CN213219493U - Plant pathogenic bacteria separation surface disinfection device - Google Patents

Abstract

The utility model discloses a plant pathogen separating surface sterilizing device, which comprises a carrying net and a cleaning device, wherein the cleaning device comprises a cleaning chamber and a waste liquid chamber, and the cleaning chamber and the waste liquid chamber are of an upper structure and a lower structure; the object carrying net comprises an object carrying frame and a net disc, the net disc is connected with the object carrying frame through a fixing buckle, a buffering device and a stirring device are arranged inside the cleaning chamber, the buffering device comprises a bearing table, a spring and a base, two ends of the spring are respectively connected with the bearing table and the base, a liquid outlet is arranged at the bottom of the waste liquid chamber, and the liquid outlet is further provided with a liquid outlet plug matched with the liquid outlet plug. The utility model discloses rational in infrastructure, the simple operation can accurate control disinfection time, avoids reagent to pollute, and guarantee test operation personnel's safety improves work efficiency.

Description

Plant pathogenic bacteria separation surface disinfection device

Technical Field

The utility model belongs to the technical field of plant protection science plant pathogenic bacteria separation and purification, concretely relates to plant pathogenic bacteria separation surface degassing unit.

Background

Plant diseases have great influence on agriculture, forestry and animal husbandry, and not only can directly influence the normal growth and development of plants, but also indirectly influence the yield and quality of products. When an unfamiliar or new disease is found, the disease is characterized clearly and can be diagnosed or identified directly, but in many cases it is difficult to identify the genus or species of the pathogenic bacterium. The Koch's Rule is a procedure for determining the etiology of invasive diseases, which states: the presence of a pathogenic microorganism is often associated with the pathogenic plants; the microorganism can be separated and purified on an isolated or artificial culture medium to obtain pure culture; inoculating pure culture to healthy plants of the same variety, and generating diseases with the same symptoms; the plants which are inoculated and diseased are separated into pure cultures, and the characters of the pure cultures are the same as those of the inocula.

After a plant disease tissue sample is collected in a laboratory, pathogenic bacteria are often separated and identified in order to diagnose the disease. In order to successfully isolate pathogenic bacteria, the surfaces of the diseased tissue need to be disinfected. Laboratory plant surface disinfection is a chemical disinfection method, which utilizes various chemical reagents, and has various steps and great time consumption. When large-batch surface disinfection is carried out, plant tissues are easy to float and accumulate in a disinfection container, and the disinfection time cannot be accurately controlled. The disinfection time directly influences whether pathogenic bacteria can be successfully separated, the disinfection time is too short, the disinfection is not thorough, and the mixed bacteria pollution can be caused; over time, the chemical agents can cause corrosive damage to the plant tissue. After the disinfection is finished, all plant tissues need to be separated from the disinfection vessel, the efficiency is low, and the plant tissues are easily damaged and polluted in the plant tissue separation process.

There are also some reports at present, for example, CN2017214708880 discloses a device for ultrasonic disinfection of surfaces of plant diseased tissues, and CN2018213451741 a device for separation and culture of plant pathogenic bacteria, the disclosed device has a relatively complex structure, cannot accurately control disinfection time, can only disinfect with one reagent in large batches, and needs to be replaced repeatedly even if the same reagent is used for disinfection. Therefore, how to rapidly sterilize plant tissues without damage and complete separation and purification of plant pathogenic bacteria is a problem to be solved at present.

SUMMERY OF THE UTILITY MODEL

The technical problem solved by the utility model is to provide a plant pathogenic bacteria separation surface degassing unit, be applicable to the surface disinfection of various plant tissues, have safety, efficient advantage.

Not enough to prior art exists, the utility model aims to provide a plant pathogenic bacteria separation surface degassing unit carries thing net, purge chamber, waste liquid chamber through setting up, and each structure combines closely, and accurate control disinfection time high efficiency is disinfected. The plant pathogenic bacteria separation surface disinfection device is operated integrally, so that reagent pollution can be avoided, and the safety of test operators is guaranteed; in addition, the device is simple in overall structure and convenient to operate, can be repeatedly used for many times, and reduces the test cost.

The utility model provides a technical scheme that its technical problem adopted is: a plant pathogenic bacteria separation surface disinfection device comprises a carrying net and a cleaning device, wherein the cleaning device comprises a cleaning chamber and a waste liquid chamber, and the cleaning chamber and the waste liquid chamber are of an upper structure and a lower structure;

the net disc comprises a buckling net and a base disc, the buckling net is connected with the base disc through a hinge, the base disc comprises a spacer rod and a positioning ring, and the positioning ring is sunken on the spacer rod;

the cleaning device comprises a cleaning chamber, a washing chamber and a cleaning device, wherein the cleaning chamber is internally provided with a buffer device and a stirring device, the buffer device comprises a bearing table, a spring and a base, two ends of the spring are respectively connected with the bearing table and the base, and the stirring device is a blade disc and is arranged between the bearing table and the base;

a liquid outlet is formed in the bottom of the waste liquid chamber, and a matched liquid outlet plug is further arranged at the liquid outlet;

furthermore, the buckling nets are divided into 20 small round buckling nets, and the number of the buckling nets is 100 meshes;

furthermore, the spacer bars are divided into transverse spacer bars and longitudinal spacer bars, and the transverse spacer bars and the longitudinal spacer bars are vertically arranged;

furthermore, the positioning ring is circular, and the diameter of the positioning ring is 6 mm;

furthermore, the net disc comprises four fastening nets and a base plate, and the hinges are fixed in series by a fixed shaft;

furthermore, the number of the springs is three, and the maximum extension range of each spring is 4-10 cm;

further, the blade disc is a three-blade;

furthermore, the inner shafts of the supporting columns and the blades are of spiral structures, and the directions of spiral threads are consistent;

further, the base is in a circular shape with a concave middle part;

further, a liquid separating plug is arranged at the joint of the cleaning chamber and the waste liquid chamber.

The beneficial effects of the utility model reside in that:

1) the device has simple integral structure, convenient operation and repeated use;

2) the device can sterilize plant tissues in batches, the plant tissues cannot float and accumulate, and the sterilization is thorough;

3) the device can accurately control the disinfection time, efficiently disinfect under the condition of not damaging plant tissues, and is time-saving and labor-saving;

4) the device is rational in infrastructure, and the operation integration avoids reagent to pollute, can ensure test operating personnel's safety.

Drawings

FIG. 1 is a schematic structural view of a plant pathogen separating surface sterilizing device of the present invention;

FIG. 2 is a schematic view of a partial structure of a carrier net of the plant pathogen separation surface disinfection device of the present invention;

FIG. 3 is a schematic view of a partial structure of a mesh plate of the plant pathogen separation surface disinfection device of the present invention;

FIG. 4 is a schematic view of a fixed shaft of the plant pathogen separation surface disinfection device of the present invention;

FIG. 5 is a schematic view of a partial structure of a hinge of the plant pathogen separation surface disinfection device of the present invention;

FIG. 6 is a schematic view of a partial structure of a stirring device of the plant pathogen separation surface disinfection device of the present invention;

FIG. 7 is a partial structure diagram of a blade disc of the surface sterilizer for separating plant pathogenic bacteria of the present invention;

in the figure:

1-carrying net, 11-air bag chamber, 12-supporting column, 121-screw thread, 13-fastening net, 14-chassis, 141-positioning ring, 142-transverse spacer bar, 143-longitudinal spacer bar, 15-hinge, 151-lower leaf plate, 152-upper leaf plate, 153-rotating shaft, 16-fixed shaft, 17-fixed buckle, 2-cleaning chamber, 21-bearing table, 22-blade disc, 221-blade inner shaft, 222-blade, 23-spring, 24-base, 25-separating plug, 3-waste liquid chamber, 31-liquid outlet, and 32-liquid outlet plug.

Detailed Description

In order to make the technical solutions and advantages of the present invention more clearly understood, the technical solutions in the embodiments of the present invention are described below in detail and completely with reference to the accompanying drawings in the specific embodiments, and it is obvious that the described embodiments are some, but not all embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by a person of ordinary skill in the art without any inventive step belong to the protection scope of the present invention. The invention will be further explained with reference to the following figures and examples:

as shown in figures 1-7, the plant pathogen separation surface disinfection device comprises a carrying net 1 and a cleaning device, wherein the cleaning device comprises a cleaning chamber 2 and a waste liquid chamber 3, the cleaning chamber 2 and the waste liquid chamber 3 are of an up-and-down structure, disinfection operation is integrated, contact between an operator and a reagent is reduced, and pollution of the reagent is avoided.

Carry thing net 1 and include carrier, net dish, the net dish passes through fixed buckle 17 with the carrier and is connected, and the carrier includes gasbag room 11, support column 12, and gasbag room 11 links to each other with support column 12 upper end, presses gasbag room top, and the support column can reciprocate. The net plate comprises four fastening nets 13 and a base plate 14, the fastening nets 13 are connected with the base plate 14 through hinges 15, a plurality of hinges 15 are fixed in series through a fixing shaft 16, a lower leaf plate 151 of each hinge 15 is connected with the fastening net 13, an upper leaf plate 152 is connected with the base plate 14, the fastening nets 13 and the base plate 14 can be opened up and down through rotating shafts on the hinges 15, the base plate 14 comprises spacer rods and positioning rings 141, the positioning rings 141 are sunken on the spacer rods, the spacer rods comprise transverse spacer rods 142 and longitudinal spacer rods 143, the transverse spacer rods 142 and the longitudinal spacer rods 143 are vertically arranged, a better supporting effect can be achieved, the positioning rings 141 are circular, the diameter of the positioning rings is 6mm, the fastening nets 13 are divided into 20 small circular fastening nets, the number of the fastening nets is 100 meshes, plant tissue (5mm) rings can be placed in the positioning rings, the fastening nets are covered, a certain space is reserved, and plant tissue blades are prevented from floating and accumulating on the surface.

A buffer device and a stirring device are arranged in the cleaning chamber 2, the buffer device comprises a bearing table 21, a spring 23 and a base 24, the base is in a circular shape with a concave middle, and the adsorption force between the base and the bottom of the cleaning chamber can be improved, so that the stability is improved; in addition, two ends of a spring 23 are respectively connected with the bearing table 21 and the base 24, the number of the springs is three, the three springs can form a stable triangle, the stability of each device is further improved, and the maximum extension range of each spring is 4-10 cm; agitating unit is blade dish 22, sets up in the middle of bearing platform and base, and blade dish 22 is three leaf blades, and support column 12, the interior axle 221 of blade are helical structure, and the spiral line direction is unanimous, and support column 12 is rotatory from top to bottom, can drive the rotation of blade dish to make the inside washing liquid of purge chamber constantly flow, disinfect to plant tissue under the circumstances of irrotational plant tissue not damaged ground, can accelerate the cleaning process.

After finishing washing, be provided with at purge chamber and waste liquid room junction and divide liquid stopper 25, take out and divide liquid stopper 25, the washing liquid can be quick flow out from the purge chamber, conveniently use same kind of reagent many times to disinfect the operation, and 3 bottoms in waste liquid room are equipped with liquid outlet 31, and liquid outlet 31 still has a assorted play liquid stopper 32, and a reagent washs the back that finishes, can open liquid outlet 31, places the washing liquid in the fixed waste liquid recovery device.

Separating test operation of pathogenic bacteria of cucumber target spot disease:

potato Dextrose Agar (PDA) medium: 200g of potatoes, 20g of glucose, 20g of agar and 1000mL of distilled water.

Preparing materials: selecting fresh leaves with obvious cucumber target spot disease symptoms, firstly washing the surfaces of the leaves clean by flowing clear water, cutting leaf tissues with the size of about 5mm at the junction of disease and health, putting the cut leaf tissues into positioning rings in a chassis 14, putting a piece of leaf tissue in each positioning ring, covering a fastening net 13 after the leaf tissues are placed, fixing the chassis 14 and the fastening net 13 together through a fixed shaft 15, and fixing a net disc on a carrier frame by using a fixed buckle 17 for later use.

And (3) disinfection: taking out the cleaning device, closing the liquid separating plug 25, covering the liquid outlet plug 32, injecting 70% alcohol into the cleaning chamber, enabling the page not to exceed the maximum extension height of the spring 23, placing the carrying net on the bearing table 21, pressing the carrying net downwards by hands, immersing the carrying net in the 70% alcohol, simultaneously pressing the air bag chamber 11 upwards and downwards by a thumb, driving the paddle disc 22 to rotate, enabling the 70% alcohol to flow, after 30s of disinfection, releasing the hands, enabling the spring 23 to rapidly eject the whole carrying net 1, opening the liquid separating plug 25 and the liquid outlet plug 32, and pouring 70% alcohol solution into a fixed waste liquid recovery device.

The above procedure was followed by sterilizing with a 70% alcohol solution replaced with a 0.1% mercury solution for 3 min.

And then changing the disinfectant into sterile water, and cleaning for 3-5 times, wherein different operations are that after each time of cleaning with the sterile water, the liquid separating plug 25 is directly opened to place the sterile water into the waste liquid chamber, the liquid separating plug 32 is intensively opened after multiple times of cleaning, and the cleaning liquid is poured into a fixed waste liquid recovery device.

After the washing is finished, leaf tissues are dried in a net disc and then placed on a PDA culture medium to be cultured in an incubator at 28 ℃.

And 3d, picking new growing colony edge hyphae, and transferring to a new PDA culture medium for multiple times of purification culture.

And (3) strawberry pathogen separation identification surface disinfection operation:

potato dextrose agar medium for use.

Preparing materials: cutting a tissue cut block with the side length of 5mm at the diseased and healthy junction of the strawberry, putting the cut tissue into positioning rings in a chassis 14, placing one tissue cut block in each positioning ring, covering a buckling net 13 after placing, fixing the chassis 14 and the buckling net 13 together through a fixed shaft 15, and fixing a net disc on a carrier by using a fixed buckle 17 for later use.

And (3) disinfection: taking out the cleaning device, closing the liquid separating plug 25, covering the liquid outlet plug 32, injecting 70% alcohol into the cleaning chamber, placing the object carrying net on the bearing table 21, pressing the object carrying net downwards by hands, immersing the object carrying net in the 70% alcohol, simultaneously pressing the air bag chamber 11 upwards and downwards by a thumb, driving the blade disc 22 to rotate, enabling the 70% alcohol to flow, after 1min of disinfection, releasing the hands, enabling the spring 23 to rapidly eject the whole object carrying net 1, opening the liquid separating plug 25 and the liquid outlet plug 32, and pouring 70% alcohol solution into the fixed waste liquid recovery device.

The above operation is followed, and the 70% alcohol solution is replaced by 2% sodium hypochlorite solution for 6 min.

Then the disinfectant is changed into sterile water to be washed for 3 times, and the different operation is that after each time of washing with the sterile water, the liquid separating plug 25 is directly opened to put the sterile water into the waste liquid chamber, after a plurality of times of washing, the liquid separating plug 32 is intensively opened, and the washing liquid is poured into a fixed waste liquid recovery device.

After cleaning, cutting the tissue into blocks, airing the tissue blocks in a net disc, placing the tissue blocks on a PDA culture medium, culturing the tissue blocks in an incubator at 28 ℃ for 3-5 days, after bacteria grow out, lightly picking a small amount of spores by using an inoculating loop to perform streak culture, streaking 10 PDA plates each time, and repeating the process for 3 times.

Surface disinfection operation for separation and identification of pathogenic bacteria of cherry trees

The tested cherry tree material was harvested in 2019 in 7 months from a tobacco terrace Laiyang cherry plantation. The material is selected by adopting a five-point sampling method, and diseased plants with integrally consistent diseases are selected.

Washing collected cherry branches and leaves with tap water, wiping, cutting the branches into 3cm small segments, and cutting the leaves into 3cm by 3cm small segments.

And (3) branch:

the obtained branch or leaf tissue is placed on the spacing rods in the chassis 14 and is placed along the transverse spacing rods 142 in the forward direction, then the buckling nets 13 are covered, the chassis 14 and the buckling nets 13 are fixed together through the fixed shafts 15, and the net disc is fixed on the carrier by the fixed buckles 17 for standby.

And (3) disinfection: taking out the cleaning device, closing the liquid separating plug 25, covering the liquid discharging plug 32, injecting 70% alcohol into the cleaning chamber, placing the object carrying net on the bearing table 21, pressing down the object carrying net by hand, immersing the object carrying net in the 70% alcohol, simultaneously pressing the air bag chamber 11 up and down by thumb, driving the blade disc 22 to rotate, making the 70% alcohol flow, disinfecting for 3min (2 min for blade), releasing the hand, the spring 23 can rapidly eject the whole object carrying net 1, opening the liquid separating plug 25 and the liquid discharging plug 32, and pouring 70% alcohol solution into the fixed waste liquid recovery device.

The sterile water is washed for 4 times, and the different operation is that after each time of washing with the sterile water, the liquid separating plug 25 is directly opened to place the sterile water into the waste liquid chamber, after multiple times of washing, the liquid separating plug 32 is intensively opened, and the washing liquid is poured into a fixed waste liquid recovery device.

The above operation is performed, and the surface of the reagent is replaced by 2% sodium hypochlorite aqueous solution for disinfection for 7min (leaf blade 4 min).

The reagent was changed to 75% ethanol and sterilized for 40s as above.

The same procedure as above was followed with 5 washes of sterile water.

And (3) inoculating the treated sample on a PDA (PDA dextrose agar) plate culture medium, placing the sample in a constant-temperature incubator at 25 ℃ for inverted culture for 3-6 days, and observing the growth condition of bacterial colonies around the tissue block. After the hyphae grow out from the cut edge, picking the tip part of the hyphae newly growing out from the cut edge and transferring the tip part to a PDA culture medium, carrying out purification culture until a pure bacterial strain is obtained, and observing and recording the growth state condition of the bacterial colony in time.

When the used sterilized tissues are not consistent with the size of the positioning ring, the sterilized tissues can be fixed in the net disc through the transverse spacer bars 142, the longitudinal spacer bars 143 and the fastening net 13 for sterilization. In addition, the above examples show that different plant tissues need different disinfection reagents, the disinfection time is also strictly limited, and the test rework can be caused when the disinfection time is too short or too long. The utility model has simple structure, convenient operation, simplified use tools, accurate control of the disinfection time, high-efficiency disinfection without damaging plant tissues, time saving and labor saving; in addition, the device can be repeatedly used after being cleaned, so that the use of test consumables is reduced, and the test cost is reduced.

The above embodiments are illustrative of specific embodiments of the present invention, and should not be construed as limitations of the present invention, and the protection scope of the present invention shall be defined by the claims, including equivalents of technical features in the technical solutions described in the claims, that is, equivalent modifications in the scope are also within the protection scope of the present invention.

Claims (10)

1. A plant pathogenic bacteria separation surface disinfection device is characterized by comprising a carrying net and a cleaning device, wherein the cleaning device comprises a cleaning chamber and a waste liquid chamber, and the cleaning chamber and the waste liquid chamber are of an upper structure and a lower structure;

the net disc comprises a buckling net and a base disc, the buckling net is connected with the base disc through a hinge, the base disc comprises a spacer rod and a positioning ring, and the positioning ring is sunken on the spacer rod;

the cleaning device comprises a cleaning chamber, a washing chamber and a cleaning device, wherein the cleaning chamber is internally provided with a buffer device and a stirring device, the buffer device comprises a bearing table, a spring and a base, two ends of the spring are respectively connected with the bearing table and the base, and the stirring device is a blade disc and is arranged between the bearing table and the base;

the bottom of the waste liquid chamber is provided with a liquid outlet, and the liquid outlet is also provided with a matched liquid outlet plug.

2. A plant pathogen separation surface disinfection apparatus as claimed in claim 1, wherein said nets are divided into 20 small circular nets with 100 mesh.

3. A phytopathogen-separating surface sanitising device as claimed in claim 1, wherein the spacer bars are divided into transverse spacer bars and longitudinal spacer bars, the transverse spacer bars being arranged perpendicular to the longitudinal spacer bars.

4. A phytopathogen separating surface sanitising device as claimed in claim 1, wherein the locating ring is circular and has a diameter of 6 mm.

5. A plant pathogen isolation surface disinfection apparatus as claimed in claim 1 wherein said mesh tray includes four clasping meshes and a base tray, each hinge being secured in series by a fixed shaft.

6. The plant pathogen separation surface disinfection apparatus of claim 1, wherein there are three springs, and the maximum extension range of each spring is 4-10 cm.

7. A phytopathogen separating surface disinfection apparatus as claimed in claim 1, wherein the paddle tray is a three-bladed paddle.

8. A plant pathogen separation surface disinfection apparatus as claimed in claim 1 wherein said support post and said inner shaft of said blade are of a helical configuration and have helical veins in the same direction.

9. A plant pathogen isolation surface disinfection apparatus as claimed in claim 1, wherein said base is in the form of a circular, centrally recessed disc.

10. The plant pathogen separation surface disinfection apparatus of claim 1, wherein a liquid separation plug is disposed at a connection of the cleaning chamber and the waste liquid chamber.

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