Integument and method for micropropagation and tissue culturing

• US 4,908,315 A
• Filed: 03/04/1987
• Issued: 03/13/1990
• Est. Priority Date: 03/04/1987
• Status: Expired due to Term

• Current Assignee: Norax Corporation
• Original Assignee: Agristar, Inc.
• Inventors: Kertz, Malcolm G.
• Primary Examiner(s): Warren, Charles F.
• Assistant Examiner(s): Fox, David T.
• Application Number:
  US07/021,408
• Time in Patent Office: 1,105 Days
• Field of Search: 47/66, 47/73, 47/84, 47/58, 47/69, 435/240.4, 435/299, 435/310, 435/311, 435/313
• US Class Current: 435/430
• CPC Class Codes: A01C 1/042 Tapes, bands or cords
• CPC Class Codes: A01G 31/04 Hydroponic culture on conveyors
• CPC Class Codes: A01H 4/002 Culture media for tissue culture
• CPC Class Codes: C12M 23/14 Bags
• CPC Class Codes: C12M 23/22 Transparent or translucent parts glassware for laboratory use B01L3/00
• CPC Class Codes: C12M 23/24 Gas permeable parts
• CPC Class Codes: Y02P 60/21 Dinitrogen oxide [N2O], e.g. using aquaponics, hydroponics or efficiency measures

Abstract

An integument and related process for the micropropagation of tissue is made of a semipermeable, translucent membrane. The integument includes a cellule for completely enclosing and sealing the tissue and growth medium from the ambient environment. Light is transmitted through the translucent membrane to the tissue. The semipermeable membrane allows the mutual penetration by diffusion and osmosis of oxygen and carbon dioxide to permit the tissue to breathe with the ambient environment. The membrane is impermeable to biological contaminants. The integument and related method provides enhanced growth of the tissue and a reduction in contamination.

Claims

1. A tissue culturing system for an atmospheric ambient environment, comprising:

plant tissue;

a tissue culturing medium for said plant tissue;

a gas permeable, liquid impermeable, and translucent membrane having interstices smaller than a virus forming a cellule for enclosing the plant tissue and tissue culturing medium, the gas mixture and pressure within said cellule and the gas mixture and pressure of the atmospheric ambient environment being permitted to equalize by gas exchange through said membrane;

said membrane being completely sealed around the plant tissue and tissue culturing medium for preventing the exposure of the plant tissue and tissue culturing medium to biological contaminants in the atmospheric ambient environment.

2. The integument of claim 1 wherein said membrane is made of high density polyethylene.

3. An integument for the micropropagation of plant tissue in the ambient environment, comprising:

a gas permeable and liquid impermeable membrane forming a cellule for enclosing the tissue, said membrane being completely sealed around the tissue for preventing the exposure of the tissue to biological contaminants in the ambient environment, and said membrane being made of Chevron high density polyethylene 9650 which will prevent oxygen and carbon dioxide to diffuse therethrough.

4. A system for plant production, comprising:

a plant;

a medium for said plant;

a gas permeable, liquid impermeable, translucent membrane made of lightly cross-linked layers of polymers and having a moisture vapor transmission rate equal to that of a high density polyethylene film with a thickness in the range of 1 to 1.5 mils, said membrane having interstices smaller than the size of a virus but large enough for gas exchange, said membrane forming a cellule for enclosing said plant and medium, said cellule being completely sealed around the plant for preventing the exposure of the plant to biological contaminants in the ambient environment.

5. An integument for the micropropagation of plant tissue in the ambient environment, comprising:

a gas permeable and liquid impermeable membrane forming a cellule for enclosing the tissue, the gas mixture and pressure within the cellule being substantially the same as the gas mixture and pressure of the ambient environment upon enclosure;

said membrane being completely sealed around the tissue for preventing the exposure of the tissue to biological contaminants in the ambient environment, said membrane having a molecular structure with interstices smaller than the size of a virus but large enough for gas exchange.

6. An integument for the micropropagation of plant tissue in the ambient environment, comprising:

a gas permeable and liquid impermeable membrane forming a cellule for enclosing the tissue, the gas mixture and pressure within the cellule being substantially the same as the gas mixture and pressure of the ambient environment upon enclosure;

said membrane being completely sealed around the tissue for preventing the exposure of the tissue to biological contaminants in the ambient environment, said membrane having interstices no greater than 0.01 micrometers in size but large enough for gas exchange.

7. An integument for containing a growing plant in a medium in an atmospheric ambient environment, comprising:

a gas permeable and liquid impermeable membrane of Chevron high density polyethylene 9650 forming a cellule for enclosing the plant, said membrane being completely sealed around the plant for preventing the exposure of the plant to contaminants in the atmospheric ambient environment, said membrane being translucent to permit light to impinge on the plant.

8. An integument free of an internal soil containment device for the growing of plants in the atmospheric ambient environment consisting of:

a medium for growing the plants; and

a translucent, gas permeable and liquid impermeable membrane of high density polyethylene forming a cellule for enclosing the plant and medium, said membrane having interstices smaller than the size of a virus and being completely sealed around the plant and medium for preventing exposure of the plant to contaminants in the atmospheric ambient environment;

said membrane transmitting therethrough light rays having a wavelength between 400 and 750 nanometers.

9. An integument for the micropropagation of plant tissue in a tissue culturing medium, comprising:

a membrane forming at least one cellule for containing and completely enclosing the plant tissue and tissue culturing medium;

said membrane being made of a single layer of high density polyethylene and having interstices smaller than a virus;

said membrane being semipermeable and allowing the diffusion of gases therethrough; and

said integument being liquid impermeable and translucent allowing the transmission of light therethrough except for any light blocked by the plant tissue and tissue culturing medium.

10. An integument for growing a plant in a medium in the atmospheric ambient environment comprising:

a membrane forming at least one cellule for containing and completely enclosing the plant and medium;

said membrane being made of a single layer of high density polyethylene and having interstices smaller than the size of a virus;

said membrane being gas permeable for allowing the diffusion of gases therethrough;

said membrane being liquid impermeable and translucent allowing the transmission of light therethrough;

said membrane sealing the plant from biological contaminants in the atmospheric ambient environment.

13. The integument of claim 10 wherein said membrane will withstand autoclaving without melting or distending.

11. A plant production system, comprising:

a plant;

medium for growing said plant;

a membrane forming at least one cellule for containing and completely enclosing the plant and medium;

said membrane being made of Chevron 9650 high density polyethylene;

said membrane being semipermeable and allowing the diffusion of gases therethrough; and

said membrane being liquid impermeable.

12. An integument for the micropropagation of plant tissue in a tissue culturing medium, comprising:

a membrane forming at least one cellule for containing and completely enclosing the plant tissue and tissue culturing medium;

said membrane being made of a single layer of high density polyethylene;

said membrane being semipermeable and allowing the diffusion of gases therethrough;

said integument being translucent and allowing the transmission of light therethrough except for any light blocked by the plant tissue and tissue culturing medium; and

said membrane having a permeability to water vapor of substantially 0.32 grams per 100 square inches per 24 hours.

14. An integument for the micropropagation of plant tissue in a tissue culturing medium, comprising:

a membrane forming at least one cellule for containing and completely enclosing the plant tissue and tissue culturing medium;

said membrane being made of a single layer of high density polyethylene;

said membrane being semipermeable and allowing the diffusion of gases therethrough;

said integument being translucent and allowing the transmission of light through the integument except for any light blocked by the plant tissue and tissue culturing medium; and

said membrane being made of Chevron high density polyethylene 9650.

15. An integument for growing plant tissue in a tissue culturing medium, comprising:

a high density polyethylene material forming at least one chamber;

said high density polyethylene material being translucent, permeable to gases and impermeable to liquids;

said high density polyethylene having interstices smaller than the size of a virus but large enough for gas exchange;

said high density polyethylene material having the characteristic of withstanding the required temperatures and pressures of an autoclave for sterilization;

said integument allowing light to pass completely therethrough; and

said chamber having an aperture for receiving the plant tissue and tissue culturing medium, said aperture being adapted for closure after said chamber receives the plant tissue and tissue culturing medium.

16. The integument of claim 15 wherein said chamber is formed by attaching at predetermined locations adjacent layers of said polyethylene material.

17. The integument of claim 16 wherein said chamber is formed by one sheet of said polyethylene folded over and heat sealed along its open sides.

18. The integument of claim 16 wherein said adjacent layers of polyethylene are heat sealed to form a plurality of chambers.

19. The integument of claim 18 wherein said chambers are expandable and permit the passage of light therebetween.

20. The integument of claim 15 wherein said material further forms a band above said chamber for suspending said chamber in the vertical position.

21. The integument of claim 20 wherein said band includes attachment means for attaching said flap to a support.

24. The integument of claim 15 further including a support for suspending said chamber in a vertical position.

22. An integument for growing plant tissue in a tissue culturing medium, comprising:

a high density polyethylene material forming at least one chamber;

said high density polyethylene material being translucent and permeable to gases and impermeable to liquids;

said high density polyethylene material having the characteristic of withstanding the required temperatures and pressures of an autoclave for sterilization;

said integument allowing light to pass completely therethrough; and

said chamber having an aperture for receiving the plant tissue and tissue culturing medium, said aperture being adapted for closure after said chamber receives the plant tissue and tissue culturing medium, and said polyethylene material being substantially 1.25 mils in thickness.

23. An integument for growing plant tissue in a tissue culturing medium, comprising:

a high density polyethylene material forming at least one chamber;

said high density polyethylene material being translucent and permeable to gases and impermeable to liquids;

said high density polyethylene material having the characteristic of withstanding the required temperatures and pressures of an autoclave for sterilization;

said integument allowing light to pass completely therethrough;

said chamber having an aperture for receiving the tissue and tissue culturing medium, said aperture being adapted for closure after said chamber receives the tissue and tissue culturing medium;

said chamber being divided into a foliage chamber and a root chamber, said root chamber accommodating the root system of the plant tissue and containing the tissue culturing medium.

25. An integument pack for growing plant tissue in a tissue culturing medium, comprising:

adjacent layers of gas permeable, liquid impermeable and translucent material being attached together at predetermined locations thereof to form a plurality of cellules for receiving the plant tissue and tissue culturing medium;

said cellules being adapted for closing upon receiving the plant tissue and tissue culturing medium whereby the plant tissue is completely enclosed from the ambient environment;

said material having light transmission properties which allow light rays having wavelengths in the range of 400 to 750 nanometers to pass through said material; and

said material having a molecular structure forming interstices sized to allow the diffusion of oxygen and carbon dioxide therethrough but preventing the passage of viral biological contaminants.

26. An integument for the micropropagation of a leafy vegetable tissue in a medium, comprising:

a gas permeable, liquid impermeable, and translucent membrane having interstices smaller than a virus forming a cellule to receive the leafy vegetable tissue and medium;

said cellule having a foliage chamber and separate root chamber made with said membrane with a reduced opening therebetween whereby the root system grown by the tissue extends from said foliage chamber, through said reduced opening and into said root chamber containing the medium.

29. The integument of claim 26 wherein said foliage chamber and root chamber are formed by attaching adjacent walls of said cellule at predetermined locations;

said attachment locations not completely extending across said cellule so as to form said opening.

30. The integument of claim 29 wherein said attachment locations are at an angle of less than 90 degrees with the central axis of said cellule so as to form a conical shaped end to said foliage chamber.

27. A system for seed germination, comprising:

a seed;

a medium for germinating said seed;

a gas permeable, liquid impermeable membrane forming a cellule for enclosing said seed and medium, said membrane having interstices smaller than a virus and being completely sealed around the seed for preventing the exposure of the seed to biological contaminants in the atmospheric ambient environment, and said membrane being a semipermeable and translucent membrane.

28. An integument for germinating a seed in a medium, comprising:

a gas permeable, liquid impermeable membrane of Chevron 9650 high density polyethylene forming a cellule for enclosing the seed, said membrane being completely sealed around the seed for sealing the seed from biological contaminants in the ambient atmosphere.

31. A support for the micropropagation of plant tissue comprising:

a frame;

a plurality of integuments having cellules containing the plant tissue and sealing the plant tissue from biological contaminants in the ambient environment, said integuments having a band above said cellules for attachment to said frame; and

said integuments being made of a gas permeable, liquid impermeable material having interstices smaller than a virus, said material being translucent such that light may pass completely through one integument and into an adjacent integument.

32. A method for growing plant tissue in an integument in an atmospheric ambient environment, comprising the steps of;

inserting the plant tissue and a tissue culturing medium in the cellule of the integument;

said integument being made of a gas permeable, liquid impermeable and translucent membrane of high density polyethylene;

completely enclosing the plant tissue within the cellule;

sealing the plant tissue from the biological contaminants in the ambient environment;

transmitting light through the translucent membrane to the plant tissue, any light not blocked by the plant tissue and tissue culturing medium passing completely through the integument; and

diffusing oxygen and carbon dioxide through said membrane to permit the plant tissue to breathe.

33. A method of micropropagation of plant tissue comprising the steps of:

placing within the cellule of an integument formed of Chevron high density polyethylene #9650 a tissue culturing medium which is suitable for establishing an initial culture of primarily undifferentiated plant tissue;

placing a plant tissue within the cellule and sealing the plant tissue and initial tissue culturing medium from biological contaminants in the ambient environment;

establishing an initial culture of primarily undifferentiated plant tissue;

transferring a portion of the plant tissue established in the initial culture to a second cellule of another unused integument having a tissue culturing medium suitable for multiplication of the plant tissue therein;

multiplying the transferred portion of the plant tissue into a mass of plant tissue of primarily undifferentiated plant tissue;

transferring a portion of multiplied plant tissue to a third cellule of another unused integument having therein a tissue culturing medium suitable for individual plant formation.

34. A method of micropropagation of plant tissue in an atmospheric ambient environment comprising the steps of:

(a) providing a pliable cellule made of a gas permeable, liquid impermeable, translucent membrane having interstices smaller than a virus but large enough for gas exchange;

(b) inserting a tissue culturing medium into the cellule for establishing an initial culture;

(c) sterilizing the cellule containing the tissue culturing medium;

(d) inserting plant tissue into the tissue culturing medium;

(e) enclosing the plant tissue within the cellule and sealing the plant tissue completely within the cellule to protect the plant tissue from biological contaminants in the ambient environment;

(f) exposing all parts of the plant tissue to the light transmitted and the gas exchanged through the membrane;

(g) transmitting light through the entire surface of the cellule to all parts of the plant tissue; and

(h) exchanging gas through the surface of the cellule to all parts of the plant tissue.

35. A method of micropropagation of plant tissue in an atmospheric ambient environment comprising the steps of:

(a) providing an unused pliable cellule made of a gas permeable, liquid impermeable, translucent membrane having interstices smaller than the size of a virus but large enough for gas exchange;

(b) inserting a tissue culturing medium for tissue multiplication into the cellule;

(c) sterilizing the cellule containing the tissue culturing medium;

(d) removing the plant tissue from another cellule containing primarily undifferentiated plant tissue;

(e) dividing the plant tissue into individual growing point tissues;

(f) inserting an individual growing point tissue in the cellule of the sterilized unused cellule containing the tissue culturing medium;

(g) enclosing the individual growing point tissue completely within the cellule and sealing the growing point tissue from biological contaminants in the ambient environment;

(h) exposing all parts of the growing point tissue to the light transmitted and gas exchanged through the membrane;

(i) transmitting light through the surface of the cellule to all parts of the growing point tissue; and

(j) exchanging gas through the surface of the cellule to all parts of the growing point tissue.

36. A method of micropropagation of plant tissue in an atmospheric ambient environment comprising the steps of:

(a) inserting a tissue culturing medium for individual plant formation into a new pliable cellule which is made of a gas permeable, liquid impermeable, translucent membrane having interstices smaller than a virus but large enough for gas exchange;

(b) sterilizing the cellule containing the tissue culturing medium;

(c) removing plant tissue from another cellule containing primarily undifferentiated plant tissue and disposing of that another cellule;

(d) inserting the plant tissue removed from the plant tissue containing cellule into the sterilized cellule containing the tissue culturing medium;

(e) enclosing the plant tissue within the sterilized cellule and sealing the plant tissue completely within the cellule to protect the tissue from biological contaminants in the ambient environment;

(f) exposing all parts of the plant tissue to the light transmitted and gas exchanged through the cellule;

(g) transmitting light through the surface of the cellule to all parts of the plant tissue; and

(h) exchanging gas through the surface of the cellule to all parts of the plant tissue.

37. A method of micropropagation of plant tissue comprising the steps of:

(a) dissolving a minimal organic medium and sucrose with distilled water to form a media solution;

(b) adjusting the pH of the media solution;

(c) transferring the media solution to a pliable cellule made of Chevron high density polyethylene #9650;

(d) autoclaving the cellule and placing the cellule under a laminar flow hood;

(e) removing differentiated tissue from a cultivar;

(f) performing surface sterilization of the plant tissue;

(g) transferring the plant tissue to the cellule under the laminar flow hood;

(h) enclosing the plant tissue within the cellule and sealing the plant tissue completely within the cellule to protect the plant tissue from biological contaminants in the ambient environment;

(i) placing the integument in a culture room;

(j) exposing all parts of the plant tissue to the light transmitted and gas exchanged through the cellule;

(k) transmitting light through the surface of the cellule to all parts of the plant tissue;

(l) exchanging gas through the surface of the cellule to allow the plant tissue to grow into a mass of primarily undifferentiated tissue; and

(m) removing the integument from the culture room upon certification that the resultant plant tissue mass is contaminant free.

38. A method of micropropagation of plant tissue comprising the steps of:

(a) dissolving a multiplication medium and sucrose with distilled water forming a media solution;

(b) adjusting the pH of the media solution;

(c) transferring the media solution to a new cellule of Chevron high density polyethylene #9650;

(d) autoclaving the new cellule and placing the cellule under a laminar flow hood;

(e) immersing a cellule containing a mass of primarily undifferentiated and certified contaminant-free plant tissue in a sterilizing agent;

(f) placing the tissue-containing cellule under a laminar flow hood;

(g) opening the tissue-containing cellule and removing the plant tissue mass from the cellule with sterilized instruments and disposing of the used cellule;

(h) dividing the plant tissue mass into individual growing point plant tissues;

(i) inserting an individual growing point plant tissue in the new cellule containing the media solution under the laminar flow hood;

(j) enclosing a growing point plant tissue within the cellule and sealing the plant tissue completely within the cellule to protect the plant tissue from biological contaminants in the ambient environment;

(k) placing the integument in a culture room;

(l) exposing all parts of the growing point plant tissue to the light transmitted and gas exchanged through the cellule;

(m) transmitting light through the surface of the cellule to all parts of the growing point plant tissue;

(n) exchanging gas through the surface of the cellule to allow the growing point plant tissue to grow into a mass of primarily undifferentiated plant tissue; and

(o) repeating steps (a) through (n) using the growing point tissue until a predetermined number of plant tissue masses have been produced.

39. A method of micropropagation of plant tissue comprising the steps of:

(a) dissolving a pretransplant medium and sucrose with distilled water forming a media solution;

(b) adjusting the pH of the media solution;

(c) transferring the media solution to a new pliable cellule of Chevron high density polyethylene #9650;

(d) autoclaving the new cellule and placing the cellule under a laminar flow hood;

(e) immersing a cellule containing a sample of primarily undifferentiated growing plant tissue in a sterilizing agent;

(f) placing the tissue-containing integument under a laminar flow hood;

(g) opening the tissue-containing cellule and removing the plant tissue from the cellule with sterilized instruments and disposing of the used cellule;

(h) inserting the plant tissue in the new cellule containing the media solution under the laminar flow hood;

(i) enclosing the plant tissue within the cellule and sealing the plant tissue completely within the cellule to protect the plant tissue from biological contaminants in the ambient environment;

(j) placing the integument in a culture room;

(k) exposing all parts of the plant tissue to the light transmitted and gas exchanged through the cellule;

(l) transmitting light through the surface of the cellule to all parts of the plant tissue;

(m) exchanging the gas through the surface of cellule to all parts of the plant tissue; and

(n) removing the plant tissue from the cellule upon the plant tissue growing into a plantlet.

40. A method of producing a plant from a seed in the atmospheric ambient environment, comprising:

(a) dissolving an organic medium and sucrose with distilled water forming a media solution;

(b) adjusting the pH of the media solution;

(c) transferring the media solution to a new pliable cellule of a gas permeable, liquid impermeable, and translucent membrane having interstices smaller than a virus but large enough for gas exchange;

(d) surface sterilizing the seed;

(e) rinsing the seed in distilled water and placing under a laminar flow hood;

(f) transferring the seed to the cellule under the laminar flow hood;

(g) enclosing the seed within the cellule and sealing the seed completely within the cellule to protect the seed from biological contaminants in the ambient environment;

(h) placing the cellule in a culture room;

(i) exposing all parts of the seed to the light transmitted and gas exchanges through the cellule;

(j) removing the plantlets from the cellule upon development of foliage and a root system;

(k) transferring the plantlet to an unused cellule; and

(l) removing the plant upon growth.

41. A combination conducive to tissue growth in an atmospheric ambient environment, comprising:

a cell; and

a gas permeable and liquid impermeable membrane of Chevron high density polyethylene #9650 forming a cellule for completely enclosing said cell and creating a closed environment for the cell, said membrane sealing said cell from biological contaminants in the atmospheric ambient environment.

42. The combination of claim 41 where said cell is alive and reproducing.

43. The combination of claim 41 further including a tissue culturing medium disposed within said cellule for supporting the growth of said cell and said membrane being translucent such that the combination includes only said cell, tissue culturing medium and membrane whereby any light not blocked by said cell and said tissue culturing medium is allowed to pass through the combination.

44. A combination conducive to cell growth in a tissue culturing medium in an atmospheric ambient environment, comprising:

a cell;

a tissue culturing medium;

a gas permeable, liquid impermeable, and translucent membrane forming a cellule for completely enclosing said cell and tissue culturing medium;

said cellule creating a closed environment for said cell, said membrane having interstices smaller than the size of a virus and sealing said cell from biological contaminants in the ambient environment; and

said membrane permitting oxygen and carbon dioxide to diffuse therethrough.

47. The combination of claim 44 wherein said membrane is made of a translucent polyethylene allowing the transmission of light therethrough.

45. A combination conducive to plant tissue growth in a tissue culturing medium in an atmospheric ambient environment comprising:

plant tissue;

a tissue culturing medium;

a container for receiving and enclosing said plant tissue and tissue culturing medium, said entire container being pliable and completely sealed around said plant tissue and said tissue culturing medium for preventing the exposure of said plant tissue and said tissue culturing medium to biological contaminants in the atmospheric ambient environment, said container being made of a single layer of translucent, liquid impermeable, and gas permeable membrane with a molecular structure having interstices smaller than a virus but large enough for gas exchange.

46. The combination of claim 45 wherein said container is made entirely of said membrane, wherein said membrane is Chevron 9650 high density polyethylene film.

48. The combination of claim 45 wherein said membrane will transmit therethrough light having a wavelength between 400 and 750 nanometers.

49. An integument pack for the micropropagation of plant tissue in a tissue culturing medium, comprising:

adjacent sheets of single layered high density polyethylene film;

said sheets being heat sealed together along the entire length thereof at a plurality of predetermined locations to form a plurality of cellules;

said cellules each having an open end adapted to receive plant tissue and tissue culturing medium;

said film being heat sealable for closing said open end and being autoclavable so as not to melt, distend, or distort;

said film being pliant and collapsible such that said cellules are flat and can be rolled;

said film being permeable to oxygen and carbon dioxide, having a water vapor transmission rate comparable to that of high density polyethylene, and impermeable to liquids; and

said film being translucent such that light can pass through the integument pack.

50. The integument pack of claim 49 wherein said cellules are made of Chevron 9650 film.

51. The integument pack of claim 49 wherein said open end includes a flap for heat sealing said cellules and suspending the integument pack.

52. The integument pack of claim 51 wherein the integument pack is disposable.

53. The integument pack of claim 49 wherein six individual cellules are formed.

54. The integument pack of claim 49 wherein the integument pack has dimensions of 12 inches wide and 6 inches high.

55. The integument pack of claim 49 wherein said film has a permeability to water vapor of 0.32 grams per 100 square inches per 24 hours.

56. The integument pack of claim 49 wherein said film has interstices no greater than 0.01 micrometers in size but great enough for gas exchange.

57. An apparatus for growing plant tissue in the atmospheric ambient environment, comprising:

a tissue culturing medium for the plant tissue;

a container for receiving and enclosing the plant tissue and tissue culturing medium;

said container being completely sealed around the plant tissue for preventing the exposure of the plant tissue to biological contaminants in the atmospheric ambient environment;

said container being made entirely of a pliable and collapsible material surrounding the plant tissue;

said material being permeable to gases, impermeable to liquids, and translucent to light;

said material having interstices smaller than a virus but large enough for gas exchange;

said container permitting gas exchange and light transmission over its entire surface area;

said container being sterilizable so as not to melt, distend, or distort; and

said container allowing all parts of the plant tissue to be exposed to gas exchanged and light transmitted through the material.

58. A method for growing plant tissue in a disposable cellule in an atmospheric ambient environment, comprising the steps of:

providing a pliable cellule made entirely of a high density polyethylene film which is impermeable to liquids;

inserting a tissue culturing medium into the cellule such that the tissue culturing medium contacts the inner surface of the polyethylene film;

sterilizing the cellule containing tissue culturing medium;

inserting and enclosing plant tissue within the cellule;

sealing the plant tissue completely within the cellule to protect the plant tissue from the biological contaminants in the atmospheric ambient environment;

allowing the gas mixture and pressure within the cellule and the gas mixture and pressure around the cellule in the ambient environment to equalize by gas exchange through said film;

exposing all parts of the plant tissue to light transmitted through the entire cellule to all parts of the plant tissue;

allowing the light to pass completely through the cellule except for any light blocked by the plant tissue and tissue culturing medium; and

diffusing oxygen and carbon dioxide through the cellule to and from all parts of the plant tissue.

59. A method of micropropagation comprising the steps of:

providing a pliable cellule made entirely of a high density polyethylene film which is impermeable to liquids;

placing a tissue culturing medium which is suitable for establishing an initial culture into the cellule;

sterilizing the cellule containing the tissue culturing medium in an autoclave;

placing a plant tissue within the cellule and sealing the plant tissue and tissue culturing medium completely within the cellule to protect the plant tissue from biological contaminants in an ambient environment similar to that of a greenhouse;

exposing all parts of the plant tissue to light transmission and gas exchange through the entire surface of the cellule;

establishing an initial culture of plant tissue;

transferring a portion of the plant tissue from the initial culture to a second cellule having a tissue culturing medium for multiplication therein and disposing of the first cellule;

exposing all parts of the plant tissue to light transmission and gas exchange through the entire surface of the cellule;

multiplying the transferred portion of the plant tissue into a mass of plant tissue;

transferring a portion of the mass of plant tissue to a third cellule having therein a tissue culturing medium suitable for individual plant formation;

exposing all parts of the plant tissue to light transmission and gas exchange through the entire surface of the cellule; and

removing the plant tissue upon plant formation.