Application of geranyl linalool in inhibiting growth of lateral roots of notopterygium incisum seedlings and / or breeding of notopterygium incisum superior seedlings

Abstract

The invention provides application of geranyl linalool to inhibition of growth of lateral roots of notopterygium incisum seedlings and / or breeding of notopterygium incisum superior seedlings, and belongs to the technical field of planting of wild notopterygium incisum seedlings. In the invention, the geranyl linalool can promote the elongation of the roots of the notopterygium incisum seedlings and inhibit the formation of lateral roots by increasing the content of auxin in the notopterygium incisum seedlings and reducing the contents of jasmonic acid, abscisic acid and 1-aminocyclopropane-1-carboxylic acid (an ethylene synthesis precursor). Moreover, the geranyl linalool can also improve the activity of polyphenol oxidase and urease in rhizosphere soil, improve the activity of nitrate reductase and glutamine synthetase in leaves, improve the content of alkali-hydrolyzable nitrogen in soil, reduce the content of total phenolic acid in soil and promote conversion of ammonium nitrogen into nitrate nitrogen, so that nitrogen absorption and utilization are promoted, and growth and shape optimization of seedlings are facilitated. In addition, the geranyl linalool can also improve the abundance of rhizosphere soil probiotic bacteria and reduce the abundance of rhizosphere soil pathogenic fungi, so that the disease resistance of the notopterygium incisum seedlings is improved.

Description

Technical Field

[0001] The invention belongs to the technical field of wild Notopterygium wilfordii seedling planting, and in particular relates to the application of geranyl linalool in inhibiting the growth of lateral roots of Notopterygium wilfordii seedlings and / or breeding Notopterygium wilfordii seedlings with superior shape and quality. Background Art

[0002] Notopterygium incisum Tingex H.T. Chang is a perennial herbaceous plant in the Apiaceae family. Its medicinal parts are the dried roots and rhizomes, which have the effects of dispelling cold, expelling wind and dampness, and tonifying joints. Artificial cultivation and simulated wild cultivation are the main methods for artificially cultivating Notopterygium incisum. However, currently cultivated Notopterygium incisum seedlings and medicinal materials generally exhibit excessive branching and overdeveloped lateral roots, which do not meet the "optimal" requirements of "silkworm Notopterygium" and "bamboo-jointed Notopterygium," significantly reducing their commercial value. Summary of the Invention

[0003] The purpose of the present invention is to provide an application of geranyl linalool in inhibiting the growth of lateral roots of Notopterygium wilfordii seedlings and / or breeding Notopterygium wilfordii seedlings with superior shape and quality.

[0004] The present invention provides an application of geranyl linalool in planting Notopterygium wilfordii seedlings, wherein the planting of Notopterygium wilfordii seedlings comprises at least one of the following:

[0005] 1) Inhibit the growth of lateral roots of Notopterygium wilfordii seedlings;

[0006] 2) Propagation of superior-shaped and / or high-quality seedlings of Notopterygium wilfordii;

[0007] 3) Increase the root length of Notopterygium wilfordii seedlings;

[0008] 4) increasing the nitrogen content of Notopterygium wilfordii seedling roots and / or increasing the absorption capacity of Notopterygium wilfordii seedling roots;

[0009] 5) Promote the growth of Notopterygium wilfordii seedlings;

[0010] 6) reducing the abundance of pathogens in Notopterygium incisum seedlings and / or increasing the abundance of probiotics in Notopterygium incisum seedlings;

[0011] 7) Improve disease resistance of Notopterygium wilfordii seedlings;

[0012] 8) improving the recycling and utilization of nitrogen in the soil where Notopterygium wilfordii seedlings are planted and / or reducing the content of autotoxic substances in the soil where Notopterygium wilfordii seedlings are planted;

[0013] 9) Reduce the number of fungal species in the rhizosphere soil of Notopterygium wilfordii seedlings.

[0014] The working concentration of the geranyl linalool is 1-3 ml / L.

[0015] The present invention also provides a geranyl linalool emulsion, comprising water and geranyl linalool at a concentration of 1 to 3 ml / L; the geranyl linalool emulsion uses Tween-80 as an emulsifier.

[0016] Preferably, the concentration of Tween-80 in the geranyl linalool emulsion is 1 ml / L.

[0017] The present invention also provides the use of the geranyl linalool emulsion described in the above scheme in planting Notopterygium wilfordii seedlings, wherein the planting of Notopterygium wilfordii seedlings comprises at least one of the following:

[0018] 1) Inhibit the growth of lateral roots of Notopterygium wilfordii seedlings;

[0019] 2) Propagation of superior-shaped and / or high-quality seedlings of Notopterygium wilfordii;

[0020] 3) Increase the root length of Notopterygium wilfordii seedlings;

[0021] 4) increasing the nitrogen content of Notopterygium wilfordii seedling roots and / or increasing the absorption capacity of Notopterygium wilfordii seedling roots;

[0022] 5) Promote the growth of Notopterygium wilfordii seedlings;

[0023] 6) reducing the abundance of pathogens in Notopterygium incisum seedlings and / or increasing the abundance of probiotics in Notopterygium incisum seedlings;

[0024] 7) Improve disease resistance of Notopterygium wilfordii seedlings;

[0025] 8) improving the recycling and utilization of nitrogen in the soil where Notopterygium wilfordii seedlings are planted and / or reducing the content of autotoxic substances in the soil where Notopterygium wilfordii seedlings are planted;

[0026] 9) Reduce the number of fungal species in the rhizosphere soil of Notopterygium wilfordii seedlings.

[0027] Preferably, the Notopterygium wilfordii includes wild Notopterygium wilfordii; the Notopterygium wilfordii includes Silkworm Notopterygium wilfordii.

[0028] Preferably, the pathogens include pathogenic fungi and / or pathogenic bacteria; the pathogenic fungi include at least one of Higgins Anthrax, Alternaria alternata, Fusarium oxysporum, Fusarium aromaticum, Colletotrichum frutescens and Alternaria dendriticum.

[0029] Preferably, the probiotics include probiotic bacteria and / or probiotic fungi; the probiotic bacteria include at least one of the genus Micromonospora, Neurospora and Bacillus.

[0030] The present invention also provides a method for inhibiting the growth of lateral roots of Notopterygium wilfordii and / or breeding superior Notopterygium wilfordii shape and / or high-quality seedlings, comprising the following steps: applying the geranyl linalool emulsion described in the above scheme during the Notopterygium wilfordii seedling stage.

[0031] Preferably, the application includes spraying and root irrigation.

[0032] Preferably, the application time is after the Notopterygium wilfordii seedlings emerge; the application frequency is once every 21 to 28 days; the total number of applications is 4 times; and the application amount for each application is 100 to 300 L / mu.

[0033] The present invention provides an application of geranyl linalool in planting Notopterygium wilfordii seedlings; the planting of Notopterygium wilfordii seedlings comprises at least one of the following: 1) inhibiting the growth of lateral roots of Notopterygium wilfordii seedlings; 2) breeding Notopterygium wilfordii seedlings with superior shape and / or high-quality Notopterygium wilfordii seedlings; 3) increasing the specific root length of Notopterygium wilfordii seedlings; 4) increasing the nitrogen content of Notopterygium wilfordii seedling roots and / or increasing the absorption capacity of Notopterygium wilfordii seedling roots; 5) promoting the growth of Notopterygium wilfordii seedlings; 6) reducing the abundance of pathogens in Notopterygium wilfordii seedlings and / or increasing the abundance of probiotics in Notopterygium wilfordii seedlings; 7) improving the disease resistance of Notopterygium wilfordii seedlings; 8) improving the circulation and utilization of nitrogen in the soil where Notopterygium wilfordii seedlings are planted and / or reducing the content of autotoxic substances in the soil where Notopterygium wilfordii seedlings are planted; 9) reducing the number of fungal species in the rhizosphere soil of Notopterygium wilfordii seedlings; the working concentration of the geranyl linalool is 1-3 ml / L.

[0034] In the present invention, geranyl linalool can promote root elongation and inhibit lateral root formation of Notopterygium incisum seedlings by increasing the auxin content of Notopterygium incisum seedlings and reducing the content of jasmonic acids, abscisic acid and 1-aminocyclopropane-1-carboxylic acid (ethylene synthesis precursor). In addition, geranyl linalool can also increase the activity of polyphenol oxidase and urease in the rhizosphere soil, increase the activity of nitrate reductase and glutamine synthetase in leaves, increase the content of soil alkaline nitrogen, reduce the total phenolic acid content in the soil and promote the conversion of ammonium nitrogen to nitrate nitrogen, thereby promoting nitrogen absorption and utilization, and contributing to the growth and good shape of seedlings. In addition, geranyl linalool can also increase the abundance of rhizosphere soil probiotic bacteria Micromonospora, Alternaria and Bacillus, and reduce the abundance of rhizosphere soil pathogenic fungi Higgins Anthracnose, Alternaria alternata, Fusarium oxysporum, Fusarium aromaticum, Colletotrichum fruiti and Alternaria tree, thereby improving the disease resistance of Notopterygium incisum seedlings. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0036] Figure 1 Analyze cladograms for bacterial LEfSe;

[0037] Figure 2 KEGG Level2 function prediction for bacterial LEfSe. DETAILED DESCRIPTION

[0038] The present invention provides an application of geranyl linalool in planting Notopterygium wilfordii seedlings, wherein the planting of Notopterygium wilfordii seedlings comprises at least one of the following:

[0039] 1) Inhibit the growth of lateral roots of Notopterygium wilfordii seedlings;

[0040] 2) Propagation of superior-shaped and / or high-quality seedlings of Notopterygium wilfordii;

[0041] 3) Increase the root length of Notopterygium wilfordii seedlings;

[0042] 4) increasing the nitrogen content of Notopterygium wilfordii seedling roots and / or increasing the absorption capacity of Notopterygium wilfordii seedling roots;

[0043] 5) Promote the growth of Notopterygium wilfordii seedlings;

[0044] 6) reducing the abundance of pathogens in Notopterygium incisum seedlings and / or increasing the abundance of probiotics in Notopterygium incisum seedlings;

[0045] 7) Improve disease resistance of Notopterygium wilfordii seedlings;

[0046] 8) improving the recycling and utilization of nitrogen in the soil where Notopterygium wilfordii seedlings are planted and / or reducing the content of autotoxic substances in the soil where Notopterygium wilfordii seedlings are planted;

[0047] 9) Reduce the number of fungal species in the rhizosphere soil of Notopterygium wilfordii seedlings.

[0048] The working concentration of the geranyl linalool is 1-3 ml / L.

[0049] In the present invention, the working concentration of geranyl linalool is 1-3 ml / L, which can effectively inhibit bacteria and obtain a higher germination index.

[0050] The present invention also provides a geranyl linalool emulsion, comprising water and geranyl linalool at a concentration of 1 to 3 ml / L; the geranyl linalool emulsion uses Tween-80 as an emulsifier.

[0051] As an embodiment, the concentration of geranyl linalool in the geranyl linalool emulsion is 3 ml / L.

[0052] As an embodiment, the concentration of Tween-80 in the geranyl linalool emulsion is 1 ml / L.

[0053] In the present invention, the Tween-80 is used to dissolve and disperse geranyl linalool and has low toxicity to Notopterygium wilfordii.

[0054] The geranyl linalool emulsion of the present invention can effectively inhibit the growth of lateral roots of Notopterygium wilfordii seedlings, increase the specific root length and root nitrogen content of Notopterygium wilfordii seedlings, improve the absorption capacity of the roots, reduce the abundance of pathogens in Notopterygium wilfordii seedlings, and promote the breeding of superior-shaped and high-quality seedlings.

[0055] The present invention also provides the use of the geranyl linalool emulsion described in the above scheme in planting Notopterygium wilfordii seedlings, wherein the planting of Notopterygium wilfordii seedlings comprises at least one of the following:

[0056] 1) Inhibit the growth of lateral roots of Notopterygium wilfordii seedlings;

[0057] 2) Propagation of superior-shaped and / or high-quality seedlings of Notopterygium wilfordii;

[0058] 3) Increase the root length of Notopterygium wilfordii seedlings;

[0059] 4) increasing the nitrogen content of Notopterygium wilfordii seedling roots and / or increasing the absorption capacity of Notopterygium wilfordii seedling roots;

[0060] 5) Promote the growth of Notopterygium wilfordii seedlings;

[0061] 6) reducing the abundance of pathogens in Notopterygium incisum seedlings and / or increasing the abundance of probiotics in Notopterygium incisum seedlings;

[0062] 7) Improve disease resistance of Notopterygium wilfordii seedlings;

[0063] 8) improving the recycling and utilization of nitrogen in the soil where Notopterygium wilfordii seedlings are planted and / or reducing the content of autotoxic substances in the soil where Notopterygium wilfordii seedlings are planted;

[0064] 9) Reduce the number of fungal species in the rhizosphere soil of Notopterygium wilfordii seedlings.

[0065] As an embodiment, the Notopterygium wilfordii includes wild Notopterygium wilfordii; the Notopterygium wilfordii includes Silkworm Notopterygium wilfordii.

[0066] As an embodiment, the pathogens include pathogenic fungi and / or pathogenic bacteria; the pathogenic fungi include at least one of Higgins Anthrax, Alternaria alternata, Fusarium oxysporum, Fusarium aromaticum, Colletotrichum frutescens and Alternaria dendriticum.

[0067] As an embodiment, the probiotics include probiotic bacteria and / or probiotic fungi; the probiotic bacteria include at least one of the genus Micromonospora, Neurospora, and Bacillus.

[0068] As an embodiment, the promoting growth of Notopterygium wilfordii seedlings is achieved by at least one of the following:

[0069] (1) Increase the auxin content of Notopterygium wilfordii seedlings;

[0070] (2) reduce the content of jasmonic acids, abscisic acid and 1-aminocyclopropane-1-carboxylic acid;

[0071] (3) increasing the activity of polyphenol oxidase and / or urease in rhizosphere soil;

[0072] (4) increasing the activity of nitrate reductase and / or glutamine synthetase in leaves;

[0073] (5) Increase soil alkaline nitrogen content

[0074] (6) Improve the soil nitrate nitrogen / ammonium nitrogen ratio;

[0075] (7) Reduce the total phenolic acid content in soil;

[0076] (8) Promote the activated carbon metabolism and nitrogen absorption and utilization of Notopterygium wilfordii seedlings.

[0077] In the present invention, geranyl linalool or geranyl linalool emulsion can effectively promote the shape of Notopterygium wilfordii seedling roots and effectively improve the absorption capacity of Notopterygium wilfordii seedling roots.

[0078] In the present invention, the circulation and utilization of soil nitrogen are improved, the content of soil autotoxic substances is reduced, and it is beneficial to the growth of Notopterygium wilfordii seedlings and the breeding of superior-shaped seedlings.

[0079] In the present invention, geranyl linalool or geranyl linalool emulsion can inhibit pathogens related to root rot, damping-off and anthracnose, and reduce diseases of Notopterygium wilfordii seedlings.

[0080] The present invention also provides a method for inhibiting the growth of lateral roots of Notopterygium wilfordii and / or breeding superior Notopterygium wilfordii shape and / or high-quality seedlings, comprising the following steps: applying the geranyl linalool emulsion described in the above scheme during the Notopterygium wilfordii seedling stage.

[0081] As an embodiment, the applying includes spraying and root irrigation.

[0082] As an embodiment, the method is implemented in fields in Gansu Province, where organic matter, nitrogen, and phosphorus are deficient and potassium is abundant, which is conducive to root growth and lateral root formation. The method of the present invention improves rhizosphere nutrient conditions and reduces lateral root formation by regulating endogenous physiological conditions.

[0083] As an embodiment, the application time is after the Notopterygium wilfordii seedlings emerge; the application frequency is once every 21 to 28 days; the total number of applications is 4 times; and the application amount for each application is 100 to 300 L / mu.

[0084] The present invention has no special limitation on the planting and management process of Notopterygium wilfordii in the method, and it can be carried out according to conventional planting and management methods in the field.

[0085] In order to further illustrate the present invention, the application of linalool provided by the present invention in inhibiting the lateral root growth of Notopterygium wilfordii seedlings and / or breeding of superior-shaped and high-quality Notopterygium wilfordii seedlings is described in detail below with reference to the accompanying drawings and examples, but they should not be understood as limiting the scope of protection of the present invention.

[0086] Geranyl linalool was purchased from Shanghai Yuanye, CAS number: 1113-21-9.

[0087] The test Notopterygium wilfordii seeds were stored in sand at low temperatures to break dormancy. The research team then propagated the seeds at the Gansu Buyun Agriculture and Animal Husbandry Technology Co., Ltd.'s Traditional Chinese Medicine Planting Base in Dangchang County (2,600 meters above sea level). The soil type is millet-calcium with a pH of 8.0. In April 2024, the land was prepared and fertilized with 1,000 kg / mu of organic fertilizer (organic matter ≥ 40%, total nutrients N+P2O5+K2O ≥ 4%). Planting was then carried out, and foliar spraying and root irrigation were carried out after the seedlings emerged.

[0088] All reagents used in the experiments were of chemically pure quality. Glutamine synthetase, glutamate synthetase, nitrite reductase, nitrate reductase, glycolate oxidase, ribulose diphosphate carboxylase, sucrose synthase, soil alkaline xylanase, soil polyphenol oxidase, soil sucrase, soil alkaline phosphatase, soil β-glucosidase, and soil urease activity assay kits, as well as assay kits for ABTS, DPPH, superoxide anion, malondialdehyde, soluble sugar, chlorophyll, carotenoids, flavonoids, nitrite, nitrate, sucrose, total phenolics, and plant tissue ammonium and nitrate nitrogen contents, were purchased from Beijing Box Biotechnology Co., Ltd.

[0089] Example 1

[0090] Preparation of geranyl linalool emulsion:

[0091] Pipette 1 mL of geranyl linalool and 1 mL of Tween 80 respectively, add them to 50 mL of water, shake well, and then dilute with water to 1 L.

[0092] Example 2

[0093] Preparation of geranyl linalool emulsion:

[0094] Separately, take 2 mL of geranyl linalool and 1 mL of Tween 80, add them to 50 mL of water, shake well, and then dilute with water to 1 L.

[0095] Example 3

[0096] Preparation of geranyl linalool emulsion:

[0097] Separately, take 3 mL of geranyl linalool and 1 mL of Tween 80, add them to 50 mL of water, shake well, and then dilute with water to 1 L.

[0098] Comparative Example 1

[0099] Preparation of geranyl linalool emulsion:

[0100] Separately, take 0.5 mL of geranyl linalool and 1 mL of Tween 80, add them to 50 mL of water, shake well, and then dilute with water to 1 L.

[0101] Comparative Example 2

[0102] Preparation of geranyl linalool emulsion:

[0103] Separately, take 4 mL of geranyl linalool and 1 mL of Tween 80, add them to 50 mL of water, shake well, and then dilute with water to 1 L.

[0104] Comparative Example 3

[0105] Preparation of geranyl linalool emulsion:

[0106] Separately, take 5 mL of geranyl linalool and 1 mL of Tween 80, add them to 50 mL of water, shake well, and then dilute with water to 1 L.

[0107] Test Example 1

[0108] Functional testing of geranyl linalool emulsion, steps are as follows:

[0109] 1) Plate antagonism experiment: The antibacterial activity was determined using the inverted plate method. The bacterial cakes of Fusarium solani and Alternaria solanacearum were inoculated in the center of the PDA plate, respectively. Sterile drug-sensitive paper sheets moistened with 0, 0.5, 1, 2, 3, 4, and 5 mL / L geranyl linalool were affixed to the center of the PDA plate. The two plates were buckled together, sealed with sealing film, and placed in an incubator at 28°C for 7 days. The growth of the pathogenic fungi was observed, and the antibacterial ability was determined based on the colony size. The relative inhibition rate was calculated. Relative inhibition rate = (colony diameter of the control - colony diameter of the treated) / colony diameter of the control. The results are shown in Table 1.

[0110] Table 1 Antibacterial experiments of different concentrations of geranyl linalool

[0111] 0mL / L 0.5mL / L 1mL / L 2mL / L 3mL / L 4mL / L 5mL / L CK colony diameter (cm) 10 10 10 10 10 10 10 Colony diameter of treatment group (cm) 0 9.5 8 6.2 4.6 4.3 4.1 Relative inhibition rate (%) 0 5 20 38 54 57 59

[0112] The antibacterial rate showed an increasing trend with the increase of the concentration of linalool emulsion. There was no significant difference among 3, 4, and 5 mL / L. Considering the cost, 1-3 mL / L was the effective concentration.

[0113] 2) Corn seed germination experiment: 100 corn seeds were treated three times. The seeds were soaked in 0.1% sodium hypochlorite for 10 minutes, rinsed with clean water, and soaked in 0, 0.5, 1, 2, 3, 4, and 5 mL / L geranyl linalool for 24 hours. The soaked seeds were placed in a moisturizing petri dish at room temperature to germinate. The average germination rate, number of roots, and average root length of the seeds were counted, and the germination index (GI) was calculated (GI = (average germination rate of treatment × average root length of treatment) / (average germination rate of control × average root length of control)). The results are shown in Table 2.

[0114] Table 2 Corn germination test with different concentrations of linalool

[0115] Germination rate (%) Average root length (cm) Number of roots (bars) Germination Index (GI) CK 86.36 11.11±2.23d 8.29±0.76b 0.5mL / L 90.91 12.38±3.06d 6.33±1.03d 1.17 1mL / L 90.91 14.58±1.91bc 8.00±1.10bc 1.38 2mL / L 90.91 19.95±3.60a 6.67±0.82d 1.89 3mL / L 95.45 20.26±4.64a 7.00±0.63cd 2.02 4mL / L 95.45 15.43±3.34b 9.75±1.49a 1.54 5mL / L 86.36 12.74±3.46cd 9.80±0.84a 1.15

[0116] High concentrations of 4 and 5 mL / L linalool emulsions showed an inhibitory effect on the germination index and a promoting effect on the root number; at 0.5-3 mL / L, as the concentration of linalool emulsion increased, the germination index increased, and the comprehensive relative inhibition rate was 1-3 mL / L as the effective concentration, and 3 mL / L was the optimal treatment.

[0117] Test Example 2

[0118] Effects of geranyllinalool emulsion treatment on root morphology and function of Notopterygium wilfordii seedlings

[0119] Set up the test group as follows:

[0120] Treatment group T: 3 mL of geranyl linalool was added to 50 mL of water, followed by 1 mL of Tween 80, which was then placed in the mixture of geranyl linalool and water. The mixture was shaken thoroughly to form a stable oil-water mixture. The mixture was then diluted to 1 L with water and sprayed on leaves and roots. The application rate was 200 L per mu, for a total of 4 applications, with an interval of 28 days.

[0121] Control group CK: 1mL Tween 80 was placed in 1L water, shaken thoroughly, sprayed on leaves and irrigated on roots, with a dosage of 200L per mu, including 20-30L for foliage spray and 170-180L for root irrigation, for a total of 4 applications, with an interval of 28 days between two adjacent applications.

[0122] The experiment adopted a single-factor completely randomized design, with two treatments, T and CK, and three replicates per treatment. The plot area was 30 m 2 Dormancy-breaking Notopterygium wilfordii seeds were sown in early October 2023 and treated four times from May to September 2024. Three days after the last treatment, 100 plants were randomly selected from each plot, Notopterygium wilfordii leaves were picked, the corresponding rhizosphere soil was collected, and the corresponding Notopterygium wilfordii rhizomes were excavated. After washing with running water, they were mixed and brought back to the laboratory for later use.

[0123] The nitrate and ammonium nitrogen contents of Notopterygium root rhizomes were determined spectrophotometrically using the kit's instructions. A root scanner was used with WinRHIZO software to scan total root length, total root surface area, total root volume, average root diameter, number of branches, and number of intersections. The roots were then shade-dried and weighed. Specific root length and root tissue density were calculated using the following formulas: Specific root length = total root length / dry weight; root tissue density = dry weight / total root volume; FC is the ratio of the corresponding data.

[0124] Table 3 Effects of linalool emulsion treatment on Notopterygium wilfordii seedlings

[0125] CK T log2FC(T / CK) Average root diameter (cm) 1.560±0.470a 1.317±0.320b -0.24 Specific root length (cm / g) 75.29±19.36b 91.24±16.02a 0.28 <![CDATA[Root tissue density (g / cm 3 )]]> 0.94±0.34 1.03±0.31 - Nitrate nitrogen (ug / g) 72.58±6.25b 107.27±13.06a 0.56 Ammonium nitrogen (ug / g) 22.78±2.55a 13.89±4.19b -0.71 Nitrate / Ammonium 3.19b 7.72a 1.28 Number of branches 433.84±274.67a 273.74±87.24b -0.67 Crossover number (times) 23(9.75,50.25) 20(11,30.25) -

[0126] Statistical analysis was performed using the t test. Different lowercase letters indicate multiple comparisons (P < 0.05).

[0127] Plant root systems are divided into "cooperative dimensions" and "conservative dimensions." The cooperative dimension explains the nutrient acquisition pattern of the plant-fungus symbiosis through root diameter and specific root length. Thicker roots rely on the symbiosis with fungi for nutrient acquisition, while longer roots acquire nutrients more autonomously. The conservative dimension reflects the balance between nutrient acquisition and storage in the root system through root tissue density and root nitrogen content. Roots with higher root tissue density tend to retain more nutrients, while roots with higher root nitrogen content exhibit a higher nutrient absorption capacity. Table 3 shows that compared with the control group CK, the group T treated with the geranyl linalool emulsion of the present invention significantly increased specific root length, nitrate nitrogen content, and nitrate / ammonium content, particularly the nitrate / ammonium ratio (log2FC) of 1.28. Average root diameter, ammonium nitrogen content, and branch number were significantly reduced. This indicates that Notopterygium incisum prefers nitrate nitrogen, while ammonium nitrogen may be toxic to it. Treatment with the geranyl linalool emulsion T of the present invention effectively promoted root morphology and significantly improved root absorption capacity.

[0128] Test Example 3

[0129] Effects of geranyllinalool emulsion treatment on endogenous hormone levels and physiological and biochemical characteristics of Notopterygium wilfordii seedlings

[0130] The experimental grouping and experimental plan are the same as those in Experimental Example 2.

[0131] The leaves were sent to a third-party testing company, and the contents of 34 related substances in nine major hormone categories, including auxin, cytokinin, gibberellin, abscisic acid, ethylene, salicylic acid, jasmonic acid, melatonin, and strigolactone, were detected by LC-MS. The most important substances (VIP>1) that effectively distinguished the differences between the T and CK groups were screened by OPLS-DA (a multivariate statistical analysis for supervised pattern recognition). The FC values ​​were used to compare the degree of difference between different substances in different groups, see Table 4.

[0132] Table 4 Endogenous hormones of Notopterygium wilfordii seedlings

[0133] VIP FDR CK T log2FC(T / CK) OPDA (ng / g) 1.003 0.000 333.243±11.790a 22.674±0.326b -3.88 GA7 (ng / g) 1.002 0.000 0.977±0.018a 0.258±0.007b -1.92 SA (ng / g) 1.002 0.000 10.242±0.407a 2.974±0.019b -1.78 JA (ng / g) 1.002 0.000 9.811±0.329a 1.993±0.069b -2.30 GA19 (ng / g) 1.002 0.001 1.000±0.000a 0.349±0.016b -1.52 SAG (ng / g) 1.001 0.000 829.959±23.109a 368.511±9.751b -1.17 JA-Ile(ng / g) 1.001 0.000 3.133±0.156a 1.007±0.012b -1.64 IAA (ng / g) 1.000 0.001 0.297±0.025b 1.012±0.010a 1.77 ABA (ng / g) 1.001 0.000 2.503±0.100a 1.120±0.054b -1.16 ACC (ng / g) 1.001 0.001 1454.357±136.797a 559.202±17.681b -1.38

[0134] Statistical analysis was performed using the t test. Different lowercase letters indicate multiple comparisons (P < 0.05).

[0135] Among the 34 substances tested, 10 had an FDR of less than 0.05 and a VIP value greater than 1: 12-oxophytodienoic acid (OPDA), gibberellin 7 (GA7), salicylic acid (SA), jasmonic acid (JA), gibberellin 19 (GA7), salicylic acid glucoside (SAG), jasmonic acid-isoleucine (JA-Ile), indoleacetic acid (IAA), abscisic acid (ABA), and 1-aminocyclopropane-1-carboxylic acid (ACC). Compared with the control group CK, the treatment group T of the present invention showed a significant increase in auxin content, while the contents of gibberellin 7, gibberellin 9, salicylic acid, salicylic acid glucoside, jasmonic acid, jasmonic acid-isoleucine, abscisic acid, and 1-aminocyclopropane-1-carboxylic acid were significantly decreased. In particular, the absolute values ​​of jasmonic acid substances and ACC were significantly decreased. The results show that the linalool emulsion described herein does not activate plant immunity; at relatively low concentrations, it increases auxin content, promoting root growth; at relatively high concentrations, it reduces jasmonic acid and ethylene content, helping to inhibit lateral root formation. Treatment with the geranyl linalool emulsion T effectively promotes root growth and shape.

[0136] Test Example 4

[0137] Effects of geranyllinalool emulsion treatment on physiological and biochemical parameters of Notopterygium wilfordii seedlings

[0138] The experimental grouping and experimental plan are the same as those in Experimental Example 2.

[0139] The activities of glutamine synthetase, glutamate synthetase, nitrite reductase, nitrate reductase, glycolate oxidase, ribulose-bisphosphate carboxylase, and peroxidase, as well as the contents of ABTS, total phenolics, flavonoids, ammonium nitrogen, nitrate nitrogen, amino nitrogen, soluble sugars, chlorophyll, sucrose, and malondialdehyde in the leaves of Notopterygium incisum seedlings were measured spectrophotometrically using the kit's instructions. OPLS-DA (a supervised pattern recognition multivariate statistical analysis) was used to identify the most important substances (VIP > 1) that effectively distinguished the T and CK groups. FC values ​​were used to compare the degree of difference between the different groups for each substance (see Table 5).

[0140] Table 5 Effects of linalool emulsion treatment on physiological and biochemical indices of Notopterygium wilfordii seedlings

[0141]

[0142] Statistical analysis was performed using the t test. Different lowercase letters indicate multiple comparisons (P < 0.05).

[0143] Among the aforementioned enzyme activities and substances, substances with FDR values ​​less than 0.05 and VIP values ​​greater than 1 included: glutamine synthetase, total chlorophyll, DPPH, and nitrate reductase, a total of four. Compared to the control group (CK), treatment with the linalool emulsion T of the present invention significantly increased glutamine synthetase and nitrate reductase activities, as well as total chlorophyll and DPPH content. This indicates that the geranyl linalool emulsion of the present invention significantly improves nitrogen supply and utilization, as well as antioxidant capacity, in Notopterygium wilfordii.

[0144] Test Example 4

[0145] The experimental grouping and experimental scheme for the effect of geranyl linalool emulsion on available nutrients, enzyme activity and autotoxic substances in the soil of Notopterygium wilfordii seedlings were the same as those in Experimental Example 2.

[0146] Soil polyphenol oxidase, sucrase, urease, β-glucosidase, xylanase, alkaline phosphatase, and xylanase activities were measured spectrophotometrically as described in the kit. Alkaline-hydrolyzed nitrogen was measured by alkaline diffusion. Available potassium was determined by flame photometry using ammonium acetate extraction. Total iron was determined by four-acid digestion-inductively coupled plasma spectrometry. Available phosphorus was determined by sodium bicarbonate extraction and molybdenum antimony colorimetry. Total phenolic acids were determined by UV-visible spectrophotometry as described in the literature (Yang Lan, Food Science, 2011). OPLS-DA (a supervised pattern recognition multivariate statistical analysis) was used to identify the most important substances (VIP>1) that effectively distinguished the T and CK groups. FC values ​​were used to compare the degree of difference between the different groups (see Table 6).

[0147] Table 6 Effects of linalool emulsion on soil nutrients, autotoxic substances content and enzyme activity

[0148]

[0149] Statistical analysis was performed using the t test. Different lowercase letters indicate multiple comparisons (P < 0.05).

[0150] Among the aforementioned enzyme activities and substances, five exhibited FDR values ​​less than 0.05 and VIP values ​​greater than 1: total phenolic acids, soil polyphenol oxidase, alkaline nitrogen, soil β-glucosidase, and urease. Compared to the control group (CK), treatment with the geranyl linalool emulsion T significantly increased soil polyphenol oxidase, β-glucosidase, and urease activities, as well as alkaline nitrogen content, and reduced total phenolic acid content. This demonstrates that the geranyl linalool emulsion described herein can enhance soil nitrogen recycling and utilization, reduce soil autotoxicants, and promote seedling growth and the development of superior seedlings.

[0151] Test Example 5

[0152] Effects of different treatments on the diversity and function of microorganisms in the rhizosphere soil of Notopterygium wilfordii seedlings

[0153] The experimental grouping and experimental plan are the same as those in Experimental Example 2.

[0154] Samples with different treatments were sent to a third-party sequencing company for microbial metagenomic analysis. α-diversity analysis, species-level microbial differential analysis, and functional prediction analysis were performed.

[0155] Alpha diversity analysis

[0156] The Chao1 and Ace indices measure species richness, or the number of species. Higher Ace and Chao1 indices indicate a greater diversity of microorganisms. The Shannon and Simpson indices measure species diversity. Higher Shannon and lower Simpson indices indicate a higher diversity of species in the sample. The microbial alpha diversity of samples treated with the inoculum described herein (T) and the control (CK) is shown in Table 7.

[0157] Table 7 Microbial α diversity

[0158]

[0159]

[0160] Different lowercase letters indicate multiple comparisons (P < 0.05)

[0161] As shown in Table 7, compared with the control group CK, the bacterial Simpson index of the geranyl linalool emulsifier T treated with the present invention was significantly reduced, indicating that the geranyl linalool emulsifier T treated with the present invention increased the bacterial diversity of the rhizosphere soil. The fungal Chao1 index and Ace index decreased significantly, indicating that the geranyl linalool emulsifier T treated with the present invention reduced the number of fungal species in the rhizosphere soil.

[0162] Bacterial genus-level LEfSe analysis

[0163] LEfSe (Line Discriminant Analysis (LDA) Effect Size) analysis is an analytical method that combines the non-parametric Kruskal-Wallis and Wilcoxon rank sum tests with the effect size of linear discriminant analysis (LDA). It can find biomarkers with statistical differences between different groups. Figure 1(LDA threshold is 3). There are 46 types of characteristic microorganisms in the rhizosphere soil of T treated with the geranyl linalool emulsifier of the present invention, and 15 genera are annotated: Microbacterium, Micromonospora, Rhodococcus, Marmoricola, Nocardioides, Streptosporangium, Conexibacter, Bacillus, Peribacillus, Priestia, Microvirga, Afipia, Sphingococcus, and Pseudomonas aeruginosa. gobium), Herbaspirilum, and Sphaerobacter; the positive control CK treatment had 25 classes of characteristic bacterial microorganisms in the rhizosphere soil, 10 of which were annotated to the genera: Streptomyces, Sphingobacterium, Scytonema, Neorhizobium, Rhizobium, Novosphingobium, Burkholderia, Cupriavidus, Paraburkholderia, and Variovorax. This showed that although the geranyl linalool emulsifier treatment of T did not activate plant immunity, it promoted the enrichment of Micromonospora, Neurospora, and Bacillus, thereby improving the disease resistance of seedlings.

[0164] Based on the KEGG Level2 functional annotation of rhizospheric soil bacteria, LEfSe analyzed the biomarkers with statistical differences in function among different groups, such as Figure 2 .

[0165] The hallmark functions of T treated with the linalool emulsion described herein were energy metabolism, other amino acid metabolism, excretion, and carbohydrate metabolism; while the hallmark functions of the control group, CK, were the immune system, chemical structure transformation profile, cell motility, digestive system, membrane transport, cofactor and vitamin metabolism, environmental adaptation, and nucleotide metabolism. This indicates that treatment of T with the linalool emulsion described herein significantly enhanced active carbon and nitrogen metabolism, promoting seedling growth and the propagation of superior-shaped seedlings.

[0166] The linalool emulsion treatment T and the control group CK were subjected to a student t-test based on the species-level fungal abundance, and the FC values ​​were calculated, as shown in Table 8.

[0167] Table 8 Fungi with significant differences in levels

[0168]

[0169] Statistical analysis was performed using the t test. Different lowercase letters indicate multiple comparisons (P < 0.05).

[0170] Compared with the control group (CK), the linalool emulsion-treated plant T significantly reduced the abundance of the pathogens Colletotrichum higginsii, Alternaria alternata, Fusarium oxysporum, Fusarium aromaticum, Colletotrichum frutescens, and Alternaria dendriticum. This suggests that while the linalool emulsion-treated plant T did not activate plant immunity, it inhibited pathogens associated with root rot, damping-off, and anthracnose, reducing seedling disease.

[0171] Although the above embodiment provides a detailed description of the present invention, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the scope of protection of the present invention.

Claims

1. The application of geranyl linalool in planting Notopterygium wilfordii seedlings is characterized in that, The planting of Notopterygium wilfordii seedlings includes at least one of the following: 1) Inhibit the growth of lateral roots of Notopterygium wilfordii seedlings; 2) Propagation of superior-shaped and / or high-quality seedlings of Notopterygium wilfordii; 3) Increase the root length of Notopterygium wilfordii seedlings; 4) increasing the nitrogen content of Notopterygium wilfordii seedling roots and / or increasing the absorption capacity of Notopterygium wilfordii seedling roots; 5) Promote the growth of Notopterygium wilfordii seedlings; 6) reducing the abundance of pathogens in Notopterygium incisum seedlings and / or increasing the abundance of probiotics in Notopterygium incisum seedlings; 7) Improve disease resistance of Notopterygium wilfordii seedlings; 8) improving the recycling and utilization of nitrogen in the soil where Notopterygium wilfordii seedlings are planted and / or reducing the content of autotoxic substances in the soil where Notopterygium wilfordii seedlings are planted; 9) Reduce the number of fungal species in the rhizosphere soil of Notopterygium wilfordii seedlings. The working concentration of the geranyl linalool is 1-3 ml / L.

2. A geranyl linalool emulsion, characterized in that: The invention comprises water and geranyl linalool with a concentration of 1-3 ml / L; the geranyl linalool emulsion uses Tween-80 as an emulsifier.

3. The geranyl linalool emulsion according to claim 2, wherein The concentration of Tween-80 in the geranyl linalool emulsion is 1 ml / L.

4. The use of the geranyl linalool emulsion according to claim 2 or 3 in planting Notopterygium wilfordii seedlings, characterized in that: The planting of Notopterygium wilfordii seedlings includes at least one of the following: 1) Inhibit the growth of lateral roots of Notopterygium wilfordii seedlings; 2) Propagation of superior-shaped and / or high-quality seedlings of Notopterygium wilfordii; 3) Increase the root length of Notopterygium wilfordii seedlings; 4) increasing the nitrogen content of Notopterygium wilfordii seedling roots and / or increasing the absorption capacity of Notopterygium wilfordii seedling roots; 5) Promote the growth of Notopterygium wilfordii seedlings; 6) reducing the abundance of pathogens in Notopterygium incisum seedlings and / or increasing the abundance of probiotics in Notopterygium incisum seedlings; 7) Improve disease resistance of Notopterygium wilfordii seedlings; 8) improving the recycling and utilization of nitrogen in the soil where Notopterygium wilfordii seedlings are planted and / or reducing the content of autotoxic substances in the soil where Notopterygium wilfordii seedlings are planted; 9) Reduce the number of fungal species in the rhizosphere soil of Notopterygium wilfordii seedlings.

5. The use according to claim 1 or 4, characterized in that The Notopterygium wilfordii includes wild Notopterygium wilfordii; the Notopterygium wilfordii includes Silkworm Notopterygium wilfordii.

6. The use according to claim 1 or 4, characterized in that The pathogens include pathogenic fungi and / or pathogenic bacteria; the pathogenic fungi include at least one of Higgins Anthrax, Alternaria alternata, Fusarium oxysporum, Fusarium aromaticum, Colletotrichum frutescens and Alternaria dendriticum.

7. The use according to claim 1 or 4, characterized in that The probiotics include probiotic bacteria and / or probiotic fungi; the probiotic bacteria include at least one of the genus Micromonospora, Neurospora and Bacillus.

8. A method for inhibiting the growth of lateral roots of Notopterygium wilfordii and / or breeding Notopterygium wilfordii seedlings of good shape and / or high quality, characterized in that: The following steps are involved: The geranyl linalool emulsion according to claim 2 or 3 is applied during the Notopterygium wilfordii seedling stage.

9. The method according to claim 8, characterized in that The application includes spraying and root irrigation.

10. The method according to claim 8, characterized in that The application time is after the Notopterygium wilfordii seedlings emerge; the application frequency is once every 21 to 28 days; the total number of applications is 4 times; and the application amount for each application is 100 to 300 L / mu.

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