New application of exogenous abscisic acid

Abstract

The invention provides novel application of exogenous abscisic acid. The novel application comprises at least one of the following applications: 1) reducing accumulation of antibiotics in vegetables; 2) the antibiotic stress of vegetables is relieved; and 3) reducing antibiotic resistance genes in the vegetables. The antibiotics comprise at least one of tetracyclines and quinolones. The invention also provides a vegetable antibiotic stress alleviator, which comprises exogenous abscisic acid. The exogenous abscisic acid treatment can enhance the antioxidant enzyme activity of the vegetables, improve the capability of removing active oxygen of a vegetable self-protection system, and is beneficial to keeping the balance of the active oxygen, thereby improving the capability of the vegetables for degrading organic pollutants such as antibiotics and the like. The exogenous abscisic acid treatment can also reduce the activity of V-ATPase and V-PPase in vegetable root system cells under antibiotic stress, reduce the absorption and accumulation of organic pollutants such as antibiotics, relieve antibiotic stress and restore the yield of vegetables and food safety quality to a certain extent, and the operation method is simple, safe and time-saving.

Description

Technical Field

[0001] The invention relates to the technical field of antibiotic pollution control, and in particular to application of exogenous abscisic acid in controlling antibiotic-contaminated vegetables. Background Art

[0002] Antibiotics are used as a treatment for disease prevention and control. Both human and veterinary antibiotics (25-75%) are excreted through urine and feces, which increases the occurrence and accumulation of antibiotics in soil. Among them, tetracyclines, macrolides, sulfonamides and quinolones are the antibiotics that accumulate the most in agricultural soils. Exogenous antibiotics accumulate in agricultural soils, and water flows through agricultural soils with antibiotic accumulation, and then the water is polluted. Usually, these polluted waters are used for crop irrigation. Therefore, crops are often continuously exposed to high concentrations of antibiotics, and the concentration of antibiotics in crop tissues increases with the increase of antibiotic concentrations in the soil. In recent years, people have paid more and more attention to the problem of antibiotic contamination of crops.

[0003] Physiological changes in plants caused by antibiotics (or their metabolic decomposition products) are intrinsically related to the ability of certain species to remove pollutants from the environment (phytoremediation). The negative effects of antibiotics on photosynthesis and the damage to antioxidant metabolism inhibit the repair ability of plants. In addition, the continued growth of antibiotics in the soil promotes the selection of antibiotic-resistant bacteria, which have stronger survival and transmission capabilities. They may enter humans and animals through the food chain, damage the human urinary system and liver and kidney functions, and even cause new infectious diseases. The problem of antibiotic contamination in food has always been taken very seriously at home and abroad, and standards for food residues and daily intake have been established. Therefore, how to reduce the accumulation of antibiotics in crops is one of the problems that need to be solved urgently. Summary of the invention

[0004] In view of this, the object of the present invention is to provide a new use of exogenous abscisic acid to reduce the accumulation of antibiotics in crops.

[0005] The purpose of the present invention is achieved through the following technical solutions: A new use of exogenous abscisic acid (ABA), including its use in at least one of the following aspects: 1) Reduce the accumulation of antibiotics in vegetables; 2) Alleviate antibiotic stress on vegetables; 3) Eliminate antibiotic resistance genes in vegetables.

[0006] The exogenous abscisic acid described in the present invention is preferably natural abscisic acid, 98%.

[0007] Furthermore, the antibiotics include at least one of tetracyclines and quinolones, such as tetracycline, ciprofloxacin, etc. The vegetables described in the present invention are cucumbers, tomatoes, eggplants, etc.

[0008] A vegetable antibiotic stress reliever includes exogenous abscisic acid with a concentration of 5 - 80 μmol / L. Among them, the concentration of the exogenous abscisic acid can be 5, 10, 20, 30, 40, 50, 60, 70, 80 μmol / L, etc.

[0009] The vegetable antibiotic stress reliever is preferably an aqueous solution, which is made by uniformly mixing each component in a light - avoiding state.

[0010] Furthermore, the vegetable antibiotic stress reliever also includes an agricultural silicone adjuvant with a concentration of 0.5 - 1 ml / L. The effective ingredient of the agricultural silicone adjuvant described in the present invention is dioxy - modified polytri - siloxane.

[0011] An application of the above - mentioned vegetable antibiotic stress reliever in at least one of the following aspects: 1) Reducing the accumulation of antibiotics in vegetables; 2) Alleviating the antibiotic stress of vegetables; 3) Eliminating antibiotic resistance genes in vegetables.

[0012] Furthermore, the application method includes: spraying the above - mentioned vegetable antibiotic stress reliever on the vegetable leaves in a light - avoiding manner. For example, spraying the above - mentioned vegetable antibiotic stress reliever at night.

[0013] Specifically, when the vegetable leaves under antibiotic stress grow to two leaves and one heart, spray the vegetable antibiotic stress reliever on the leaves of the vegetables in a light - avoiding manner, and conduct the spraying treatment once every 3 days. Preferably, spray at least 2 times.

[0014] In the vegetable planting system, a large amount of organic fertilizer is usually applied as the base fertilizer, and a large amount of unmetabolized veterinary antibiotics are contained in the organic fertilizer. Exogenous abscisic acid treatment can enhance the activity of antioxidant enzymes in vegetables, ensure the normal progress of photosynthetic and respiratory electron transport chains, reduce the oxidative stress induced by the disorder of electron flow in vegetables under antibiotic stress, thereby improving the ability of the vegetable self - protection system to scavenge reactive oxygen species, being beneficial to maintaining the reactive oxygen species balance, and further improving the ability of vegetables to degrade organic pollutants such as antibiotics. Exogenous abscisic acid treatment can also reduce the activities of V - ATPase and V - PPase in the root cells of vegetables under antibiotic stress, reduce the absorption and accumulation of organic pollutants such as antibiotics, alleviate the antibiotic stress, and to a certain extent restore the yield and food safety quality of vegetables. The operation method is simple, safe, time - saving, and provides a new idea and method for efficiently using medium - and low - concentration antibiotic - contaminated soil in the region to produce safe agricultural products. Description of the Drawings

[0015] Figure 1 Phenotype diagrams of cucumber under different concentrations of exogenous ABA treatment under the stress of antibiotics ciprofloxacin hydrochloride (CIP) and tetracycline (TC); Figure 2 For the contents of H2O2 and O 2.- in leaves after treatment with 20 μmol / L exogenous ABA; Figure 3 Diagrams showing the effects of 20 μmol / L exogenous ABA treatment on the activities and gene expressions of APX, CAT, POD, and SOD; Figure 4 Diagrams showing the effects of 20 μmol / L exogenous ABA treatment on the activities of V-PPase and V-ATPase in cucumber roots; Figure 5 For the CIP contents in the shoots and roots of cucumber after treatment with 20 μmol / L exogenous ABA; Figure 6 For the TC contents in the shoots and roots of cucumber after treatment with 20 μmol / L exogenous ABA; Figure 7 Phenotype diagrams of cucumber treated with 20 μmol / L exogenous ABA under the combined stress of CIP and TC. Detailed implementation manners

[0016] To make the objectives, technical solutions, and advantages of the present invention clearer, the technical solutions of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention.

[0017] In the ranges disclosed in the present invention, the endpoints and any values are not limited to the precise ranges or values. These ranges or values should be understood to include values close to these ranges or values. For numerical ranges, between the endpoint values of each range, between the endpoint values of each range and individual point values, and between individual point values, they can be combined with each other to obtain one or more new numerical ranges, and these numerical ranges should be regarded as specifically disclosed in the present invention.

[0018] Unless otherwise specified, the terms used in the present invention are all common terms in the art. For the preparation processes, test methods, etc. used in each implementation manner without special instructions, they are all conventional means well-known to those skilled in the art, and the raw materials and equipment used can be obtained from public commercial channels.

[0019] Exogenous abscisic acid (natural, 98%), purchased from Aladdin Biochemical Technology Co., Ltd., product number: A100492.

[0020] Agricultural silicone adjuvant: dioxy-modified polytrioxane, purchased from Shijiazhuang Jiunongfeng Chemical Technology Co., Ltd.

[0021] The indoor hydroponic experiment used cucumber variety Jinyan No. 4 as the test material, and the seeds were purchased from Xiangyun Seed Industry Co., Ltd., Xintai City, Shandong Province.

[0022] In the indoor hydroponic experiment, Hoagland nutrient solution was prepared with distilled water. Antibiotics were added to the hydroponic solution during the experiment. The light cycle of the experiment was 14 / 10 h (day / night), the temperature was 27°C / 25°C (day / night), the average relative humidity was 65 - 75%, and the light intensity was 300 µmolm -2 s -1 。

[0023] For the quantitative determination of hydrogen peroxide (H2O2) and superoxide anion (O2 .– ), and the detection methods of antioxidant enzyme activities such as ascorbate peroxidase (APX), catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD) were carried out using commercial kits (Suzhou Mengxi Biomedical Technology Co., Ltd.).

[0024] Examples 1 - 4 Each of Examples 1 - 4 provides a vegetable antibiotic stress reliever, which includes uniformly mixed exogenous ABA, agricultural organosilicon adjuvant, and water. Among them, the concentration of the agricultural organosilicon adjuvant is 0.8 ml / L, and the concentrations of exogenous ABA in Examples 1 - 4 are 10, 20, 40, and 80 µmol / L respectively.

[0025] Each example also provides a method for regulating exogenous abscisic acid to relieve cucumber antibiotic stress and reduce cucumber antibiotic accumulation by using the above - mentioned vegetable antibiotic stress reliever, which specifically includes the following steps: S1. Seedling raising: Vermiculite was evenly filled in a 50 - mesh seedling tray. One germinated cucumber seed of Jinyan No. 4 was sown in each hole, covered with a thin layer of vermiculite, gently pressed, and 500 ml of water was poured at the bottom of each hole tray to ensure the temperature and humidity required for cucumber seed germination. During this period, the seedling tray was always kept moist.

[0026] S2. Hydroponics: When the cucumber seedlings grew to "one leaf and one heart", select uniform and well - growing seedlings, remove the seedlings together with the whole roots from the hole tray, wash them clean with distilled water, cut and leave 4 cm of roots with a blade, and let them grow in a 4 L plastic box (4 plants per box). The box was filled with 3.5 L of Hoagland nutrient solution (1 / 2 working concentration), which was changed every three days. In all experiments, the growth conditions of the seedlings were: light cycle 14 / 10 h (day / night), temperature 27°C / 25°C (day / night), average relative humidity 65 - 75%, light intensity 300 µmol m -2 s -1. During the planting period, observe the growth of the plants daily, promptly remove the diseased seedlings and weak seedlings with soft and rotten roots, and stir the nutrient solution in the plastic box with a glass rod to ensure the uniformity of the nutrient solution in the box and prevent the occurrence of pathogen contamination among the plants.

[0027] S3. Treatment: The cucumber seedlings grow in the hydroponic solution, the wounds of the plant roots heal and grow normally. When the plants grow to the stage of "two leaves and one heart", select the uniform seedlings, spray two treatments of the vegetable antibiotic stress reliever (+ABA) provided in Examples 1-4 and distilled water (-ABA) on the leaves, and transplant both treatments into the hydroponic nutrient solutions respectively added with antibiotics CIP and TC, so that the final concentrations of CIP and TC in them are both 1 mg / L. Among them, in the distilled water treatment, an agricultural organic silicon adjuvant with a concentration of 0.8 ml / L is added to the pure distilled water. Set a blank control of spraying distilled water and not adding antibiotics (CK), with three replicates.

[0028] S4. Sampling: On the 7th day of the treatment, detect the phenotypes of the cucumber seedling plants as Figure 1 shown, and take the middle-section roots and above-ground parts samples of the cucumber plants at this 7th day.

[0029] S5. Statistical data: Statistically analyze the growth data and chlorophyll content of the cucumber plants on the 7th day of the treatment, and the results are shown in Tables 1-4.

[0030] Table 1 Effects of 10 µmol / L ABA on the growth and chlorophyll content of cucumber plants under antibiotic stress Treatment Plant height (cm) Root length (cm) Fresh weight (g) Dry weight (g) <![CDATA[Chlorophyll content (mg∙g -1 ).]]> CK 29.33±1.96a 27.13±0.14a 24.41±1.10a 1.33±0.08a 19.72±1.24a CIP 19.78±0.84c 15.10±1.37c 12.42 ± 0.51 cd 0.71±0.06c 8.86±1.43c CIP + ABA 23.96±1.42b 16.96±0.84b 14.71±1.06b 0.89±0.06b 12.94±1.05b TC 18.48±0.91d 14.81±1.09c 11.09±1.07d 0.62±0.09c 6.14±0.65d TC + ABA 23.21±0.50b 16.51±0.87b 13.58±0.98c 0.85±0.07b 9.78±0.97c Table 2 Effects of 20 µmol / L ABA on the growth and chlorophyll content of cucumber plants under antibiotic stress Treatment Plant height (cm) Root length (cm) Fresh weight (g) Dry weight (g) <![CDATA[Chlorophyll content (mg∙g -1 ).]]> CK 30.28±1.86a 28.52±0.46a 25.13±1.29a 1.21±0.09a 21.58±1.81a CIP 21.94±0.47d 12.39±1.57c 13.93±0.72c 0.69±0.07c 6.14±1.53c CIP + ABA 25.60±0.71b 18.90±0.48b 17.72±1.19b 1.02±0.04b 15.96±0.89b TC 20.94±0.48d 11.48±1.23c 12.70±0.97c 0.61±0.05c 4.90±0.62c TC + ABA 24.42±0.99c 18.50±0.96b 16.61±1.07b 0.93±0.07b 15.54±1.63b Table 3 Effects of 40 µmol / L ABA on the growth and chlorophyll content of cucumber plants under antibiotic stress Treatment Plant height (cm) Root length (cm) Fresh weight (g) Dry weight (g) <![CDATA[Chlorophyll content (mg∙g -1 ).]]> CK 31.48±1.21a 28.14±0.27a 24.87±0.84a 1.32±0.07a 21.78±1.80a CIP 19.38±0.88c 14.88±1.53c 12.46±0.69c 0.73±0.06c 6.34±1.41c CIP + ABA 21.67±1.33b 16.61±0.51b 16.15±0.49b 0.88±0.10b 12.76±1.11b TC 18.78±0.87c 14.29±1.45c 11.92±1.13c 0.67±0.05c 5.56±0.19c TC + ABA 21.01±1.34b 15.57 ± 1.09 bc 15.69±1.12b 0.82±0.08b 12.18±1.64b Table 4 Effects of 80 µmol / L ABA on the growth and chlorophyll content of cucumber plants under antibiotic stress Treatment Plant height (cm) Root length (cm) Fresh weight (g) Dry weight (g) <![CDATA[Chlorophyll content (mg∙g -1 ).]]> CK 30.28±1.86a 27.74±0.61a 24.03±1.02a 1.17±0.06a 22.58±1.13a CIP 19.58±0.54b 14.36±1.19b 11.82±0.97b 0.64±0.03b 7.32±1.47c CIP + ABA 19.90±0.36b 15.08±0.49b 11.79±0.56b 0.65±0.04b 11.73±1.82b TC 18.32±0.35c 14.40±1.23b 11.65±1.23b 0.46±0.07c 6.04±0.87c TC + ABA 17.58±0.34c 14.14±0.81b 10.62±0.79b 0.53±0.06c 10.67±1.79b It can be seen from Tables 1-4 that: 20 µmol / L of exogenous ABA (Example 2) can significantly relieve the morphological indexes of cucumbers under CIP and TC stress, and the plant height, root length, fresh weight and dry weight are all significantly increased, second only to the control group CK (the blank control group without adding antibiotics).

[0031] Conduct the following detections on the cucumber plant samples obtained in Example 2 respectively: Take 0.2 g of leaf tissue for H2O2 and O2 .–Quantitative determination and antioxidant enzyme activity detection, the results are as follows Figure 2 and 3 As shown. Figure 2 It can be seen that the H2O2 and O2 induced by CIP and TC stress .– Excessive precipitation and exogenous ABA treatment significantly reduced H2O2 and O2 .– The accumulation of Figure 3 It can be seen that CIP and TC stress significantly increased the activities of APX, CAT, SOD and POD in cucumber leaves. Under CIP and TC stress, compared with the treatment without exogenous ABA, exogenous ABA treatment further significantly increased the above physiological parameters and significantly increased the mRNA levels of the above parameters, indicating that exogenous ABA can improve the antibiotic resistance of cucumber seedlings by regulating the antioxidant system.

[0032] The cucumber root samples treated in Example 2 were subjected to V-ATPase (vacuolar H + -ATPase) and V-PPase (vacuolar H + -pyrophosphatase) content test (He Peng (Shanghai) Biotechnology Co., Ltd.), the results are as follows Figure 4 As shown. Figure 4 It can be seen that the activities of V-ATPase and V-PPase in cucumber roots were significantly increased under CIP and TC stress compared with the CK phase (distilled water treatment). Under CIP and TC stress, the activities of V-ATPase and V-PPase were significantly reduced when treated with exogenous ABA.

[0033] The cucumber plant samples treated in Example 2 were tested for antibiotic content (Suzhou Mengxi Biopharmaceutical Technology Co., Ltd.), and the results were as follows: Figure 5 and 6 As shown. Figure 5 It can be seen that under CIP stress, the CIP accumulation in the cucumber root system treated with 20 μmol / L exogenous ABA provided in Example 2 was reduced by 54.17%, and the CIP accumulation in the aboveground part was reduced by 41.92%; Figure 6 It shows that under TC stress, the TC accumulation in the cucumber root system and the TC accumulation in the aboveground part were reduced by 73.52% and 54.76% respectively when 20 µmol / L exogenous ABA was sprayed as provided in Example 2.

[0034] In summary, exogenous ABA can reduce the accumulation of antibiotics in the cultivation process of Jinyan No. 4 cucumber and alleviate the stress of antibiotics on the plants. The concentration of exogenous ABA is preferably 10-80 µmol / L, more preferably 10-40 µmol / L. Among them, 20 µmol / L of exogenous ABA has the best effect on antibiotic accumulation and alleviating antibiotic stress on cucumbers.

[0035] Examples 5 - 7 Examples 5 - 7 provide an exogenous abscisic acid regulation method for the vegetable antibiotic stress reliever provided in Example 2 to relieve the combined stress of antibiotics CIP and TC in cucumbers. The specific method is basically the same as the method provided in Example 2, and the main differences are as follows: The mass ratios of the antibiotic combination CIP:TC in Examples 5 - 7 are 1:1, 1:2, and 2:1 in sequence, and the total antibiotic concentrations therein are 2 mg / L, 3 mg / L, and 3 mg / L in sequence; The phenotypes of cucumber seedling plants were detected on the 7th day of treatment as Figure 7 shown; The growth data and chlorophyll content of cucumber plants on the 7th day of treatment were statistically analyzed, and the results are shown in Table 5.

[0036] Table 5 Effects of exogenous ABA on the growth and chlorophyll content of cucumber plants under combined antibiotic stress Treatment Plant height (cm) Root length (cm) Fresh weight (g) Dry weight (g) <![CDATA[Chlorophyll content (mg∙g -1 ).]]> CK 13.88±0.62a 28.03±1.16a 30.66±0.35a 1.42±0.29a 26.16±2.18a 1:1 8.99 ± 0.74 de 18.83±1.35e 13.82±0.17c 0.65±0.50c 7.81 ± 0.95 de 1:1 + ABA 9.79±1.24b 20.53±1.64b 14.99±0.33b 0.78±0.29b 11.98±1.73b 1:2 8.63±0.85e 17.78±1.43f 11.26 ± 0.95 ef 0.45±0.29e 6.73±1.25e 1:2 + ABA 9.28 ± 1.01 cd 19.41±1.81c 12.26±0.65d 0.50±0.36d 9.5±1.47c 2:1 7.95±0.66f 17.187±1.02g 9.75±0.75f 0.35±0.33f 5.4±0.75f 2:1 + ABA 9.4±1.02c 19.11±1.29d 11.58 ± 0.33 de 0.46 ± 0.25 de 8.18±1.5d From Figure 7 and Table 5, it can be seen that: The vegetable antibiotic stress reliever provided in the embodiments of the present invention can significantly relieve the morphological indexes of cucumbers under the combined stress of CIP and TC, and the plant height, root length, fresh weight, and dry weight are all significantly increased, second only to the control group CK.

[0037] Therefore, using exogenous natural abscisic acid in the embodiments of the present invention can, to a certain extent, reduce the harm degree of antibiotic stress in the cucumber planting system, inhibit the accumulation of antibiotics in cucumbers, relieve antibiotic stress, and restore the yield and food safety quality of cucumbers to a certain extent. The method is simple and time - saving.

[0038] Finally, it should be noted that: The above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them; Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that: It is still possible to modify the specific implementation manners of the present invention or perform equivalent replacements for some technical features; Without departing from the spirit of the technical solutions of the present invention, they should all be covered within the scope of the technical solutions claimed by the present invention.

Claims

1. A new use of exogenous abscisic acid, characterized in that, including its application in at least one of the following aspects: 1) Reducing the accumulation of antibiotics in vegetables; 2) Alleviating the antibiotic stress of vegetables; 3) Eliminating antibiotic resistance genes in vegetables.

2. The new use according to claim 1, characterized in that, The antibiotics include at least one of tetracyclines and quinolones.

3. The new use according to claim 1 or 2, characterized in that, The vegetables are cucumbers, tomatoes or eggplants.

4. The new use according to claim 1 or 2, characterized in that, The alleviation of the antibiotic stress of the vegetables is manifested as: enhancing the activity of vegetable antioxidant enzymes and reducing the activities of V-ATPase and V-PPase in vegetable root cells.

5. A vegetable antibiotic stress reliever, characterized in that, It includes exogenous abscisic acid with a concentration of 5 - 80 μmol / L.

6. The vegetable antibiotic stress reliever according to claim 5, characterized in that The concentration of the exogenous abscisic acid is 10 - 40 μmol / L.

7. The vegetable antibiotic stress reliever according to claim 5 or 6, characterized in that, It also includes an agricultural silicone adjuvant with a concentration of 0.5 - 1 ml / L.

8. Application of the vegetable antibiotic stress reliever according to any one of claims 5 - 7 in at least one of the following aspects: 1) Reducing the accumulation of antibiotics in vegetables; 2) Alleviating the antibiotic stress of vegetables; 3) Eliminating antibiotic resistance genes in vegetables.

9. The application according to claim 8, wherein, It includes: Spraying the vegetable antibiotic stress reliever on the vegetable leaves in the dark.

10. The application according to claim 9, characterized in that, When the vegetable leaves under antibiotic stress grow to two leaves and one heart, spray the vegetable antibiotic stress reliever on the leaves of the vegetables in the dark, and perform the spraying treatment once every 3 days.

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