Method for increasing nervonic acid content in acer truncatum seed oil

Abstract

The invention discloses a method for increasing the content of nervonic acid in acer truncatum seed oil, and belongs to the technical field of plant quality-improving and efficiency-improving cultivation. According to the invention, the prepared foliar fertilizer containing monopotassium phosphate is sprayed on the leaves of acer truncatum. According to the method, the monopotassium phosphate foliar fertilizer with an optimized proportion is sprayed for multiple times in the growth and development fast-growing period of acer truncatum fruits, field trials and grease component analysis show that the spraying of the foliar fertilizer can effectively promote the biosynthesis of unsaturated fatty acid in acer truncatum seeds, and the content of nervonic acid is obviously increased; meanwhile, the composition proportion of unsaturated fatty acids such as alpha-linolenic acid and erucic acid is also remarkably optimized, the quality of the acer truncatum grease is improved, and the acer truncatum grease has important practical guidance value for promoting efficient development and utilization of acer truncatum nervonic acid resources.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of plant quality improvement and synergistic cultivation, and particularly relates to a method for increasing nervonic acid content in Pterocarya stenoptera seed oil. BACKGROUND

[0002] With people's increasing attention to the nutritional value of food, the content of unsaturated fatty acids has become an important standard for measuring the quality of healthy edible oil, so the content of unsaturated fatty acids directly affects the nutritional value and market competitiveness of Pterocarya stenoptera seed oil, and optimizing the composition of unsaturated fatty acids in Pterocarya stenoptera seed oil, especially increasing the content of nervonic acid, has important significance for developing Pterocarya stenoptera brain health products and high-end functional edible oil.

[0003] A large number of domestic and foreign studies have proved that the supplementation of mineral nutrients plays a significant role in promoting high-quality and high-yield of economic tree species, and appropriate application can achieve the purpose of regulating plant growth and development, which is beneficial to increasing the content of nutritional components in plant seeds and improving their economic value.

[0004] The characteristics of Pterocarya stenoptera seeds are greatly affected by ecological environment, and serious germplasm differentiation occurs in long-term natural selection and species evolution, so there are certain differences in seed oil characteristics, fatty acid composition ratio and content, which limits the maximization of their economic value. At present, there are few reports on increasing the content of nervonic acid in Pterocarya stenoptera through exogenous intervention technology, therefore, how to improve the fatty acid ratio in Pterocarya stenoptera seed oil, especially the content of nervonic acid, has become a hot research topic. Increasing the content of nervonic acid in Pterocarya stenoptera seeds has important significance for supplying nervonic acid raw materials needed for the prevention and treatment of brain diseases. SUMMARY

[0005] The technical problem to be solved by the present application is to provide a method for increasing the content of nervonic acid in Pterocarya stenoptera seed oil, which effectively promotes the biosynthesis of unsaturated fatty acids in Pterocarya stenoptera seeds, and has important practical guiding value for promoting the efficient development and utilization of Pterocarya stenoptera nervonic acid resources.

[0006] In order to solve the above technical problems, the technical scheme adopted by the present application is as follows:

[0007] A method for increasing the content of nervonic acid in Pterocarya stenoptera seed oil, wherein a leaf fertilizer containing potassium dihydrogen phosphate is sprayed on the leaves of Pterocarya stenoptera.

[0008] Further, the Pterocarya stenoptera is selected from 10-year-old healthy plants.

[0009] Further, the concentration of potassium dihydrogen phosphate is 1.5 g / L to 7.5 g / L.

[0010] Further, the spraying frequency is 1 to 3 times.

[0011] Further, the spraying method is: about 5L of the use amount per tree is uniformly sprayed on the leaves of Acer truncatum Bunge, the spraying period is from the middle of June to the middle of August, the best spraying period is to spray once in the middle of June and the middle of July, a total of 2 times, and the spraying is re-performed when it rains.

[0012] Further, 6.0g / L of potassium dihydrogen phosphate is prepared to spray on the leaves of Acer truncatum Bunge once in the middle of June and the middle of July, a total of 2 times.

[0013] A method for improving the content of oleic acid in Acer truncatum Bunge seed oil, 1.5g / L of potassium dihydrogen phosphate is sprayed on the leaves of Acer truncatum Bunge once in the middle of June, the middle of July and the middle of August, a total of 3 times.

[0014] A method for improving the content of linoleic acid in Acer truncatum Bunge seed oil, 7.5g / L of potassium dihydrogen phosphate is sprayed on the leaves of Acer truncatum Bunge once in the middle of June, the middle of July and the middle of August, a total of 3 times.

[0015] A method for improving the content of alpha-linolenic acid in Acer truncatum Bunge seed oil, 4.5g / L of potassium dihydrogen phosphate is sprayed on the leaves of Acer truncatum Bunge once in the middle of June, the middle of July and the middle of August, a total of 3 times.

[0016] A method for improving the content of gamma-linolenic acid in Acer truncatum Bunge seed oil, 4.5g / L of potassium dihydrogen phosphate is sprayed on the leaves of Acer truncatum Bunge once in the middle of June, the middle of July and the middle of August, a total of 3 times.

[0017] Compared with the prior art, the present application has the following advantages:

[0018] (1) The present application sprays the leaf fertilizer of the optimized proportion of potassium dihydrogen phosphate on the fast-growing period of Acer truncatum Bunge fruit growth and development, and through field tests and oil component analysis, it is found that the spraying of the leaf fertilizer can effectively promote the biosynthesis of unsaturated fatty acids in Acer truncatum Bunge seeds, and the content of nervonic acid is particularly obvious, which is 8.04%, and the composition ratio of unsaturated fatty acids such as alpha-linolenic acid and erucic acid is also significantly optimized, and the quality of Acer truncatum Bunge oil is improved.

[0019] (2) The technical scheme of the present application is simple and convenient to operate, and the cost is controllable, which provides an important theoretical basis and technical support for the high-quality and efficient cultivation of Acer truncatum Bunge, an important woody oil tree species, has important practical guiding value for promoting the efficient development and utilization of nervonic acid resources of Acer truncatum Bunge, and provides a new research idea for the quality control of woody oil crops. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1Figure 1 is a gas chromatogram of main fatty acid components of Acer truncatum Bunge seeds under different spraying treatments in the present application; in the figure, 1, palmitic acid; 2, heptadecanoic acid; 3, stearic acid; 4, oleic acid; 5, linoleic acid; 6, a-linolenic acid; 7, γ-linolenic acid; 8, arachidonic acid; 9, erucic acid; 10, nervonic acid. DETAILED DESCRIPTION

[0021] The present application will be further illustrated below in combination with specific examples, which are implemented on the premise of the technical solutions of the present application, and it should be understood that these examples are only used to illustrate the present application and not used to limit the scope of the present application.

[0022] The test site of the present research is the Acer truncatum Bunge test forest in Yangcang Village, Tengzhou City, Shandong Province (117°15'E, 35°08'N), which belongs to a temperate monsoon continental climate with sufficient light and abundant precipitation. The average annual temperature is 14.6°C, the average annual ground temperature is 16.3°C, the average annual precipitation is about 773.1 mm, the frost-free period is about 210 days, and the annual sunshine duration is 2383 hours.

[0023] The Acer truncatum Bunge used in the following examples is from the Acer truncatum Bunge test forest in Yangcang Village, Tengzhou City, Shandong Province, and 10-year-old healthy Acer truncatum Bunge (Acer truncatum Bunge) with consistent site conditions, good growth, healthy tree body, no diseases and insect pests, and normal flowering and fruiting is selected as the test tree. The average tree height is (6.5±0.5) m, the average diameter at breast height is (15.5±0.8) cm, the tree spacing is 8 m x 8 m, and the conventional management is adopted.

[0024] The potassium dihydrogen phosphate used in the following examples is "Guangwang brand power potassium" (KH2PO4≧98%, P2O5≧51%, K2O≧33.8%) produced by Sichuan Runer Technology Co., Ltd.; the gibberellin is "Sanliu brand" gibberellic acid (effective ingredient content 75%) produced by Shanghai Tongrui Biological Technology Co., Ltd.; petroleum ether (analytical pure, boiling range 60-90°C) and methanol (HPLC) are both purchased from National Pharmaceutical Group Chemical Reagent Co., Ltd.; and sodium hydroxide (analytical pure) is purchased from Xilong Chemical Co., Ltd.

[0025] The spraying method of Acer truncatum Bunge in the following examples is as follows: about 5 L of each tree is uniformly sprayed on the leaf surface of Acer truncatum Bunge. The spraying period is from the middle of June to the middle of August, and the best spraying period is to spray once in the middle of June and the middle of July, a total of 2 times. It is necessary to choose a sunny and windless day for spraying, and if it rains after spraying, it is necessary to re-spray.

[0026] Example 1

[0027] In the fast-growing period of Acer truncatum fruit growth and development from June to August in 2023, 10-year-old healthy plants in Yangcangcun Acer truncatum test forest were selected for different times of potassium dihydrogen phosphate and gibberellin leaf spraying test every month, and each month was sprayed once to screen the best exogenous substance type. The exogenous substances used were 3 g / L potassium dihydrogen phosphate aqueous solution and 50 mg / L GA3 aqueous solution, and three spraying times were set, i.e. 1 time, 2 times and 3 times, with spraying the same amount of water as the control. In early November 2023, when the fruits of Acer truncatum were mature, the alate fruits that developed normally and were complete were collected, and the pericarp and seed coat were removed to determine the content of each fatty acid component of Acer truncatum seeds, and the results are shown in Tables 1-2.

[0028] The test method of each fatty acid component content: accurately take 1.5 g of Acer truncatum kernel sample and grind it into fine powder in a mortar, quantitatively transfer it to a 10 mL centrifuge tube, add 2 mL of analytical pure petroleum ether (boiling range 60-90°C), and place it in a 40 kHz ultrasonic cleaner at 25°C for 2 h, then add 2 mL of 0.4 mol / L methanol-sodium hydroxide solution (preparation method: accurately weigh 1.6 g of sodium hydroxide solid and dissolve it in 100 mL of HPLC methanol), and ultrasonic methyl esterification under the same conditions for 1 h. After the reaction is completed, add distilled water to make up to 10 mL, mix well by vortex oscillator for 30 s, and then centrifuge in a table centrifuge at 5000 r / min for 5 min. Take the supernatant and put it into a brown sample bottle for testing. Use a Japanese Shimadzu gas chromatograph-mass spectrometer QP2010 ultra equipped with a TR-WAX MS capillary column (30.00 m x 0.25 mm x 0.25 μm) for fatty acid component analysis. The carrier gas is high-purity helium, and the constant flow mode is 1.0 mL / min with a split ratio of 10. The program temperature conditions are: initial temperature 80°C, hold for 2.5 min, increase to 210°C at 15°C / min, then increase to 230°C at 2°C / min and hold for 10 min, and the injection port temperature is 200°C. Qualitative analysis is performed by comparing with known fatty acid methyl ester standards, and quantitative analysis is performed by peak area normalization method to calculate the relative content of each fatty acid, as shown in Figure 1

[0029] Table 1 Effect of different types of exogenous substances on the content of saturated fatty acid components in Acer truncatum seeds in 2023

[0030]

[0031]

[0032] Table 2 Effect of different types of exogenous substances on the content of unsaturated fatty acid components in Acer truncatum seeds in 2023

[0033] ​

[0034] From the results of Table 1 and Table 2, it can be seen that the spraying of potassium dihydrogen phosphate and gibberellin has different degrees of influence on the content of each saturated and unsaturated fatty acid component of Acer truncatum seeds. Compared with the other, the promotion of unsaturated fatty acid component content by potassium dihydrogen phosphate is more favorable. Among them, when T1 treatment, i.e. spraying 1 time with 3 g / L potassium dihydrogen phosphate, the content of unsaturated fatty acid such as a-linolenic acid, g-linolenic acid, erucic acid and nervonic acid in Acer truncatum seeds reaches a high level. Therefore, the potassium dihydrogen phosphate mixed aqueous solution is selected as the best exogenous substance type for improving the content of nervonic acid in Acer truncatum seeds and improving the quality of seed oil.

[0035] Example 2

[0036] Based on the results of the spraying test in 2023, in the fast-growing period of 6-8 months of Acer truncatum fruit growth and development in 2024, 10-year-old healthy plants in Yangcangcun Acer truncatum test forest were selected, and different concentrations (1.5-7.5 g / L) and times of potassium dihydrogen phosphate foliar spraying test were carried out monthly, each month was sprayed once, in order to determine the best spraying concentration and times of potassium dihydrogen phosphate, three spraying times were set, and spraying the same amount of water was used as a control. In early November 2024, when the fruits of Acer truncatum were mature, the alar fruits that developed normally and were complete were collected, the pericarp and seed coat were removed, and the content of each fatty acid component in the seeds was determined. The results are shown in Tables 3-4.

[0037] Test method: In early November 2024 (fruit ripening period), several alar fruits that developed normally were collected from the east, south, west and north directions of the test Acer truncatum, and were put into marked net bags, then were taken back to the laboratory for drying. Several Acer truncatum seed kernels with pericarp and seed coat removed were randomly selected from each treatment for determination of the content of each fatty acid component in the seeds. The specific test method of each fatty acid component content is the same as that of Example 1.

[0038] Table 3 Influence of different concentrations and times of potassium dihydrogen phosphate spraying on the content of saturated fatty acid components in Acer truncatum seeds in 2024

[0039]

[0040]

[0041] Table 4 Influence of different concentrations and times of potassium dihydrogen phosphate spraying on the content of unsaturated fatty acid components in Acer truncatum seeds in 2024

[0042]

[0043]

[0044]

[0045] From the results of Table 3 and Table 4, it can be seen that for the content of each component of the fatty acid of the Acer truncatum Bunge seed, the content of the four fatty acids (heptadecanoic acid, stearic acid, arachidonic acid, and erucic acid) is basically consistent among different treatment groups; the content of the other six fatty acids (palmitic acid, oleic acid, linoleic acid, α-linolenic acid, γ-linolenic acid, and nervonic acid) shows significant differences, indicating that the spraying of potassium dihydrogen phosphate can cause significant changes in the content, but the increasing and decreasing effects are different.

[0046] From the results of the content of each unsaturated fatty acid of the Acer truncatum Bunge seed, it can be seen that the content of oleic acid reaches the highest level in the A3 treatment, i.e., 1.5 g / L of potassium dihydrogen phosphate is sprayed for three times, which is significantly increased by 1.5% compared with the control group. The content of linoleic acid reaches the highest level in the E3 treatment, i.e., 7.5 g / L of potassium dihydrogen phosphate is sprayed for three times, and the effect of spraying 2 times of 3.0 g / L of potassium dihydrogen phosphate is the second. When 4.5 g / L, 6.0 g / L, or 7.5 g / L of potassium dihydrogen phosphate is sprayed for three times, the content of α-linolenic acid reaches a relatively high level, which is significantly increased by 0.38%, 0.33%, and 0.30% compared with the control group, respectively. The content of γ-linolenic acid is relatively high in the D1, D3, E2, and E3 treatments, which is significantly increased compared with the control. For the content of nervonic acid, except for the spraying of 1.5 g / L of potassium dihydrogen phosphate for two or three times or the spraying of 7.5 g / L of potassium dihydrogen phosphate for three times, due to insufficient or excessive intensity, the content of nervonic acid is increased but the effect is not significant; the remaining spraying treatments can significantly increase the content of nervonic acid in the Acer truncatum Bunge seed oil, which shows that the spraying of potassium dihydrogen phosphate with an appropriate concentration and number of times has a very obvious positive promoting effect on the increase of the content of nervonic acid, and the content of nervonic acid reaches the highest level of 8.04% in the D2 treatment, i.e., 6.0 g / L of potassium dihydrogen phosphate is sprayed for two times, which is increased by 2.14% compared with the control group, and the effects of the C3 and B2 treatments are the second, and the content of nervonic acid is 7.80% and 7.49%, respectively, which is increased by 1.90% and 1.59% compared with the control group, respectively.

[0047] The above only describes the preferred embodiments of the present application, and it should be noted that for ordinary skilled persons in the technical field, some improvements and refinements can be made without departing from the principles of the present application, and these improvements and refinements should also be considered as the protection scope of the present application.

Claims

1. A method for increasing the content of nervonic acid in Acer truncatum seed oil, characterized in that: Foliar fertilizer containing potassium dihydrogen phosphate was prepared and sprayed on the leaves of Acer truncatum.

2. The method for increasing the content of nervonic acid in Acer truncatum seed oil according to claim 1, characterized in that: The maple is selected from 10-year-old healthy plants.

3. The method for increasing the content of nervonic acid in Acer truncatum seed oil according to claim 1, characterized in that: The concentration of the potassium dihydrogen phosphate is 1.5 g / L to 7.5 g / L.

4. The method for increasing the content of nervonic acid in Acer truncatum seed oil according to claim 1, characterized in that: The spraying times are 1 to 3 times.

5. The method for increasing the content of nervonic acid in Acer truncatum seed oil according to claim 1, characterized in that: The spraying method is: spray about 5L per tree evenly on the leaves of the maple tree. The spraying period is from mid-June to mid-August. The best spraying period is once in mid-June and mid-July, for a total of 2 sprays. Re-spray in case of rain.

6. The method for increasing the content of nervonic acid in Acer truncatum seed oil according to claim 1, characterized in that: Prepare a solution containing 6.0 g / L potassium dihydrogen phosphate and spray it on the leaves of Acer truncatum once in mid-June and mid-July, for a total of 2 sprays.

7. A method for increasing the oleic acid content in Acer truncatum seed oil, characterized by: Potassium dihydrogen phosphate with a concentration of 1.5g / L was sprayed on the leaves of Acer truncatum once in mid-June, mid-July and mid-August, for a total of three times.

8. A method for increasing the linoleic acid content in Acer truncatum seed oil, characterized by: The potassium dihydrogen phosphate with a concentration of 7.5 g / L was sprayed on the leaves of Acer truncatum once in mid-June, mid-July and mid-August, for a total of three sprayings.

9. A method for increasing the α-linolenic acid content in Acer truncatum seed oil, characterized by: The potassium dihydrogen phosphate with a concentration of 4.5 g / L was sprayed on the leaves of Acer truncatum once in mid-June, mid-July and mid-August, for a total of three sprayings.

10. A method for increasing the content of γ-linolenic acid in Acer truncatum seed oil, characterized by: The potassium dihydrogen phosphate with a concentration of 4.5 g / L was sprayed on the leaves of Acer truncatum once in mid-June, mid-July and mid-August, for a total of three sprayings.

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