APPLICATION OF SYRINGIC ACID IN PROMOTING NITRIFICATION INHIBITION ACTIVITY OF DECANEDIOL

Abstract

An application of syringic acid in promoting the inhibition effect of decanediol on the nitrosification activity of nitrosomonas is provided. By inhibiting the activity of a nitrosomonas, the combined use of syringic acid and 1,9-decanediol can more efficiently inhibit a nitrification process in soil and at the plant rhizosphere, increase the nitrogen use efficiency, and reduce leaching of nitrate nitrogen, thereby reducing the loss of nitrogen and discharge of the greenhouse gas nitrous oxide in a denitrification process. The addition of syringic acid in the present invention can reduce the concentration at which 1,9-decanediol functions under the equivalent condition of nitrification inhibition activity, and thus save the relative input cost.

Claims

1. An application of syringic acid to promote an inhibition effect of 1,9-decanediol on the nitrosification activity of a nitrosomonas.

2. The application of syringic acid of claim 1, wherein the nitrosomonas is Nitrosomonas europaea (NBRC 14298) and Nitrosomonas stercoris (NBRC 110753).

3. The application of syringic acid of claim 1, wherein an addition concentration of syringic acid is 10-500 g.Math.mL.sup.1.

4. An application of syringic acid to prepare a formulation for promoting an inhibition effect of 1,9-decanediol on the nitrosification activity of a nitrosomonas.

5. An application of syringic acid to prepare a composition for reducing nitrogen loss in farmland.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Various additional features and advantages of the invention will become more apparent to those of ordinary skill in the art upon review of the following detailed description of one or more illustrative embodiments taken in conjunction with the accompanying drawings. The accompanying drawings, which are incorporated in and constitutes a part of this specification, illustrate one or more embodiments of the invention and, together with the general description given above and the detailed description given below, explain the one or more embodiments of the invention.

[0015] FIG. 1 is a graphical plot showing a GC/MS analysis spectrum of the components in the root exudate of the rice variety Wuyunjing 7 according to one embodiment of the present invention, showing that 1,9-decanediol is a substance which has a peak at 20.04 min, and syringic acid is a substance which has a peak at 27.44 min.

[0016] FIG. 2 is a graphical bar chart showing the effect of syringic acid on the nitrification inhibition activity of 1,9-decanediol to the Nitrosomonas europaea (meanSE, n=3) according to embodiments of the invention, where different lowercase letters indicate significant differences between groups (P<0.05).

[0017] FIG. 3 is a graphical bar chart showing the effect of syringic acid on the nitrification inhibition activity of 1,9-decanediol to the Nitrosomonas stercoris (meanSE, n=3) according to embodiments of the invention, where different lowercase letters indicate significant differences between groups (P<0.05).

DETAILED DESCRIPTION

[0018] The following clearly and completely describes the technical solutions in the embodiments of the present invention with reference to the accompanying drawings in the embodiments of the present invention. To make objectives, features, and advantages of the present invention clearer, the following describes embodiments of the present invention in more detail with reference to accompanying drawings and specific implementations.

[0019] Embodiment 1: The Effect of Syringic Acid on the Nitrification Inhibition Activity of 1,9-Decanediol to the Nitrosomonas europaea

1.1 Experimental Design

[0020] Standard: a syringic acid standard was purchased from Sigma-Aldrich (St. Louis, Mo., USA), and the solid powder thereof was weighed and dissolved in DMSO. A 1,9-decanediol standard was purchased from WuXi AppTec, and the solid powder thereof was dissolved in DMSO while frozen over the dry ice (the 1,9-decanediol standard was a viscous liquid substance under normal temperature).

[0021] Microbial strains: Nitrosomonas europaea (NBRC 14298) was purchased from Biological Resource Center, NITE, Japan.

[0022] Microbial culture medium: a HEPES medium, which contained 2.5 g of (NH.sub.4).sub.2SO.sub.4, 0.5 g of KH.sub.2PO.sub.4, 11.92 g of HEPES, 0.5 g of NaHCO.sub.3, 100 mg of MgSO.sub.4.7H.sub.2O, 5 mg of CaCl.sub.2.2H.sub.2O, and 75 mg of Fe-EDTA per 1 L liquid medium, at pH 7.8-8.0.

[0023] Culture of microorganisms: Nitrosomonas europaea was inoculated in the HEPES medium and cultured at 30 C. and 200 rpm under a dark condition (aerobic), and the Nitrosomonas europaea enters a stable period 7-9 days after transfer each time.

[0024] Nitrification activity inhibition experiment: The bacterial solution cultured for 7 days was collected, centrifuged at 5000 g for 20 min, and resuspended in a fresh and sterile HEPES medium until the OD600 reached about 1.0, with a concentration multiple of 40-50 times. Taken was a 1.5 mL sterile centrifuge tube, and it was sequentially added with 195 L of sterile water, 5 L of different concentrations of solutions of syringic acid and 1,9-decanediol in DMSO, 100 L of the fresh and sterile HEPES medium, and 200 L of a resuspended bacterial solution, and then cultured in a water bath at 25 C. under a dark condition for 2 hours. Then the mixed system was added with 20 L of 0.1M Allylthiourea to terminate the nitrosification. 200-400 pL of the reacted mixture was added into a 10 mL colorimetric tube, diluted to about 5 mL with deionized water, added with 1 mL of a sulfanilic acid solution, shaken well and then subjected to standing for 2-8 min, then added with 1 mL of a hydrochloric acid N-(1-naphthyl)-ethylenediamine solution, shaken well, diluted to 10 mL with water to obtain a constant volume. Deionized water was used as a reference, and the absorbances were determined at a wavelength of 540 nm. The reticle of NO.sub.2.sup. was made in the same manner to quantify the NO.sub.2.sup. generated in the sample system, and the sample inhibition rate was calculated by the following formula. This determination method was an improved Griess method, which could be referred to the national standard determination of nitrite in the wet precipitation -N-(1-naphthyl)-1,2-diaminoethane spectrophotometry.

[0025] Nitrosification inhibition rate (%)=(1-the produced amount of NO.sub.2.sup. of the sample/the produced amount of NO.sub.2.sup. of a blank)100%

1.2. Experimental Result

[0026] The experimental result as shown in FIG. 2 was that syringic acid had a significant synergistic effect on the nitrification inhibition activity of 1,9-decanediol. When 1,9-decanediol was added alone, its inhibition rate for the nitrosification process of Nitrosomonas europaea was 41.2%, and after syringic acid each at the concentrations of 10, 100 and 500 .Math.mL.sup.1 syringic acid was added on this basis, the nitrification inhibition effect of 1,9-decanediol was significantly improved in such a manner that the nitrification inhibition rates were increased to 51.7%, 61.1% and 69.2%, respectively.

[0027] Embodiment 2: The effect of syringic acid on the nitrification inhibition activity of 1,9-decanediol to the Nitrosomonas stercoris 1.1 Experimental Design

[0028] The selection and formulation of the standard were the same as those of Embodiment 1.

[0029] Microbial Strains: Nitrosomonas stercoris (NBRC 110753) was purchased from Biological Resource Center, NITE, Japan.

[0030] The microbial culture medium and microbial culture were the same as those of Embodiment 1.

[0031] Nitrification activity inhibition experiment: The bacterial solution of Nitrosomonas stercoris cultured for 7 days was collected, centrifuged at 5000 g for 20 min, and resuspended in a fresh and sterile HEPES medium until the OD600 reached about 1.0, with a concentration multiple of 40-50 times. Taken was a 1.5 mL sterile centrifuge tube, and it was sequentially added with 195 L of sterile water, 5 L of different concentrations of solutions of syringic acid and 1,9-decanediol in DMSO, 100 L of the fresh and sterile HEPES medium, and 200 L of a resuspended bacterial solution, and then cultured in a water bath at 25 C. under a dark condition for 2 hours. Then the mixed system was added with 20 L of 0.1M Allylthiourea to terminate the nitrosification. The determination method for the NO.sub.2.sup. generated in the system after the reaction was the same as that of Embodiment 1.

1.2. Experimental Result

[0032] The experimental result as shown in FIG. 3 was that, syringic acid also promoted the inhibition effect of 1,9-decanediol on Nitrosomonas stercoris, where when 10, 100 and 500 g.Math.mL.sup.1 of syringic acid were added, the nitrification inhibition effect of 1,9-decanediol was significantly increased, with the inhibition rate increased from 51.5% to 60.5%, 70.2% and 89.9% accordingly. Therefore, syringic acid has a good application prospect in promoting the inhibition activity of 1,9-decanediol on the Nitrosomonas.

[0033] The embodiments described above are only descriptions of preferred embodiments of the present invention, and do not intended to limit the scope of the present invention. Various variations and modifications can be made to the technical solution of the present invention by those of ordinary skills in the art, without departing from the design and spirit of the present invention. The variations and modifications should all fall within the claimed scope defined by the claims of the present invention.