A method of drip irrigation and fertilizer coupling microbial agent suitable for improving the soil microenvironment of edible rose planting land in Yunnan red soil area and improving quality and yield

Abstract

The application provides a method for improving the soil microenvironment of edible rose and increasing yield by drip irrigation and fertilizer coupling microbial agent. Through the test of applying microbial agent and water-soluble fertilizer of macroelement, the change rule of soil environment and yield and quality of rose under different microbial agent and fertilizer coupling treatment is explored. Four different microbial agents are set in the specific test, which are no microbial agent (CK), bacillus subtilis (B), trichoderma harzianum (T) and compound microbial agent (C); three inorganic fertilizer application levels are set, which are low fertilizer (FD: 505 kg·ha ‑2 ·year ‑1 ), medium fertilizer (FZ: 905 kg·ha ‑2 ·year ‑1 ) and high fertilizer (FG: 1300 kg·ha ‑2 ·year ‑1 ), totally 12 complete combination treatments. It is proposed that the comprehensive income of the method of applying trichoderma harzianum at medium fertilizer level (FZT) is the largest, which can effectively improve the soil environment of edible rose planting area, promote soil health, improve the yield and quality of edible rose, and realize the green and efficient production of edible rose.

Description

Technical Field

[0001] This invention belongs to the field of agricultural planting and relates to a method for drip irrigation and fertilization coupled with microbial agents that is suitable for improving the soil microenvironment and increasing the yield of edible roses in the red soil region of Yunnan. Background Technology

[0002] Roses, belonging to the Rosaceae family, are widely cultivated due to their high nutritional and economic value. Edible roses are extensively grown in Yunnan Province, China. As of 2020, Yunnan Province had 193,000 mu (approximately 12,533 hectares) of edible flower cultivation area, with edible roses accounting for the largest share at 95,000 mu (approximately 6,333 hectares), making it a major supplier of raw materials for edible rose processing nationwide. However, many challenging problems remain in the cultivation of edible roses in Yunnan. These include a lack of scientific cultivation methods to guide production, and farmers' pursuit of maximum economic benefits through excessive and uncontrolled application of chemical fertilizers. This leads to continuous deterioration of the soil microenvironment, reduced fertility and quality, decreased organic matter content, soil compaction, secondary salinization, and nutrient imbalances. Ultimately, this results in slow growth, declining development, and continuously deteriorating yield and quality of edible roses in Yunnan's red soil regions.

[0003] Therefore, there is an urgent need to develop a green cultivation model and planting method that can both improve the soil microenvironment of edible rose planting areas in the red soil region of Yunnan and promote the quality and yield of edible roses. Summary of the Invention

[0004] Currently, the water and fertilizer requirements of edible rose cultivation are not fully understood. In pursuit of maximum economic benefits, excessive or unbalanced application of chemical fertilizers leads to the deterioration of soil microbial flora, reduced fertility and quality, ultimately resulting in slow plant growth, decreased vigor, and inferior quality. To address these issues, we propose combining microbial inoculants with inorganic fertilizers in the cultivation of edible roses, exploring a drip irrigation and fertilization method coupled with microbial inoculants to improve the soil microenvironment and enhance yield and quality in edible rose cultivation.

[0005] To achieve the above-mentioned objectives, this application provides the following technical solution:

[0006] First aspect:

[0007] 1. A method for drip irrigation and fertilization coupled with microbial agents suitable for improving the soil microenvironment and increasing yield of edible roses in the red soil region of Yunnan, comprising the following steps:

[0008] (1) Select one-year-old edible roses as seedlings, with a spacing of 20×25cm between seedlings;

[0009] (2) Adaptive drip irrigation was used for irrigation. The drip inlet was located 3 cm from the root of the edible rose, the drip water output was 1.5 L / h, and the drip irrigation cycle was 6 days.

[0010] (3) Microbial agents were applied during irrigation, namely Bacillus subtilis (B), Bacillus harzianum (T) and compound microbial agent (C).

[0011] (4) During the rose growth process, four types of microbial inoculant treatments were set up: no microbial inoculant (CK), Bacillus subtilis (B), Trichoderma harzianum (T), and compound inoculant (C); three inorganic fertilizer application levels were set up: low fertilizer (FD: 505 kg·ha). -2 ·year -1 ), medium fertilizer (FZ: 905kg·ha) -2 ·year -1 ) and high fertilizer (FG: 1300 kg·ha) -2 ·year -1 The effective viable count of Bacillus subtilis was 20.53 billion g. -1 The effective viable count of Trichoderma harzianum is 21.5 billion g. -1 The effective live bacteria count of the compound microbial agent is 7.5 billion g. -1 .

[0012] Furthermore, the drip irrigation fertilization and microbial agent method for improving the soil microenvironment and increasing the quality and yield of edible roses as described in (2) is characterized in that a 7cm deep circular trench is dug at a distance of 6cm from each edible rose seedling, and then the microbial agent is poured on at 5-6 pm, and the soil is covered after even irrigation, with the soil covering height 2cm higher than the root zone of the plant.

[0013] Further, as described in (3), the Bacillus subtilis is diluted with water at a ratio of 1:1200, added to warm water at 35.6 degrees Celsius to dissolve, and after it is fully dissolved, a mixture of glucose and brown sugar (the ratio of glucose to brown sugar is 1.5:1) is added again, and the mixture is left to stand in the dark for 2 to 4 hours until it is fully mixed.

[0014] Further, as described in (3), the characteristic is that the water ratio of Trichoderma harzianum is 1:1500, which is added to warm water at 37.5 degrees Celsius to dissolve. After it is fully dissolved, a mixture of glucose and brown sugar (the ratio of glucose to brown sugar is 2:1) is added again, and the mixture is left to stand in the dark for 1.5 hours. During this period, it is stirred 2-3 times until it is fully mixed.

[0015] Further, as described in (3), the compound microbial agent is characterized by a water dilution ratio of 1:1000, a water temperature of 33 degrees Celsius, and after it is fully dissolved, a mixture of glucose and sucrose (glucose to sucrose ratio of 3:1) is added again, and the mixture is left to stand in the dark for 2 hours until it is fully mixed.

[0016] The second aspect:

[0017] This invention also provides a method for improving the soil microenvironment and increasing the yield of edible roses through drip irrigation and fertilization coupled with microbial agents. This method is used to increase rose yield and improve the content of active ingredients in roses. Its key feature is that by applying the method described above to cultivate edible roses, the activity of the roses can be improved. Compared with not applying the microbial agent, applying the microbial agent can improve the content of total polyphenols, total flavonoids, soluble sugars, and anthocyanins in rose petals. Simultaneously, compared with low fertilizer (FD: 505 kg·ha)... -2 ·year -1 ) and high fertilizer (FG: 1300 kg·ha) -2 ·year -1 ), using medium fertilizer (FZ: 905 kg·ha) -2 ·year -1 The amount of fertilizer applied is also more conducive to increasing the content of total polyphenols, total flavonoids, soluble sugars and anthocyanins in rose petals.

[0018] The beneficial effects of this invention are:

[0019] (1) This invention provides a cultivation method for edible roses suitable for the red soil region of Yunnan. Through the scientific application of inorganic fertilizers, the soil environment deterioration caused by the excessive application of chemical fertilizers in the current rose planting area can be changed. At the same time, by combining different microbial agents, the life activities of microorganisms can be used to improve the soil microenvironment, repair the soil degradation area, and ultimately achieve the goal of promoting the sustainable development of edible rose planting land in Yunnan.

[0020] (2) This invention achieves the effect of increasing the yield of edible roses in the red soil area of ​​Yunnan by innovating and optimizing the application methods of three different microbial agents and combining them with the scientific application amount of inorganic fertilizer, thereby improving the economic benefits of growers.

[0021] (3) This invention combines microbial agents with inorganic fertilizer drip irrigation. On the one hand, microbial agents can be used to improve the soil and promote the growth of edible roses. On the other hand, the advantages of drip irrigation in saving water and fertilizer can be fully utilized. The organic combination of the two can improve the quality of edible roses (increase the content of total polyphenols, total flavonoids, soluble sugars and anthocyanins), improve the soil, and ultimately achieve the effect of improving quality and increasing yield. Attached Figure Description

[0022] To more clearly illustrate the technical solutions of the embodiments of this application, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the accompanying drawings illustrate the embodiments of this application and should therefore be regarded as a limitation of the scope. However, for those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0023] Figure 1 This is a technical roadmap for Example 1;

[0024] Figure 2 This is a cultivation and management layout diagram for Example 1;

[0025] Figure 3 A map showing soil environmental indicators for edible roses;

[0026] Figure 4 A graph showing the yield of edible roses;

[0027] Figure 5 This is a quality image of edible roses. Detailed Implementation

[0028] Example 1:

[0029] A method for drip irrigation and fertilization coupled with microbial agents, suitable for improving the soil microenvironment and increasing the yield of edible roses in the red soil region of Yunnan, includes the following steps:

[0030] (1) Select one-year-old edible roses as seedlings, with a spacing of 20×25cm between seedlings;

[0031] (2) Irrigation is carried out by drip irrigation. The drip inlet is located 3cm from the root of the edible rose, the drip irrigation flow rate is 1.5L / h, and the drip irrigation cycle is 6 days. A 7cm deep circular trench is dug 6cm away from each edible rose seedling with a small shovel. Then, the fungicide is applied at 5-6 pm. After even irrigation, the soil is covered and the soil height is 2cm higher than the root zone of the plant.

[0032] (3) Microbial agents were applied during irrigation, namely Bacillus subtilis (B), Trichoderma harzianum (T), and a compound microbial agent (C). Bacillus subtilis was diluted with water at a ratio of 1:1200 and dissolved in 35.6°C warm water. After complete dissolution, a mixture of glucose and brown sugar (1.5:1 ratio) was added, and the mixture was left to stand in the dark for 2-4 hours until fully mixed. Trichoderma harzianum was diluted with water at a ratio of 1:1500 and dissolved in 37.5°C warm water. After complete dissolution, a mixture of glucose and brown sugar (2:1 ratio) was added, and the mixture was left to stand in the dark for 1.5 hours, stirring thoroughly 2-3 times during this period until fully mixed. When applying the compound microbial agent, the dilution ratio with water is 1:1000, and the water temperature is 33 degrees Celsius. After it is fully dissolved, add the mixture of glucose and sucrose (the ratio of glucose to sucrose is 3:1), and let it stand in the dark for 2 hours until it is fully mixed.

[0033] (4) During the rose growth process, four types of microbial inoculant treatments were set up: no microbial inoculant (CK), Bacillus subtilis (B), Trichoderma harzianum (T), and compound inoculant (C); three inorganic fertilizer application levels were set up: low fertilizer (FD: 505 kg·ha). -2 ·year -1 ), medium fertilizer (FZ: 905kg·ha) -2 ·year -1 ) and high fertilizer (FG: 1300 kg·ha) -2 ·year -1 The number of viable Bacillus subtilis spores was 20.53 billion g. -1 The effective viable count of Trichoderma harzianum is 21.5 billion g. -1 The effective live bacteria count of the compound microbial agent is 7.5 billion g. -1 .

[0034] Example 1

[0035] To compare the advantages of this invention in edible rose cultivation technology, we conducted targeted experiments according to the following methods to verify the practical effects of this invention.

[0036] A method for drip irrigation and fertilization coupled with microbial agents, suitable for improving the soil microenvironment and increasing the yield of edible roses in the red soil region of Yunnan, specifically includes the following steps:

[0037] (1) Application target: Select one-year-old edible roses as seedlings, with a spacing of 20×25cm between seedlings;

[0038] (2) The specific steps of the method are as follows:

[0039] The verification experiment was conducted from July 2021 to November 2023 in a plastic greenhouse at the College of Modern Agricultural Engineering, Kunming University of Science and Technology, Yunnan Province, in southwestern China (24°84′N, 102°86′E, altitude 1978.9m). In the experimental area for edible rose cultivation, the average annual temperature was maintained at 23.5℃, and the relative humidity was 35-66%.

[0040] This experiment simultaneously set up three adaptive drip irrigation fertilization rates and four different microbial inoculant application rates for comparison. The three adaptive drip irrigation fertilization rates were low fertilizer (FD: 505 kg·ha). -2 ·year -1 ), medium fertilizer (FZ: 905kg·ha) -2 ·year -1 ) and high fertilizer (FG: 1300 kg·ha) -2 ·year -1The amount of water irrigated was then controlled by a water meter, with an irrigation rate of 1.5 L / h during the experiment. Drip irrigation fertilization (equal amounts applied each time) was conducted on July 11, 2021, September 13, 2021, November 19, 2021, March 15, 2022, May 17, 2022, July 19, 2022, September 23, 2022, November 19, 2022, March 15, 2023, May 23, 2023, July 3, 2023, and September 15, 2023. Four microbial agents were used: a sterile agent (CK), Bacillus subtilis (B), Trichoderma harzianum (T), and a compound agent (C). The design consisted of 12 treatments: FDCK, FDB, FDT, FDC, FZCK, FZB, FZT, FZC, FGCK, FGB, FGT, and FGC, with each treatment replicated three times. The types of microbial inoculants and water-soluble fertilizers used are shown in Table 1.

[0041] Table 1. Types of Microbial Inoculants and Water-Soluble Fertilizers

[0042]

[0043] The methods for determining the indicators in this verification experiment are as follows:

[0044] (1) Methods for determining soil fertility: Nitrate nitrogen in the soil was extracted with KCl solution and then determined by spectrophotometry; available phosphorus was extracted with NaHCO3 solution and determined by molybdenum antimony colorimetric method; available potassium was extracted with NaH4Ac and then determined by flame photometry.

[0045] (2) Methods for determining soil enzyme activity: Catalase activity was determined by potassium permanganate titration; urease activity was determined by sodium phenolate-sodium hypochlorite colorimetric method; and phosphatase activity was determined by disodium phenyl phosphate colorimetric method.

[0046] (3) Edible roses were harvested in 2022 and 2023, and the yield of edible roses (t·ha) was determined for two consecutive years using a weighing method. -2 When calculating yield, the actual planting area is used. The specific procedure for yield measurement is as follows: Half-open roses are picked, meaning the rose is bowl-shaped with the outer 3-4 layers of petals open and the remaining petals closed, and the stamens not yet exposed. The fresh weight of the rose is then measured. The fresh flowers are then vacuum freeze-dried (using an LGJ-12 freeze dryer), with the vacuum level set to 7-10 kPa. The refrigeration is then turned on, and the freezing temperature is adjusted to -65°C and maintained for 24 hours. Finally, the dried flowers are removed, weighed, and the moisture content is calculated. After freezing, the flowers are packaged into different sealed bags according to different treatment methods, ground into powder, and ready for measurement.

[0047] (4) The method for determining total flavonoids is as follows: Accurately weigh 0.20 g of freeze-dried rose powder and place it in a 10 ml Erlenmeyer flask. Then add 5.5 ml of 73% ethanol solution, seal the flask with plastic wrap, and perform constant temperature ultrasonic-assisted extraction for 39 min under the conditions of ultrasonic power 413 W and temperature 53 °C. After the extraction process, remove the Erlenmeyer flask and allow it to cool naturally to room temperature. Then centrifuge and filter the extract. Next, use a pipette to add 1.0 ml of the extract to a 10 ml volumetric flask, add various reagents in sequence, and after color development, use a UV spectrophotometer to measure the absorbance at a wavelength of 510 nm. Calculate the total flavonoid content based on the measured absorbance.

[0048] (5) The method for determining total polyphenols is referred to "T / AHFIA005-2018 Determination of total polyphenol content in plant extracts and their products - spectrophotometry".

[0049] (6) Total antioxidant capacity was determined using a kit.

[0050] (7) The method for determining soluble sugars is in accordance with GB / T36056-2018, and is performed using liquid chromatography.

[0051] (8) The anthocyanin content in roses was determined by ultraviolet spectrophotometry, and the citrate-disodium hydrogen phosphate buffer method was used for comparison and verification.

[0052] The final results of the soil environmental indicators show that, in terms of improving soil nitrogen, phosphorus, and potassium, the compound microbial agent treatment resulted in higher nitrate nitrogen content. However, *Trichoderma harzianum* was the most effective in promoting the increase of available phosphorus content. The soil with *Trichoderma harzianum* applied also had higher available potassium content, increasing by 52.84% and 24.91% respectively compared to the control (CK). The application of microbial agents also increased the activities of urease, phosphatase, and catalase in the soil, with *Trichoderma harzianum* showing the best promoting effect on the activities of these three enzymes, increasing them by 26.01%, 31.25%, and 14.32% respectively.

[0053] The final yield results show that, under the FD treatment, compared with no inoculant application, the yield of roses treated with Bacillus subtilis and the compound inoculant increased by 13.08% and 31.05%, respectively. Under the FZ treatment, the total annual yield of roses treated with Trichoderma harzianum was higher than that of the treatments treated with Bacillus subtilis and the compound inoculant, reaching its maximum value. The total yield of roses treated with the compound inoculant was higher than that of roses treated with Bacillus subtilis. Under the FG treatment, the yield of roses treated with Trichoderma harzianum was the highest, followed by the yield of roses treated with the compound inoculant. The yield of roses varied significantly under different fertilization amounts. Under the Trichoderma harzianum treatment, the yield of edible roses gradually decreased with increasing fertilization amount, exhibiting a parabolic change. Overall, under the same inoculant treatment conditions, the maximum yield was achieved at FZ.

[0054] The final quality results show that, under FD treatment, compared with no fungicide application, the total flavonoid and soluble sugar content of roses treated with Bacillus subtilis were higher, increasing by 6.94% and 2.92% respectively. The total polyphenol and anthocyanin content of rose petals treated with Trichoderma harzianum were also higher, increasing by 46.55% and 46.23% respectively. Under FZ treatment, the total flavonoid, total polyphenol, anthocyanin content, and total antioxidant capacity of rose petals treated with Trichoderma harzianum all reached their maximum values. Under FG treatment, the total polyphenol, total flavonoid, and total antioxidant capacity of rose petals treated with Trichoderma harzianum were also higher, but lower than those treated with FZ. Under the same microbial agent treatment, different fertilization amounts had varying effects on rose quality indicators, but overall showed a trend of first increasing and then decreasing. Under Bacillus subtilis treatment, the contents of total polyphenols, total flavonoids, soluble sugars, and anthocyanins gradually decreased with increasing fertilization amount, exhibiting a parabolic change. Under Trichoderma harzianum treatment, the changes in rose petal quality indicators were similar to those observed with Bacillus subtilis application.

[0055] This invention uses the TOPSIS method to evaluate the comprehensive effects of different combinations of microbial agents and water-soluble fertilizers on edible roses. By quantifying the distance between the evaluation objects and indicators, the relative advantages of each strategy are further determined. The TOPSIS comprehensive score and ranking results are shown in Table 2.

[0056] Table 2

[0057]

[0058] As shown in Table 2, considering the final results of soil environmental, yield, and quality indicators, the evaluation results using the entropy weight-TOPSISI method show that the top three treatments, ranked from highest to lowest score, are FZT, FZC, and FDT, with scores of 0.949, 0.766, and 0.715, respectively. This indicates that applying Trichoderma harzianum (FZT) at a moderate fertilization level yields the best results. It has the best effect on improving the soil environment, yield, and quality of edible roses, and is most beneficial for promoting the quality and efficiency of edible roses in Yunnan Province.

[0059] The specific embodiments of the present invention have been described in detail above. The present invention is limited to the above embodiments, but simple changes can be made within the scope of knowledge possessed by those skilled in the art without departing from the spirit of the present invention.

Claims

1. A method for improving the soil microenvironment of the planting site of edible roses in Yunnan red soil areas and improving the quality and yield of the planting site by coupling drip irrigation and microbial inoculants, characterized in that, It comprises the following steps: (1) irrigation is carried out by using self-adaptive drip irrigation method, dripper is located at 3 cm from the root of the edible rose, drip irrigation water output is 1.5 L / h, drip irrigation irrigation period is 6 days; (2) microbial inoculant is applied during irrigation, the inoculant is Trichoderma harzianum; (3) In the process of rose growth, H. zeae and medium fertilizer are set up, and the medium fertilizer is 905 kg·ha -2 ·year -1 ; the effective viable count of H. zeae is 21.5 billion·g -1 .

2. The method for improving the soil microenvironment and quality and yield of edible rose planting in Yunnan red soil area by drip irrigation and fertilizer coupling microbial agent according to claim 1, characterized in that, The application method of step (2) is as follows: a 7 cm deep annular ditch is dug around the edible rose seedling with a plant spacing of 20*25 cm and at a distance of 6 cm from each edible rose seedling by using a self-made fertilizer shovel, then the inoculant is poured at 5-6 pm on time, the soil is covered after uniform irrigation, and the soil covering height is 2 cm higher than the root zone of the plant.

3. The method for improving the soil microenvironment and quality and yield of edible rose planting in Yunnan red soil area by drip irrigation and fertilizer coupling microbial agent according to claim 1, characterized in that, The water mixing ratio of Trichoderma harzianum is 1:1500, it is added to 37.5 degrees Celsius warm water for dissolution, after complete dissolution, a mixed solution of glucose and brown sugar is added again, the ratio of glucose and brown sugar is 2:1, it is placed in dark for 1.5 hours, and stirring is carried out 2-3 times during this period until complete mixing.

4. The method for improving the soil microenvironment and quality and yield of edible rose planting in Yunnan red soil area by drip irrigation and fertilizer coupling microbial agent according to claim 1, characterized in that, It is used for improving the yield of rose flowers and improving the content of active ingredients of rose flowers.

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