A cultivation method for enriching lithium element in pilularia fontana schkuhr

Abstract

The present application relates to the field of agricultural planting, in particular to a kind of cultivation method of enriching lithium element of Pteroxygonum giraldii. The cultivation method comprises the following steps: (1) seedling raising: Pteroxygonum giraldii seeds are sown in the substrate for seedling raising; (2) transplanting: when Pteroxygonum giraldii seedlings grow to 4-5 true leaves, the Pteroxygonum giraldii seedlings are transplanted into a water culture box containing clean water; after 3 days of transplanting, the clean water in the water culture box is replaced with 1 / 8 concentration nutrient solution and exogenous selenium, and then the nutrient solution and exogenous selenium are replaced every 10 days; after 45-60 days of transplanting, 1 / 4 concentration nutrient solution is replaced; after 2 days of water culture with 1 / 4 concentration nutrient solution, Li source is added to the culture solution, and after 6-12 days of continuous water culture, it is harvested. By adding lithium source to the culture solution, Pteroxygonum giraldii rhizomes and leaves are enriched with lithium element, thereby providing a safer raw material source for the production of bio-lithium-rich active ingredients.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of agricultural planting, in particular to a cultivation method for enriching lithium elements in Cardamine violifolia. BACKGROUND

[0002] Cardamine violifolia belongs to the genus Cardamine of the family Brassicaceae, mainly distributed in Hubei Enshi and Huping Mountain in Hunan, and is a perennial plant. The tender stems and leaves of Cardamine violifolia are edible and delicious, and are a kind of wild vegetable resources for local villagers. Cardamine violifolia has strong tolerance and enrichment capacity for trace elements, especially selenium.

[0003] Lithium is a chemical element, and its beneficial effects on the human body include treating bipolar disorder, preventing cognitive decline, anti-inflammatory effect, and influence on cardiovascular health. Lithium salt is a common drug for treating bipolar disorder, which can reduce symptoms such as mood swings, insomnia, anxiety, etc. Studies have shown that lithium salt helps prevent neurodegenerative diseases such as cognitive decline in the elderly and Alzheimer's disease. Lithium also has an inhibitory effect on inflammation, which can reduce the symptoms of some inflammatory diseases. Lithium can lower blood pressure and improve heart health, and has a preventive effect on cardiovascular diseases. However, current medical lithium salts are mostly synthesized from purified industrial lithium, and there is no report on biological lithium raw materials. And lithium is not a necessary nutrient element for plants, and improper concentration can easily cause plant poisoning.

[0004] At present, Cardamine violifolia is cultivated as a selenium-enriched crop, and is mostly produced on a large scale by soil or substrate cultivation. There is no related report on water culture and lithium enrichment, so it has become a technical problem to be solved in the field to develop a precise and quantitative method for enriching lithium elements in Cardamine violifolia with less soil pollution. SUMMARY

[0005] Based on the above, the present application provides a cultivation method for enriching lithium elements in Cardamine violifolia.

[0006] To achieve the above-mentioned purpose, the present application provides the following solutions:

[0007] The present application provides a cultivation method for enriching lithium elements in Cardamine violifolia, comprising the following steps:

[0008] (1) Seedling raising: spread the seeds of Cardamine violifolia in the substrate for seedling raising;

[0009] (2) Transplanting: when the Cardamine violifolia seedlings grow to 4-5 true leaves, transplant the Cardamine violifolia seedlings into a water culture box filled with clean water;

[0010] After 3 days of transplanting, the water in the hydroponic box is replaced with 1 / 8 concentration nutrient solution and exogenous selenium is added, and then the nutrient solution and exogenous selenium are replaced every 10 days; after 45-60 days of transplanting, 1 / 4 concentration nutrient solution is replaced;

[0011] After 2 days of hydroponics with 1 / 4 concentration nutrient solution, Li source is added to the culture solution, and after 6-12 days of continuous hydroponics, the collection is completed, that is, the enrichment of lithium element of the Gonostegia polilifolia is completed.

[0012] In a preferred embodiment of the present application, in step (1), the environmental temperature for seedling is 18±1℃, and the relative humidity is 70%-80%.

[0013] In a preferred embodiment of the present application, in step (1), the light condition for seedling is: light 100 μmol / m 2 / s, day / night 14 / 10 hours.

[0014] In a preferred embodiment of the present application, in step (2), the temperature after transplanting is 22±1℃ day / 16±1℃ night, and the environmental relative humidity is 70%-80%.

[0015] In a preferred embodiment of the present application, in step (2), the light condition after transplanting is: 200 μmol / m 2 / s, day / night 16 / 8 hours.

[0016] In a preferred embodiment of the present application, in step (2), the nutrient solution is Hoggland nutrient solution.

[0017] In a preferred embodiment of the present application, the exogenous selenium is sodium selenite; the concentration of the exogenous selenium in the 1 / 8 concentration nutrient solution is 1.5 μmol / L.

[0018] In a preferred embodiment of the present application, the Li source is LiCl; the concentration of the Li source in the 1 / 4 concentration nutrient solution is 50-200 mg / L; further preferably, 200 mg / L.

[0019] In a preferred embodiment of the present application, the time for continuing hydroponics after adding the Li source in step (2) is 9d.

[0020] The present application discloses the following technical effects:

[0021] The present application utilizes the characteristics of Gonostegia polilifolia that is easy to enrich trace elements, adds lithium source in the culture solution, so that the Gonostegia polilifolia rhizome and leaf blade enrich lithium element, and further provides a safer raw material source for producing bio-lithium-rich active ingredients.

[0022] The cultivation method of this invention not only promotes the vigorous growth of *Cephalotaxus fortunei*, but also significantly increases the total lithium content of *Cephalotaxus fortunei*, thereby enhancing its nutritional and commercial value.

[0023] This invention utilizes hydroponics to cultivate lithium-enriched Corydalis yanhusuo. The operation is simple, low-cost, labor-saving, and material-saving, with minimal residue. It has high application value for the large-scale cultivation of lithium-enriched Corydalis yanhusuo and can be used for the large-scale production of lithium-enriched Corydalis yanhusuo. Attached Figure Description

[0024] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0025] Figure 1 The fresh weight of hydroponically grown *Violet spp.* in different concentrations of Li after 12 days is shown in Example 1.

[0026] Figure 2 The total lithium content of the upper part and root system of hydroponically grown *Vigna arvense* with different concentrations of Li was measured in Example 1 after 12 days.

[0027] Figure 3 The lithium content of different cell components in the upper part of *Vigna oleifera* hydroponically grown for 12 days using different concentrations of Li in Example 1 is shown.

[0028] Figure 4 The figures in Example 1 show the different dissolved lithium contents in the upper part of hydroponically grown Viola yedoensis leaves after 12 days using different concentrations of Li.

[0029] Figure 5 The fresh weight of *Violet spp.* hydroponically grown at different growth days using 200 mg / L Li in Example 2 is shown.

[0030] Figure 6 The total lithium content of the aboveground parts and roots of *Vigna arvense* hydroponically grown at different growth days using 200 mg / L Li in Example 2 is shown.

[0031] Figure 7 This refers to the lithium content of different cell components in the aboveground parts of *Vigna arvense* hydroponically grown at different growth days using 200 mg / L Li in Example 2. Detailed Implementation

[0032] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0033] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0034] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0035] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0036] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0037] This invention utilizes hydroponics, first cultivating *Cephalotaxus fortunei* in a nutrient solution supplemented with exogenous selenium. Once the seedlings reach a certain growth stage, a lithium source is added to the nutrient solution. A suitable hydroponic concentration for lithium-enriched cultivation of *Cephalotaxus fortunei* seedlings was determined, resulting in a complete hydroponic method for *Cephalotaxus fortunei*. Using this cultivation method, not only is the growth of *Cephalotaxus fortunei* robust, but the total lithium content is also significantly increased, enhancing its nutritional and commercial value.

[0038] This invention provides a cultivation method for enriching lithium in *Violet-leaved Water Chestnut*, comprising the following steps:

[0039] (1) Seedling raising: Sow the seeds of Viola yedoensis in the substrate for seedling raising;

[0040] (2) Transplanting: When the seedlings of Viola yedoensis have grown to 4-5 true leaves, transplant them into a hydroponic box filled with clean water.

[0041] Since *Calamus odoratus* is a hyper-selenium-rich plant, a certain amount of selenium can effectively promote its healthy growth and improve its resistance to adverse conditions. Therefore, 3 days after transplanting, replace the water in the hydroponic box with 1 / 8 concentration nutrient solution and add exogenous selenium at the same time. Then, replace the nutrient solution and exogenous selenium every 10 days. This step is very important for subsequent Li-enriched cultivation. 45-60 days after transplanting, replace the nutrient solution with 1 / 4 concentration.

[0042] After hydroponically cultivating the plant for two days using a 1 / 4 concentration nutrient solution, a lithium source is added to the culture medium. Since lithium is not an essential nutrient for plants and can easily cause toxicity, it must be added at the appropriate time. At this stage, *Cephalotaxus fortunei* is growing vigorously, has strong resistance, and a robust root system. Adding exogenous lithium allows for rapid absorption and conversion, accumulating in the leaves, while also promoting healthy growth. Adding exogenous lithium too early results in small, weak plants that are prone to toxicity; adding it too late leads to decreased root vitality and difficulty in accumulating a high lithium content. After adding exogenous lithium, continue hydroponics for 6-12 days before harvesting, thus completing the lithium enrichment cultivation of *Cephalotaxus fortunei*.

[0043] Unless otherwise specified, the technical solutions described in this invention are all conventional solutions in the field, and the reagents or raw materials used are all purchased from commercial channels or are publicly available unless otherwise specified.

[0044] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0045] Example 1

[0046] Seedling cultivation of *Violet-leaved Watercress*: Seedlings were cultivated in 32-cell seed trays filled with river sand. On July 23, 2023, seeds were sown on the sand in the seed trays. The seedling environment was maintained at 18℃, relative humidity 70%-80%, and light intensity 100 μmol / m². 2 / s, day and night are 14 hours / 10 hours.

[0047] Transplanting: When the seedlings of *Cephalotaxus fortunei* have grown to four to five true leaves, carefully remove them from the seedling trays on September 9, 2023. Wash off the river sand from the roots and transplant them into a hydroponic box, securing them in the planting basket with a sponge. At this time, fill the hydroponic box with clean water, without adding nutrient solution, to allow the transplanted seedlings to adapt to the hydroponic environment. Simultaneously, place an aeration pump in the hydroponic box (air output 1.5 liters / minute, aeration 14 hours per day, consistent with the photoperiod). The light conditions after transplanting are 200 μmol / m². 2 / s, day / night is 16 / 8 hours.

[0048] Three days after transplanting, the water in the hydroponic box was replaced with 1 / 8 Hoagland nutrient solution, and sodium selenite was added to bring the final concentration to 1.5 μmol / L. The nutrient solution and sodium selenite were then replaced every 10 days. Forty-five days after transplanting, the water was replaced with 1 / 4 Hoagland nutrient solution, and no more sodium selenite was added. After continuing hydroponics for two days, LiCl was added to the culture solution to bring the final concentrations of LiCl to 0 mg / L, 50 mg / L, 100 mg / L, 200 mg / L, and 400 mg / L, respectively. After continuing hydroponics for 12 days, the plants were harvested, thus completing the lithium enrichment cultivation of *Cephalotaxus fortunei*.

[0049] Figure 1 The figure shows the fresh weight of hydroponically grown *Vigna arvensis* at different concentrations of LiCl (in the figure, CK represents the final concentration of LiCl as 0 mg / L, Li 50 represents the final concentration of LiCl as 50 mg / L, and so on, with Li 400 representing the final concentration of LiCl as 400 mg / L). The results show that LiCl concentrations of 50-200 mg / L had no significant effect on the growth of *Vigna arvensis*, while 400 mg / L significantly inhibited its growth.

[0050] Figure 2 The figure shows the total lithium content in the aboveground parts and roots of hydroponically grown *Vigna oleifera* at different concentrations of LiCl. From... Figure 2 It can be seen that the Li absorbed by *Cephalotaxus fortunei* mainly accumulates in the aboveground parts, with less Li accumulating in the roots. Furthermore, the Li content in *Cephalotaxus fortunei* increases significantly with increasing LiCl concentration. The Li content in the aboveground parts of *Cephalotaxus fortunei* treated with 100 mg / L and 200 mg / L LiCl was quite similar, and both exceeded 100 mg / kg fresh weight.

[0051] Figure 3 The figure shows the lithium content of different cell components in the aerial parts of *Vigna oleifera* hydroponically cultured at different Li concentrations. As can be seen from the figure, Li in the aerial parts of *Vigna oleifera* mainly accumulates in the cell wall and soluble components. This indicates that Li in *Vigna oleifera* should primarily be bio-extracted from the cell wall and soluble components.

[0052] Figure 4 The figure shows the dissolved lithium content in the aboveground parts of *Vigna oleifera* hydroponically grown at different concentrations of Li. As can be seen from the figure, Li in the aboveground parts of *Vigna oleifera* is mainly found in alcohol-soluble (Li-ethanol) and water-soluble (Li-H2O) forms. This indicates that Li in *Vigna oleifera* should primarily be extracted biologically using ethanol and water.

[0053] Example 2

[0054] Based on the results of Example 1, a LiCl concentration of 200 mg / L was determined to be the preferred concentration. Example 2 was then conducted based on these results.

[0055] Seedling cultivation of *Violet-leaved Watercress*: Seedlings were cultivated in 32-cell seed trays filled with river sand. On March 10, 2024, *Violet-leaved Watercress* seeds were sown on the sand in the seed trays. The seedling environment was maintained at a temperature of 18℃, a relative humidity of 70%-80%, and a light intensity of 100 μmol / m². 2 / s, day and night are 14 hours / 10 hours.

[0056] Transplanting: When the seedlings of *Violet-leaved Water Chestnut* have grown to four to five true leaves, carefully remove them from the seedling trays on April 21, 2024. Wash off the river sand from the roots and transplant them into a hydroponic box, securing them in the planting basket with a sponge. At this time, fill the hydroponic box with clean water, without adding nutrient solution, to allow the transplanted seedlings to adapt to the hydroponic environment. Simultaneously, place an aeration pump in the hydroponic box (air output 1.5 liters / minute, aeration 14 hours per day, consistent with the photoperiod). The light conditions after transplanting are 200 μmol / m². 2 / s, day / night is 16 / 8 hours).

[0057] Three days after transplanting, the water in the hydroponic container was replaced with 1 / 8 Hoagland nutrient solution, and sodium selenite was added to achieve a final concentration of 1.5 μmol / L. The nutrient solution and sodium selenite were then replaced every 10 days. Forty-five days after transplanting, the solution was replaced with 1 / 4 Hoagland nutrient solution, and no more sodium selenite was added. Hydroponics continued for two days, and then LiCl was added to the culture medium to achieve a final LiCl concentration of 200 mg / L. Hydroponics continued, and harvesting was carried out in batches on days 3, 6, 9, and 12 after Li addition. The Li accumulation of *Corydalis violaceus* at different growth stages was investigated at the optimized concentration.

[0058] Figure 5 The figure shows the fresh weight of *Vigna arvensis* hydroponically grown in 200 mg / L Li solution at different growth days. As can be seen from the figure, *Vigna arvensis* continued to grow in this Li concentration nutrient solution, reaching its maximum fresh weight on day 9.

[0059] Figure 6 The figure shows the total lithium content in the aboveground parts and roots of *Vigna oleifera* hydroponically cultured with 200 mg / L Li for different growth days. As the figure shows, the Li content in the aboveground parts and roots of *Vigna oleifera* significantly increased with prolonged culture time, with the aboveground content being significantly higher than that in the roots. A considerable content was observed as early as day 9.

[0060] Figure 7 The lithium content of different cell components in the aerial parts of *Vigna oleifera* hydroponically cultured with 200 mg / L Li was determined. Similar to Example 1, Li in the aerial parts of *Vigna oleifera* mainly accumulated in the cell walls and soluble components.

[0061] The results above show that this invention utilizes hydroponics, first pre-culturing *Cephalotaxus fortunei* in a selenium-containing nutrient solution to ensure its healthy growth. Once it reaches a certain stage, lithium is added to the nutrient solution to enrich it. Suitable hydroponic concentrations and harvesting times for lithium-enriched *Cephalotaxus fortunei* seedling cultivation were determined, resulting in a complete hydroponic method for *Cephalotaxus fortunei*. Using the cultivation method described in this application, the total lithium content of *Cephalotaxus fortunei* is significantly increased, enhancing its nutritional and commercial value. Hydroponic cultivation of *Cephalotaxus fortunei* for lithium enrichment is simple, low-cost, labor-saving, and produces minimal residue. It has high application value for the large-scale cultivation of lithium-enriched *Cephalotaxus fortunei* and can be used for the large-scale production of lithium-enriched *Cephalotaxus fortunei*.

[0062] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. A cultivation method for enriching lithium in *Violet-leaved Watercress*, characterized in that, Includes the following steps: (1) Seedling raising: Sow the seeds of Viola yedoensis in the substrate for seedling raising; (2) Transplanting: When the seedlings of Viola yedoensis have grown to 4-5 true leaves, transplant them into a hydroponic box filled with clean water. Three days after transplanting, replace the water in the hydroponic box with 1 / 8 concentration nutrient solution and add exogenous selenium. Then replace the nutrient solution and exogenous selenium every 10 days. 45-60 days after transplanting, replace the nutrient solution with 1 / 4 concentration nutrient solution. After hydroponically cultivating with 1 / 4 concentration nutrient solution for 2 days, add Li source to the culture solution and continue hydroponics for 6-12 days before harvesting, thus completing the lithium enrichment cultivation of Viola yedoensis. The nutrient solution is Hoagland nutrient solution; The exogenous selenium is sodium selenite; the concentration of the exogenous selenium in the 1 / 8 concentration nutrient solution is 1.5 μmol / L; The Li source is LiCl; the concentration of the Li source in the 1 / 4 concentration nutrient solution is 50-200 mg / L.

2. The cultivation method according to claim 1, characterized in that, In step (1), the ambient temperature for seedling cultivation is 18±1℃ and the relative humidity is 70%-80%.

3. The cultivation method according to claim 1, characterized in that, In step (1), the light conditions for seedling cultivation are: 100 μmol / m² light intensity. 2 / s, day / night is 14 / 10 hours.

4. The cultivation method according to claim 1, characterized in that, In step (2), the temperature after transplanting is 22±1℃ day / 16±1℃ night, and the relative humidity is 70%-80%.

5. The cultivation method according to claim 1, characterized in that, In step (2), the light conditions after transplanting are: 200 μmol / m². 2 / s, day / night is 16 / 8 hours.

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