Application of uniconazole and prohexadione-calcium in regulating growth and metabolite accumulation of ginkgo biloba

Abstract

The application of uniconazole and alminotran in regulating the growth and metabolite accumulation of ginkgo biloba is disclosed, and relates to the technical field of plant growth promotion. The application of uniconazole and alminotran in regulating the growth and metabolite accumulation of ginkgo biloba is disclosed, and takes 2-year-old ginkgo biloba seedlings as materials, and applies 30-60 mg / L uniconazole or 50-200 mg / L concentration of alminotran, and the best uniconazole and alminotran concentrations for promoting the growth of ginkgo biloba, increasing the chlorophyll content, increasing the primary metabolite activity and increasing the secondary metabolite content are 50 mg / L and 100 mg / L respectively. Taking 3-year-old grafted ginkgo biloba seedlings as materials, the comprehensive indexes of ginkgo biloba leaves are best after 25 d of spraying 50 mg / L uniconazole and 100 mg / L alminotran.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of plant growth promotion, and more particularly relates to application of uniconazole and prohexadione-calcium in regulating growth and metabolite accumulation of Ginkgo biloba. BACKGROUND

[0002] Ginkgo biloba L. is the oldest surviving plant in the gymnosperms, and is known as a living fossil. Ginkgo leaves are rich in flavonoids and terpene lactones, and have important medicinal value. EGb761 (Ginkgo biloba extract 761) is a standardized extract of Ginkgo biloba leaves, and its preparations are widely used in the fields of medicine, health care products and food additives. Flavonoids are the most critical active ingredients (about 24% of the components) in EGb761, and are mainly composed of quercetin, isorhamnetin and kaempferol [2] Flavonoids help protect plants from biological and abiotic stress by functions such as anti-pathogen, antioxidant, and active oxygen scavenging, and also act as regulatory and signaling compounds. Flavonoids also play an important role in treating age-related cognitive decline, tinnitus, dizziness and Alzheimer's disease in humans. The chemical synthesis of flavonoids from Ginkgo biloba leaves is difficult, and currently, flavonoids are mainly extracted from Ginkgo biloba leaves. Therefore, increasing the content of flavonoids is beneficial to the survival of Ginkgo biloba and the development of the pharmaceutical industry of Ginkgo biloba leaf extract, and has great practical value and broad application prospects.

[0003] Uniconazole and prohexadione-calcium are new plant growth regulators that play a role in plant growth, metabolic regulation and response to adversity stress. Spraying exogenous plant growth regulators to improve the content of secondary metabolites in Ginkgo biloba leaves has always been a research focus. The research on the application of new plant growth regulators is of great significance to improve the yield and quality of Ginkgo biloba leaves. Moreover, so far, no study has disclosed the mechanism of uniconazole and prohexadione-calcium in Ginkgo biloba. SUMMARY

[0004] In view of the above problems existing in the prior art, the technical problems to be solved by the present application are to provide application of uniconazole and prohexadione-calcium in regulating the growth of Ginkgo biloba seedlings, such as seedling height, ground diameter and biomass. Another technical problem to be solved by the present application is to provide application of uniconazole and prohexadione-calcium in regulating metabolite accumulation of Ginkgo biloba, such as soluble protein, soluble sugar, antioxidant, endogenous hormone, total flavonoids, terpene lactones and the content of each component.

[0005] In order to solve the above technical problems, the technical scheme adopted by the present application is as follows:

[0006] The application of uniconazole in regulating the growth of Ginkgo biloba seedlings, wherein the growth is regulated in terms of seedling height, ground diameter and biomass.

[0007] The concentration of uniconazole is less than 60 mg / L, which promotes the growth of Ginkgo seedling height; the concentration of uniconazole is not less than 30 mg / L, which promotes the growth of Ginkgo diameter; the concentration of uniconazole is 40 mg / L or 60 mg / L, which promotes the increase of Ginkgo stem biomass.

[0008] The concentration of uniconazole is 60 mg / L, which inhibits the growth of Ginkgo seedling height and diameter.

[0009] The application of uniconazole in regulating the accumulation of metabolites in Ginkgo seedlings, including chlorophyll, soluble protein, soluble sugar, total flavonoids, ginkgolide, lactone A, lactone B, lactone C, peroxidase, superoxide dismutase, glutathione, gibberellin, abscisic acid, zeatin riboside, auxin, abscisic acid.

[0010] The concentration of uniconazole is 50-60 mg / L, which promotes the increase of chlorophyll content in Ginkgo leaves; the concentration of uniconazole is 30-60 mg / L, which promotes the increase of total terpene lactone, ginkgolide, lactone A, lactone B, lactone C content in Ginkgo leaves; the concentration of uniconazole is 50 mg / L, which promotes the increase of soluble sugar content, soluble protein content, total flavonoids content, peroxidase activity, superoxide dismutase activity, glutathione content, auxin content in Ginkgo leaves; uniconazole inhibits the increase of gibberellin and zeatin riboside content.

[0011] The application of madiaxanthin in regulating the growth of Ginkgo seedlings, including the regulation of height, diameter, biomass growth.

[0012] The concentration of madiaxanthin is 50-200 mg / L, which promotes the growth of Ginkgo seedling height; the concentration of madiaxanthin is not less than 150 mg / L, which promotes the growth of Ginkgo diameter; the concentration of madiaxanthin is greater than 50 mg / L, which promotes the increase of Ginkgo root biomass; the concentration of madiaxanthin is 50-200 mg / L, which promotes the increase of Ginkgo stem biomass and single plant biomass; the concentration of madiaxanthin is less than 200 mg / L, which promotes the increase of Ginkgo aboveground biomass; the concentration of madiaxanthin is 150 mg / L, which promotes the increase of Ginkgo leaf biomass.

[0013] The application of madiaxanthin in regulating the accumulation of metabolites in Ginkgo seedlings, including chlorophyll, soluble protein, soluble sugar, total flavonoids, ginkgolide, lactone A, lactone B, lactone C, peroxidase, superoxide dismutase, glutathione, gibberellin, abscisic acid, zeatin riboside, auxin, abscisic acid.

[0014] The amine ester concentration is 150-200 mg / L, which promotes the increase of the chlorophyll content of the ginkgo leaf; the amine ester concentration is 100 mg / L, which promotes the increase of the soluble sugar content, the soluble protein content, the total flavonoid content, the peroxidase activity, the superoxide dismutase activity, the glutathione content and the auxin content of the ginkgo leaf; the amine ester concentration is 50-200 mg / L, which promotes the increase of the total terpene lactone content, the ginkgolide content, the lactone A content, the lactone B content and the lactone C content of the ginkgo leaf.

[0015] The amine ester inhibits the increase of the gibberellin content, the abscisic acid content and the zeatin riboside content.

[0016] Compared with the prior art, the present application has the following beneficial effects:

[0017] The present application takes ginkgo seedling as the material, applies different concentrations of uniconazole and amine ester, and screens the optimal concentration for promoting the growth of ginkgo, the increase of the chlorophyll content, the increase of the primary metabolite activity and the increase of the secondary metabolite content. The present application takes 3-year-old ginkgo grafted seedlings as the material, screens the optimal treatment time of S-3307 and DTA-6 spraying, and the results of the examples show that:

[0018] 1) The 60 mg / L uniconazole treatment inhibits the growth of the ginkgo seedling height and ground diameter, and the overall uniconazole treatment inhibits the accumulation of the single-plant biomass. The amine ester treatment promotes the growth of the ginkgo seedling height and has a promoting effect on the single-plant biomass.

[0019] 2) After the 50 mg / L uniconazole and 100 mg / L amine ester treatment, the soluble sugar content and the soluble protein content reach the maximum values.

[0020] 3) The uniconazole treatment inhibits the POD activity, but the amine ester treatment improves the POD activity. Under the treatment of the two plant growth regulators, the SOD activity and the GSH content are generally increased.

[0021] 4) Under the treatment of the uniconazole and the amine ester, the IAA content is generally increased. Under the uniconazole treatment, the ABA content is generally increased, but the GA3 and ZR contents are decreased. Under the amine ester treatment, the IAA content is generally increased, but the GA3, ABA and ZR contents are decreased.

[0022] 5) When the uniconazole concentration is 50 mg / L or the amine ester concentration is 100 mg / L, the total flavonoid content and the terpene lactone and each component content are increased. BRIEF DESCRIPTION OF DRAWINGS

[0023] Figure 1 It is a diagram for the effect of different hormone treatments on the chlorophyll content of the 2-year-old ginkgo seedling leaf;

[0024] Figure 2Figure 1 is a graph showing the effect of different hormone treatments on the soluble sugar content of the leaves of 2-year-old Ginkgo seedling;

[0025] Figure 3 Figure 2 is a graph showing the effect of different hormone treatments on the soluble protein content of the leaves of 2-year-old Ginkgo seedling;

[0026] Figure 4 Figure 3 is a graph showing the effect of different hormone treatments on the total flavonoid content of the leaves of 2-year-old Ginkgo seedling;

[0027] Figure 5 Figure 4 is a graph showing the effect of different hormone treatments on the total terpene lactone content of the leaves of 2-year-old Ginkgo seedling;

[0028] Figure 6 Figure 5 is a graph showing the effect of different hormone treatments on the ginkgolide content of the leaves of 2-year-old Ginkgo seedling;

[0029] Figure 7 Figure 6 is a graph showing the effect of different hormone treatments on the lactone A content of the leaves of 2-year-old Ginkgo seedling;

[0030] Figure 8 Figure 7 is a graph showing the effect of different hormone treatments on the lactone B content of the leaves of 2-year-old Ginkgo seedling;

[0031] Figure 9 Figure 8 is a graph showing the effect of different hormone treatments on the lactone C content of the leaves of 2-year-old Ginkgo seedling;

[0032] Figure 10 Figure 9 is a graph showing the effect of different hormone treatments on the chlorophyll content of the leaves of 3-year-old Ginkgo grafted seedling;

[0033] Figure 11 Figure 10 is a graph showing the effect of different hormone treatments on the soluble sugar content of the leaves of 3-year-old Ginkgo grafted seedling;

[0034] Figure 12 Figure 11 is a graph showing the effect of different hormone treatments on the soluble protein content of the leaves of 3-year-old Ginkgo grafted seedling;

[0035] Figure 13 Figure 12 is a graph showing the effect of different hormone treatments on the peroxidase (POD) activity of the leaves of 3-year-old Ginkgo grafted seedling;

[0036] Figure 14 Figure 13 is a graph showing the effect of different hormone treatments on the superoxide dismutase (SOD) activity of the leaves of 3-year-old Ginkgo grafted seedling;

[0037] Figure 15 Figure 14 is a graph showing the effect of different hormone treatments on the glutathione (GSH) activity of the leaves of 3-year-old Ginkgo grafted seedling;

[0038] Figure 16 Figure 15 is a graph showing the effect of different hormone treatments on the IAA content of the leaves of 3-year-old Ginkgo grafted seedling;

[0039] Figure 17 Figure 3 is a graph showing the effect of different hormone treatments on the GA3 content of 3-year-old ginkgo grafted seedlings;

[0040] Figure 18 Figure 4 is a graph showing the effect of different hormone treatments on the ABA content of 3-year-old ginkgo grafted seedlings;

[0041] Figure 19 Figure 5 is a graph showing the effect of different hormone treatments on the ZR content of 3-year-old ginkgo grafted seedlings;

[0042] Figure 20 Figure 6 is a graph showing the effect of different hormone treatments on the total flavonoid content of 3-year-old ginkgo grafted seedlings. DETAILED DESCRIPTION

[0043] In order to make the objectives, technical solutions and advantages of the present application clearer, the present application will be further described below in combination with specific examples. In the following examples, if not specifically described, the technical means used are all conventional means well known to those skilled in the art.

[0044] The 2-year-old ginkgo seedlings and the 3-year-old ginkgo grafted seedlings used in the present application are purchased from the Germplasm Resource Garden of the National Ginkgo Variety Base in Pizhou City, Jiangsu Province (scion: 15-year-old 'Tancheng No. 1' ginkgo in the National Ginkgo Variety Base in Pizhou City).

[0045] Example 1: Effect of different hormone treatments on 2-year-old ginkgo seedlings

[0046] The 2-year-old ginkgo seedlings are treated with enechlorazole (S 30 , 30 mg / L; S 40 , 40 mg / L; S 50 , 50 mg / L; S 60 , 60 mg / L) and dimethomorph (D 50 , 50 mg / L; D 100 , 100 mg / L; D 150 , 150 mg / L; D 200 , 200 mg / L), with pure water treatment as the control (CK), a total of 9 treatments. The plant growth regulator powder is first dissolved in anhydrous ethanol, and then diluted with pure water to the required concentration. Among them, the ethanol accounts for 1% (v / v), and Tween 80 accounts for 0.1% (v / v). CK is sprayed with an equal amount of pure water. After the solution is prepared, a 2L sprayer is used to uniformly spray on the ginkgo leaves, ensuring that the treated ginkgo leaves are completely wet on both sides. After spraying, samples are taken at 5, 10, 15, 20 and 25 days, and the height, ground diameter, biomass of roots, stems and leaves are measured after 25 days.

[0047] 1. Effects of different hormone treatments on height and ground diameter of 2-year-old Ginkgo seedlings

[0048] The results are shown in Table 1. The effects of the two plant growth regulators on the growth of the height and ground diameter of the Ginkgo seedlings did not reach a significant difference level. 30 -S 50 The height of the Ginkgo seedlings treated with the plant growth regulators was higher than that of CK, but the difference was not significant. 60 The treatments inhibited the growth of the height of the Ginkgo seedlings. Except for the S 30 The treatments promoted the growth of the ground diameter of the Ginkgo seedlings, especially the S 60 treatment, with a growth of 0.17 mm.

[0049] The treatments with the dimethylammonium herbicide promoted the growth of the height of the Ginkgo seedlings, especially the D 100 treatment, with a growth of 2.44 cm. The effects of the dimethylammonium herbicide on the growth of the ground diameter of the Ginkgo seedlings showed a rule of inhibition at a low concentration and promotion at a high concentration, and the ground diameter of the Ginkgo seedlings treated with the D 200 treatment had the largest growth.

[0050] Table 1. Effects of different hormone treatments on the height and ground diameter of 2-year-old Ginkgo seedlings and multiple comparison results

[0051]

[0052] Note: S is uniconazole, and D is dimethylammonium herbicide. The subscripted numbers represent the concentration of the plant growth regulator, in mg / L. Different lowercase letters represent significant differences (P<0.05) among different treatments.

[0053] 2. Effects of different hormone treatments on the biomass of 2-year-old Ginkgo seedlings

[0054] The results are shown in Table 2. The effects of the two plant growth regulators on the biomass of the Ginkgo seedlings did not reach a significant difference level. Overall, the leaf biomass, root biomass, aboveground biomass, and single-plant biomass of the Ginkgo seedlings treated with uniconazole at different concentrations were lower than those of CK, and the stem biomass was higher than that of CK only in the S 40 and S 60 treatments.

[0055] After treatment with the dimethylammonium herbicide, the stem and single-plant biomass of the Ginkgo seedlings treated at different concentrations were higher than those of CK, and the root biomass and aboveground biomass were higher than those of CK except for the D 50 and D 200 treatments. The D 150 treatment had higher leaf, stem, root, aboveground, and single-plant biomass than CK, with an increase of 10.08%, 22.12%, 13.1%, 16.12%, and 15.20% compared to CK.

[0056] Table 2. Effects of different hormone treatments on the biomass of 2-year-old Ginkgo seedlings and results of multiple comparisons.

[0057]

[0058] 3. Effects of different hormone treatments on chlorophyll content in leaves of 2-year-old Ginkgo seedlings

[0059] The results are as follows Figure 1 As shown, after treatment with tebuconazole, the chlorophyll content of all concentrations was significantly lower than that of the control (CK) at 10 days. At 10-20 days, S... 50 S 60 The chlorophyll content in the treatments gradually accumulated and was significantly higher than that in the control at 20 days, being 1.23 and 1.61 times that of the control, respectively.

[0060] After treatment with aminoethyl ester, the chlorophyll content at all locations was significantly lower than that in the control (CK) at 5-10 days. 150 During treatment, chlorophyll content gradually increased from 5 to 15 days, and was significantly higher than the control (CK) at 15 days, reaching 3.474 mg / g. 200 During the treatment period of 10-25 days, the chlorophyll content gradually increased, and at 20 days and 25 days, the chlorophyll content was 24.69% and 11.32% higher than that of the control, respectively.

[0061] 4. Effects of different hormone treatments on the soluble sugar content in the leaves of 2-year-old Ginkgo seedlings

[0062] The results are as follows Figure 2 As shown, after treatment with uniconazole, the soluble sugar content (excluding S) decreased after 5-10 days. 40 The soluble sugar content in all treatments (d5) was significantly lower than that in the control (CK). The soluble sugar content in each treatment showed a gradual accumulation trend from 5 to 15 days, with S... 30 The soluble sugar content in the treatment group was significantly higher than that in the control group (CK) at 15 days, being 1.15 times higher. 40 (Except for 15d), S 60 The soluble sugar content of the treated samples was lower than that of the control (CK). 50 Soluble sugar content gradually accumulated from 5 to 20 days, and began to decrease at 25 days, but was still significantly higher than CK, increasing by 8.40% compared to CK.

[0063] After treatment with aminoethyl ester, at 5d, 10d, and 20d (except for D at 5d) 50 D 150 The soluble sugar content of the (external) treatments was significantly lower than that of the control (CK). Except for D 200 Except for the treatment, the soluble sugar content of the other treatments gradually increased from 5 to 15 days, and D increased at 15 days. 100 D 150The soluble sugar content in the treatment group was significantly higher than that in the control group (CK), being 1.12 and 1.18 times higher, respectively. 200 The soluble sugar content of the treatment group generally showed an increasing trend, and was significantly lower than that of the control group (CK) from 5 to 20 days, decreasing by 14.48%, 35.16%, 39.05%, and 31.05% respectively. However, it was significantly higher than that of the CK at 25 days, reaching 21.5 mg / g.

[0064] 5. Effects of different hormone treatments on the soluble protein content in the leaves of 2-year-old Ginkgo seedlings

[0065] The results are as follows Figure 3 As shown, after treatment with uniconazole, the soluble protein content of the treated group was significantly higher than that of the control group (CK) at 10 days. 30 The content of soluble protein in the treatment group generally showed an accumulation trend in the first 20 days, with significantly higher levels than the control (CK) at days 5, 10, and 20, increasing by 28.77%, 24.00%, and 4.11%, respectively. 40 The soluble protein content in the treatment group was significantly higher than that in the control group (CK) at 10 days, but significantly lower than that in the control group at 20 days. 50 S 60 The soluble protein content showed an increasing trend from 5 to 20 days, reaching its maximum at 20 days, with soluble protein contents of 1.42 and 1.54 mg / g, respectively. 60 The soluble protein content was significantly higher than that of CK at 10 and 20 days of treatment, being 1.27 and 1.05 times that of CK, respectively.

[0066] After treatment with aminoethyl esters, there were no significant differences between the treatments and the control (CK) at 5 days. However, at 10 days, the soluble protein content of the treatments was significantly higher than that of the CK, being 1.57, 1.19, 2.13, and 1.57 times that of the CK, respectively. 50 The content of soluble protein in the treatment showed an increasing trend from 5 to 15 days, and the content at 10 and 15 days was significantly higher than that in the control (CK), increasing by 57.33% and 29.06% respectively. 100 The soluble protein content in the treatment group was significantly higher than that in the control group (CK) at day 10, reaching 0.89 mg / g. There were no significant differences between the treatment group and the CK group at other time points. 150 D 200 The content of the treatment on days 10 and 20 was significantly higher than that of the control (CK), especially on day 20 when it reached the maximum value, which was 17.12% and 21.23% higher than that of the control, respectively.

[0067] 6. Effects of different hormone treatments on the total flavonoid content in the leaves of 2-year-old Ginkgo seedlings

[0068] The results are as follows Figure 4 As shown, after treatment with uniconazole, at 5 days, only S 50The treatment was significantly lower than the control (CK). At 10 days, there were no significant differences between any treatment and the CK. At 15 days, only the S treatment showed significant differences. 50 The total flavonoid content in the treatment was significantly higher than that in the control (CK), reaching 15.31 mg / g, which was 1.06 times that of the CK. At 20 days, the total flavonoid content in all treatments was lower than that in the CK, except for S. 40 All treatments were significantly lower than the control (CK). At 25 days, only S... 30 The treatment was significantly lower than the control (CK), while other treatments showed no significant difference compared to the control (CK).

[0069] After treatment with amino esters, D 50 D 100 D 150 The total flavonoid content in all treatments showed a trend of first increasing and then decreasing over 5-25 days. 50 and D 100 All treatments reached their maximum values ​​on day 15, with total flavonoid contents of 14.90 and 16.08 mg / g, respectively, representing increases of 2.69% and 10.82% compared to the control (CK). Meanwhile, D... 150 The total flavonoid content reached its maximum at 20 days, at which point it was significantly higher than the control (CK), being 1.09 times higher. At 20 days, D... 100 The total flavonoid content in the treated group was also significantly higher than that in the control group (CK), reaching 15.59 mg / g, which was 1.06 times that of CK. At 25 days, only D... 50 The treatment was significantly lower than the control (CK), while other treatments showed no significant difference compared to the control (CK).

[0070] 7. Effects of different hormone treatments on the total terpene lactone content in the leaves of 2-year-old Ginkgo seedlings

[0071] The results are as follows Figure 5 As shown, after treatment with uniconazole, S 30 S 50 During treatments of 15-25 days, the total content of Ginkgo total terpenoid lactones was higher than that of the control (CK), reaching its maximum on day 20. 50 The total terpene lactone content of Ginkgo biloba was significantly lower than that of the control group (CK) at 5-10 days, but significantly higher than that of the control group at 15-25 days, showing an effect of first inhibiting and then promoting.

[0072] After treatment with amino esters, D 50 D 200 The total terpene lactone content of Ginkgo biloba showed a trend of first increasing and then decreasing, reaching its maximum on day 20, at which point it was 35.31% and 22.64% higher than the control (CK), respectively. At day 20 of treatment, D... 100 The content of ginkgolide lactones reached its maximum value at each treatment period, which was 5.16 mg / g.

[0073] 8. Effects of different hormone treatments on the content of ginkgo lactone in the leaves of 2-year-old Ginkgo seedlings

[0074] Results as shown in Figure 6 , with the extension of time after spraying, each treatment of uniconazole white kernel lactone content did not show a consistent change, but reached a maximum in 20 d after spraying, this time S 30 and S 50 treatment of white kernel lactone content was significantly higher than CK, respectively, 1.33, 1.49 times of CK.

[0075] After the treatment of fresh amine, D 50 and D 200 treatment of white kernel lactone content showed a first increase and then decrease, in 5-10 d, were lower than CK, in 20 d, were significantly higher than CK, white kernel lactone content was 1.68, 1.35 mg / g. Each concentration treatment in 25 d were significantly higher than CK, respectively, 2.23, 2.37, 1.54 and 1.49 times of CK.

[0076] 9、Different hormone treatment on 2 years old ginkgo seedling leaf lactone A content

[0077] Results as shown in Figure 7 , after the treatment of uniconazole, S 60 treatment in 5, 10, 25 d were significantly higher than CK, 15-20 d were lower than CK, indicating that the same concentration of uniconazole treatment in different time period may play different roles. S 50 treatment of ginkgo lactone A showed a first increase and then decrease, in 20 d, reached a maximum, was significantly higher than CK, 1.99 mg / g. In addition, S 30 , S 40 and other treatments were different, in 25 d, ginkgo lactone A reached the maximum of all periods, respectively, 1.99, 1.65 times of CK.

[0078] After the treatment of fresh amine, 20 d, each treatment reached the maximum. At this time, except for D 150 treatment, other treatments of ginkgo lactone A content were significantly higher than CK, respectively, increased by 49.03%, 47.74%, 50.32%. In 25 d, each treatment of ginkgo lactone A content were significantly higher than CK, respectively, 1.59, 1.47, 1.07, 1.16 mg / g.

[0079] 10、Different hormone treatment on 2 years old ginkgo seedling leaf lactone B content

[0080] Results as shown in Figure 8 , after 10 d of uniconazole treatment, S 30 , S 40 treatment of ginkgo lactone B content was significantly lower than CK, respectively, decreased by 21.74%, 17.39%. Except for 5 d after spraying, S50 S 60 The content of ginkgolide B in the treatment group was significantly higher than that in the control group, and reached its maximum value on day 20, at which time the content of ginkgolide B was 0.69 and 0.58 mg / g, respectively.

[0081] After treatment with aminoethyl esters, the results of each treatment group (except D) 200 The content of ginkgolide B in all treatments was significantly higher than that in the control group (CK) on day 20, reaching the maximum value at each time point. At this time, the content of ginkgolide B in each treatment was 1.24, 2.02, and 1.29 times that of the CK, respectively.

[0082] 11. Effects of different hormone treatments on the content of lactone C in the leaves of 2-year-old Ginkgo seedlings

[0083] The results are as follows Figure 9 As shown, after treatment with uniconazole, except for 5 days, S 50 S 60 The content of ginkgolide C in the treatment group was significantly higher than that in the control group (CK) at other time points, especially reaching a maximum on day 20, where it was 41.94% and 22.58% higher than that in the control group, respectively. Meanwhile, the S group... 30 S 40 The content of ginkgolide C in the treated group was lower than that in the control group (CK) from 15 to 20 days, especially at 20 days. 40 The treatment was significantly lower than the control (CK), decreasing by 22.58% compared to the CK.

[0084] After treatment with aminoethyl esters, the content of ginkgolide C in all treatments was lower than that in the control (CK) on day 5, but higher than that in all treatments from day 15 to day 25, indicating that aminoethyl esters began to promote the accumulation of ginkgolide C over time. Overall, the content of ginkgolide C in all treatments reached its maximum on day 20, increasing by 54.84%, 22.58%, 12.90%, and 38.71% compared to the CK, respectively.

[0085] Example 2: Effects of two plant growth regulators on 3-year-old grafted ginkgo seedlings

[0086] Using the results of the experiment on 2-year-old Ginkgo seedlings as a reference, a total of 3 treatments were applied: a control (CK), and two treatments: 50 mg / L uniconazole and 100 mg / L aminoethyl ester, which had the highest comprehensive scores in the global principal component analysis of the 2-year-old seedlings. Fifteen seedlings were selected for each treatment, for a total of 45 seedlings. The spraying time, sampling method, and sampling frequency were basically consistent with those for the 2-year-old Ginkgo seedlings.

[0087] 1. Effects of different hormone treatments on chlorophyll content in leaves of 3-year-old grafted ginkgo seedlings

[0088] The results are as follows Figure 10As shown, after treatment with S, the chlorophyll content showed a trend of first decreasing and then increasing, reaching its maximum value of 1.28 mg / g at 20 days. Throughout the fluctuation of chlorophyll content, the chlorophyll content of treatment with S at 10 and 20 days was significantly higher than that of the control (CK), while no significant difference was found between treatments with S and CK at other times.

[0089] After treatment D, chlorophyll content showed a trend of first increasing and then decreasing. The chlorophyll content at 10 and 15 days was significantly higher than that at other time points. The chlorophyll content at 5 and 20 days of treatment D was significantly lower than that of the control (CK), but the difference was not statistically significant. The chlorophyll content at 10, 15, and 25 days was significantly higher than that of the CK, increasing by 33.68%, 20.18%, and 5.77%, respectively.

[0090] 2. Effects of different hormone treatments on the soluble sugar content in the leaves of 3-year-old grafted ginkgo seedlings

[0091] The results are as follows Figure 11 As shown, the soluble protein content generally increased after treatment with S and D. The soluble protein content at 25 days after S treatment was significantly higher than that at 5-20 days, increasing by 80.51%, 34.34%, 32.56%, and 28.34% respectively compared to other time periods. At 5 days, the soluble sugar content in the S treatment was significantly lower than that in the control (CK). At 10 days, the soluble sugar content began to significantly exceed that of the CK, reaching 1.27 times that of the CK. From 15 to 25 days, the soluble sugar content in the S treatment was consistently significantly higher than that of the CK, increasing by 46.66%, 27.32%, and 15.05% respectively.

[0092] After treatment D, at 5 days, the soluble sugar content in treatment D was significantly lower than that in control (CK). The soluble protein content at 15 and 25 days was significantly higher than at other time points, at 24.65 and 24.72 mg / g, respectively. At 15 days, the soluble sugar content in treatment D was significantly higher than that in control (CK), being 1.92 times that of control. From 20 to 25 days, the soluble sugar content in treatment D was higher than that in control (CK), but the difference was not statistically significant.

[0093] 3. Effects of different hormone treatments on the soluble protein content in the leaves of 3-year-old grafted ginkgo seedlings

[0094] The results are as follows Figure 12 As shown, after S and D treatments, the soluble protein content generally showed an increasing trend before 20 days, reaching its maximum value at 20 days, and decreasing to varying degrees at 25 days. After S treatment, the soluble protein content significantly increased at 5-10 days and 15-20 days, respectively. The soluble protein content of S treatment was significantly lower than CK at 5 days, decreasing by 10.00% compared to CK; at 10 days, the soluble protein content was equal to CK; and at 20 days and 25 days, the soluble protein content was significantly higher than CK, increasing by 18.81% and 22.68% compared to CK, respectively.

[0095] After D treatment, the soluble protein content increased significantly at 10-20 d, and the soluble protein content at 20 d was 1.29 times that at 10 d. The soluble protein content of D treatment was significantly lower than that of CK at 10 d, which was 10.19% lower than that of CK. With the gradual increase of soluble protein content, the soluble protein content of D treatment was significantly higher than that of CK at 20 d, which was 13.86% higher than that of CK.

[0096] 4. Effects of different hormone treatments on peroxidase (POD) activity in leaves of 3-year-old ginkgo grafting seedlings

[0097] The results are shown in Figure 13 After S treatment, the POD activity showed a general downward trend. The POD activity at 25 d was significantly lower than that at 5 d, which was 30.32% lower than that at 5 d. The POD activity at 20 and 25 d was significantly lower than that of CK, and the POD activity at 10 and 15 d was higher than that of CK. The POD activity of S treatment at 15 d was significantly higher than that of CK, which was 55.30% higher than that of CK.

[0098] After D treatment, the POD activity showed a general upward trend, and the POD activity at 25 d reached the maximum value of 258.77 U·g-1. The POD activity at 25 d was significantly higher than that at 10 d, which was 1.54 times that at 10 d. The POD activity of D treatment at 5 d was significantly lower than that of CK, and the POD activity at other periods was higher than that of CK. The POD activity at 15 and 25 d was significantly higher than that of CK, which was 6.51% and 33.95% higher than that of CK, respectively.

[0099] 5. Effects of different hormone treatments on superoxide dismutase (SOD) activity in leaves of 3-year-old ginkgo grafting seedlings

[0100] The results are shown in Figure 14 After S and D treatments, the SOD activity showed a general upward trend, and the SOD activity at 25 d reached the maximum value of 685.05 and 729.80 U·g-1, respectively. The SOD activity of S treatment at 10 d was lower than that of CK, but did not reach a significant difference level. The SOD activity at other periods was higher than that of CK, and the SOD activity at 5 d was significantly higher than that of CK, which was 1.07 times that of CK.

[0101] After D treatment, the SOD activity at 25 d was significantly higher than that at 20 d, which was 1.13 times that at 20 d. The SOD activity of D treatment at 10 d was lower than that of CK, but did not reach a significant difference level. The SOD activity of D treatment at other periods was higher than that of CK. The SOD activity at 5, 15 and 25 d was significantly higher than that of CK, which was 10.44%, 6.36% and 6.19% higher than that of CK, respectively.

[0102] 6、Different hormone treatments on the content of glutathione (GSH) in leaves of 3-year-old ginkgo grafting seedlings

[0103] The results are shown in Figure 15 After S and D treatments, the GSH content showed a trend of first increasing and then decreasing. At 10 d, the GSH content reached the maximum value of each treatment period, which was 1.31 and 1.35 μmol·g-1, respectively. After S treatment, the GSH content at 10 d was significantly higher than that at other periods, which was 1.18, 1.41, 1.36, and 1.17 times of that at other periods, respectively. The GSH content at 15 and 20 d after S treatment was significantly lower than that of CK, which decreased by 6.06% and 15.04%, respectively. The GSH content at 5, 10, and 25 d was significantly higher than that of CK, especially at 10 d, which was 1.52 times of that of CK.

[0104] After D treatment, the GSH content at 10 d was significantly higher than that at other periods, which was 1.42, 1.35, 1.42, and 1.17 times of that at other periods, respectively. The GSH content at 5 and 20 d after D treatment was significantly lower than that of CK, which decreased by 9.52% and 15.93%, respectively. The GSH content at 15 d was higher than that of CK, but did not reach a significant difference level. The GSH content at 10 and 25 d was significantly higher than that of CK, especially at 10 d, which was 1.57 times of that of CK.

[0105] 7、Different hormone treatments on the content of indole-3-acetic acid (IAA) in leaves of 3-year-old ginkgo grafting seedlings

[0106] The results are shown in Figure 16 After S and D treatments, the IAA content showed an overall upward trend. After S treatment, the IAA content showed a fluctuating upward trend, and the IAA content at 25 d reached the maximum value of each period, which was 72.04 ng·g-1 and significantly higher than that at other periods. The IAA content at 5 d was higher than that of CK, and the IAA content at 10-25 d was significantly lower than that of CK, which decreased by 39.79%, 22.17%, 28.83%, and 10.79%, respectively.

[0107] After D treatment, the IAA content increased significantly at 10-20 d, and the IAA content at 20 d was significantly higher than that at other periods. The IAA content at 10 d after D treatment was lower than that of CK, but did not reach a significant difference level. The IAA content at other periods was higher than that of CK, especially at 15 and 20 d, which was 1.12 and 1.43 times of that of CK, respectively.

[0108] 8、Different hormone treatments on the content of gibberellin (GA3) in leaves of 3-year-old ginkgo grafting seedlings

[0109] The results are shown in Figure 17As shown, after treatment with S, the GA3 content showed a decreasing trend from 5 to 20 days, and the GA3 content at 20 days was significantly lower than that at other time points. At 5 days after treatment with S, the GA3 content was significantly higher than that of CK, and from 10 to 20 days, as the GA3 content decreased, it began to be significantly lower than that of CK, decreasing by 14.29%, 22.46%, and 26.12% compared to CK, respectively.

[0110] After treatment D, the GA3 content showed a trend of first increasing and then rapidly decreasing. The GA3 content at 10 and 15 days was significantly higher than that at other times. The GA3 content of treatment D from 5 to 20 days was significantly higher than that of the control (CK), being 1.05, 1.63, 1.44, and 1.26 times that of CK, respectively. With the rapid decrease in GA3 content, the GA3 content at 25 days was significantly lower than that of CK, decreasing by 34.99% compared to CK.

[0111] 9. Effects of different hormone treatments on abscisic acid (ABA) content in leaves of 3-year-old grafted ginkgo seedlings

[0112] The results are as follows Figure 18 As shown, after treatment with S, the ABA content generally showed an upward trend, reaching its maximum value at 20 days, and was significantly higher than at other times. From 5 to 15 days after treatment with S, the ABA content was lower than the control (CK), especially at 10 and 15 days. With increasing ABA content, from 20 to 25 days, it began to be significantly higher than the CK, reaching 1.06 and 1.67 times that of the CK, respectively.

[0113] After treatment D, the ABA content generally showed a trend of first increasing and then decreasing, reaching the maximum value of 150.16 ng·g⁻¹ at 15 days. The content of D at 5 and 15 days was significantly higher than that of CK, but as the ABA content decreased, the ABA content at 20 and 25 days was significantly lower than that of CK, decreasing by 21.38% and 19.00% respectively.

[0114] 10. Effects of different hormone treatments on the content of zeatin (ZR) in the leaves of 3-year-old grafted Ginkgo biloba seedlings

[0115] The results are as follows Figure 19 As shown, the ZR content generally showed a decreasing trend after S treatment. The ZR content was significantly higher than CK at 5 days of S treatment. As the ZR content decreased, the content at other time points was significantly lower than CK, decreasing by 20.92%, 41.37%, 44.96%, and 20.62% respectively.

[0116] After treatment D, the ZR content generally showed a trend of first increasing and then decreasing, reaching its maximum value at 10 days, which was significantly higher than at other times. The ZR content of treatment D differed from that of control (CK) at different times: it was significantly higher than CK from 5 to 10 days, lower than CK from 15 to 25 days, and significantly lower than CK only at 15 and 25 days.

[0117] 11. Effects of different hormone treatments on the total flavonoid content in the leaves of 3-year-old grafted Ginkgo seedlings

[0118] The results are as follows Figure 20 As shown, after treatment with S, the total flavonoid content did not show a regular change. The total flavonoid content at 10d and 25d was significantly higher than that at other times. The total flavonoid content at 5d and 20d was lower than that at CK, but the difference was not statistically significant. The total flavonoid content at 10d and 15d was higher than that at CK, and the total flavonoid content at 15d was significantly higher than that at CK, being 1.07 times that of CK.

[0119] After treatment D, the total flavonoid content generally showed an upward trend for the first 20 days, reaching its maximum value of 15.77 mg / g at 20 days. The total flavonoid content at 20 days was significantly higher than that at 10 days, being 1.09 times higher. Except for 25 days, the total flavonoid content was higher than that of the control (CK) at all other time points, especially at 15 and 20 days, where the total flavonoid content was significantly higher than that of the CK, increasing by 20.97% and 11.69% respectively.

[0120] 12. Effects of different hormone treatments on the total terpene lactone content in the leaves of 3-year-old grafted Ginkgo seedlings

[0121] The results are shown in Table 3. After S and D treatments, no significant differences were observed between the different time periods or between each treatment and the control (CK). After S and D treatments, the total terpene lactone content of Ginkgo biloba generally showed a trend of first increasing and then decreasing, reaching its maximum at 20 days, at which point it was 1.04 and 1.03 times that of the CK, respectively. The total terpene lactone content of Ginkgo biloba in the S treatment was higher than that in the CK at 10, 20, and 25 days, while the total terpene lactone content of Ginkgo biloba in the D treatment was higher than that in the CK except at 5 days.

[0122] Table 3. Effects of different hormone treatments on the total terpene lactone content of 3-year-old grafted Ginkgo biloba seedlings and results of multiple comparisons.

[0123]

[0124] 13. Effects of different hormone treatments on the content of ginkgolide in the leaves of 3-year-old grafted ginkgo seedlings

[0125] The results are shown in Table 4. After treatment S, the content of ginkgolide in ginkgo biloba showed a fluctuating upward trend, with higher contents at 10 and 25 days, at 1.88 and 1.94 mg / g, respectively. The ginkgolide content at 25 days of treatment S was significantly higher than that of the control (CK), being 1.10 times that of CK. After treatment D, the content of ginkgolide generally showed an upward trend, reaching its maximum value of 1.89 mg / g at 25 days. The ginkgolide content at 25 days of treatment D was significantly higher than that at 5 and 10 days, increasing by 6.18% and 8.00%, respectively. The ginkgolide content at 20 and 25 days of treatment D was significantly higher than that of CK, being 1.04 and 1.07 times that of CK, respectively.

[0126] Table 4 Effects of different hormone treatments on the content of ginkgolides in 3-year-old ginkgo grafted seedlings and multiple comparison results

[0127]

[0128] 14. Effects of different hormone treatments on the content of ginkgolide A in leaves of 3-year-old ginkgo grafted seedlings

[0129] The results are shown in Table 5. After S and D treatments, the content of ginkgolide A in each period was not significantly different from that of CK. After S treatment, the content of ginkgolide A gradually increased at 10-20 d and reached a maximum value of 1.96 mg / g at 20 d. The content of ginkgolide A in S-treated seedlings was equal to that of CK at 5 d, lower than that of CK at 10 d and 25 d, and higher than that of CK at 15 d and 20 d. After D treatment, the content of ginkgolide A gradually increased but was lower than that of CK at 20 d and 25 d.

[0130] Table 5 Effects of different hormone treatments on the content of ginkgolide A in 3-year-old ginkgo grafted seedlings and multiple comparison results

[0131]

[0132] 15. Effects of different hormone treatments on the content of ginkgolide B in leaves of 3-year-old ginkgo grafted seedlings

[0133] The results are shown in Table 6. After S and D treatments, the content of ginkgolide B generally increased, but did not show a regular change in each period compared with that of CK. After S treatment, the content of ginkgolide B was significantly higher than that of CK at 10 d and 20 d, which was 1.17 times and 1.09 times that of CK, respectively. After D treatment, the content of ginkgolide B was higher than that of CK at 10 d and 20 d, but was lower than that of CK at the other times, but did not reach a significant difference level.

[0134] Table 6 Effects of different hormone treatments on the content of ginkgolide B in 3-year-old ginkgo grafted seedlings and multiple comparison results

[0135]

[0136] 16. Effects of different hormone treatments on the content of ginkgolide C in leaves of 3-year-old ginkgo grafted seedlings

[0137] The results are shown in Table 7. After S treatment, the content of ginkgolide C did not show a regular change and reached a maximum value of 0.73 mg / g at 15 d. After S treatment, the content of ginkgolide C was significantly higher than that of CK at 20 d, which was increased by 10.94% compared with that of CK. After D treatment, the content of ginkgolide C generally showed an upward trend and reached a maximum value of 0.72 mg / g at 25 d. After D treatment, the content of ginkgolide C was higher than that of CK at 10 d, 20 d, and 25 d, but did not reach a significant difference level.

[0138] Table 7 Effects of different hormone treatments on the content of endocrocl C in 3-year-old ginkgo grafted seedlings and multiple comparison results

[0139]

[0140] The above description is only illustrative and is not restrictive; many modifications, changes and equivalents can become apparent to those skilled in the art without departing from the spirit and scope of the application as defined by the appended claims.

Claims

1. The application of uniconazole in regulating the accumulation of metabolites in Ginkgo seedlings, characterized in that, The metabolites include chlorophyll, soluble protein, soluble sugar, total flavonoids, ginkgolide, ginkgolide A, ginkgolide B, ginkgolide C, peroxidase, superoxide dismutase, glutathione, gibberellin, zeatin nucleoside, auxin, and abscisic acid.

2. The application according to claim 1, characterized in that, The uniconazole promotes the increase of chlorophyll content, ginkgolide content, ginkgolide A content, ginkgolide B content, ginkgolide C content, soluble sugar content, soluble protein content, total flavonoid content, peroxidase activity, superoxide dismutase activity, glutathione content, auxin content, and abscisic acid content in Ginkgo biloba leaves, while inhibiting the increase of gibberellin and zeatin nucleoside content.

3. The application of aminoethyl esters in regulating the accumulation of metabolites in Ginkgo seedlings, characterized in that, The metabolites include chlorophyll, soluble protein, soluble sugar, total flavonoids, ginkgolide, ginkgolide A, ginkgolide B, ginkgolide C, peroxidase, superoxide dismutase, glutathione, gibberellin, zeatin nucleoside, auxin, and abscisic acid.

4. The application according to claim 3, characterized in that, The amino acid ester promotes the increase of chlorophyll content, soluble sugar content, soluble protein content, total flavonoid content, peroxidase activity, superoxide dismutase activity, glutathione content, auxin content, ginkgolide content, ginkgolide A content, ginkgolide B content, and ginkgolide C content in ginkgo leaves, while inhibiting the increase of gibberellin, abscisic acid, and zeatin nucleoside content.

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