Application of uniconazole and diethyl aminoethyl hexanoate in regulation and control of ginkgo growth and metabolite accumulation

Abstract

The invention discloses application of uniconazole and diethyl aminoethyl hexanoate in regulation and control of ginkgo growth and metabolite accumulation, and relates to the technical field of plant growth promotion. According to the application of uniconazole and diethyl aminoethyl hexanoate in regulation and control of growth of ginkgo biloba and metabolite accumulation, biennial ginkgo biloba seedlings are used as materials, uniconazole with the concentration of 30-60 mg / L or diethyl aminoethyl hexanoate with the concentration of 50-200 mg / L is applied, the optimal uniconazole concentration and the optimal diethyl aminoethyl hexanoate concentration which can promote the growth of the ginkgo biloba, increase the chlorophyll content, increase the primary metabolite activity and increase the secondary metabolite content are screened out and are 50 mg / L and 100 mg / L respectively. Three-year-old gingko grafted seedlings are used as materials, and after 50 mg / L uniconazole and 100 mg / L diethyl aminoethyl hexanoate are sprayed respectively, the comprehensive index of gingko leaves is optimal 25 days later.

Description

Technical Field

[0001] The invention belongs to the technical field of plant growth promotion, and more specifically, relates to the application of uniconazole and diethylaminoethyl ester in regulating the growth of ginkgo and the accumulation of metabolites. Background Art

[0002] Ginkgo biloba L. is the oldest relict plant among the existing gymnosperms and is known as a "living fossil". Ginkgo biloba leaves are rich in flavonoids and terpene lactone compounds and have important medicinal value. EGb761 (Ginkgo bilobaextract 761) is a standardized ginkgo leaf extract, and its preparations are widely used in the fields of medicine, health products and food additives. Flavonoids are the most critical active ingredients (accounting for about 24% of its components), mainly composed of quercetin, isorhamnetin and kaempferol. [2] . Flavonoids help protect the plant from biotic and abiotic stresses through their anti-pathogen, anti-oxidant, and reactive oxygen scavenging functions, and act as regulatory and signaling compounds. They also play an important role in treating age-related cognitive decline, tinnitus, vertigo, and Alzheimer's disease in humans. Ginkgo flavonoids are difficult to chemically synthesize, and are currently mainly extracted from ginkgo leaves. Therefore, increasing the content of flavonoids is beneficial to the survival of ginkgo itself and the development of the ginkgo leaf extract pharmaceutical industry, and has great practical value and broad application prospects.

[0003] Uniconazole and diethylaminoethyl are new plant growth regulators that play a role in plant growth, metabolic regulation, and response to adverse stress. Increasing the content of secondary metabolites in ginkgo leaves by spraying exogenous plant growth regulators has always been a research focus. Conducting research on the application of new plant growth regulators is of great significance for improving the yield and quality of ginkgo leaves. In addition, no research has disclosed the mechanism of action of the two plant growth regulators, uniconazole and diethylaminoethyl, in ginkgo. Summary of the invention

[0004] In view of the above problems existing in the prior art, the technical problem to be solved by the present invention is to provide the application of uniconazole and diethylaminoethyl in regulating the growth of ginkgo seedling height, ground diameter or biomass. Another technical problem to be solved by the present invention is to provide the application of uniconazole and diethylaminoethyl in regulating the accumulation of ginkgo metabolites, which is used to regulate the content of ginkgo soluble protein, soluble sugar, antioxidants, endogenous hormones, total flavonoids, terpene lactones and various components.

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

[0006] The invention discloses an application of uniconazole in regulating the growth of ginkgo seedlings, wherein the growth regulation includes regulating the seedling height, ground diameter and biomass growth.

[0007] When the concentration of uniconazole is less than 60 mg / L, the growth of ginkgo seedling height is promoted; when the concentration of uniconazole is not less than 30 mg / L, the growth of ginkgo ground diameter is promoted; when the concentration of uniconazole is 40 mg / L or 60 mg / L, the increase of ginkgo stem biomass is promoted.

[0008] When the concentration of uniconazole is 60 mg / L, the growth of the height and the ground diameter of the ginkgo seedlings is inhibited.

[0009] The invention discloses an application of uniconazole in regulating the accumulation of metabolites in ginkgo seedlings, wherein the metabolites include chlorophyll, soluble protein, soluble sugar, total flavonoids, bilobalide, lactone A, lactone B, lactone C, peroxidase, superoxide dismutase, glutathione, gibberellin, abscisic acid, zeatin riboside, auxin and abscisic acid.

[0010] When the concentration of the uniconazole is 50-60 mg / L, the chlorophyll content of the ginkgo leaf is increased; when the concentration of the uniconazole is 30-60 mg / L, the total terpene lactone, bilobalide, lactone A, lactone B, and lactone C content of the ginkgo leaf are increased; when the concentration of the uniconazole is 50 mg / L, the soluble sugar content, soluble protein content, total flavonoid content, peroxidase activity, superoxide dismutase activity, glutathione content, auxin content, and abscisic acid content of the ginkgo leaf are increased; the uniconazole inhibits the increase of gibberellin and zeatin riboside content.

[0011] The invention discloses an application of diethylaminoethyl hexanoate in regulating the growth of ginkgo seedlings, wherein the growth regulation includes regulating the seedling height, ground diameter and biomass growth.

[0012] When the concentration of aminoethyl hexanoate is 50-200 mg / L, it promotes the growth of ginkgo seedling height; when the concentration of aminoethyl hexanoate is not less than 150 mg / L, it promotes the growth of ginkgo ground diameter; when the concentration of aminoethyl hexanoate is greater than 50 mg / L, it promotes the increase of ginkgo root biomass; when the concentration of aminoethyl hexanoate is 50-200 mg / L, it promotes the increase of ginkgo stem biomass and single plant biomass; when the concentration of aminoethyl hexanoate is less than 200 mg / L, it promotes the increase of ginkgo aboveground biomass; when the concentration of aminoethyl hexanoate is 150 mg / L, it promotes the increase of ginkgo leaf biomass;

[0013] The invention discloses an application of aminoethyl hexanoate in regulating the accumulation of metabolites in ginkgo seedlings, wherein the metabolites include chlorophyll, soluble protein, soluble sugar, total flavonoids, bilobalide, lactone A, lactone B, lactone C, peroxidase, superoxide dismutase, glutathione, gibberellin, abscisic acid, zeatin riboside, auxin and abscisic acid.

[0014] When the concentration of aminoethyl hexanoate is 150-200 mg / L, it promotes the increase of chlorophyll content in ginkgo leaves; when the concentration of aminoethyl hexanoate is 100 mg / L, it promotes the increase of soluble sugar, soluble protein, total flavonoid content, peroxidase activity, superoxide dismutase activity, glutathione content, and auxin content in ginkgo leaves; when the concentration of aminoethyl hexanoate is 50-200 mg / L, it promotes the increase of total terpene lactones, bilobalide, lactone A, lactone B, and lactone C content in ginkgo leaves;

[0015] The aminoethyl hexadecene inhibits the increase of the contents of gibberellins, abscisic acid and zeatin riboside.

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

[0017] The present invention uses ginkgo seedlings as materials, applies different concentrations of uniconazole and diethylaminoethyl to screen out the optimal concentrations that promote ginkgo growth, increase chlorophyll content, increase primary metabolite activity, and increase secondary metabolite content. Using 3-year-old ginkgo grafted seedlings as materials, the optimal treatment time for spraying S-3307 and DTA-6 is screened out. The results of the embodiment show that:

[0018] 1) The 60 mg / L uniconazole treatment inhibited the growth of Ginkgo biloba seedling height and ground diameter, and all concentrations of treatments generally inhibited the accumulation of Ginkgo biloba plant biomass. Diethylaminoethyl hexanoate treatment promoted the growth of Ginkgo biloba seedling height and had a promoting effect on the biomass of the plant.

[0019] 2) After treatment with 50 mg / L fenthione and 100 mg / L ampicillin, the soluble sugar content and soluble protein content reached the maximum.

[0020] 3) POD activity was inhibited by the treatment of clofosinate, but increased by the treatment of ampicillin. Under the treatment of the two plant growth regulators, SOD activity and GSH content increased overall.

[0021] 4) Under the treatments of ethylenediaminetetrachloridone and anthracene, the IAA content generally increased; under the treatment of ethylenediaminetetrachloridone, the ABA content generally increased, while the GA3 and ZR contents decreased; under the treatment of anthracenetetrachloridone, the IAA content generally increased, while the GA3, ABA, and ZR contents decreased.

[0022] 5) When the concentration of clofosamide was 50 mg / L or the concentration of aminoethyl hexanoate was 100 mg / L, the contents of total flavonoids, terpene lactones and each component increased. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 This is the effect of different hormone treatments on the chlorophyll content in the leaves of two-year-old Ginkgo seedlings;

[0024] Figure 2This is the effect of different hormone treatments on the soluble sugar content in the leaves of two-year-old Ginkgo seedlings;

[0025] Figure 3 This is the effect of different hormone treatments on the soluble protein content in the leaves of two-year-old Ginkgo seedlings;

[0026] Figure 4 This is the effect of different hormone treatments on the total flavonoids content in the leaves of two-year-old Ginkgo seedlings;

[0027] Figure 5 This is the effect of different hormone treatments on the total terpene lactone content in the leaves of two-year-old Ginkgo seedlings;

[0028] Figure 6 This is the effect of different hormone treatments on the bilobalide content in the leaves of two-year-old Ginkgo seedlings;

[0029] Figure 7 This is the effect of different hormone treatments on the content of lactone A in leaves of two-year-old Ginkgo seedlings;

[0030] Figure 8 This is the effect of different hormone treatments on the content of lactone B in leaves of two-year-old Ginkgo seedlings;

[0031] Fig. 9 This is the effect of different hormone treatments on the lactone C content in the leaves of two-year-old Ginkgo seedlings;

[0032] Fig.10 This is the effect of different hormone treatments on the chlorophyll content in the leaves of three-year-old grafted ginkgo seedlings;

[0033] Fig.11 This is the effect of different hormone treatments on the soluble sugar content in leaves of three-year-old grafted ginkgo seedlings;

[0034] Fig.12 This is the effect of different hormone treatments on the soluble protein content in the leaves of three-year-old grafted ginkgo seedlings;

[0035] Fig.13 This is the effect of different hormone treatments on the peroxidase (POD) activity in the leaves of three-year-old grafted Ginkgo seedlings;

[0036] Fig.14 This is the effect of different hormone treatments on the activity of superoxide dismutase (SOD) in the leaves of three-year-old grafted ginkgo seedlings;

[0037] Fig.15 This is the effect of different hormone treatments on glutathione (GSH) activity in leaves of three-year-old grafted ginkgo seedlings;

[0038] Fig.16 This is the effect of different hormone treatments on the IAA content in the leaves of three-year-old grafted ginkgo seedlings;

[0039] Fig.17 This is the effect of different hormone treatments on GA3 content in three-year-old grafted ginkgo seedlings;

[0040] Fig.18 This is the effect of different hormone treatments on ABA content in leaves of three-year-old grafted Ginkgo seedlings;

[0041] Fig.19 This is the effect of different hormone treatments on ZR content in leaves of three-year-old grafted Ginkgo seedlings;

[0042] Fig. 20 This is a graph showing the effects of different hormone treatments on the total flavonoids content in the leaves of three-year-old grafted Ginkgo seedlings. DETAILED DESCRIPTION

[0043] In order to make the purpose, technical scheme and advantages of the present invention clearer, the present invention is further described below in conjunction with specific embodiments. Unless otherwise specified in the following embodiments, the technical means used are conventional means well known to those skilled in the art.

[0044] The 2-year-old ginkgo seedlings and 3-year-old ginkgo grafted seedlings used in this application were purchased from the germplasm resource garden of the National Ginkgo Seedling Base in Pizhou City, Jiangsu Province (the scion was the 15-year-old 'Tancheng No. 1' ginkgo in the germplasm resource garden of the National Ginkgo Seedling Base in Pizhou City).

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

[0046] Using succinimide (S 30 , 30mg / L; S 40 , 40mg / L; S 50 , 50mg / L; S 60 , 60mg / L) and aminoethyl hexanoate (D 50 , 50mg / L; D 100 , 100mg / L; D 150 , 150mg / L; D 200 , 200mg / L;) foliar spraying was performed on 2-year-old ginkgo seedlings, with pure water treatment as the control (CK), for a total of 9 treatments. Use anhydrous ethanol to dissolve the plant growth regulator powder first, and then dilute it with pure water to the required concentration. Among them, ethanol accounts for 1% (v / v) and Tween80 accounts for 0.1% (v / v). CK is sprayed with an equal amount of pure water. After the solution is prepared, use a 2L sprayer to evenly spray it on the ginkgo leaves, ensuring that the front and back of the treated ginkgo leaves are completely wet. After spraying, samples are collected at 5, 10, 15, 20, and 25 days. After 25 days, the seedling height and ground diameter are measured, and the biomass is measured by roots, stems, and leaves.

[0047] 1. Effects of different hormone treatments on the height and ground diameter of two-year-old ginkgo seedlings

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

[0049] The treatment with ammonium ethyl hexachloride promoted the growth of Ginkgo biloba seedlings, especially D 100 The height of ginkgo seedlings increased by 2.44 cm. The effect of aminoethyl ester on the growth of ginkgo seedlings showed a pattern of low concentration inhibition and high concentration promotion. 200 The treatment had the largest increase in ground diameter.

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

[0051]

[0052] Note: S is succinonitrile, D is ampicillin; the subscript numbers represent the concentration of plant growth regulators, unit: mg / L. Different lowercase letters indicate significant differences among different treatments (P<0.05), the same below.

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

[0054] The results are shown in Table 2. The effects of the two plant growth regulators on the biomass of each part of Ginkgo biloba did not reach a significant difference level. In general, the leaf biomass, root biomass, aboveground biomass, and individual plant biomass of the treatments with each concentration of uniconazole were all lower than those of CK. Only S 40 , S 60 treatment was higher than CK.

[0055] After the treatment with diethylaminoethyl ester, the stem and plant biomass of ginkgo in each concentration treatment were higher than those in CK, and the root biomass was 50 Treatment, aboveground biomass except D 200 The treatments were higher than CK. 150 The biomass of leaves, stems, roots, aboveground parts and individual plant biomass of the treatment were higher than those of CK, increasing by 10.08%, 22.12%, 13.1%, 16.12% and 15.20% respectively compared with CK.

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

[0057]

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

[0059] The results are as follows Figure 1 As shown in the figure, after the treatment with uniconazole, the chlorophyll content of each concentration treatment was significantly lower than that of CK at 10 days. 50 , S 60 The chlorophyll contents of the treatments gradually accumulated and were significantly higher than those of CK at 20 days, which were 1.23 and 1.61 times that of CK, respectively.

[0060] After treatment with aminoethyl hexanoate, the chlorophyll content at all sites was significantly lower than that of CK at 5-10 days. 150 During the 5-15 days of treatment, the chlorophyll content gradually increased and was significantly higher than CK at 3.474 mg / g on the 15th day. 200 During the 10-25 days of treatment, the chlorophyll content gradually increased, and at 20 days and 25 days, the chlorophyll content increased by 24.69% and 11.32% compared with CK, respectively.

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

[0062] The results are as follows Figure 2 As shown in the figure, after treatment with uniconazole, the soluble sugar content (except S 40 The soluble sugar content of each concentration treatment showed a gradual accumulation trend from 5 to 15 days, among which S 30 The soluble sugar content of the treatment was significantly higher than that of CK at 15 days, which was 1.15 times that of CK. 40 (Except 15d), S 60 The soluble sugar content of the treatments was lower than that of CK. 50 The soluble sugar content of the treatment gradually accumulated during 5-20 days, and began to decrease at 25 days, but was still significantly higher than that of CK, an increase of 8.40% over CK.

[0063] After treatment with aminoethyl hexanoate, the D 50 , D 150 The soluble sugar content of all treatments except D was significantly lower than that of CK. 200 Except for the treatment, the soluble sugar content of the other treatments gradually increased from 5 to 15 days, and at 15 days, D 100 , D 150The soluble sugar content of the treatment was significantly higher than that of CK, which was 1.12 and 1.18 times that of CK respectively. 200 The soluble sugar content of the treatment showed an overall upward trend, and was significantly lower than that of CK at 5-20 days, decreasing by 14.48%, 35.16%, 39.05% and 31.05% respectively compared with CK. However, it was significantly higher than CK at 25 days, reaching 21.5 mg / g.

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

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

[0066] After treatment with aminoethyl hexanoate, there was no significant difference between each treatment and CK at 5 days, but the soluble protein content of each treatment was significantly higher than that of CK at 10 days, which were 1.57, 1.19, 2.13, and 1.57 times that of CK, respectively. 50 The soluble protein content of the treated samples showed an increasing trend from 5 to 15 days, and the content at 10 and 15 days was significantly higher than that of CK, increasing by 57.33% and 29.06% respectively. 100 The soluble protein content of the treatment was significantly higher than that of CK at 10 days, reaching 0.89 mg / g, and there was no significant difference compared with CK at other periods. 150 , D 200 The contents at 10 and 20 days of treatment were significantly higher than those at CK, especially at the 20th day, when they reached the maximum value, increasing by 17.12% and 21.23% respectively compared with CK.

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

[0068] The results are as follows Figure 4 As shown, after treatment with uniconazole, only S 50The treatments were significantly lower than CK. At 10 days, there were no significant differences between the treatments and CK. At 15 days, only S 50 The total flavonoid content of the treatment was significantly higher than that of CK, and the total flavonoid content at this time was 15.31 mg / g, which was 1.06 times that of CK. At 20 days, the total flavonoid content of each treatment was lower than that of CK, except for S 40 At 25 days, only S 30 The treatment was significantly lower than CK, and there was no significant difference between other treatments and CK.

[0069] After treatment with aminoethyl hexanoate, D 50 , D 100 , D 150 The total flavonoids content showed a trend of increasing first and then decreasing during 5-25 days. 50 and D 100 The maximum values ​​of the total flavonoids in the treatments were reached on the 15th day, when the total flavonoids contents were 14.90 and 16.08 mg / g, respectively, which were 2.69% and 10.82% higher than those in CK. 150 The treatment reached its maximum value at 20 days, when the total flavonoids content was significantly higher than that of CK, 1.09 times that of CK. 100 The total flavonoid content of D was also significantly higher than that of CK, reaching 15.59 mg / g, which was 1.06 times that of CK. 50 The treatment was significantly lower than CK, and there were no significant differences between other treatments and CK.

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

[0071] The results are as follows Figure 5 As shown, after treatment with uniconazole, S 30 , S 50 At 15-25 days of treatment, the content of total terpene lactones in Ginkgo was higher than that in CK and reached the maximum value at the 20th day. 50 The content of total terpene lactones in Ginkgo biloba was significantly lower than that in CK at 5-10 days, and significantly higher than that in CK at 15-25 days, showing an effect of first inhibition and then promotion.

[0072] After treatment with aminoethyl hexanoate, D 50 , D 200 The content of total terpene lactones in Ginkgo biloba treated with D showed a trend of first increasing and then decreasing, and reached the maximum value on the 20th day, which was 35.31% and 22.64% higher than that of CK, respectively. 100 The content of terpene lactones in treated Ginkgo biloba reached the maximum value at each period, which was 5.16 mg / g.

[0073] 8. Effects of different hormone treatments on bilobalide content in leaves of two-year-old Ginkgo seedlings

[0074] The results are as follows Figure 6 As shown in Figure 2, with the extension of time after spraying, the content of bilobalide in each treatment of uniconazole did not show a consistent change pattern, but all reached the maximum value on the 20th day after spraying. 30 and S 50 The content of bilobalide in the treated group was significantly higher than that in CK, which was 1.33 and 1.49 times that of CK respectively.

[0075] After treatment with aminoethyl hexanoate, D 50 and D 200 The content of bilobalide in the treatments showed a pattern of first increasing and then decreasing. It was lower than CK at 5-10 days, and significantly higher than CK at 20 days, with bilobalide contents of 1.68 and 1.35 mg / g, respectively. The treatments of each concentration were significantly higher than CK at 25 days, which were 2.23, 2.37, 1.54 and 1.49 times that of CK, respectively.

[0076] 9. Effects of different hormone treatments on the content of lactone A in leaves of two-year-old Ginkgo seedlings

[0077] The results are as follows Figure 7 As shown, after treatment with uniconazole, S 60 The treatment was significantly higher than CK at 5, 10, and 25 days, and lower than CK at 15-20 days, indicating that the same concentration of succinimide treatment may play different roles in different time periods. 50 The content of Ginkgolide A in the treatment showed a trend of first increasing and then decreasing, reaching a maximum value at 20 days, which was significantly higher than CK, at 1.99 mg / g. 30 , S 40 Different from other treatments, Ginkgolide A reached its maximum value at 25 days, which was 1.99 and 1.65 times that of CK respectively.

[0078] After treatment with aminoethyl hexanoate, all treatments reached their maximum values ​​at 20 days. 150 Except for the treatment, the contents of Ginkgolide A in other treatments were significantly higher than that in CK, increasing by 49.03%, 47.74% and 50.32% respectively. At 25 days, the contents of Ginkgolide A in each treatment were significantly higher than that in CK, which were 1.59, 1.47, 1.07 and 1.16 mg / g respectively.

[0079] 10. Effects of different hormone treatments on the content of lactone B in leaves of two-year-old Ginkgo seedlings

[0080] The results are as follows Figure 8 As shown, 10 days after the treatment with uniconazole, S 30 , S 40 The content of ginkgolide B in the treatment was significantly lower than that in CK, decreasing by 21.74% and 17.39% respectively.50 , S 60 The contents of Ginkgolide B in the treatments were significantly higher than those in CK and reached the maximum value on the 20th day, when the contents of Ginkgolide B were 0.69 and 0.58 mg / g, respectively.

[0081] After treatment with aminoethyl hexanoate, all treatment groups (except D 200 The contents of ginkgolide B in the treatments were significantly higher than those in CK on the 20th day, and reached the maximum value in each period. At this time, the contents of ginkgolide B in each treatment were 1.24, 2.02 and 1.29 times that of CK, respectively.

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

[0083] The results are as follows Fig. 9 As shown in the figure, after treatment with uniconazole, except for 5 days, S 50 , S 60 The content of Ginkgolide C in the treatment was significantly higher than that in CK at other stages, especially reaching the maximum value on the 20th day, which was 41.94% and 22.58% higher than that in CK, respectively. 30 , S 40 The content of Ginkgolide C in treatment was lower than that in CK at 15-20 days, especially at 20 days. 40 The treatment was significantly lower than CK, reduced by 22.58% compared with CK.

[0084] After treatment with aminoethyl hexanoate, the content of ginkgolide C in each treatment was lower than that in CK on the 5th day, and higher than that in CK on the 15th to 25th day, indicating that aminoethyl hexanoate began to promote the accumulation of ginkgolide C over time. In general, the content of ginkgolide C in each treatment reached the maximum on the 20th day, which was 54.84%, 22.58%, 12.90% and 38.71% higher than that in CK, respectively.

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

[0086] The test results of 2-year-old ginkgo seedlings were used as a reference, with a total of 3 treatments. They were control (CK), 50 mg / L nitropropane and 100 mg / L diethylaminoethyl, which had the highest comprehensive scores in the global principal component analysis of the 2-year-old seedling test. Fifteen seedlings were selected for each treatment, for a total of 45 seedlings. The spraying time, sampling method and sampling times were basically the same as those of the 2-year-old ginkgo seedlings.

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

[0088] The results are as follows Fig.10As shown in the figure, after S treatment, the chlorophyll content showed a trend of first decreasing and then increasing, and the chlorophyll content at 20 days reached the maximum value of each period, which was 1.28 mg / g. During the fluctuation of the chlorophyll content, the chlorophyll content at 10 and 20 days of S treatment was significantly higher than that of CK, and no significant difference was reached in other periods compared with CK.

[0089] After D treatment, the 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 periods. The chlorophyll content at 5 and 20 days of D treatment was significantly lower than that of CK, but did not reach a significant difference level. The chlorophyll content at 10, 15 and 25 days was significantly higher than that of CK, which was 33.68, 20.18 and 5.77% higher than that of CK, respectively.

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

[0091] The results are as follows Fig.11 As shown in the figure, after S and D treatments, the soluble protein content showed an overall increasing trend. The soluble protein content at 25 days after S treatment was significantly higher than that at 5-20 days, and increased by 80.51%, 34.34%, 32.56% and 28.34% respectively compared with each period. The soluble sugar content of S treatment was significantly lower than that of CK at 5 days, and began to be significantly higher than that of CK at 10 days, which was 1.27 times that of CK. The soluble sugar content of S treatment was significantly higher than that of CK at 15-25 days, which was 46.66%, 27.32% and 15.05% higher than that of CK respectively.

[0092] After D treatment, at 5 days, the soluble sugar content of D treatment was significantly lower than that of CK. The soluble protein content at 15 and 25 days was significantly higher than that of the other periods, which were 24.65 and 24.72 mg / g, respectively. The soluble sugar content of D treatment was significantly higher than that of CK at 15 days, which was 1.92 times that of CK. At 20-25 days, the soluble sugar content of D treatment was higher than that of CK but did not reach a significant difference level.

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

[0094] The results are as follows Fig.12 As shown in the figure, after S and D treatments, the soluble protein content generally showed an increasing trend before 20 days, reached the maximum value of each treatment period at 20 days, and decreased to varying degrees at 25 days. After S treatment, the soluble protein content increased significantly at 5-10 days and 15-20 days, respectively. The soluble protein content of S treatment was significantly lower than that of CK at 5 days, which was 10.00% lower than that of CK. The soluble protein content was equal to CK at 10 days, and the soluble protein content was significantly higher than CK at 20 days and 25 days, which was 18.81% and 22.68% higher than that of CK, respectively.

[0095] After D treatment, the soluble protein content increased significantly from 10 to 20 days, and the soluble protein at 20 days was 1.29 times that at 10 days. The soluble protein content of D treatment was significantly lower than that of CK at 10 days, which was 10.19% lower than that of CK. As the soluble protein content gradually increased, the soluble protein content of D treatment was significantly higher than that of CK at 20 days, which was 13.86% higher than that of CK.

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

[0097] The results are as follows Fig.13 As shown in the figure, after S treatment, POD activity showed a downward trend overall. The POD activity at 25 days was significantly lower than that at 5 days, and decreased by 30.32% compared with 5 days. The POD activity at 20 and 25 days was significantly lower than that of CK, and the POD activity at 10 and 15 days was higher than that of CK. Only at 15 days of S treatment was the POD activity significantly higher than that of CK, which was 55.30% higher than that of CK.

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

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

[0100] The results are as follows Fig.14 As shown in the figure, after S and D treatment, SOD activity showed an overall upward trend, reaching the maximum value of each treatment period at 25 days, which was 685.05 and 729.80 U·g-1, respectively. Only at 10 days in S treatment, SOD activity was lower than that in CK, but it did not reach a significant difference level. SOD activity in other periods was higher than that in CK, and SOD activity was significantly higher than CK at 5 days, which was 1.07 times that of CK.

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

[0102] 6. Effects of different hormone treatments on glutathione (GSH) content in leaves of three-year-old grafted ginkgo seedlings

[0103] The results are as follows Fig.15 As shown in the figure, after S and D treatments, the GSH content showed an overall trend of first increasing and then decreasing, reaching the maximum values ​​of each treatment period at 10 days, which were 1.31 and 1.35 μmol·g-1, respectively. After S treatment, the GSH content at 10 days was significantly higher than that of other periods, which was 1.18, 1.41, 1.36, and 1.17 times that of other periods, respectively. The GSH content at 15 and 20 days after S treatment was significantly lower than that of CK, which was 6.06 and 15.04% lower than that of CK, and was significantly higher than that of CK at 5, 10, and 25 days, especially at 10 days, which was 1.52 times that of CK.

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

[0105] 7. Effects of different hormone treatments on the content of auxin (IAA) in leaves of three-year-old grafted ginkgo seedlings

[0106] The results are as follows Fig.16 As shown in the figure, 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 reached the maximum value of each period at 25 days, which was 72.04 ng·g-1, significantly higher than other periods. Except for 5 days, the IAA content was higher than CK, and it was significantly lower than CK at 10-25 days, which was 39.79%, 22.17%, 28.83% and 10.79% lower than CK, respectively.

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

[0108] 8. Effects of different hormone treatments on the content of gibberellin (GA3) in leaves of three-year-old grafted ginkgo seedlings

[0109] The results are as follows Fig.17As shown in the figure, after S treatment, GA3 content showed a downward trend from 5 to 20 days, and the GA3 content at 20 days was significantly lower than that at other periods. GA3 content was significantly higher than that of CK at 5 days after S treatment, and as GA3 content decreased from 10 to 20 days, it began to be significantly lower than that of CK, which was 14.29, 22.46, and 26.12% lower than that of CK, respectively.

[0110] After D treatment, the GA3 content showed a trend of first increasing and then decreasing sharply. The GA3 content at 10 and 15 days was significantly higher than that at other periods. The GA3 content at 5-20 days of D treatment was significantly higher than that of CK, which was 1.05, 1.63, 1.44, and 1.26 times that of CK, respectively. With the rapid decrease of GA3 content, the GA3 content at 25 days was significantly lower than that of CK, which was 34.99% lower than that of CK.

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

[0112] The results are as follows Fig.18 As shown in the figure, after S treatment, ABA content showed an overall upward trend, reaching the maximum value of each period at 20 days, and significantly higher than other periods. ABA content was lower than CK at 5-15 days after S treatment, especially at 10 and 15 days. With the increase of ABA content, it began to be significantly higher than CK at 20-25 days, which was 1.06 and 1.67 times that of CK, respectively.

[0113] After D treatment, 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 treatment at 5 and 15 days was significantly higher than that of CK, but with the decrease of ABA content, the ABA content at 20 and 25 days was significantly lower than that of CK, which was 21.38 and 19.00% lower than that of CK, respectively.

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

[0115] The results are as follows Fig.19 As shown in the figure, the ZR content showed a downward trend after S treatment. The ZR content was significantly higher than that of CK at 5 days after S treatment. As the ZR content decreased, the content at other periods was significantly lower than that of CK, which was 20.92%, 41.37%, 44.96% and 20.62% lower than that of CK, respectively.

[0116] After D treatment, the ZR content showed a general trend of first increasing and then decreasing. The ZR content reached the maximum value at 10 days and was significantly higher than that at other periods. The ZR content in D treatment showed different performances compared with CK at different periods. It was significantly higher than CK at 5-10 days, lower than CK at 15-25 days, and only significantly lower than CK at 15 and 25 days.

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

[0118] The results are as follows Fig. 20 As shown in the figure, after S treatment, the total flavonoid content did not show regular changes, and the total flavonoid content at 10d and 25d was significantly higher than that at other periods. The total flavonoid content at 5d and 20d was lower than that at CK, but did not reach a significant difference level. 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, which was 1.07 times that of CK.

[0119] After D treatment, the total flavonoid content showed an overall upward trend in the first 20 days, and reached a maximum of 15.77 mg / g at 20 days. The total flavonoid content at 20 days was significantly higher than that at 10 days, 1.09 times that at 10 days. Except for 25 days, the total flavonoid content in the remaining periods was higher than that in CK, especially at 15 and 20 days, which was significantly higher than that in CK, increasing by 20.97 and 11.69% respectively compared with CK.

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

[0121] The results are shown in Table 3. After S and D treatments, there was no significant difference between each period and between each treatment and CK. After S and D treatments, the content of total terpene lactones in Ginkgo biloba generally showed a trend of first increasing and then decreasing, reaching the maximum value at 20 days, which was 1.04 and 1.03 times that of CK, respectively. The content of total terpene lactones in Ginkgo biloba treated with S was higher than that in CK at 10, 20, and 25 days, and the content of total terpene lactones in Ginkgo biloba treated with D was higher than that in CK except for 5 days.

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

[0123]

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

[0125] The results are shown in Table 4. After S treatment, the content of bilobalide in Ginkgo biloba showed a fluctuating upward trend. The bilobalide content at 10 and 25 days was relatively high, 1.88 and 1.94 mg / g, respectively. The bilobalide content at 25 days after S treatment was significantly higher than that of CK, 1.10 times that of CK. After D treatment, the bilobalide content showed an overall upward trend, reaching a maximum of 1.89 mg / g at 25 days. The bilobalide content at 25 days after D treatment was significantly higher than that at 5 and 10 days, increasing by 6.18% and 8.00%, respectively. The bilobalide content at 20 and 25 days after D treatment was significantly higher than that of CK, 1.04 and 1.07 times that of CK, respectively.

[0126] Table 4 Effects of different hormone treatments on bilobalide content in three-year-old grafted Ginkgo seedlings and multiple comparison results

[0127]

[0128] 14. Effects of different hormone treatments on the content of lactone A in leaves of three-year-old grafted Ginkgo seedlings

[0129] The results are shown in Table 5. After S and D treatments, no significant differences were achieved between each period and between each treatment and CK. After S treatment, the content of Ginkgolide A gradually increased from 10 to 20 days, reaching a maximum of 1.96 mg / g at 20 days. The content of Ginkgolide A in S treatment was equal to CK at 5 days, lower than CK at 10 and 25 days, and higher than CK at 15 and 20 days. After D treatment, the content of Ginkgolide A gradually increased, but was lower than CK at 20 and 25 days.

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

[0131]

[0132] 15. Effects of different hormone treatments on the content of lactone B in leaves of three-year-old grafted Ginkgo seedlings

[0133] The results are shown in Table 6. After S and D treatments, the content of Ginkgolide B increased overall, but compared with CK, there was no regular change in each period. The S treatment was significantly higher than CK only at 10d and 20d, which was 1.17 times and 1.09 times that of CK. The D treatment was higher than CK at 10d and 20d, and lower than CK at other periods, but did not reach a significant difference level.

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

[0135]

[0136] 16. Effects of different hormone treatments on the content of lactone C in leaves of three-year-old grafted Ginkgo seedlings

[0137] The results are shown in Table 7. After S treatment, the C content in Ginkgo biloba did not show regular changes. The Ginkgolide C content reached a maximum of 0.73 mg / g at 15 days. The Ginkgolide C content in S treatment was significantly higher than that in CK only at 20 days, which was 10.94% higher than that in CK. After D treatment, the C content in Ginkgo biloba showed an overall upward trend, reaching a maximum of 0.72 mg / g at 25 days. The D treatment was higher than CK at 10, 20, and 25 days, but did not reach a significant difference level.

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

[0139]

[0140] The above description is only illustrative rather than restrictive of the present invention. Those skilled in the art will understand that many modifications, changes or equivalents may be made without departing from the spirit and scope defined by the appended claims, but all will fall within the scope of protection of the present invention.

Claims

1. Application of clofoconazole in regulating the growth of Ginkgo seedlings.

2. The use according to claim 1, characterized in that: The growth regulation is to regulate the seedling height, ground diameter and biomass growth.

3. Application of clofoconazole in regulating the accumulation of metabolites in Ginkgo seedlings.

4. The use according to claim 3, characterized in that: The metabolites include chlorophyll, soluble protein, soluble sugar, total flavonoids, bilobalide, lactone A, lactone B, lactone C, peroxidase, superoxide dismutase, glutathione, gibberellin, abscisic acid, zeatin riboside, auxin, and abscisic acid.

5. The use according to claim 4, characterized in that: The uniconazole promotes the increase of chlorophyll content, total terpene lactone content, biloba lactone content, lactone A content, lactone B content, lactone C content, soluble sugar content, soluble protein content, total flavonoids content, peroxidase activity, superoxide dismutase activity, glutathione content, auxin content, and abscisic acid content in ginkgo leaves, and inhibits the increase of gibberellin and zeatin riboside content.

6. Application of aminoethyl ester in regulating the growth of Ginkgo seedlings.

7. The use according to claim 6, characterized in that: The growth regulation is to regulate the seedling height, ground diameter and biomass growth.

8. Application of aminoethyl ester in regulating the accumulation of metabolites in Ginkgo seedlings.

9. The use according to claim 8, characterized in that: The metabolites include chlorophyll, soluble protein, soluble sugar, total flavonoids, bilobalide, lactone A, lactone B, lactone C, peroxidase, superoxide dismutase, glutathione, gibberellin, abscisic acid, zeatin riboside, auxin, and abscisic acid.

10. The use according to claim 9, characterized in that: The aminoethyl hexadecene promotes the increase of chlorophyll content, soluble sugar content, soluble protein content, total flavonoids content, peroxidase activity, superoxide dismutase activity, glutathione content, auxin content, total terpene lactone content, biloba lactone content, lactone A content, lactone B content, and lactone C content of ginkgo leaves, and inhibits the increase of gibberellin, abscisic acid, and zeatin riboside content.

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