% higher than that of the control group.

[0026] Moreover, the present invention has made a new discovery in the research: unlike the sowing of traditional Fagaceae plants, the sowing of thin-sliced ​​Quercus glauca requires sowing with the cupules attached.

[0027] Finally, this invention also resolves the contradiction between the nutrients provided by the fruit shell and the rot pathogens through the treatment of the embedding agent. It can ensure that the fruit shell provides nutrients for seed germination and improve the germination rate, while also preventing harmful pathogens from the rotting of the fruit shell, thereby further improving the survival rate of seedlings. Attached Figure Description

[0028] Figure 1 Top view, side view, and bottom view of a thin slice of oak fruit;

[0029] Figure 2 A side sectional view of a thin slice of oak fruit after half of the husk has been removed. Detailed Implementation

[0030] This invention provides a method for the long-term preservation or long-distance transportation of thin slices of Quercus glauca seeds, specifically comprising the following steps:

[0031] (1) After collecting thin slices of oak fruit, wash off the surface dirt and stains, and then soak them in an aqueous solution containing 25-35 mg / L of 5-ALA (5-aminolevulinic acid) and 0.2 mg / L of ABA (abscisic acid) for 20-30 minutes.

[0032] (2) Remove the fruit, drain the water, and spread it out in a dark place to dry for 12-24 hours.

[0033] (3) After checking that the fruit surface is free of moisture, place the fruit in a sealed space (such as a sealed box, a well-sealed room, etc.), calculate the internal volume of the space, and introduce 1-MCP sodium salt (1-methylcyclopropene) at a ratio of 1.2 μL / L for fumigation treatment. The specific fumigation method is as follows: Take a glass, add purified water at a weight ratio of water to 1-methylcyclopropene sodium salt ≥ 1:30 (1-methylcyclopropene sodium salt can release 1-MCP by reacting with a small amount of water), then quickly pour in the weighed 1-MCP sodium salt, seal the space, and fumigate for 16 to 24 hours, followed by ventilating for 4 hours.

[0034] (4) Prepare the fruit treatment agent by weight ratio of quicklime: sodium dithionite: dried sphagnum moss = 20:1:5~25:1.5:10. To ensure effectiveness, the compressed sphagnum moss should be soaked in water beforehand, then broken up and dried before use. Mix the fruit treatment agent according to the ratio, and then mix it with the sliced ​​oak fruit at a ratio of 10:1 (10kg fruit: 1kg fruit treatment agent), and seal for storage.

[0035] Preferably, after inhibiting fruit germination and preserving fruit, the present invention may further include methods to improve fruit germination rate and seedling survival rate:

[0036] (5) Pre-sowing treatment

[0037] After transportation or storage, open the packaging and sift out the fruit. Rinse the fruit with clean water to remove any residual pesticide from the surface. Then, soak the fruit in a gibberellin aqueous solution with a concentration of 40–50 ppm and a water temperature of 28–35°C for 10–15 hours. After soaking, remove the fruit and drain the water to relieve the inhibitory effect of the pesticide on the seeds.

[0038] Mix the thin slices of Quercus glauca seeds with the growth promoter. The growth promoter formula, by weight, is: difenoconazole: azoxystrobin: fludioxonil: thiamethoxam: water-retaining agent (starch grafting acrylate) = 1:3:2:5:15~1:4:5:7:25. Mix the fruit and promoter at a rate of 1 kg of promoter per 50 kg of fruit, ensuring all fruit is coated with the promoter.

[0039] (6) The treated fruit can be sown.

[0040] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Unless otherwise specified, the technical means used in the embodiments are conventional means well known to those skilled in the art, and the data obtained are all average values ​​obtained after at least three repetitions, and each repetition yields valid data.

[0041] Example 1: Effect of cupules on the germination rate of thin-shelled oak seeds

[0042] Thin-sliced ​​oak seeds look like Figure 1 , 2 As shown. Figure 1 Photos of the fruit with its complete cupule, from top to bottom: Top view Figure 1 Looking down Figure 2 Top view and side view. Figure 2 The images show side and cross-sectional views of the fruit with half of the cupule removed. It can be seen that most of the thin-sliced ​​oak fruit is enclosed by the cupule, which is quite thick. Under natural growth and budding conditions, the cupule does not fall off naturally.

[0043] Healthy, plump, and intact *Quercus glauca* fruits collected from a *Quercus glauca* forest in Hanmi Village, Beibeng Township, Motuo County, Tibet, were selected. One hundred fruits were randomly chosen, with the incisors completely removed but the pericarps retained, to form the experimental group. Another 100 fruits with the incisors retained served as the control group. Seeds from each group were washed with clean water and then placed in a seedling tray. The temperature was controlled at 22℃ and the humidity at 70–85%. Seed germination was observed, and the germination rate was recorded over time. The results are shown in Table 1.

[0044] Table 1. Comparison of the effects of cupule shells on seed germination rate

[0045]

[0046] The results showed that removing the cupule significantly reduced the germination rate of *Quercus glauca* seeds, with no new seedlings emerging after about 10 days. Dissection of ungerminated seeds 12 days after sowing revealed that the seeds were internally rotten and inactive. This may be because the cupule contains certain unknown components that help maintain seed viability and promote germination. This is in stark contrast to other *Quercus* seeds, which require cupule removal (in some varieties, the cupule may detach naturally) or even removal of the pericarp / pericarp to promote germination; this characteristic is unique to *Quercus stenoptera*. Therefore, the cupule should be retained in the seedling cultivation of *Quercus stenoptera*.

[0047] Example 2: Effects of germination inhibitors and anti-fruit rot agents on seed germination

[0048] Seeds of *Quercus acutissima* will begin to germinate naturally about 3-5 days after harvesting or falling to the ground. However, if the growing environment is not ideal at this time, or if sowing has not yet occurred, the semi-germinated seeds will quickly become inactive and cannot be transported or stored. Therefore, it is necessary to treat them with an appropriate concentration of seed germination inhibitor to extend the transportation and storage time. However, due to the characteristics of *Quercus acutissima* seeds, the type and concentration of seed inhibitors will affect seed germination, and may even cause the seeds to fail to germinate or even die.

[0049] 1. Effect of ABA concentration on seed germination

[0050] Healthy, plump, and intact Quercus glauca seeds from the same batch were selected and grouped into sets of 100. After being thoroughly washed with clean water, the cupules were preserved. Each seed was then soaked individually for 4 hours in ABA at the concentrations shown in Table 2. Subsequently, all seeds were placed in a seedling tray at a controlled temperature of 22℃ and humidity of 70–85%. Seed germination was observed, and the germination rate was recorded over time. The results are shown in Table 2. The group with an ABA concentration of 0 in Table 2 serves as the control group in Table 1 of Example 1.

[0051] Table 2. Effects of ABA concentration on seed germination rate (Comparison table)

[0052]

[0053] The results showed that when the ABA concentration was 0.2 mg / L, the germination time of 90% of the seeds could be extended from 6-8 days to about 20 days, and the overall germination rate was relatively high, meeting transportation requirements. As the ABA concentration continued to increase, the germination time of most seeds was further prolonged, but the overall germination rate decreased significantly, possibly due to irreversible inhibition and inactivation of the seeds by ABA. Therefore, an ABA concentration of 0.2 mg / L was chosen.

[0054] 2. Effects of 5-ALA concentration on fruit storage time and bud break

[0055] Two hundred fruits were selected for each group, and the treatment method was the same as in step 1. The fruits were soaked for 4 hours using a combination of 0.2 mg / L ABA and different concentrations of 5-ALA. The concentration of 5-ALA in each group is shown in Table 3. After treatment, each group was divided into four portions of 50 fruits each and placed in a polyethylene sealed bag for storage at 20–28°C (room temperature). The preservation status of the fruits with shells in the bags was observed and recorded at the time points shown in Table 3. After being stored in the sealed bags for the corresponding time, one portion was taken out for a sowing and germination experiment. The sowing and germination experiment method was as follows: the corresponding fruits were taken out of the sealed bags and rinsed with clean water 5 times to wash off the residual agent on the surface of the fruits. The fruits were then soaked in gibberellin aqueous solutions with a concentration of 50 ppm and a temperature of 30°C for 10 hours, and then removed and drained. The fruits and growth promoters were mixed at a ratio of 50:1 (50 kg fruits: 1 kg agent) to ensure that all fruits were coated with the agent. The growth-promoting agent formulation (active ingredient weight ratio) is: difenoconazole: azoxystrobin: fludioxonil: thiamethoxam: starch-grafted acrylate = 1:3:2:5:15. Unless otherwise specified below, the sowing and germination experiment method is the same as described here.

[0056] Table 3. Effects of 5-ALA concentration on fruit storage time and germination rate (Comparison Table)

[0057]

[0058] Table 2 shows germination experiments conducted in a laboratory seedling tray under optimal temperature and humidity conditions. Only the fruit germination rate was recorded, without further observation. Table 3, however, shows fruits treated and stored at room temperature in sealed bags, more closely resembling the transportation and storage conditions of fruits in a real environment. It can be seen that under real-world conditions, a 5-ALA concentration of 35 mg / L can increase the seed germination rate to 75% after 10 days. As the concentration of the herbicide continues to increase, the seed germination rate decreases sharply, possibly due to the herbicide effect caused by excessively high concentrations, leading to seed inactivation.

[0059] 3. The impact of anti-rot agents on fruit storage time and germination rate

[0060] After extensive preliminary testing, three agents that can delay the decay of thin-sliced ​​oak fruits were selected from various agents: 5-ALA, 1-MCP, and a mixed treatment agent made by mixing quicklime, sodium dithionite, and dried sphagnum moss in a specific ratio (quicklime: sodium dithionite: dried sphagnum moss = 20:1:5). The fruits were treated according to the methods shown in Table 4, as detailed below:

[0061] Two hundred fruits were selected from each group and treated with different agents and combinations thereof, using 0.2 mg / L ABA. The treatment methods are as described in step 2. After treatment, each experimental group was divided into four portions of 50 fruits each, and each portion was placed in a sealed polyethylene bag and stored at 20–28°C (room temperature). The preservation status of the fruits with inverted shells was observed and recorded at the time points listed in Table 4. One portion was then taken out sequentially for a sowing and germination experiment to observe the fruit condition and germination rate.

[0062] Table 4. Comparison of the effects of different agents and combinations of agents on fruit storage time and germination rate.

[0063]

[0064]

[0065] The results showed that both 5-ALA and 1-MCP significantly prolonged the shelf life of thin-sliced ​​oak fruits, but their effect on improving seed germination rate was relatively limited. The combined use of 5-ALA and 1-MCP significantly improved seed germination rate compared to using either agent alone. The mixed treatment was not ideal for fruit preservation, but it was better at maintaining seed germination rate, especially during the first 10 days of storage. The combined use of 5-ALA, 1-MCP, and the mixed treatment significantly improved both fruit preservation and seed germination rate compared to using either agent alone.

[0066] Therefore, when the agents are used in combination, the fruits can be transported and stored normally at room temperature for more than 20 days, effectively solving the problem of difficult transportation and storage of seeds after collection in remote areas.

[0067] Example 3: Effect of growth-promoting agents on seedling survival rate

[0068] One hundred thin-sliced ​​oak seeds (Group 6, Table 4, Example 2) that had been treated and stored for 20 days were subjected to a growth-promoting treatment. The specific treatment method was as follows: the fruits were rinsed with clean water to remove any residual agent from the surface. Then, they were soaked in a gibberellin aqueous solution with a concentration of 50 ppm and a water temperature of 28–35°C for 10 hours, after which they were removed and drained. Finally, the thin-sliced ​​oak seeds were mixed thoroughly with the growth-promoting agent.

[0069] The growth-promoting agent formula, by weight ratio, is: difenoconazole: azoxystrobin: fludioxonil: thiamethoxam: water-retaining agent (starch grafting acrylate) = 1:3:2:5:15. Mix the fruit and agent thoroughly according to a dosage of 1 kg of agent per 50 kg of fruit, ensuring all fruit is coated. Sow the treated fruit. Results showed that the seed germination rate was over 90%, and the seedling survival rate after three months was 94%.

[0070] Meanwhile, 100 seeds were kept under identical conditions except that they were not treated with growth promoters or any other fungicides, serving as control group 1; and seeds soaked in 1000 times diluted carbendazim for 4 hours were sown, serving as control group 2. The seed germination rate of control group 1 was 90%, but the seedling survival rate was only 44% after 3 months; the seed germination rate of control group 2 was 90%, and the seedling survival rate after three months was 48%.

[0071] In addition, the experiment found that adding starch-grafted acrylate to the growth promoter, when mixed with the moist cupules (washed fruit), will turn into a jelly-like colloid when it gets wet in the soil, thus encapsulating the cupules. This prevents the growth promoter from being lost quickly, but releases it slowly, adhering to the fruit surface for a long time and stably exerting its antibacterial effect.

[0072] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Any modifications, alterations, substitutions, or variations made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention shall fall within the protection scope defined by the claims of the present invention.

Claims

1. A composition for extending the shelf life of Cyclocarya paliurus fruits, characterized in that: The composition comprises 5-aminolevulinic acid and abscisic acid. ​ 2. The composition of claim 1, wherein: The composition further comprises 1-methylcyclopropene.

3. A method for extending the shelf life of a thin slice of a blueberry fruit, the method comprising: The method comprises the following steps: ​ (1) collecting the fruits of Cyclobalanopsis meijsnieri, and soaking them in an aqueous solution containing 5-aminolevulinic acid and abscisic acid for 20-30 min; (2) taking out the fruits, draining the water, and airing them in a dark place for 12-24 h; (3) placing the fruits in a sealed space, and fumigating them with 1-methylcyclopropene for 16-24 h.

4. The method of claim 3, wherein: The method further comprises: (4) preparing a fruit treatment agent comprising slaked lime, sodium hydrosulfite, and dry water moss, and uniformly mixing the fruit treatment agent with the fruits of Cyclobalanopsis meijsnieri, and sealing and storing them.

5. A method of increasing the germination rate and / or the survival rate of seedlings of Cyclobalanopsis gilva, characterized by: The method comprises the following steps: (1) collecting the fruits of Cyclobalanopsis meijsnieri, and soaking them in an aqueous solution containing 5-aminolevulinic acid and abscisic acid for 20-30 min; (2) taking out the fruits, draining the water, and airing them in a dark place for 12-24 h; (3) placing the fruits in a sealed space, and fumigating them with 1-methylcyclopropene for 16-24 h; (4) preparing a fruit treatment agent comprising slaked lime, sodium hydrosulfite, and dry water moss, and uniformly mixing the fruit treatment agent with the fruits of Cyclobalanopsis meijsnieri, and sealing and storing them; (5) after the storage is completed, washing the fruits with clean water to clean the residual agents on the surface of the fruits, and sowing the fruits.

6. The method of claim 5, wherein: After the fruits are cleaned in step (5), before sowing, the method further comprises: (6) placing the fruits in an aqueous gibberellin solution with a concentration of 40-50 ppm and a water temperature of 28-35℃, soaking them for 10-15 h, and then taking them out and draining the water.

7. The method of claim 6, wherein: The method further comprises: (7) uniformly mixing the fruits with a growth-promoting agent; The growth-promoting agent comprises difenoconazole, azoxystrobin, fludioxonil, and thiamethoxam.

8. A composition for improving the survival rate of seedlings of Cyclobalanopsis gilva, characterized by comprising: The composition comprises difenoconazole, azoxystrobin, fludioxonil, and thiamethoxam. ​ 9. The composition of claim 8, wherein: The composition further comprises a water-retaining agent.

10. A method of increasing the survival rate of seedlings of Cyclobalanopsis gilva, characterized by: The method comprises the following steps: uniformly mixing the seeds of Cyclobalanopsis meijsnieri with a composition capable of improving the survival rate of Cyclobalanopsis meijsnieri seedlings, the composition comprising difenoconazole, azoxystrobin, fludioxonil, and thiamethoxam.

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