CN112753559B

CN112753559B - Bioreactor for culturing hairy roots

Info

Publication number: CN112753559B
Application number: CN202110008591.7A
Authority: CN
Inventors: 黄丽萍; 郭洁茹; 刘力宁; 矫金航; 李帅华; 喻敏
Original assignee: Foshan University
Current assignee: Foshan University
Priority date: 2021-01-05
Filing date: 2021-01-05
Publication date: 2022-06-17
Anticipated expiration: 2041-01-05
Also published as: CN112753559A

Abstract

The invention discloses a bioreactor for culturing hairy roots, which comprises a box body, an air inlet hole, a first exhaust hole, an object carrying piece and a spraying mechanism, wherein the air inlet hole, the first exhaust hole, the object carrying piece and the spraying mechanism are arranged on the box body, a plurality of meshes are arranged on the object carrying piece, the object carrying piece is arranged in a curved surface mode, and the vertical section of the object carrying piece is in a conical shape. According to the invention, plants are placed on the object carrying member through the meshes, the vertical section of the object carrying member is conical, so that a large enough growth interval exists between adjacent hairy roots, the condition that the hairy roots are agglomerated is reduced, water drops formed by the spraying mechanism for a period of time can flow down along the surface of the object carrying member, the condition that the hairy roots are soaked in the nutrient solution for a long time is reduced, and the normal growth and development of the hairy roots are facilitated.

Description

Bioreactor for hairy root culture

Technical Field

The invention relates to a bioreactor, in particular to a bioreactor for hairy root culture.

Background

The hairy root is a pathological phenomenon generated by the infection of a whole plant or a certain organ, tissue, single cell or even protoplast by agrobacterium rhizogenes, is a phenotype obtained by inserting T-DNA of plasmid in a cell into a nuclear genome of a host cell, mainly can generate a large amount of by-products, namely the hairy root, on or near an infected part, and can show the synthetic capacity of secondary metabolites under the condition of in vitro culture.

At present, the cultivation of the hairy roots can use a cultivation net in an atomization bioreactor to adopt a planar design, and the phenomenon that the hairy roots are gathered into a conglobate is increased because the self spraying of a sprayer is uneven and easily aggregated into water drops, so that the internal hairy roots are aged and accelerated and even die, and the cultivation of the hairy roots is finally influenced.

Disclosure of Invention

The technical problem to be solved by the invention is as follows: to solve one or more of the problems of the prior art and to provide at least one advantageous alternative or creation condition.

The solution of the invention for solving the technical problem is as follows:

the utility model provides a bioreactor is cultivateed to hairy root, includes the box, set up in inlet port, first exhaust hole, year article and the spraying mechanism of box, be provided with a plurality of meshs on carrying the article, it is the curved surface setting to carry the article, the vertical cross-section who carries the article is the toper.

The invention has the beneficial effects that: the plants are placed on the object carrying bodies through the meshes, the vertical sections of the object carrying bodies are conical, a large enough growth interval exists between the adjacent hairy roots, the conglobation situation of the hairy roots is reduced, water drops formed by the spraying mechanism for a period of time can flow down along with the surfaces of the object carrying bodies, the situation that the hairy roots are soaked in the nutrient solution for a long time is reduced, and the normal growth and development of the hairy roots are facilitated.

As a further improvement of the technical scheme, the spraying mechanism comprises a nutrient solution input pipe and a spraying head, the nutrient solution input pipe is arranged in the box body, the spraying head is arranged on the bottom surface of the object carrier, and the nutrient solution input pipe and the spraying head are communicated with each other.

The plant inserts through the through-hole and locates carrying the article, corresponds the growth of hairy root towards keeping away from the direction growth that carries the thing piece bottom surface, connects and to carry the atomising head of thing piece bottom surface can be more accurate spout nutrient solution to hairy root, and thereby reduces the nutrient solution and gathers the condition that soaks hairy root, helps the normal growth and development of hairy root.

As a further improvement of the above technical solution, the highest point of the bottom surface of the object carrying piece is located at the center of the bottom surface of the object carrying piece, and the spray head is arranged at the highest point of the bottom surface of the object carrying piece.

Through the connecting position who sets up the atomising head to the spraying range of injecing the spraying process, it is more even to the spraying of the corresponding hairy root in position on the same thing spare of carrying.

As a further improvement of the technical scheme, the object carrying body is provided with a plurality of objects along the vertical direction, the object carrying body is detachably connected with the box body, the spraying mechanism is provided with a plurality of objects, and the object carrying bodies are in one-to-one correspondence with the spraying mechanisms.

The plant is placed and can put into the box with it at once after the thing net that carries and cultivates, and greatly reduced plant exposes in external situation, and spraying mechanism and carry thing piece one-to-one, reduces the condition that hairy root is desiccated, helps the normal growth development of hairy root, and a plurality of year objects can the rational utilization space in the box.

As a further improvement of the technical scheme, a nutrient solution inlet pipe is arranged in the box body and detachably connected with the spraying mechanism.

The nutrient solution input amount of the plurality of spraying mechanisms can be directly adjusted by controlling the nutrient solution input amount of the nutrient solution liquid inlet pipe, and the operation is convenient.

As a further improvement of the above technical solution, a plurality of guide rails are arranged in the box body and are evenly arranged along the circumferential direction of the box body, a plurality of pulleys are arranged on the side of the object carrying member and are slidably connected to the guide rails, and a plurality of limiting grooves are arranged on the guide rails along the up-down direction and are clamped in the corresponding limiting grooves corresponding to the pulleys.

Through the mutual cooperation of guide rail and pulley, be convenient for put into the box with carrying the thing piece.

As a further improvement of the technical scheme, the top surface of the object carrying piece is provided with a plurality of drainage catheters which are arranged obliquely downwards.

The drainage catheter can collect water drops formed by spray aggregation of the spray mechanism above the drainage catheter, and the effect of convenient collection is achieved.

As a further improvement of the technical scheme, a space is reserved between the outer side edge of the object carrying piece and the inner side wall of the box body, a nutrient solution storage box is arranged in the box body and located below the object carrying piece, and a liquid outlet is formed in the nutrient solution storage box.

Water drops formed by spray aggregation of the spraying mechanism can be guided into the nutrient solution storage box through the overload objects and the inner side wall of the box body, so that the condition that the nutrient solution aggregates to soak the hairy roots is reduced, and the normal growth and development of the hairy roots are facilitated.

As a further improvement of the technical scheme, a filter screen is arranged in the box body and is positioned between the object carrying object and the nutrient solution storage box.

Particle impurities are removed through the filter screen, so that the purity of the nutrient solution in the nutrient solution storage tank is higher.

As a further improvement of the technical scheme, the inner side wall of the box body is provided with a heat insulation layer, a temperature measuring mechanism is arranged in the box body, a pH measuring mechanism is arranged in the nutrient solution storage box, an electronic display screen is arranged outside the box body, and the electronic display screen is connected with the temperature measuring mechanism and the pH measuring mechanism.

Temperature measurement mechanism and pH survey mechanism feed back the reading to electronic display screen on, the condition in the worker observation box of being convenient for can in time adjust the temperature that is fit for hairy root growth, guarantees hairy root's normal growth and development.

Drawings

In order to more clearly illustrate the technical solution in the embodiments of the present invention, the drawings used in the description of the embodiments will be briefly described below. It is clear that the described figures are only some embodiments of the invention, not all embodiments, and that a person skilled in the art can also derive other designs and figures from them without inventive effort.

FIG. 1 is a sectional view of a hairy root culture bioreactor of the present invention taken along a vertical section;

FIG. 2 is a cross-sectional view of a hairy root culture bioreactor of the present invention taken along a transverse section.

In the figure, 1, a box body; 11. a box door; 12. an air inlet; 13. a first exhaust port; 14. a second vent hole; 15. a guide rail; 16. a filter screen; 17. a nutrient solution storage tank; 18. a liquid discharge port; 2. carrying an object; 21. a drainage catheter; 3. a spraying mechanism; 31. a nutrient solution infusion tube; 32. a spray head; 33. a nutrient solution inlet pipe; 4. a limiting mechanism; 41. connecting columns; 42. a pulley; 5. a temperature measuring mechanism; 6. a pH measuring mechanism; 7. an electronic display screen.

Detailed Description

The conception, the specific structure, and the technical effects produced by the present invention will be clearly and completely described below in conjunction with the embodiments and the accompanying drawings to fully understand the objects, the features, and the effects of the present invention. It is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments, and those skilled in the art can obtain other embodiments without inventive effort based on the embodiments of the present invention, and all embodiments are within the protection scope of the present invention. In addition, all the coupling/connection relationships mentioned herein do not mean that the components are directly connected, but mean that a better coupling structure can be formed by adding or reducing coupling accessories according to specific implementation conditions. All technical characteristics in the invention can be interactively combined on the premise of not conflicting with each other.

Referring to FIGS. 1 to 2, the following examples are made of the hairy root culture bioreactor of the present invention:

including box 1, box 1 is vertical setting, and the top of box 1 is provided with chamber door 11, and 11 one end of chamber door is connected through the pivot with box 1, and the rotatable angle scope of 11 rotatable angles of chamber door is 120 to 180.

Be provided with inlet port 12 and first exhaust hole 13 on the box 1, inlet port 12 and first exhaust hole 13 are located box 1 top and adjacent setting, it is concrete, be provided with the filter on the box 1 and correspond inlet port 12 department, can guarantee under the sufficient circumstances of oxygen suppliment in keeping box 1 that the vegetation environment is sterile state, and the bottom that is close to box 1 on the box 1 is provided with second exhaust hole 14, second exhaust hole 14 and first exhaust hole 13 set up relatively, thereby guarantee that the interior upper and lower part of box 1 internal gas homoenergetic realizes discharging, it is provided with one-way gas filter screen to correspond first exhaust hole 13 and second exhaust hole 14 department on the box 1, can prevent harmful gas in the inflow air, thereby can release the gaseous pressure that reaches stable in the box 1 simultaneously.

Set up in the box 1 and carry article 2 and spraying mechanism 3, carry the area of 2 cross sections of article the area less than or equal to box 1 open-top's cross section area, it is provided with a plurality of meshes that supply the plant to place on the article 2 to carry, it is the curved surface setting to carry article 2, the vertical cross section of carrying article 2 is the toper, the plant passes through the mesh and places in carrying article 2, the growing direction of hairy root is towards the bottom of box 1, hairy root is located the bottom surface of carrying article 2, because the vertical cross section that carries article 2 is the toper, then there is sufficient distance between the hairy root of different trunk and supplies its growth, reduce the situation that hairy root conglobation.

The spraying mechanism 3 comprises a nutrient solution input pipe 31 and a spraying head 32, the nutrient solution input pipe 31 is arranged in the box body 1, the spraying head 32 is arranged on the bottom surface of the carrier 2, the nutrient solution input pipe 31 is communicated with the spraying head 32, in order to enable the spraying range to be more uniform, the highest point of the bottom surface of the carrier 2 is located at the center of the bottom surface of the carrier 2, the spraying head 32 is arranged at the highest point of the bottom surface of the carrier 2, the spraying range of the spraying head 32 is circular, and nutrient solution sprayed by the spraying head 32 is directly sprayed to hairy roots.

Carry article 2 and be provided with a plurality of along upper and lower direction, a plurality of carries 2 equidistance distributions of article, carries article 2 and can dismantle with box 1 and be connected, and spraying mechanism 3 is provided with a plurality of, and a plurality of carries article 2 and a plurality of spraying mechanism 3 one-to-one, and then the hairy root on every carries article 2 all not influenced by adjacent spraying mechanism 3.

A nutrient solution inlet pipe 33 is arranged in the box body 1, and the nutrient solution inlet pipe 33 is detachably connected with the nutrient solution input pipe 31.

A plurality of guide rails 15 are arranged in the box body 1, the guide rails 15 are uniformly arranged along the circumferential direction of the box body 1, the side edge of the object carrying body 2 is connected with pulleys 42 through connecting columns 41, the number of the pulleys 42 on the same object carrying body 2 is the same as that of the guide rails 15, the pulleys 42 are connected on the corresponding guide rails 15 in a sliding way, a plurality of limiting grooves are arranged along the up-and-down direction of the same guide rail 15 and are horizontally arranged, the limiting grooves are communicated with the guide rails 15, the side surfaces of the limiting grooves, which are close to the object carrying body 2, are communicated with the inside of the box body 1, the corresponding pulleys 42 are clamped in the corresponding limiting grooves, specifically, the object carrier 2 needs to rotate, the nutrient solution input pipe 31 is arranged as a corrugated hose, when the object carrying member 2 slides to a proper position along the up-down direction, the object carrying member 2 is rotated to enable the pulley 42 on the object carrying member 2 to slide into the limiting groove, so that the height of the object carrying member 2 in the vertical direction is limited.

Carry and leave the interval between the outside limit of article 2 and the inside wall of box 1, when spraying mechanism 3 sprayed the plant, unnecessary nutrient solution can flow down along the top surface that carries article 2, thereby prevent that the nutrient solution from stopping the situation of soaking the plant, the top surface that carries article 2 is provided with a plurality of drainage pipe 21, drainage pipe 21 is the slope and sets up downwards, be provided with nutrient solution storage box 17 in the box 1, nutrient solution storage box 17 is located the below of carrying article 2 of lower, nutrient solution storage box 17 is provided with leakage fluid dram 18, leakage fluid dram 18 and box 1 intercommunication. A filter screen 16 is arranged in the box body 1, and the filter screen 16 is positioned between the lowest object carrying part 2 and a nutrient solution storage tank 17.

The inside wall of box 1 is provided with the heat preservation, box 1 outer joint has electronic display screen 7, be provided with temperature measurement mechanism 5 in the box 1, electronic display screen 7 is connected with temperature measurement mechanism 5, read the temperature in the box 1 through temperature measurement mechanism 5, real-time temperature reaches electronic display screen 7, and temperature measurement mechanism 5 can change the temperature in the box 1, nutrient solution storage box 17 is provided with pH survey mechanism 6, pH survey mechanism 6 is connected with electronic display screen 7, feed back the supply volume and the supply quality of nutrient solution through pH survey mechanism 6.

When the bioreactor for cultivating the hairy roots is used, the temperature measuring mechanism 5 is adjusted according to the characteristics of plants, so that the proper temperature is kept in the box body 1, the plants are placed on the object carrying member 2 through the through holes, the top surface of the object carrying member 2 has certain gradient due to the particularity of the shape of the object carrying member 2, the sufficient distance exists between the hairy roots of adjacent plants of the same object carrying member 2, the conglobation of the hairy roots is reduced, the space for growing the hairy roots is large enough, the dissolved oxygen is sufficient, the growth and development of the hairy roots are facilitated, the object carrying member 2 is placed in the box body 1 through the matching of the pulley 42 and the guide rail 15, the object carrying member 2 is rotated after sliding to the proper position so that the pulley 42 enters the limiting groove, the nutrient solution input pipe 31 is connected with the nutrient solution inlet pipe 33, and the spray head 32 sprays the hairy roots, if necessary, the operation is repeated, the spraying of the cultivated plants is not affected by the adjacent spraying mechanisms, the normal growth and development of the hairy roots are ensured, the water discharged from the spraying head 32 is in a mist shape, the water mist is sprayed to the hairy roots and is rarely accumulated to form a water droplet shape, and when the situation occurs, the water droplet can slide down along with the top surface of the carrier 2 or slide down along the drainage conduit 21 to the nutrient solution storage box 17 because the through holes of the carrier 2 are inserted by the plants.

While the preferred embodiments of the present invention have been illustrated and described, it will be understood by those skilled in the art that the present invention is not limited to the details of the embodiments shown and described, but is capable of numerous modifications and substitutions without departing from the spirit of the present invention and within the scope of the appended claims.

Claims

1. A bioreactor for culturing hairy roots comprises a box body (1), an air inlet (12) arranged on the box body (1), a first exhaust hole (13), a carrying piece (2) and a spraying mechanism (3), and is characterized in that the carrying piece (2) is provided with a plurality of meshes, the carrying piece (2) is arranged in a curved surface manner, and the vertical section of the carrying piece (2) is in a conical shape; the spraying mechanism (3) comprises a nutrient solution input pipe (31) and a spraying head (32), the nutrient solution input pipe (31) is arranged in the box body (1), the spraying head (32) is arranged on the bottom surface of the carrier (2), and the nutrient solution input pipe (31) and the spraying head (32) are communicated with each other; the highest point of the bottom surface of the object carrying body (2) is positioned at the center of the bottom surface of the object carrying body (2), and the spray head (32) is arranged at the highest point of the bottom surface of the object carrying body (2); the plurality of the object carrying bodies (2) are arranged along the vertical direction, the object carrying bodies (2) are detachably connected with the box body (1), the plurality of spraying mechanisms (3) are arranged, and the plurality of the object carrying bodies (2) correspond to the plurality of spraying mechanisms (3) one by one; the novel box is characterized in that a plurality of guide rails (15) are arranged in the box body (1), the guide rails (15) are evenly arranged along the circumferential direction of the box body (1), a plurality of pulleys (42) which are connected to the guide rails (15) in a sliding mode are arranged on the side edge of the carrier (2), a plurality of limiting grooves are formed in the guide rails (15) along the vertical direction, and the limiting grooves correspond to the pulleys (42) which are tightly clamped in the corresponding limiting grooves.

2. The bioreactor for cultivating hairy roots according to claim 1, wherein a nutrient solution inlet pipe (33) is arranged in the box body (1), and the nutrient solution inlet pipe (33) is detachably connected with the spraying mechanism (3).

3. A bioreactor for cultivating hairy roots according to claim 1, wherein the top surface of the carrier member (2) is provided with a plurality of drainage tubes (21), and the drainage tubes (21) are arranged obliquely downward.

4. A bioreactor for hairy root cultivation according to claim 1, wherein a distance is left between the outer side edge of the carrier (2) and the inner side wall of the tank (1), a nutrient solution storage tank (17) is arranged in the tank (1), the nutrient solution storage tank (17) is located under the carrier (2), and the nutrient solution storage tank (17) is provided with a drainage port (18).

5. A bioreactor for hairy root cultivation according to claim 4, wherein a filter screen (16) is arranged in the tank (1), said filter screen (16) being located between the carrier (2) and the nutrient solution storage tank (17).

6. The bioreactor for cultivating hairy roots according to claim 5, wherein the inner side wall of the box body (1) is provided with a heat insulation layer, the box body (1) is internally provided with a temperature measuring mechanism (5), the nutrient solution storage tank (17) is provided with a pH measuring mechanism (6), the box body (1) is externally provided with an electronic display screen (7), and the electronic display screen (7) is connected with the temperature measuring mechanism (5) and the pH measuring mechanism (6).