Systems and methods for agroinfiltration for recombinant protein production

Abstract

Provided herein is an agroinfiltration system including an agroinfiltrator comprising a vacuum chamber, a top cover, vacuum and hydraulic pumps box, hydraulic controller, plant height adjustment, electric box and one or more sub-chambers to infiltrate plants; and one or more tray assemblies configured to hold soil for plant growth, wherein the agroinfiltrator is configured to receive the one or more tray assemblies.

Description

SYSTEMS AND METHODS FOR AGROINFILTRATION FOR RECOMBINANT PROTEIN PRODUCTIONCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Application Serial No. 63 / 659,904, filed June 14, 2024, the entire contents of which are incorporated herein by reference.STATEMENT OF FEDERALLY FUNDED RESEARCH

[0002] None.TECHNICAL FIELD OF THE INVENTION

[0003] The present invention relates in general to recombinant protein production and more particularly, to agroinfiltration for recombinant protein production.BACKGROUND OF THE INVENTION

[0004] Without limiting the scope of the invention, its background is described in connection with particular embodiments of the present invention that permit agroinfiltration of Nicotiana benthamiana and other leafy plants for the production of recombinant proteins.

[0005] Conventional infiltrators for agroinfiltration for the production of recombinant proteins typically use soil pots and can only infiltrate fewer plants at a time, precluding efficient mid-scale production of recombinant proteins.

[0006] One such conventional system in taught in U.S. Patent Publication No. 20140290135, filed by Carraro, et al., entitled “Method and System for the Vacuum Infiltration of Plants”. These applicants are said to teach a system for infiltrating plant tissue with macromolecules or microorganisms including an infiltration chamber having at least one sealable hatch for receiving plant tissue, means for reducing pressure within the infiltration chamber; a first release valve in fluid communication with the infiltration chamber, a controller for operating coordinately the first release valve and the pressure reducing means such that the pressure in the infiltration chamber is (i) reduced from a start pressure to a target pressure that is less than or equal to 300 mbar in a first time period, (ii) maintained at the target pressure for a second time period, and (iii) increased from the target pressure to a final pressure of at least approximately 1 bar within a third time period of less than or equal to five (5) seconds.

[0007] Despite these advances, efficient systems and methods for the mid-scale production of recombinant proteins are desirable.SUMMARY OF THE INVENTION

[0008] As embodied and broadly described herein, an aspect of the present disclosure relates to an agroinfdtration system comprising: an agroinfdtrator comprising a vacuum chamber, a top cover, and one or more sub-chambers to infdtrate plants; one or more tray assemblies configured to hold soil for plant growth, wherein the agroinfiltrator is configured to receive the one or more tray assemblies; and a vacuum inlet, a pressure gauge, and a drain pipe in communication with the vacuum chamber. In one aspect, each of the one or more tray assemblies comprises a tray top having a plurality of holes for plants and a tray bottom configured to receive the tray top. In another aspect, the agroinfiltrator comprising metal or plastic. In another aspect, the one or more tray assemblies comprise metal or plastic. In another aspect, the agroinfiltration system further comprises one or more growth chambers for post-infiltration incubation configured to receive plants infiltrated by the agroinfiltration system. In another aspect, the one or more growth chambers comprise metal or plastic. The present invention can also be used with any of the following non-limiting list of additional plant species: Nicotiana benthamiana, Medicago truncatula; Lettuce (Lactuca sativa); Arabidopsis thaliana; Nicotiana tabacunr, Spinach (Spinacia oleracea),' Zucchini (Cucurbita pepo var. cylindrical and / or Squash (Cucurbita pepo).

[0009] As embodied and broadly described herein, an aspect of the present disclosure relates to an agroinfiltration system kit, comprising: an agroinfiltrator comprising a vacuum chamber, a top cover, and one or more sub-chambers to infiltrate plants; one or more tray assemblies configured to hold soil for plant growth, wherein the agroinfiltrator is configured to receive the one or more tray assemblies; and one or more fasteners or tools or both for assembling the agroinfiltrator and the one or more tray assemblies. In one aspect, the agroinfiltrator comprises a vacuum inlet, a pressure gauge, and a drainpipe. In another aspect, each of the one or more tray assemblies comprises a tray top having a plurality of holes for plants and a tray bottom configured to receive the tray top. In another aspect, the agroinfiltrator comprising metal or plastic. In another aspect, the one or more tray assemblies comprise metal or plastic. In another aspect, the agroinfiltration system kit further comprises one or more growth chambers for post-infiltration incubation configured to receive plants infiltrated by the agroinfiltration system. In another aspect, one or more growth chambers comprise metal or plastic. The present invention can also be used with any of the following non-limiting list of additional plant species: Nicotiana benthamiana, Medicago truncatukr, Lettuce (Lactuca sativa), Arabidopsis thaliana, Nicotiana tabacunr, Spinach (Spinacia oleracea),' Zucchini (Cucurbita pepo var. cylindrical and / or Squash (Cucurbita pepo).

[0010] As embodied and broadly described herein, an aspect of the present disclosure relates to a method of agroinfiltration comprising: providing an agroinfiltration system comprising: an agroinfiltrator comprising a vacuum chamber, a top cover, and one or more sub-chambers to infiltrate plants; and one or more tray assemblies configured to hold soil for plant growth, wherein the agroinfiltrator is configured to receive the one or more tray assemblies; and infiltrating a plurality of plants using the agroinfiltration system. In one aspect, agroinfiltrator comprises a vacuum inlet, a pressure gauge, and a drain pipe. Inanother aspect, each of the one or more tray assemblies comprises a tray top having a plurality of holes for plants and a tray bottom configured to receive the tray top. In another aspect, the agroinfiltrator comprising metal or plastic. In another aspect, the one or more tray assemblies comprise metal or plastic. In another aspect, the agroinfiltration system further comprises one or more growth chambers for post-infiltration incubation, configured to receive plants infiltrated by the agroinfiltration system; and incubating the plants in the one or more growth chambers. In another aspect, the one or more growth chambers comprise metal or plastic. The present invention can also be used with any of the following non-limiting list of additional plant species: Nicotiana benihamiana. Medicago truncatulcr, Lettuce (Lactuca sativa); Arabidopsis thaliancr, Nicotiana tabacunr, Spinach (Spinacia oleracea)~ Zucchini (('ucurbiia pepo var. cylindrical and / or Squash (('ucurbiia pepo).BRIEF DESCRIPTION OF THE DRAWINGS

[0011] For a more complete understanding of the features and advantages of the present invention, reference is now made to the detailed description of the invention along with the accompanying figures and in which:

[0012] FIG. 1A shows a side view of an embodiment of the present invention. FIG. IB shows a front view of the embodiment. FIG. 1C shows an isometric view of the embodiment with a tray detached. FIG. ID shows a top view of the tray. FIG. IE shows an isometric view of the embodiment with the tray in place.

[0013] FIGS. 2A and 2B show anisometric views of components of a tray of another embodiment detached and assembled, respectively. FIGS. 2C and 2D show isometric views of the embodiment of FIGS. 2A and 2B with the tray detached and in place, respectively.

[0014] FIG. 3 A shows an isometric view of another embodiment of the invention with one tray detached. FIG. 3B shows a top view of two trays in place in the embodiment of FIG. 3A. FIGS. 3C and 3D show a front view and an isometric view of the embodiment with two trays in place, respectively.

[0015] FIG. 4 shows a flowchart for a method embodiment of the present invention.DETAILED DESCRIPTION OF THE INVENTION

[0016] While the making and using of various embodiments of the present invention are discussed in detail below, it should be appreciated that the present invention provides many applicable inventive concepts that can be embodied in a wide variety of specific contexts. The specific embodiments discussed herein are merely illustrative of specific ways to make and use the invention and do not delimit the scope of the invention.

[0017] To facilitate the understanding of this invention, a number of terms are defined below. Terms defined herein have meanings as commonly understood by a person of ordinary skill in the areas relevantto the present invention. Terms such as “a”, “an” and “the” are not intended to refer to only a singular entity, but include the general class of which a specific example may be used for illustration. The terminology herein is used to describe specific embodiments of the invention, but their usage does not delimit the invention, except as outlined in the claims.

[0018] The materials required for infiltration using the present invention, systems and methods for agroinfiltration for recombinant protein production, include Nicotiana benthamiana plants that are 4-6 weeks old; and an engineered agrobacterium that contains a plasmid for recombinant protein production. The materials also include an agroinfiltration buffer that includes 10 mM MES (2-(N-morpholino) ethane sulfonic acid), pH 5.6; 10 mM MgCL (Magnesium chloride); and 200 pM Acetosyringone. The agrofiltration buffer is prepared fresh or stored at 4°C for no more than 24 hours before use. The materials further include a centrifuge for bacterial pelleting; and a spectrophotometer for ODeoo measurement.

[0019] Preparation of the engineered agrobacterium includes cultivation of an engineered agrobacterium strain harboring a plasmid for recombinant protein production in a Luria-Bertani (LB) medium including appropriate antibiotics at 28 °C with shaking at 200 rpm for two days. The cultivation further includes harvesting of agrobacterial cultures by centrifugation at 5000g for 15 minutes at room temperature to produce a bacterial pellet. The cultivation also includes resuspending the bacterial pellet in the agrofiltration buffer to achieve an ODeoo of 1.0.

[0020] System embodiments of the present invention, agroinfiltrator systems, include an agroinfiltrator with a vacuum chamber and a top cover and a plurality of sub-chambers to infiltrate plants; one or more tray assemblies or growth chambers (1, 2, 3, or more trays or growth chambers) configured to hold soil for plant growth, each with a tray top and a tray bottom configured to receive the tray top and / or, optionally, one or more growth chambers for post-infiltration incubation, which are configured to receive plants infiltrated by the agroinfiltration system. The agroinfiltrator, the 1, 2 or 3 tray assemblies, and the one or more growth chambers or both can include metal or plastic. The trays for the equipment are specifically designed for nicotiana benthamiana plants, keeping in view the height and plant to plant space required for optimal growth. The present invention includes kits that include the components of agroinfiltration systems disclosed herein in a partially or completely disassembled state, with one or more fasteners or tools or both for assembling the components.

[0021] Referring to the drawings, FIG. 1A shows a side view of an embodiment of the present invention, the agroinfiltration system 100, including an agroinfiltrator 105 with atop cover 110. The agroinfiltration system 100 is shown mounted on a wheeled cart. The agroinfiltrator has the hydraulic control for the top cover 110. The electric box controller 107 can include one or more processors that control turning off and off the vacuum pumps, hydraulic pump, self-cleaning pumps. The agroinfiltrator has one or more pumps. In one example, the agroinfiltrator 100 has three pumps: (1) for the hydraulic system, (2) for self-cleaning, and (3) for creating a vacuum 109.

[0022] FIG. IB shows a front view of the agroinfiltration system 100 with the agroinfiltrator 105 and the top cover 110. The agroinfiltrator 105 also includes a vacuum inlet 115, a pressure gauge 120, vacuum and hydraulic pumps box 109, electric box controller 107 and a drain pipe 125. The agroinfiltration system 100 is shown, in this example, mounted on the wheeled cart but can also be fixed permanently or semipermanently to the floor or wall.

[0023] FIG. 1C shows an isometric view of the agroinfiltration system 100 with a tray assembly 130 detached, including a tray bottom 135, vacuum and hydraulic pump 109, hydraulic controller 106, electric box controller 107, plant height adjustor 111 and a tray top 140. The tray assembly 130 is configured to fit into the top of the agroinfiltrator 105, under the top cover 110.

[0024] FIG. ID shows a top view of the tray assembly 130, with the tray bottom 135, the tray top 140, and a plurality of holes 145 in the tray top 140 for plants.

[0025] FIG. IE shows an isometric view of the agroinfiltration system 100 with the agroinfiltrator 105 and the top cover 110, vacuum and hydraulic pumps box 109, electric box 107, plant height adjustor 111, and with the tray assembly 130 in place can be moved up or down depending on the height of the plant being infiltrated. The height of the tray can be adjusted manually or automatically in relation to the solution that will contain the infiltrating material used to modify the plants. The agroinfiltration system 100 permits for the adjustment of the portions of the plant to be infiltrated, such as the top of the leaves, the top and middle portion of the leaves and portions of the stems, the entire surface of the leaves and most or all of the stem, or even a portion of the roots.

[0026] FIGS. 2A and 2B show isometric views of components of a tray assembly 230 of another embodiment of the present invention, the agroinfiltration system 200 (not shown, see FIGS. 2C and 2D), detached and assembled, respectively. The tray assembly is configured to fit into the top of the agroinfiltrator 205, under a top cover 210 (not shown). FIGS. 2A and 2B show the tray assembly 230 with a tray bottom 235, a tray top 240, and a plurality of holes 245 in the tray top 140 for plants.

[0027] FIGS. 2C and 2D show isometric views of the agroinfiltration system 200 of FIGS. 2A and 2B with the tray assembly 230 detached and in place, respectively. FIGS. 2C and 2D also show the agroinfiltrator 205. The agroinfiltrator 205 has a vacuum inlet, a pressure gauge, and a drain pipe, but these are not shown in FIGS. 2C and 2D. Multiple tray assemblies 230 can be positioned within the agroinfiltrator 205. The agroinfiltrator 205 can also be sized such that one or more tray assemblies 230 (see, e.g., FIG. 3B) can be raised or lowered within the agroinfiltrator 205 to match the height of the plants (not depicted) in the tray assembly 230.

[0028] FIG. 3A shows an isometric view of another embodiment of the present invention, the agroinfiltration system 300, which is sized to accommodate two tray assemblies 330a, 330b (330b not shown in FIG. 3A, see FIG. 3B) with one tray assembly 330a detached. FIG. 3A shows the agroinfiltrator 305 and the tray assembly 330a with a tray bottom 335a, a tray top 340a, and a plurality of holes 345a inthe tray top 340a for plants. FIG. 3B shows a top view of the two tray assemblies 330a, 330b in place in the agroinfiltrator 305, with the tray top 340b and the holes 345b indicated. FIG. 3C shows a front view and FIG. 3D an isometric view, respectively, of the agroinfiltration system 300 with the two tray assemblies 330a, 330b in place. FIGS. 3C and 3D also show the agroinfdtrator 305 in relation with drainpipe 325. The agroinfiltrator also has a vacuum inlet and a pressure gauge, but these are not shown in FIGS. 3A-3D. The agroinfiltration system 300 can be configured to accommodate more than two tray assemblies similar to the tray assemblies 330a, 330b, that can be side-by-side and form an array of tray assemblies, such as 2x2, 3x3, 4x4, 2x3, 2x4, 3x4, etc.

[0029] Method embodiments of the present invention include providing an agroinfiltration system as disclosed herein and pouring resuspended agrobacterium at an ODeoo of 1.0 into the agroinfiltrator of the agroinfiltration system. Whole Nicotiana benthamiana plants, which are grown in the one or more tray assemblies, are carefully inverted and submerged upside-down in the infiltration solution within the agroinfiltrator. A vacuum of 25-27 in-Hg is applied to the agroinfiltrator using a vacuum pump. A data of vacuum pressure and time are provided in Table 1. The vacuum is maintained for 1-3 minutes, depending how many trays are used and on the leaf texture and thickness to ensure thorough infiltration. The vacuum is gradually released to avoid damaging the plants, and the plant leaves are checked for successful infiltration. The infiltrated plants are removed from the agroinfiltrator and gently blotted to remove excess infiltration solution. The infiltrated plants are placed in a controlled growth chamber with the following conditions: a temperature of 22-24 °C; a light cycle of 16-hour light and 8-hour dark; and a humidity of 70-80%. The infiltrated plants are monitored daily for signs of infiltration success and overall health. This protocol ensures the efficient introduction of agrobacterium carrying the recombinant protein plasmid into Nicotiana benthamiana, facilitating high levels of protein expression within the plant tissues.

[0030] FIG. 4 shows a flowchart for a method embodiment of the present disclosure. Method 400 includes block 405, providing an agroinfiltration system including an agroinfiltrator comprising a vacuum chamber, a top cover, and plurality of sub-chambers to infiltrate plants; and one or more tray assemblies configured to hold soil for plant growth, wherein the agroinfiltrator is configured to receive the one or more tray assemblies. Block 410 includes infiltrating a plurality of plants using the agroinfiltration system.

[0031] The following are characteristics of the method of operation of one embodiment of the present disclosure. To achieve the maximum vacuum of rated value 25 inHg. Should be able to handle 1, 2, and 3 tray(s) of specified size. Adjustable elevation of the tray for each chamber from the bottom.

[0032] Analysis. Data was taken during the experiment at different vacuum values as 15, 20, 22, and 25 inHg. Main chamber by placing three trays and half filled with water. Sub-chamber 1 by placing two trays half filled with water. Sub-chamber 2 by placing one tray half filled with water.

[0033] Chamber lid opening time 22 seconds. Chamber lid closing time 16 seconds.

[0034] Table 1. Data of vacuum pressure and time.

[0035] Although embodiments of the present invention disclosed herein are discussed in terms of specifics measures, quantities, dimensions, and the like, one skilled in the art of infiltration systems will recognize that other such measures, quantities, dimensions, and the like can be used for the present invention. Those disclosed are given as non-limiting examples. And although the use of Nicotiana benihamiana. one skilled in the art of infiltration systems for recombinant protein production will recognize that other plants suitable for recombinant protein production can be used with the present invention. Nicotiana benthamiana is disclosed as non-limiting example.

[0036] It is contemplated that any embodiment discussed in this specification can be implemented with respect to any method, kit, reagent, or composition of the invention, and vice versa. Furthermore, compositions of the invention can be used to achieve methods of the invention.

[0037] It will be understood that particular embodiments described herein are shown by way of illustration and not as limitations of the invention. The principal features of this invention can be employed in various embodiments without departing from the scope of the invention. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, numerous equivalents to the specific procedures described herein. Such equivalents are considered to be within the scope of this invention and are covered by the claims.

[0038] All publications and patent applications mentioned in the specification are indicative of the level of skill of those skilled in the art to which this invention pertains. All publications and patent applications are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.

[0039] The use of the word “a” or “an” when used in conjunction with the term “comprising” in the claims and / or the specification may mean “one,” but it is also consistent with the meaning of “one or more,” “at least one,” and “one or more than one.” The use of the term “or” in the claims is used to mean “and / or” unless explicitly indicated to refer to alternatives only or the alternatives are mutually exclusive, although the disclosure supports a definition that refers to only alternatives and “and / or.” Throughout this application, the term “about” is used to indicate that a value includes the inherent variation of error for the device, the method being employed to determine the value, or the variation that exists among the study subjects.

[0040] As used in this specification and claim(s), the words “comprising” (and any form of comprising, such as “comprise” and “comprises”), “having” (and any form of having, such as “have” and “has”), “including” (and any form of including, such as “includes” and “include”) or “containing” (and any form of containing, such as “contains” and “contain”) are inclusive or open-ended and do not exclude additional, unrecited elements or method steps. In embodiments of any of the compositions and methods provided herein, “comprising” may be replaced with “consisting essentially of’ or “consisting of’. As used herein, the phrase “consisting essentially of’ requires the specified integer(s) or steps as well as those that do not materially affect the character or function of the claimed invention. As used herein, the term “consisting” is used to indicate the presence of the recited integer (e.g., a feature, an element, a characteristic, a property, a method / process step or a limitation) or group of integers (e.g., feature(s), element(s), characteristic(s), propertie(s), method / process steps or limitation(s)) only.

[0041] The term “or combinations thereof’ as used herein refers to all permutations and combinations of the listed items preceding the term. For example, “A, B, C, or combinations thereof’ is intended to include at least one of: A, B, C, AB, AC, BC, or ABC, and if order is important in a particular context, also BA, CA, CB, CBA, BCA, ACB, BAC, or CAB. Continuing with this example, expressly included are combinations that contain repeats of one or more item or term, such as BB, AAA, AB, BBC, AAABCCCC,CBBAAA, CABABB, and so forth. The skilled artisan will understand that typically there is no limit on the number of items or terms in any combination, unless otherwise apparent from the context.

[0042] As used herein, words of approximation such as, without limitation, “about”, "substantial" or "substantially" refers to a condition that when so modified is understood to not necessarily be absolute or perfect but would be considered close enough to those of ordinary skill in the art to warrant designating the condition as being present. The extent to which the description may vary will depend on how great a change can be instituted and still have one of ordinary skilled in the art recognize the modified feature as still having the required characteristics and capabilities of the unmodified feature. In general, but subject to the preceding discussion, a numerical value herein that is modified by a word of approximation such as “about” may vary from the stated value by at least ±1, 2, 3, 4, 5, 6, 7, 10, 12 or 15%.

[0043] Additionally, the section headings herein are provided for consistency with the suggestions under 37 CFR 1.77 or otherwise to provide organizational cues. These headings shall not limit or characterize the invention(s) set out in any claims that may issue from this disclosure. Specifically and by way of example, although the headings refer to a “Field of Invention,” such claims should not be limited by the language under this heading to describe the so-called technical field. Further, a description of technology in the “Background of the Invention” section is not to be construed as an admission that technology is prior art to any invention(s) in this disclosure. Neither is the “Summary” to be considered a characterization of the invention(s) set forth in issued claims. Furthermore, any reference in this disclosure to “invention” in the singular should not be used to argue that there is only a single point of novelty in this disclosure. Multiple inventions may be set forth according to the limitations of the multiple claims issuing from this disclosure, and such claims accordingly define the invention(s), and their equivalents, that are protected thereby. In all instances, the scope of such claims shall be considered on their own merits in light of this disclosure, but should not be constrained by the headings set forth herein.

[0044] All of the compositions and / or methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the compositions and / or methods and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit and scope of the invention. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.

[0045] To aid the Patent Office, and any readers of any patent issued on this application in interpreting the claims appended hereto, applicants wish to note that they do not intend any of the appended claims to invoke paragraph 6 of 35 U.S.C. § 112, U.S.C. § 112 paragraph (f), or equivalent, as it exists on the date of filing hereof unless the words “means for” or “step for” are explicitly used in the particular claim.

[0046] For each of the claims, each dependent claim can depend both from the independent claim and from each of the prior dependent claims for each and every claim so long as the prior claim provides a proper antecedent basis for a claim term or element.

Claims

WHAT IS CLAIMED IS:

1. An agroinfiltration system comprising: an agroinfiltrator comprising a vacuum chamber, a top cover, vacuum and hydraulic pumps box, electric box, plant height adjustment, and one or more sub-chambers to infiltrate plants; and one or more tray assemblies or growth chambers configured to hold soil for plant growth, wherein the agroinfiltrator is configured to receive the one or more tray assemblies; and a vacuum inlet, a pressure gauge, and a drain pipe in communication with the vacuum chamber.

2. The agroinfiltration system of claim 1, wherein each of the one or more tray assemblies comprises a tray top having a plurality of holes for plants and a tray bottom configured to receive the tray top.

3. The agroinfiltration system of claim 1, wherein the agroinfiltrator comprising metal or plastic.

4. The agroinfiltration system of claim 1, wherein the one or more tray assemblies, the growth chambers, or both comprise metal or plastic.

5. The agroinfiltration system of claim 1, further comprising one or more growth chambers for postinfiltration incubation configured to receive plants infiltrated by the agroinfiltration system.

6. The agroinfiltration system of claim 5, wherein the agroinfiltrator is self-cleaning and drains off one or more solutions used in the agroinfiltrator.

7. An agroinfiltration system kit, comprising: an agroinfiltrator comprising a vacuum chamber, a top cover, vacuum and hydraulic pumps box, electric box, hydraulic controller, plant height adjustment, and one or more sub-chambers to infiltrate plants; one or more tray assemblies or growth chambers configured to hold soil for plant growth, wherein the agroinfiltrator is configured to receive the one or more tray assemblies; and one or more fasteners or tools or both for assembling the agroinfiltrator and the one or more tray assemblies.

8. The agroinfiltration system kit of claim 7, wherein the agroinfiltrator comprises a vacuum inlet, a pressure gauge, and a drain pipe.

9. The agroinfiltration system kit of claim 7, wherein each of the one or more tray assemblies comprises a tray top having a plurality of holes for plants and a tray bottom configured to receive the tray top.

10. The agroinfiltration system kit of claim 7, wherein the agroinfiltrator comprising metal or plastic.

11. The agroinfiltration system kit of claim 7, wherein the one or more tray assemblies, the growth chambers, or both comprise metal or plastic.

12. The agroinfiltration system kit of claim 7, further comprising one or more growth chambers for post-infiltration incubation configured to receive plants infiltrated by the agroinfiltration system.

13. The agroinfiltration system kit of claim 12, wherein the agroinfiltrator is self-cleaning and drains off one or more solutions used in the agroinfiltrator .

14. A method of agroinfiltration comprising: providing an agroinfiltration system comprising: an agroinfiltrator comprising a vacuum chamber, a top cover, vacuum and hydraulic pumps box, electric box, plant height adjustment, and one or more sub-chambers to infiltrate plants; and one or more tray assemblies configured to hold soil for plant growth, wherein the agroinfiltrator is configured to receive the one or more tray assemblies; and infiltrating a plurality of plants using the agroinfiltration system.

15. The method of agroinfiltration of claim 14, wherein the agroinfiltrator comprises a vacuum inlet, a pressure gauge, and a drain pipe.

16. The method of agroinfiltration of claim 14, wherein each of the one or more tray assemblies comprises a tray top having a plurality of holes for plants and a tray bottom configured to receive the tray top.

17. The method of agroinfiltration of claim 14, wherein the agroinfiltrator comprising metal or plastic.

18. The method of agroinfiltration of claim 14, wherein the one or more tray assemblies comprise metal or plastic.

19. The method of agroinfiltration of claim 14, wherein the agroinfiltration system further comprises one or more growth chambers for post-infiltration incubation, configured to receive plants infiltrated by the agroinfiltration system; and incubating the plants in the one or more growth chambers.

20. The method of agroinfiltration of claim 19, wherein the one or more growth chambers comprise metal or plastic.

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