Method for separating water plant viruses and application thereof

Abstract

The invention relates to the technical field of water plant virus separation, and discloses a water plant virus separation method and application thereof, and the method comprises the step of mixing an amino-functionalized carbon nanomaterial and a water body containing plant viruses. The amino-functionalized carbon nanomaterial adopted by the method can be combined with plant viruses in a water body, so that the adsorption capacity of the amino-functionalized carbon nanomaterial to the plant viruses is improved, and the method is simple to operate, efficient and low in cost, does not generate toxic substances, avoids secondary pollution possibly caused by chemical disinfection, and is suitable for industrial production. The invention provides an economic and sustainable solution for solving the problem of plant virus pollution, and shows a wide application prospect in the field of water plant virus removal.

Description

Technical Field

[0001] The present invention relates to the technical field of carbon nanomaterials, and in particular to a method for separating aquatic plant viruses and an application thereof. Background Art

[0002] Pepper Mild Mottle Virus (PMMoV) is a common plant virus that primarily infects peppers and other Solanaceae crops. PMMoV is highly environmentally stable, surviving in water for extended periods and spreading over long distances through irrigation systems. PMMoV-infected crops exhibit symptoms such as significant growth inhibition, leaf mottles, and fruit deformities, severely impacting agricultural yield and value.

[0003] Currently, chemical disinfection technologies (including sodium hypochlorite treatment and ozone oxidation), ultraviolet disinfection, and membrane separation are the main approaches for reducing PMMoV in water. However, these methods all have limitations in practical application. Dissolved organic matter in water can compete with disinfectants, reducing the effective concentration of the oxidant, decreasing the inactivation efficiency of plant viruses, and producing toxic byproducts. Furthermore, ultraviolet light is susceptible to the optical properties of water; increased turbidity significantly reduces its penetration, reducing its inactivation efficiency. Although membrane separation technology has a certain degree of retention for plant virus particles, the extremely small (nanoscale) particle size of plant viruses makes them difficult to capture or adsorb by membranes, making membrane separation of plant viruses less effective. Furthermore, in practical operation, membrane fouling is a serious challenge. This not only increases operating costs but also potentially shortens the lifespan of the membrane material. Therefore, the development of new, efficient, cost-effective, and environmentally friendly materials for plant virus removal is of great significance.

[0004] Carbon nanotubes (CNTs) offer significant advantages in pollutant removal due to their unique tubular structure, large surface area, and tunable surface chemical properties. Therefore, effectively functionalizing CNTs to efficiently separate and remove plant viruses from aquatic environments has become a pressing issue in agricultural water treatment technology. Summary of the Invention

[0005] The purpose of the present invention is to overcome the problems of poor removal effect of plant viruses in water bodies, high energy consumption, and environmental pollution (such as the generation of disinfection by-products and membrane pollution) in the existing technology, and to provide a method for separating plant viruses in water bodies and its application. The amino-functionalized carbon nanomaterial used in this method can combine with plant viruses in water bodies, thereby improving the adsorption capacity of the amino-functionalized carbon nanomaterial for plant viruses.

[0006] In order to achieve the above objectives, the first aspect of the present invention provides a method for separating plant viruses from water, wherein the method comprises mixing amino-functionalized carbon nanomaterials with water containing plant viruses.

[0007] The second aspect of the present invention provides the use of the method of the present invention in separating aquatic plant viruses.

[0008] Through the above technical solution, the present invention achieves the following technical effects: (1) The amino-functionalized carbon nanotubes used in the present invention can bind to plant viruses, thereby improving the adsorption capacity of the amino-functionalized carbon nanomaterial for plant viruses in water. In a preferred embodiment, the amino-functionalized carbon nanomaterial has a removal rate of more than 81% for plant viruses in water (especially pepper mild mottle virus, PMMoV). This method shows broad application prospects in the field of plant virus removal in water. (2) When the method of the present invention is used to separate and remove plant viruses from water, the rapid separation of plant viruses in water can be achieved by simple filtration or centrifugation. The operation method is simple, efficient, and low-cost. In addition, the method of the present invention does not produce toxic substances, avoiding the secondary pollution that may be caused by chemical disinfection, and provides an economical and sustainable solution to the problem of plant virus contamination. (3) The method of the present invention is simple and efficient, does not require complex equipment throughout the process, and is suitable for large-scale production. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] Figure 1 This is a scanning electron microscope (SEM) image of the amino-functionalized carbon nanomaterial prepared in Example 1 of the present invention. DETAILED DESCRIPTION

[0010] The endpoints of the ranges and any values disclosed herein are not limited to the precise ranges or values, and these ranges or values should be understood to include values close to these ranges or values. For numerical ranges, the endpoints of each range, the endpoints of each range and individual point values, and the individual point values can be combined with each other to obtain one or more new numerical ranges, which should be considered to be specifically disclosed herein.

[0011] A first aspect of the present invention provides a method for separating plant viruses from water, wherein the method comprises mixing an amino-functionalized carbon nanomaterial with water containing the plant viruses.

[0012] According to some embodiments of the present invention, the mass ratio of the amino-functionalized carbon nanomaterial to the water containing the plant virus is 1:(100-500).

[0013] In the present invention, the process of mixing the amino-functionalized carbon nanomaterial and the water containing the plant virus is the process of the amino-functionalized carbon nanomaterial adsorbing the plant virus. The entire adsorption process is carried out at room temperature, such as around 25°C.

[0014] In order to improve the adsorption efficiency of amino-functionalized carbon nanomaterials on plant viruses, preferably, the mixing conditions include: a stirring speed of 200-300 rpm and a time of 30-60 min.

[0015] In the present invention, the method further comprises, after the amino-functionalized carbon nanomaterial and the water containing the plant virus are mixed, performing solid-liquid separation on the mixed liquid to achieve rapid separation and removal of the plant virus in the water. Preferably, the solid-liquid separation method comprises filtration or centrifugation, more preferably filtration.

[0016] Preferably, the pore size of the filtration membrane is 0.45-0.8 μm.

[0017] In the present invention, the amino-functionalized carbon nanomaterial refers to a method in which multi-walled carbon nanotubes are carboxylated (carboxyl groups are introduced onto the surface of the carbon nanotubes) using a strong oxidizing acid (such as sulfonic acid or a mixture of concentrated sulfuric acid and concentrated nitric acid) to obtain carboxylated multi-walled carbon nanotubes; the carboxylated multi-walled carbon nanotubes are then chemically reacted (amide reaction) with nitrogen-containing functional groups to introduce amino groups onto the surface of the carbon nanotubes in the form of amide bonds to obtain amino-modified carbon nanomaterials.

[0018] According to some embodiments of the present invention, the method for preparing the amino-functionalized carbon nanomaterial comprises the following steps: (1) mixing carboxylated multi-walled carbon nanotubes and water and performing ultrasonication to obtain a carboxylated multi-walled carbon nanotube suspension; (2) The carboxylated multi-walled carbon nanotube suspension and an amine compound are mixed to undergo an amide reaction to obtain amino-functionalized carbon nanomaterials.

[0019] In the present invention, there is no particular limitation on the source of the carboxylated multi-walled carbon nanotubes; they can be homemade or commercially available. In a specific embodiment provided herein, the carboxylated multi-walled carbon nanotubes are commercially available, for example, from Shanghai MacLean Biochemical Technology Co., Ltd., product number 308068-56-6. The carboxylated multi-walled carbon nanotubes have a purity of >95%, an inner diameter (ID) of 2-5 nm, an outer diameter (OD) of <8 nm, a length of 0.5-2 μm, and a carboxyl group content of approximately 3.9 wt%.

[0020] The inventors of the present invention have discovered that the surface of the amino-functionalized carbon nanomaterial prepared by the method of the present invention has abundant amino groups and carries a certain amount of charge (the potential value satisfies a specific range), which enables the amino-functionalized carbon nanomaterial to efficiently combine with plant viruses, thereby improving the adsorption capacity of the amino-functionalized carbon nanomaterial for plant viruses in water bodies.

[0021] According to some embodiments of the present invention, the plant virus is selected from at least one of pepper mild mottle virus (PMMoV), tobacco mosaic virus (TMV), cucumber mosaic virus (CMV) and potato virus Y (PVY virus).

[0022] Preferably, the plant virus is pepper mild mottle virus.

[0023] According to some embodiments of the present invention, the Zeta potential value of the amino-functionalized carbon nanomaterial is 12-25 mV, preferably 17-24 mV, and more preferably 17.5-21 mV.

[0024] In the present invention, the Zeta potential value of the amino-functionalized carbon nanomaterial is measured by using a Zeta potential and particle size analyzer (the instrument is purchased from Malvern, product model: Zetasizer Nano ZS).

[0025] In the present invention, the pore size of the amino-functionalized carbon nanomaterial is measured using an electron microscope scanner (the electron microscope scanner is purchased from JEOL Ltd., and the instrument model is JSM-7800F).

[0026] According to some embodiments of the present invention, in step (1), based on the mass of 1 g of the carboxylated multi-walled carbon nanotubes, the amount of water used is 100-1000 mL, preferably 200-500 mL.

[0027] According to some embodiments of the present invention, in step (2), based on the mass of 1 g of the carboxylated multi-walled carbon nanotubes, the amount of the amine compound used is 15-100 g, preferably 20-80 g, and more preferably 20-40 g.

[0028] According to some embodiments of the present invention, the amine compound is selected from at least one of polyethyleneimine, ethylenediamine and hexamethylenediamine, and is preferably polyethyleneimine.

[0029] Preferably, the weight average molecular weight of the polyethyleneimine is 4-6 g / mol. In the present invention, the polyethyleneimine is commercially available, for example, from Shanghai Yien Chemical Technology Co., Ltd., with product model number 9002-98-6.

[0030] According to some embodiments of the present invention, in step (1), the ultrasonic conditions include: an ultrasonic frequency of 40-80 kHz, and an ultrasonic time of 30-60 min.

[0031] According to some embodiments of the present invention, in step (2), the conditions of the amide reaction include: temperature of 50-60° C. and time of 12-15 h.

[0032] According to some embodiments of the present invention, the method further comprises pretreating the carboxylated multi-walled carbon nanotubes.

[0033] Preferably, the pretreatment method includes washing the carboxylated multi-walled carbon nanotubes with an organic alcohol and water, respectively. The number of washing times is not particularly limited and can be adjusted according to actual conditions, such as 1-3 times.

[0034] More preferably, the organic alcohol is selected from at least one of methanol, ethanol and propanol.

[0035] According to some embodiments of the present invention, the method further comprises washing the amino-functionalized carbon nanomaterial with water and drying the solid particles obtained by solid-liquid separation. Preferably, the washing times are 5-7 times.

[0036] Preferably, the solid-liquid separation method is selected from filtration and / or centrifugation, more preferably filtration.

[0037] Preferably, the pore size of the filtration membrane is 0.45-0.8 μm.

[0038] Preferably, the drying conditions include: a temperature of 50-60° C. and a drying time of 12-15 hours.

[0039] In the present invention, the water is ultrapure water; and the ethanol refers to anhydrous ethanol reagent.

[0040] According to a particularly preferred embodiment of the present invention, a method for isolating aquatic plant viruses is provided, the method comprising the following steps: (1) washing the carboxylated multi-walled carbon nanotubes with ethanol and ultrapure water for 3-5 times, respectively, adding the washed carboxylated multi-walled carbon nanotubes to ultrapure water, and ultrasonicating them at an ultrasonic frequency of 65-80 kHz for 45-60 minutes to obtain a carboxylated multi-walled carbon nanotube suspension; Wherein, based on the mass of 1g of the carboxylated multi-walled carbon nanotubes, the amount of water used is 100-200mL; Wherein, the mass ratio of the amino-functionalized carbon nanomaterial to the water containing the plant virus is 1:(100-200); (2) Adding polyethyleneimine to the above suspension, heating and stirring at 55-60°C for 13-15 hours, washing the synthesized material with ultrapure water 5-7 times after the reaction, filtering (filter membrane pore size is 0.45 μm), and drying the obtained solid particles at 55-60°C for 13-15 hours to prepare amino-functionalized carbon nanomaterials (the Zeta potential value of the amino-functionalized carbon nanomaterials is 17.5-20 mV); Wherein, based on the mass of 1g of the carboxylated multi-walled carbon nanotubes, the amount of the polyethyleneimine used is 20-30g; (3) The prepared amino-functionalized carbon nanomaterials were mixed with water containing plant viruses and stirred at room temperature (25°C) for 30-60 min at a stirring speed of 200-300 rpm; the mixed solution was filtered (the pore size of the filter membrane was 0.45-0.8 μm), and the supernatant was collected into a centrifuge tube to determine the content of pepper mild mottle virus in the water; Wherein, the mass ratio of the amino-functionalized carbon nanomaterial to the water containing the plant virus is 1:(100-500).

[0041] The present invention will be described in detail below through examples.

[0042] Carboxylated multi-walled carbon nanotubes were purchased from Shanghai MacLean Biochemical Technology Co., Ltd., product model 308068-56-6. The purity of the carboxylated multi-walled carbon nanotubes was >95%, the inner diameter (ID) was 2-5 nm, the outer diameter (OD) was <8 nm, the length was 0.5-2 μm, and the carboxyl content was approximately 3.9 wt%.

[0043] The Zeta potential value of the amino-functionalized carbon nanomaterial was measured using a Zeta potential and particle size analyzer (the instrument was purchased from Malvern, product model: Zetasizer Nano ZS).

[0044] The pore size of the amino-functionalized carbon nanomaterial was measured using an electron microscope scanner (the electron microscope scanner was purchased from JEOL Ltd., instrument model: JSM-7800F).

[0045] Polyethyleneimine was purchased from Shanghai Yien Chemical Technology Co., Ltd. with a weight-average molecular weight of 4-6 g / mol.

[0046] In the following examples, ethanol refers to anhydrous ethanol.

[0047] Example 1 (1) Carboxylated multi-walled carbon nanotubes (100 mg) were washed three times with ethanol and ultrapure water, respectively. The washed carboxylated multi-walled carbon nanotubes were added to 100 mL of ultrapure water and ultrasonicated at an ultrasonic frequency of 80 kHz for 60 min to obtain a carboxylated multi-walled carbon nanotube suspension (mass fraction 1 mg / mL). (2) 2 g of polyethyleneimine was added to the above suspension, and the mixture was heated and stirred at 60°C for 15 h. After the reaction, the synthesized material was washed 7 times with ultrapure water, filtered (with a pore size of 0.45 μm), and the solid particles obtained were dried at 60°C for 15 h to prepare amino-functionalized carbon nanomaterials (the Zeta potential value of the amino-functionalized carbon nanomaterials was 17.8 mV); The prepared amino-functionalized carbon nanomaterials were scanned by transmission electron microscopy (the results are shown in Figure 1 ). Figure 1 It can be seen that a network-like structure is formed inside the amino-functionalized carbon nanomaterial prepared in Example 1, which increases its specific surface area and is beneficial to the adsorption of plant viruses.

[0048] Example 2 (1) Carboxylated multi-walled carbon nanotubes (100 mg) were washed twice with ethanol and ultrapure water, respectively. The washed carboxylated multi-walled carbon nanotubes were added to 100 mL of ultrapure water and ultrasonicated at an ultrasonic frequency of 60 kHz for 40 min to obtain a carboxylated multi-walled carbon nanotube suspension (mass fraction 1 mg / mL). (2) 4 g of polyethyleneimine was added to the above suspension, and the mixture was heated and stirred at 50 °C for 12 h. After the reaction, the synthesized material was washed five times with ultrapure water and filtered (with a pore size of 0.8 μm). The solid particles obtained were dried at 50 °C for 12 h to prepare amino-functionalized carbon nanomaterials (the Zeta potential value of the amino-functionalized carbon nanomaterials was 20.8 mV).

[0049] Example 3 According to the method of Example 1, except that in step (2), the amount of polyethyleneimine used is 8 g, and the other steps and conditions are the same as in Example 1, amino-functionalized carbon nanomaterials are prepared (the Zeta potential value of the amino-functionalized carbon nanomaterials is 24 mV).

[0050] Example 4 The method of Example 1 was followed, except that polyethyleneimine in step (2) was replaced by ethylenediamine. The other steps and conditions were the same as those in Example 1 to prepare amino-functionalized carbon nanomaterials (the Zeta potential value of the amino-functionalized carbon nanomaterials was 12.3 mV).

[0051] Example 5 The method of Example 1 was followed, except that in step (1), the amount of ultrapure water used was 600 mL, and the other steps and conditions were the same as in Example 1, to prepare amino-functionalized carbon nanomaterials (the Zeta potential value of the amino-functionalized carbon nanomaterials was 9.2 mV).

[0052] Example 6 The method of Example 1 was followed, except that in step (1), the ultrasonic frequency was 20 kHz, and the other steps and conditions were the same as those of Example 1, to prepare amino-functionalized carbon nanomaterials (the Zeta potential value of the amino-functionalized carbon nanomaterials was 11.5 mV).

[0053] Example 7 The method of Example 1 was followed, except that in step (2), the temperature of the amide reaction was 40° C. and the time was 4 h. The other steps and conditions were the same as those of Example 1, and an amino-functionalized carbon nanomaterial (the Zeta potential value of the amino-functionalized carbon nanomaterial was 8.4 mV) was prepared.

[0054] Example 8 The method of Example 1 was followed, except that in step (2), the amount of polyethyleneimine used was 0.5 g, and the other steps and conditions were the same as in Example 1, to prepare amino-functionalized carbon nanomaterials (the Zeta potential value of the amino-functionalized carbon nanomaterials was 9.7 mV).

[0055] Example 9 According to the method of Example 1, except that in step (2), the amount of polyethyleneimine used is 1 g, and the other steps and conditions are the same as in Example 1, amino-functionalized carbon nanomaterials are prepared (the Zeta potential value of the amino-functionalized carbon nanomaterials is 11.8 mV).

[0056] Example 10 The method of Example 1 was followed, except that in step (2), the amount of polyethyleneimine used was 16 g, and the other steps and conditions were the same as in Example 1, to prepare amino-functionalized carbon nanomaterials (the Zeta potential value of the amino-functionalized carbon nanomaterials was 50.7 mV).

[0057] Example 11 The method of Example 1 was followed, except that in step (2), the amount of polyethyleneimine used was 32 g, and the other steps and conditions were the same as in Example 1, to prepare amino-functionalized carbon nanomaterials (the Zeta potential value of the amino-functionalized carbon nanomaterials was 57.7 mV).

[0058] Comparative Example 1 The method of Example 1 was followed, except that the polyethyleneimine in step (2) was replaced by ascorbic acid. The other steps and conditions were the same as those in Example 1, to prepare a hydroxyl-functionalized carbon nanomaterial (the Zeta potential value of the hydroxyl-functionalized carbon nanomaterial was -12 mV).

[0059] Test Case The amino-functionalized carbon nanomaterials prepared in the above examples and comparative examples were added to water containing plant viruses (prepared in-house), and the removal rate of pepper mild mottle virus (PMMoV) in the water was measured.

[0060] The method for preparing water containing plant viruses includes: (1) Take 300 g of diseased leaves infected with pepper mild mottle virus, grind them with liquid nitrogen, add an equal amount of 0.02 mol / LPB (pH = 7.4) buffer, homogenize, filter through 3 layers of gauze or centrifuge at low speed for 5 minutes, and collect the supernatant; (2) Add 8% (w / v) n-butanol to the supernatant, stir magnetically for 20 min, clarify, centrifuge at 4000 rpm, 4°C for 20 min, and collect the supernatant; (3) Add NaCl and polyethylene glycol 6000 to the supernatant to a final concentration of 5.8% and 4% (w / v), respectively. Stir at 4°C to fully dissolve, place in a refrigerator at 4°C overnight, centrifuge at 4000 rpm, 4°C for 30 min, and collect the precipitate. (4) The precipitate was fully suspended in 50 mL of 0.01 mol / L PB (pH = 7.4) buffer, stirred at 4°C to fully dissolve, centrifuged at 4000 rpm at 4°C for 20 min, and the supernatant was collected; (5) Add 20% (w / v) sucrose cushion, centrifuge at 36,000 rpm, 4°C for 1.5 h, and collect the precipitate; resuspend the precipitate in 4 mL of 0.01 mol / L PB (pH = 7.4) buffer; (6) Use a 30% (w / v) sucrose linear gradient centrifugation column, centrifuge horizontally at 25,000 rpm and 4°C for 2 h; place each 1 mL into a centrifuge tube and measure OD260 / 280 using a UV spectrophotometer; (7) Collect the virus-rich fraction, centrifuge at 36,000 rpm, 4°C for 2 h, and collect the precipitate; (8) Add 0.01 mol / L PB (pH = 7.4) buffer to dissolve the precipitate, centrifuge at 10,000 rpm and 4°C for 20 min, take the supernatant (i.e., the purified PMMoV solution), and store at -20°C for later use.

[0061] The preparation method for a sucrose linear gradient centrifugal column includes: preparing 30%, 20%, and 10% (w / v) sucrose solutions using 0.02 mol / LPB (pH 7.4). Then, using a syringe, draw up 10 mL of the 10% (w / v) sucrose solution and inject it along the wall of the centrifuge tube. Then, draw up 10 mL of the 20% (w / v) sucrose solution and gently inject it along the wall of the centrifuge tube underneath the 10% (w / v) sucrose cushion. Similarly, draw up 10 mL of the 30% (w / v) sucrose solution and gently inject it along the wall of the centrifuge tube underneath the 20% (w / v) sucrose cushion. The column is then allowed to equilibrate in a refrigerator at 4°C for 24 hours to form a natural sucrose gradient column with a 30%-10% (w / v) sucrose gradient from bottom to top.

[0062] Method for determining the removal rate of pepper mild mottle virus in water: 100 mg of amino-functionalized carbon nanomaterials was added to 50 mL of water containing plant viruses (the content of pepper mild mottle virus was 41 ng / L), and stirred at room temperature (25°C) (stirring speed was 200 rpm). After stirring for 30 min and 60 min, the mixed solution was filtered (the pore size of the filter membrane was 0.8 μm), and then 1 mL of the supernatant was taken into a centrifuge tube. The content of pepper mild mottle virus in the water was determined according to the following detection method, and the removal rate of pepper mild mottle virus in the water was calculated according to the following formula. Three replicates were set for each treatment. The measurement results are shown in Table 1.

[0063] The content of pepper mild mottle virus in water was determined by enzyme-linked immunosorbent assay (ELISA).

[0064] The removal rate of pepper mild mottle virus in water (%) = 100% × (the content of pepper mild mottle virus in the original water - the pepper mild mottle virus in the treated water) / (the content of pepper mild mottle virus in the original water).

[0065] Table 1

[0066] By comparing Examples 1-4 with Examples 5-11, it can be seen that the preferred method of the present invention can further improve the adsorption capacity of amino-functionalized carbon nanomaterials for pepper mild mottle virus.

[0067] Compared with Example 1, in Comparative Example 1, polyethyleneimine is replaced with ascorbic acid, and the potential value of the prepared hydroxyl-functionalized carbon nanomaterial drops sharply, resulting in a significant decrease in the adsorption capacity of the hydroxyl-functionalized carbon nanomaterial for pepper mild mottle virus.

[0068] In summary, the amino-functionalized carbon nanomaterial prepared by the embodiment of the present invention has a high removal effect on pepper mild mottle virus in water.

[0069] The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited thereto. Within the technical concept of the present invention, various simple variations of the technical solution of the present invention may be made, including combining the various technical features in any other appropriate manner. These simple variations and combinations should also be regarded as disclosed in the present invention and fall within the scope of protection of the present invention.

Claims

1. A method for separating aquatic plant viruses, characterized in that: The method includes mixing amino-functionalized carbon nanomaterials and water containing plant viruses.

2. The method according to claim 1, wherein The plant virus is selected from at least one of pepper mild mottle virus (PMMoV), tobacco mosaic virus (TMV), cucumber mosaic virus (CMV) and potato virus Y (PVY virus). Preferably, the plant virus is pepper mild mottle virus. And / or, the Zeta potential value of the amino-functionalized carbon nanomaterial is 12-25 mV, preferably 17-24 mV, more preferably 17.5-21 mV.

3. The method according to claim 1 or 2, wherein: The mass ratio of the amino-functionalized carbon nanomaterial to the water containing the plant virus is 1:(100-500); Preferably, the mixing conditions include: a stirring speed of 200-300 rpm and a time of 30-60 min.

4. The method according to any one of claims 1 to 3, wherein: The preparation method of the amino-functionalized carbon nanomaterial comprises the following steps: (1) mixing carboxylated multi-walled carbon nanotubes and water and performing ultrasonication to obtain a carboxylated multi-walled carbon nanotube suspension; (2) The carboxylated multi-walled carbon nanotube suspension and an amine compound are mixed to undergo an amide reaction to obtain amino-functionalized carbon nanomaterials.

5. The method according to claim 4, wherein In step (1), based on the mass of 1 g of the carboxylated multi-walled carbon nanotubes, the amount of water used is 100-1000 mL, preferably 200-500 mL; And / or, in step (2), based on the mass of 1g of the carboxylated multi-walled carbon nanotubes, the amount of the amine compound used is 15-100g, preferably 20-80g, and more preferably 20-40g.

6. The method according to claim 4 or 5, wherein: The amine compound is selected from at least one of polyethyleneimine, ethylenediamine and hexamethylenediamine, preferably polyethyleneimine; Preferably, the weight average molecular weight of the polyethyleneimine is 4-6 g / mol.

7. The method according to any one of claims 4 to 6, wherein: In step (1), the ultrasonic conditions include: ultrasonic frequency of 40-80 kHz, ultrasonic time of 30-60 min; And / or, in step (2), the conditions of the amide reaction include: temperature of 50-60° C. and time of 12-15 h.

8. The method according to any one of claims 4 to 7, wherein: The method further comprises pretreating the carboxylated multi-walled carbon nanotubes; Preferably, the pretreatment method includes: washing the carboxylated multi-walled carbon nanotubes with organic alcohol and water respectively; More preferably, the organic alcohol is selected from at least one of methanol, ethanol and propanol.

9. The method according to any one of claims 4 to 8, wherein: The method further comprises washing the amino-functionalized carbon nanomaterial with water, and drying the solid particles obtained by solid-liquid separation; Preferably, the solid-liquid separation method is selected from filtration and / or centrifugation; Preferably, the drying conditions include: a temperature of 50-60° C. and a drying time of 12-15 hours.

10. Use of the method according to any one of claims 1 to 9 in isolating aquatic plant viruses.

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