Patent Application
20090005245
The present disclosure relates generally to artificial seeds and, more particularly, to coatings for a live end seal attached to an artificial seed.
Asexual propagation of plants has been shown for some species to yield large numbers of genetically identical embryos, each having a capacity to develop into a normal plant. Such embryos are usually further cultured under laboratory conditions until they reach an autotrophic “seedling” state characterized by an ability to produce its own food via photosynthesis, resist desiccation, produce roots able to penetrate soil, and fend off soil microorganisms. Some researchers have experimented with the production of artificial seeds, known as manufactured seeds, in which individual plant somatic or zygotic embryos are encapsulated in a seed coat. Examples of such manufactured seeds are disclosed in U.S. Pat. No. 5,701,699, issued to Carlson et al., the disclosure of which is hereby expressly incorporated by reference.
Typical manufactured seeds include a seed shell, synthetic gametophyte and a plant embryo. A manufactured seed that does not include the plant embryo is known in the art as a “seed blank.” The seed blank typically is a cylindrical capsule having a closed end and an open end. The synthetic gametophyte is placed within the seed shell to substantially fill the interior of the seed shell. A longitudinally extending hard porous insert, known as a cotyledon restraint, may be centrally located within one end of the seed shell, surrounded by the synthetic gametophyte, and includes a centrally located cavity extending partially through the length of the cotyledon restraint.
The cavity is sized to receive the plant embryo therein. The well-known plant embryo includes a radicle end and a cotyledon end. The plant embryo is deposited within the cavity of the cotyledon restraint, cotyledon end first. The plant embryo is then sealed within the seed blank by an end seal. There is a weakened spot in the end seal to allow the radicle end of the plant embryo to penetrate the end seal.
In the past, the end seat is attached to the manufactured seed by either stretching a wax base film, such as Parafilm®, or forming a wax seal to enclose the embryo within the manufactured seed. Although such types of end seals are successful in sealing the embryo within the manufactured seed, they are not without their problems. As a non-limiting example, such end seals work well in laboratory conditions but can prematurely break when placed in more rigorous handling environments, such as agricultural sowers. Additionally, to protect against microbial invasion, such end seals have been treated with a tribiotic ointment. Such a treatment further reduces the strength of the end seal. Because the tertiary seal structurally degrades when exposed to predetermined environmental conditions, (e.g., it swells when hydrated) it allows the tertiary seal to become penetrable, thereby facilitating germination through both the primary and secondary end seals. Additionally, the tertiary seal is suitable as a carrier for pesticides that further protect the embryo prior to and during germination. As germination occurs through the tertiary seal, the pesticides remain functional as the tertiary seal is penetrated.
Currently, the artificial seed may be coated with an anti-microbial. In such cases, the anti-microbial applies an anti-microbial coating to the germinate as it penetrates the tertiary seal. Although effective, it has been discovered by the inventors of the present disclosure that such artificial seeds having an anti-microbial coating provide limited benefit in production.
As a non-limiting example, artificial seeds are often subjected to daily irrigation either in the form of rain or crop irrigation. In such instances, much of the available anti-microbial is lost to the surrounding soil long before the embryo begins germination. Even if an effective amount of anti-microbial remains after irrigation, the germinate often penetrates the tertiary seal at a non-perpendicular angle to the surface of the tertiary seal. Such penetration results in a gap between the germinate and the tertiary seal. This is undesirable as gapping creates a spacing wherein no anti-microbial is applied to the germinate, thereby defeating the purpose of an anti-microbial coating. Thus, there exists a need to inhibit bacterial growth on an end seal of an artificial seed.
An artificial seed is provided. In accordance with one embodiment of the present disclosure, such an artificial seed includes a seed shell having a cavity sized and configured to receive an embryo. The artificial seeds also includes a live end seal and a tertiary seal. The live end seal is attached to the seed shell and is positioned to substantially seal the embryo within the seed shell. The tertiary seal is attached to the live end seal and includes a pesticide additive to inhibit at least bacterial growth on the tertiary seal.
An artificial seed formed in accordance with the various embodiments of the present disclosure have several advantages over currently available manufactured seeds. In that regard, the pesticide additive inhibits bacterial growth on the tertiary seal thereby increasing survivability of the germinant.
The foregoing aspects and many of the attendant advantages of this disclosure will become better understood by reference to the following detailed description, when taken in conjunction with the accompanying drawings, wherein:
In other embodiments, the seed shell 24 is formed from a tubular section of biodegradable, plastic material. One such material is a utilized polylatic acid (“PLA”) and is sold by NAT-UR of Los Angeles, Calif. Another material within the scope of the disclosure is a polycaprolactone (“PCL”) mixture, such as Dow Tone P-787 (Dow Chemical Co., Midland, Mich. 48647) with a 1% Tegomer H SI6440 plasticizer (Degussa Goldschmidt Chemical Corp, 914 East Randolph Road, Hopewell, Va. 23860).
Such biodegradable plastic tubes are similarly sectioned into appropriate lengths for a manufactured seed. Further, such biodegradable plastic tubes do not require a wax coating as such tubes are already resistive to environmental elements. It should be apparent that although sectioning tubes is preferred, other embodiments, such as obtaining tubes of appropriate size for use as manufactured seeds, are also within the scope of the present disclosure.
The cylcap 22, also known as a restraint, is suitably manufactured from a porous material having a hardness strong enough to resist puncture or fracture by a germinating embryo, such as a ceramic or porcelain material, and includes an end seal portion 30 and a cotyledon restraint portion 32. The cotyledon restraint portion 32 is suitably integrally or unitarily formed with the end seal portion 30. The cylcap 22 also includes a longitudinally extending cavity 34 extending through the end seal portion 30 and partially through one end of cotyledon restraint portion 32. The open end of the cavity 34 is known as a cotyledon restraint opening 36. The cavity 34 is sized to receive a plant embryo 42 therein.
In certain embodiments, as the cylcap 22 is suitably manufactured from a porous material, it may be desirable to coat the cylcap 22 with a barrier material to reduce the rate of water loss and restrict or reduce microbial entry. Such barriers include wax, polyurethane, glaze, nail polish, and a coating sold by Airproducts Airflex 4514.
The end seal portion 30 is suitably circular when viewed in a top planar view and includes sidewalls 38. Although circular is the preferred embodiment of the end seal portion 30, other embodiments and shapes, such as polygonal, square, triangular, oval and other shapes, are also within the scope of the present disclosure.
In the embodiment of
The sidewalls 38 may include a tapered portion 40. The tapered portion 40 may be a chamfer of one end of the end seal portion 30. The tapered portion 40 assists in assembling the cylcap 22 to the seed coat 24 during manufacturing. Although a tapered portion 40 is preferred, other embodiments, such as a cylcap that does not include a tapered portion, are also within the scope of the present disclosure. An embryo 42 is disposed within the cavity 34 and is suitably sealed therein by a live end seal 43.
The live end seal 43 includes a primary end seal 44 and a secondary end seal 21. The primary end seal 44 is suitably formed from a PCL material described above and includes a centrally located opening 50. The opening 50 is sized to correspond to diameter of the cavity 34 of the cylcap 22 to permit a germinating embryo 42 to pass therethrough. The primary end seal 44 is suitably attached to the end seal portion 30 by a variety of methods, including glue or heat bonding.
As a non-limiting example, the primary end seal 44 is mated to a pre-heated cylcap 22, such that the opening 50 is located above the cavity 34. The heat welds or bonds the primary end seal 44 to the cylcap 22. It should be apparent that the primary end seal 44 may be attached to the cylcap 22 before or after the cylcap 22 is attached to the seed shell 24. Also, if the seed shell 24 is constructed from PCL, it is desirable but not necessary that the melt temperature of the primary end seal 44 and the seed shell 24 be similar.
As another non-limiting example of attaching the primary end seal 44 to the cylcap 22, includes an adhesive gasket. In this example, the primary end seal 44 is heat sealed or bonded to the cylcap 22 with the opening 50 co-axially aligned with the cavity 34. In this process, a form is used to bend edges of the primary end seal 44 around the perimeter of the end seal portion 30 of the cylcap 22. If the melt temperature of the primary end seal 44 and the seed shell 24 are different, then a low bloom cyanoacrylate is used as an adhesive gasket to bond the primary end seal 44 and the seed shell 22.
Heat is applied after the glue and is done so as to thin the glue seal by melting incongruities that typically occur when manufacturing the seed shell 24 and forming the adhesive joint. Thereafter, the cylcap 22, including the primary end seal 44, is attached to the seed shell 24. As noted above, this method is also suitable to a cylcap 22 that is already attached to the seed shell 24. Finally, the foregoing method of attaching a primary end seal 44 to a seed shell 24 may be used for heat weld compatible or incompatible materials.
The secondary end seal 21 will now be described in greater detail. In that regard, the secondary end seal 21 is suitably formed from a well-known sealing material, such as Parafilm®. The secondary end seal 21 is formed and attached to the primary end seal 44 by a well-known method, such as heat bonding or gluing. In some embodiments, a sealing wax may be used to facilitate bonding between the PCL and the Parafilm. The secondary end seal 21 also includes a predetermined burst strength to permit a germinating embryo 42 to penetrate through the live end seal 44.
Still referring to
The combination of the tertiary seal 60 and live end seal 43 creates a sealing surface, wherein the sealing layer, defined by the tertiary seal 60, is made from a predetermined material that degrades in structural integrity after a predetermined exposure to environmental conditions. The tertiary seal 60 also serves as an anti-microbial sealant to seal and protect around the embryo as the embryo germinates and emerges from within the seed shell 24 and protects the cotyledon restraint cavity. Suitable materials used to manufacture the tertiary seal 60 include water soluble materials, wax, environmentally degradable materials, and biodegradable materials. Thus, such materials, as well as materials equivalent in structure and properties, are within the scope of the present disclosure.
If the material used to manufacture the tertiary seal 60 is water soluble, it may include anti-microbial agents. As an example, a water soluble glue having prills of controlled release anti-microbial agents may be applied to the secondary end seal 21. The water soluble glue having flowable or wettable powder pesticides held in suspension within the glue is also within the scope of the present disclosure. In still yet another embodiment, the glue may include an adsorptive agent or carrier on which pesticides are adsorbed (e.g., charcoal or lignin). Also, a water soluble glue without an anti-microbial agent is within the scope of the present disclosure.
If a wax is used as the tertiary seal, it is desirable that the wax be of the type that is solid at sowing temperatures, and melts when exposed to a predetermined temperature, such as a mid-day seed zone temperature of between 25°-35° C. As still yet another example, the tertiary seal 60 may be manufactured from a polymer glue, such as H. B. Fuller PD 120, with filler or controlled release agent dispersed within. Such fillers within the scope of the present disclosure include activated charcoal, powdered lignin, fine sand and talc.
The tertiary seal 60 is also suitably manufactured from a hydroxypropylmethylcellulose. Other types of hydrophilic materials and cellulose-based coatings include cellulose acetate phthalate, hydroxypropylethylcellulose, ethylcellulose, methylcellulose, microcrystalline cellulose, and carrageenan. Such materials have the desired properties of having a relatively high structural integrity when dry and such structural integrity degrades when exposed to environmental conditions, such as water.
In certain embodiments, it is desirable to add an anti-microbial agent, such as Thiram 42S. Any anti-microbial agent that is substantially non-phytotoxic at the desired concentration is also within the scope of the present disclosure. As is described in greater detail below, a tertiary seal 60 treated with an anti-microbial agent is suitable as a carrier for pesticides to protect the embryo 42 prior to and during germination.
The break-through strength of the tertiary seal 60 is a function of the polymer used and the amount of it used to create the tertiary seal 60. As a non-limiting example, breaking strength was tested using a tertiary seal 60 manufactured from hydroxypropylmethylcellulose (HPMC) treated with Thiram 50WP as the anti-microbial agent. A test was conducted to determine the breaking strength of various mixtures. In that regard, a total of six treatments, as set forth below, were tested for break-through strength. A mixture of 2.64 g of HPMC 120 and 0.36 g HPMC 4000 was created for use in treatments 1 and 2.
Treatment 1 used a 0.91 g HPMC mix plus 0.4823 g Thiram and 8.61 ml of water, resulting in a 9.1% HPMC mix by weight.
Treatment 2 used 1.25 g HPMC mix plus 0.4823 g Thiram and 8.27 ml of water, resulting in 12.5% HPMC mix by weight.
Treatment 3 included 0.91 g HPMC 4000 plus 0.4823 g Thiram and 8.61 ml of water, resulting in 9.1% HPMC 4000 by weight.
Treatment 4 utilized 0.86 g HPMC 4000 plus 0.4823 g Thiram and 8.66 ml of water.
Treatment 5 utilized a mechanically disturbed lid attached to the seed.
Treatment 6 used a mechanically disturbed lid attached to the seed and then coated with a tribiotic ointment and left for 24 hours before testing. In this case, the secondary end seal has been slightly disturbed with an abrasion pad scrubber to allow the tertiary seal to be glued to the primary end seal.
Treatments 1-4 were done on top of the seed made as in treatment 6.
Twelve seeds per treatment were tested after coating and drying, and another twelve were tested 1 to 1.5 hours after they were rewetted with water. Table 1, set forth below, sets forth the results.
As may be best seen by referring to
When the artificial seed 20 is handled and sowed, the tertiary seal 60 protects the live end seal 43 from damage associated with such activities. The tertiary seal 60 softens during irrigation following sowing to allow the live end seal 43 to break at the desired level during germination. The tertiary seal 60 softens when exposed to water due to the hydrophilic properties of the materials used to manufacture the tertiary seal 60. As a result, the structural integrity of the tertiary seal 60 degrades when exposed to various environmental conditions, while initially maintaining its structural integrity during handling and sowing.
Referring to
An artificial seed 220 constructed in accordance with still yet another embodiment of the present disclosure may be best understood by referring to
Artificial seeds 220 having microencapsulated pesticide additives are also within the scope of the present disclosure. As an example, microencapsulating silver oxide is one method of creating a controlled release chemical to provide a desired release of the pesticide additive during the germination period of an embryo 42 dispose within the artificial seed 220. One such controlled released pattern is illustrated in
The inclusion of a pesticide additive to the tertiary seal chemistry has been discovered by the current inventors to inhibit fungal and bacterial growth on the tertiary seal 260. This is distinguished from the previous embodiments, wherein the anti-microbial coats the germinates during germination to apply a pesticide coating to the germinating embryo. The present inclusion of a pesticide additive to the tertiary seal chemistry inhibits actual fungal and bacterial growth on the tertiary seal, such that the germinates need not be necessarily coated with the pesticide to enhance survivability. The current inventors conducted an experiment to measure the inhibition of the invasion of tertiary seal material by microbes from soil or a potting mix.
In the experiment, potato dextrose agar was made up with either soil suspension using nursery soil or a mixture of peat/vermiculite/perlite potting mix commonly used in container growing plants. The medium included 19 g of potato dextrose broth and 12 g agar dissolved in 800 ml of water in an autoclave for 20 minutes and, thereafter, cooled in a water bath to 38° C. The soil agar suspension was made by adding 0.5 g of soil or potting mix to 80 ml sterilized water agar (1 g agar in 1,000 ml water) and shaken until it is well suspended. A 0.4 ml aliquot of soil suspension is placed onto a sterile Petri plate and then cooled.
The chemistry, including a pesticide additive, used to make the tertiary seal was microencapsulated and thereafter placed on a wax paper sheet and dried. They were then lifted off the wax paper as dried disks and placed near the center of each Petri dish containing the inoculated medium. These Petri dishes were then placed in an incubator at 24° C. Periodically, the progression of infection on the plates and the tertiary seal material was scored.
Thus, it has been surprisingly discovered by the inventors of the present disclosure that the addition of a pesticide additive to the tertiary seal chemistry resulted in an inhibition of fungal and bacteria overgrowth on the resulting tertiary seal 260. The benefit of keeping inoculums from growing on the tertiary end seal material results in a germinant having enhanced survivability as it is isolated from potential infection. Although mixing a pesticide additive into the tertiary seal chemistry is preferred, it should be apparent that other mixings, such as mixing the pesticide additive into the nutritive media 26, are also within the scope of the present disclosure.
While the preferred embodiment of the disclosure has been illustrated and described, it will be appreciated that various changes can be made therein without departing from the spirit and scope of the disclosure.
The present application claims the benefit of U.S. Provisional Application No. 60/947,032, filed Jun. 29, 2007.
| Number | Date | Country | |
|---|---|---|---|
| 60947032 | Jun 2007 | US |
