FreshPatents.com Logo
stats FreshPatents Stats
1 views for this patent on FreshPatents.com
2012: 1 views
Updated: December 22 2014
newTOP 200 Companies filing patents this week


Advertise Here
Promote your product, service and ideas.

    Free Services  

  • MONITOR KEYWORDS
  • Enter keywords & we'll notify you when a new patent matches your request (weekly update).

  • ORGANIZER
  • Save & organize patents so you can view them later.

  • RSS rss
  • Create custom RSS feeds. Track keywords without receiving email.

  • ARCHIVE
  • View the last few months of your Keyword emails.

  • COMPANY DIRECTORY
  • Patents sorted by company.

Your Message Here

Follow us on Twitter
twitter icon@FreshPatents

somatic embryogenesis and embryo harvesting and method and apparatus for preparing plant embryos for plant production

last patentdownload pdfdownload imgimage previewnext patent

20120276634 patent thumbnailZoom

somatic embryogenesis and embryo harvesting and method and apparatus for preparing plant embryos for plant production


Described herein are methods and media for facilitating somatic embryogenesis and for collecting, conditioning, and transferring the washed embryos onto a substrate and into an environment suitable for conditioning the embryos for a desired period of time so they become germination-competent for plant production. The described plant embryo cleaning apparatus and method are used for preparing multiple plant embryos for plant production. The apparatus and method can use a cleaning fluid source, a fluid-conditioning system, a fluid-delivery structure, a cleaning station, an outlet mechanism, a negative pressure source, and a controller.
Related Terms: Embryo Somatic Embryogenesis

Browse recent Arborgen Inc. patents - ,
Inventors: John Joseph CLARK, Narender Singh NEHRA, Mark Russell RUTTER, Jessica S. SAGE, Sydney Keith SEYMOUR, Timothy Joel STOUT, George SURRITTE, Ronald W. WINKLES
USPTO Applicaton #: #20120276634 - Class: 435422 (USPTO) - 11/01/12 - Class 435 
Chemistry: Molecular Biology And Microbiology > Plant Cell Or Cell Line, Per Se (e.g., Transgenic, Mutant, Etc.); Composition Thereof; Process Of Propagating, Maintaining, Or Preserving Plant Cell Or Cell Line; Process Of Isolating Or Separating A Plant Cell Or Cell Line; Process Of Regenerating Plant Cells Into Tissue, Plant Part, Or Plant, Per Se, Where No Genotypic Change Occurs; Medium Therefore >Culture, Maintenance, Or Preservation Techniques, Per Se >Involving Conifer Cell Or Tissue (e.g., Pine, Spruce, Fir, Cedar, Etc.)



view organizer monitor keywords


The Patent Description & Claims data below is from USPTO Patent Application 20120276634, somatic embryogenesis and embryo harvesting and method and apparatus for preparing plant embryos for plant production.

last patentpdficondownload pdfimage previewnext patent

This application is a continuation application of U.S. application Ser. No. 12/511,548, filed on Jul. 29, 2009 (U.S. Pat. No. 8,216,841) which is a divisional of U.S. application Ser. No. 11/413,105, filed on Apr. 28, 2006 (U.S. Pat. No. 7,665,243), both of which claim priority to U.S. Provisional Application Ser. No. 60/675,949, filed on Apr. 29, 2005. All applications are incorporated herein in their entirety by reference.

FIELD OF THE INVENTION

Described herein are methods and media for facilitating somatic embryogenesis and for collecting, conditioning, and storing of large numbers of plant embryos prior to germination. Also described herein are a method and apparatus for preparing plant embryos for plant production.

BACKGROUND

Collecting, storing, and conditioning plant embryos, especially somatic embryos, prior to germination are key processes in many aspects of the agriculture industry. The activities necessary for performing these processes, however, are usually performed by hand. For instance, individual embryos are typically transferred to and from various media and vessels and must be plated onto gel media, one by one using forceps and often with the guidance of a dissecting microscope.

Such “hand harvesting” methods are burdensome, time-consuming, costly, and susceptible to contamination. Not only that, but only a limited number of embryos can be collected and treated by a single person during a given period of time. Accordingly, any attempt to increase the number of embryos that can be harvested and subsequently conditioned for germination necessarily requires an increase in manpower, which itself can be costly and often impractical.

An added concern is the inclusion of polyethylene glycol in embryo development media as a osmotic agent. Polyethylene glycol has been incorporated into various media to boost embryogenic development because it is thought to help trigger embryo development. See Fowke et al., Somatic Cell Genetics and Molecular Genetics of Trees, Quebec City, Canada, Aug. 12-16, 1997, which is incorporated herein by reference.

A problem with polyethylene glycol, however, is that it adheres to embryos, possibly interfering with embryo germination. Traditionally, removal of polyethylene glycol is accomplished by storing polyethylene glycol (PEG)-treated embryos on a gel medium without PEG in the cold for a number of weeks. The polyethylene glycol eventually diffuses into the medium away from the embryos. Not surprisingly, this is a time-consuming and burdensome treatment and removal strategy, which imparts an oftentimes unacceptable delay in the overall harvesting and conditioning process.

The agricultural industry and, in particular, the forestry sciences, therefore, are faced with a laborious, expensive, and inefficient method for making, gathering and preparing plant embryos. Such factors prove to be obstacles when operating at commercial levels. And still, hand harvesting is a typically routine practice.

As explained below, however, the present invention provides a robust “Mass Harvesting” method that is rapid and inexpensive. Since Mass Harvesting (MH) minimizes human intervention, it is less susceptible to contamination. Furthermore, the present invention also provides a new way for removing polyethylene glycol. Moreover, the Mass Harvesting method is highly efficient, allowing the simultaneous collection of thousands and hundreds of thousands of plant embryos during a period of time, and can be readily scaled-up for commercial purposes.

In this respect, the present invention also provides a combinatorial approach to exploiting and optimizing genotype-by-treatment interactions of multiple steps in the somatic embryogenesis process.

SUMMARY

In one aspect of the invention, a method for preparing embryos for plant production is provided, which comprises (i) washing multiple plant embryos simultaneously, and (ii) transferring the washed embryos onto a substrate and into an environment suitable for conditioning the embryos for a desired period of time so they become germination-competent for plant production. The method may further comprise retrieving one or more of the embryos at any time point during the desired period of time.

In one embodiment, the plant embryos are somatic embryos. In another embodiment, the embryos are washed on a porous surface. In yet another embodiment, no single embryo has been individually placed by hand onto the porous surface.

In one embodiment, the substrate that is suitable for storing the embryos is a gel, which comprises maltose, glutamine, and abscisic acid. The gel also may contain other ingredients, such as inorganic nutrients. The person of skill in the art of embryo storage and development knows what other ingredients are useful for maintaining and manipulating plant embryos. In another embodiment, the substrate is a filter paper saturated with a volume of liquid media, which comprises maltose, glutamine, and abscisic acid. The gel also may contain other ingredients, such as inorganic nutrients. In another embodiment, the volume of the liquid media that is added to the substrate is 1 ml or 2 ml.

Other conditioning embodiments include, but are not limited to, the following: embryos stored on a gelled medium in cold (1° C. to 12° C., optimally 3 to 6° C.) for varying time (1 day to 24 weeks, optimally from 3 to 12 weeks). During this cold storage the embryos can be placed on a polyester or paper membrane to facilitate subsequent transfer. Embryos on the polyester or paper membrane are then transferred as an entire unit to a vessel and sealed with Nescofilm™, or optionally are placed on top of a dry filter paper within the vessel and sealed with Nescofilm™. Embryos in the sealed vessel are held at room temperature (15 to 30° C., ideally 20 to 28° C.) for varying time (1 to 12 weeks, optimally from 2 to 5 weeks depending on the temperature to which the embryos were exposed during either of the above steps of this conditioning method. That is during: a. cold on a gelled medium and, b. warm in sealed vessel).

In one embodiment, the embryos are stored for about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks, about 12 weeks, about 13 weeks, about 14 weeks, about 15 weeks, about 16 weeks, about 17 weeks, about 18 weeks, about 19 weeks, about 20 weeks, about 21 weeks, about 22 weeks, about 23 weeks, about 24 weeks, or more than about 24 weeks.

Another aspect of the present invention is a liquid medium for growing embryonic tissue that comprises a high concentration of casein. A high concentration of casein may be about 900 mg/l, about 1000 mg/l, about 1100 mg/l, about 1200 mg/l, about 1300 mg/l, about 1400 mg/l, about 1500 mg/l, about 1600 mg/l, about 1700 mg/l, about 1800 mg/l, about 1900 mg/l, about 2000 mg/l, about 2100 mg/l, about 2200 mg/l, about 2300 mg/l, about 2400 mg/l, about 2500 mg/l, about 2600 mg/l, about 2700 mg/l, about 2800 mg/l, about 2900 mg/l, about 3000 mg/l, or more than 3000 mg/l. In one embodiment the concentration of casein is between 1100 mg/l and 3000 mg/l.

In one embodiment, the embryonic tissue is from a conifer. In a preferred embodiment, the conifer is pine. In a more preferred embodiment, the pine is Loblolly pine.

In another embodiment, the coniferous tree is selected from the group consisting of Eastern white pine, Western white, Sugar pine, Red pine, Pitch pine, Jack pine, Longleaf pine, Shortleaf pine, Loblolly pine, Slash pine, Virginia pine, Ponderosa pine, Jeffrey pine, Pond pine, and Lodgepole pine, Radiata pine and hybrid crosses thereof. In another preferred embodiment, the coniferous tree is selected from the group consisting of, but not limited to, Abies alba, Abies amabilis, Abies balsamea, Abies bornmuelleriana, Abies concolor, Abies fraseri, Abies grandis, Abies koreana, Abies lasiocarpa, Abies nordmanniana, Abies procera, Araucaria angustifolia, Araucaria araucana, Araucaria bidwillii, Araucaria cunninghamii, Cedrus atlantica, Cedrus deodara, Chamaecyparis lawsoniana, Chamaecyparis pisifera, Cryptomeria japonica, Cuppressocyparis leylandii, Larix decidua, Larix occidentalis, Metasequoia glyptostroboides, Picea abies, Picea engelmannii, Picea glauca, Picea mariana, Picea pungens, Picea rubens, Picea sitchensis, Pinus banksiana, Pinus caribaea, Pinus contorta, Pinus echinata, Pinus edulis, Pinus elliotii, Pinus jeffreyi, Pinus korariensis, Pinus lambertiana, Pinus merkusii, Pinus monticola, Pinus nigra, Pinus palustris, Pinus pinaster, Pinus ponderosa, Pinus rigida, Pinus radiata, Pinus resinosa, Pinus serotina, Pinus strobus, Pinus sylvestris, Pinus taeda, Pinus virginiana, Pseudotsuga menziesii, Sequoia sempervirens, Sequoiadendron giganteum, Taxodium ascends, Taxodium distichum, Taxus baccata, Taxus brevifolia, Taxus cuspidata, Thuja occidentalis, Thuja plicata, Tsuga canadensis, Tsuga heterophylla, and hybrid crosses thereof.

Specific examples of each of such coniferous tree includes: Abies alba, European silver fir; Abies amabilis, Pacific silver fir; Abies balsamea, Balsam fir; Abies bornmuelleriana, Turkish fir; Abies concolor, White fir; Abies fraseri, Fraser fir; Abies grandis, Grand fir; Abies koreana, Korean fir; Abies lasiocarpa, Alpine fir; Abies nordmanniana, Nordman fir; Abies procera, Noble fir; Araucaria angustifolia, Parana pine; Araucaria araucana, Monkeypuzzle tree; Araucaria bidwillii, Bunya pine; Araucaria cunninghamii, Hoop pine; Cedrus atlantica, Atlas cedar; Cedrus deodara, Deodar cedar; Chamaecyparis lawsoniana, Port-Orford-cedar; Chamaecyparis pisifera, Sawara cypress; Cryptomeria japonica, Japanese cedar (Japanese cryptomeria); Cuppressocyparis leylandii, Leyland Cypress; Larix decidua, European larch; Larix occidentalis, Western larch; Metasequoia glyptostroboides, Dawn redwood; Picea abies, Norway spruce; Picea engelmannii, Englemann spruce; Picea glauca, White spruce; Picea mariana, Black spruce; Picea pungens, Colorado blue spruce; Picea rubens, Red spruce; Picea sitchensis, Sitka spruce; Pinus banksiana, Jack pine; Pinus caribaea, Caribbean pine; Pinus contorta, lodgepole pine; Pinus echinata, Shortleaf pine; Pinus edulis, Pinyon pine; Pinus elliotii, Slash pine; Pinus jeffreyi, Jeffrey Pine; Pinus korariensis, Korean pine; Pinus lambertiana, Sugar pine; Pinus merkusii, Sumatran pine; Pinus monticola, Western white pine; Pinus nigra, Austrian pine; Pinus palustris, Longleaf pine; Pinus pinaster, Maritime pine; Pinus ponderosa, Ponderosa pine; Pinus rigida, Pitch pine; Pinus radiata, Radiata pine; Pinus resinosa, Red pine; Pinus serotina, Pond pine; Pinus strobus, Eastern white pine; Pinus sylvestris, Scots (Scotch) pine; Pinus taeda, Loblolly pine; Pinus virginiana, Virginia pine; Pseudotsuga menziesii, Douglas-fir; Sequoia sempervirens, Redwood; Sequoiadendron giganteum, Sierra redwood; Taxodium ascends, Pond cypress; Taxodium distichum, Bald cypress; Taxus baccata, European yew; Taxus brevifolia, Pacific or Western yew; Taxus cuspidata, Japanese yew; Thuja occidentalis, Northern white-cedar; Thuja plicata, Western red cedar; Tsuga canadensis, Eastern hemlock; Tsuga heterophylla, Western hemlock.

In another embodiment, the coniferous plant tissue is a Southern Yellow pine. In yet another embodiment, the Southern Yellow pine is selected from the group consisting of Pinus taeda, Pinus serotina, Pinus palustris, and Pinus elliottii.

The present invention contemplates the Mass Harvesting of somatic embryos from any of these coniferous trees. The present invention is not limited, however, to the Mass Harvesting of only coniferous tree tissues and somatic embryos.

In another embodiment, therefore, the plant tissue, such as embryogenic tissue or a somatic embryo is from a tree selected from the group consisting of chestnut, ash, beech, basswood, birch, black cherry, black walnut/butternut, chinkapin, cottonwood, elm, eucalyptus, hackberry, hickory, holly, locust, magnolia, maple, oak, poplar, red alder, royal paulownia, sassafras, sweetgum, sycamore, tupelo, willow, and yellow-poplar, and intra- and inter-species hybrid crosses thereof. A particularly preferred chestnut for use in the present invention is the American Chestnut.

In one embodiment, the concentration of casein in the liquid medium is about 400, about 500, about 600, about 700, about 800, about 900, about 1000, about 1100, about 1200, about 1300, about 1400, about 1500, about 1600, about 1700, about 1800, about 1900, about 2000, about 2100, about 2200, about 2300, about 2400, about 2500, about 2600, about 2700, about 2800, about 2900, or about 3000 mg/l or any integer in between these concentrations.

In one embodiment, the casein is casein hydrolysate.

Another aspect of the present invention is a method for obtaining germinating embryos, comprising (i) placing embryogenic cultures from cryostorage onto cryoretrieval medium for a period of time and thereafter growing the embryogenic tissue in liquid medium, (ii) transferring the embryogenic tissue to embryo development medium to generate embryos, (iii) washing a mass of the generated embryos with water, (iv) placing the washed mass of embryos on a substrate that is saturated with conditioning medium, and (v) germinating embryos therefrom, wherein (a) the cryoretrieval medium comprises at least one of a high concentration of casein or an amount of Brassinolide, (b) the liquid medium has a high concentration of casein, (c) the embryo development medium has a desired amount of polyethylene glycol, and (d) the conditioning medium is liquid.

In this method, the liquid medium comprises a concentration of casein which is about 1100, about 1200, about 1300, about 1400, about 1500, about 1600, about 1700, about 1800, about 1900, about 2000, about 2100, about 2200, about 2300, about 2400, about 2500, about 2600, about 2700, about 2800, about 2900, or about 3000 mg/l or any integer in between these concentrations.

In another embodiment, the percentage of polyethylene glycol in the embryo development medium is 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20%. In one embodiment, the percentage of polyethylene glycol in the embryo development medium is 7%. In another embodiment, the percentage of polyethylene glycol in the embryo development medium is 13%.

In one embodiment, the cryoretrieval medium comprises an amount of Brassinolide. In one embodiment, the amount of Brassinolide is 0.01 μM, 0.02 μM, 0.03 μM, 0.04 μM, 0.05 μM, 0.06 μM, 0.07 μM, 0.08 μM, 0.09 μM, 0.10 μM, 0.11 μM, 0.12 μM, 0.13 μM, 0.14 μM, 0.15 μM, 0.16 μM, 0.17 μM, 0.18 μM, 0.19 μM, 0.20, or 0.50 μM. In one embodiment, the concentration of Brassinolide is 0.10 μM.

In another aspect, a method for identifying optimal genotype-specific conditions for embryogenic tissue growth is provided, comprising (i) growing embryogenic tissue that has been retrieved from cryostorage on a medium that comprises an amount of Brassinolide and (ii) comparing the growth of the embryogenic tissue to the growth of embryogenic tissue from the same genotype on media that comprises at least one different amount of Brassinolide.

In another aspect, a method for identifying optimal genotype-specific conditions for embryo production is provided, comprising (i) growing embryogenic cultures on an embryo development medium that comprises an amount of polyethylene glycol and (ii) comparing the growth of the embryogenic cultures into embryos to the growth of embryos from the same genotype on embryo development media that comprises at least one different amount of polyethylene glycol.

In another aspect of the methods disclosed herein are combined to produce a method for identifying optimal genotype-specific conditions for embryogenic tissue growth and embryo production for a particular plant genotype.

In one embodiment, after Mass Harvesting according to any one of these methods, embryos are placed onto a substrate that has been saturated with a volume of liquid conditioning medium, which contains nutrients necessary to prepare the embryos for germination. The substrate may be a filter paper.

In one embodiment, the saturated filter paper onto which the embryos are placed is retained within a dish, such as a Petri dish. In another embodiment, the dish is wrapped with tape or porous wrapping material to control the loss of moisture from the dish. In another embodiment, the dish, which contains the filter paper and the embryos thereon is stored in the cold for a period of time.

The length of time a Mass Harvested somatic embryo can be stored in the cold is from 1 to 5 weeks, for at least 5 weeks, for at least 8 weeks, for at least 10 weeks, for at least 12 weeks, for at least 13 weeks, for at least 14 weeks, for at least 15 weeks, for at least 16 weeks, 18 weeks, 20 weeks, 22 weeks, 24 weeks, or for more than 24 weeks.

For instance, a Mass Harvested somatic embryo may be stored in the cold under the conditions described herein for 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 weeks, 49 weeks, 50 weeks, 51 weeks, or 52 weeks, or beyond 52 weeks.

In one aspect of the present invention is a combinatorial method for optimizing somatic embryogenesis, comprising (i) initiating embryogenesis of a plant embryogenic tissue on an initiation medium that comprises a high concentration of casein, (ii) maintaining the initiated embryogenic tissue on a maintenance medium that comprises a high concentration of casein prior to cryostorage, (iii) recovering the embryogenic tissue from cryostorage on a medium that comprises at least one of (a) high concentration of casein or (b) an amount of Brassinolide, and (iv) developing embryos from the recovered embryogenic tissue on an embryo development medium that comprises a percentage of polyethylene glycol that is optimal for the genotype of the embryogenic tissue from which embryos are to developed.

In one embodiment of this method, the percentage of polyethylene glycol in the embryo development medium is 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20%. In one embodiment, the percentage of polyethylene glycol in the embryo development medium is 7%. In another embodiment, the percentage of polyethylene glycol in the embryo development medium is 13%.

In another embodiment, the medium onto which the embryogenic tissue is recovered after cryostorage comprises a high concentration of casein and an amount of Brassinolide.

In one embodiment, the amount of Brassinolide is 0.01 μM, 0.02 μM, 0.03 μM, 0.04 μM, 0.05 μM, 0.06 μM, 0.07 μM, 0.08 μM, 0.09 μM, 0.10 μM, 0.11 μM, 0.12 μM, 0.13 μM, 0.14 μM, 0.15 μM, 0.16 μM, 0.17 μM, 0.18 μM, 0.19 μM, or 0.20 μM. In one embodiment, the concentration of Brassinolide is about 0.10 μM.

In another embodiment, the initiation medium further comprises a low concentration of maltose. In one embodiment, the concentration of maltose is about 1 g/l, 2 g/l, 3 g/l, 4 g/l, 5 g/l, 6 g/l, 7 g/l, 8 g/l, 9 g/l, 10 g/l, 11 g/l, 12 g/l, 13 g/l, 14 g/l, 15 g/l, 16 g/l, 17 g/l, 18 g/l, 19 g/l, or 20 g/l. In one embodiment, the concentration of maltose is about 15 g/l.

In another aspect of the present invention is a method for preparing embryos for storage, comprising (i) simultaneously washing multiple plant embryos, and (ii) transferring the washed embryos onto a substrate suitable for conditioning the embryos for storage in a vessel for a desired period of time. In one embodiment, wherein the plant embryos are somatic embryos. In one embodiment, the plant embryos are washed onto a mesh that permits passage of cellular debris and liquid but not the passage of the embryos. Hence, in one embodiment, the embryos are washed on a porous surface and wherein no embryo is placed by hand onto the porous surface. In one embodiment, the step of transferring the washed embryos comprises inverting the mesh on which the embryos were washed directly onto the substrate, wherein the substrate is either already in the vessel or is subsequently moved to a vessel or environment for suitable conditioning and storage. Hence, the embryos may be inverted from the washing mesh and onto a conditioning substrate.

In another embodiment, the conditioning substrate is a gel comprising maltose, glutamine, and abscisic acid. In another embodiment, the conditioning substrate is a filter paper saturated with a volume of liquid media, which comprises maltose, glutamine, and abscisic acid. In one embodiment, the volume of the liquid media is 1 ml or 2 ml.

In one embodiment, conditioning takes place in a high relative humidity environment without cold storage. In another embodiment, conditioning comprises storing the embryos on a gelled medium in the cold for a period of time. In another embodiment, the method further comprises placing the embryos onto a polyester or paper membrane, transferring the membrane to a vessel, which is then sealed, maintaining the vessel at a warm temperature for a period of time.

An aspect of the present invention relates to an apparatus for preparing multiple plant embryos for plant production. The apparatus includes a fluid-delivery structure for delivering input liquid to the multiple plant embryos, a cleaning station in fluid communication with the fluid-delivery structure and configured to hold the multiple plant embryos to receive input liquid from the fluid-delivery structure to clean cellular debris from the multiple plant embryos, an outlet mechanism in fluid communication with the cleaning station and configured to receive output liquid from the cleaning station, and a controller configured to control at least one of the fluid-delivery structure, the cleaning station, and the outlet mechanism.

In an embodiment, the fluid-delivery structure can include a spray mechanism for spraying the multiple plant embryos.

In another embodiment, the cleaning station can include a wash unit for washing the multiple plant embryos, and a rinse unit for rinsing the multiple plant embryos.

In yet another embodiment, the rinse unit can include a porous material configured to hold the multiple plant embryos and having a pore size within a range of 15 microns to 65 microns. The porous material can be configured to hold the multiple plant embryos, the porous material being removable to remove the multiple plant embryos from the rinse unit.

In yet another embodiment, the cleaning station can include a holding unit that transports the multiple plant embryos from the wash unit to the rinse unit. The holding unit can include a porous material in which the pore size can be within the range of 400 microns to 900 microns. The holding unit can include a first porous material configured to hold the multiple plant embryos and having a first pore size. The rinse unit can include a second porous material configured to hold the multiple plant embryos and having a second pore size. Preferably, the second pore size is smaller than the first pore size.

In yet another embodiment, at least one of the fluid delivery structure, wash unit, rinse unit, and holding unit includes a substantially transparent housing to permit monitoring of at least one of washing and rinsing through the substantially transparent housing.

In yet another embodiment, the apparatus includes structure controlled by the controller to move the holding unit from the wash unit to the rinse unit.

In yet another embodiment, the outlet mechanism can include a first outlet in fluid communication with the wash unit and configured to receive output liquid from the wash unit, and a second outlet in fluid communication with the rinse unit and configured to receive output liquid from the rinse unit.

In yet another embodiment, the apparatus can include a negative pressure source in fluid communication with the outlet mechanism to provide a negative pressure. The negative pressure source can include a vacuum system comprising an electronic valve connected to a vacuum pump. The negative pressure source can include a check valve in fluid communication with the cleaning station and configured to operate as a function of output liquid weight and a force of the negative pressure.

In another embodiment, preferably, the controller is configured to control the flow of input liquid through the fluid-delivery structure. The controller can be configured to control the pressure of input liquid delivered by the fluid-delivery structure. The controller can be configured to maintain the impingement of the input liquid within a range of 0.00506 to 0.027 pounds per square inch at a normalized standard distance of twelve inches.

In yet another embodiment, the apparatus can include a negative pressure source in fluid communication with the outlet mechanism, wherein the controller is configured to control a pressure of input liquid delivered by the fluid-delivery structure and to control a pressure supplied by the negative pressure source to the outlet mechanism.

In yet another embodiment, the cleaning station can include a wash unit, and a rinse unit configured to hold the multiple plant embryos. The outlet mechanism can include a first outlet in fluid communication with the wash unit and configured to receive first output liquid from the wash unit, and a second outlet in fluid communication with the rinse unit and configured to receive second output liquid from the rinse unit. The apparatus can further include a negative pressure source in fluid communication with the first and second outlets to supply negative pressure to the first and second outlets, wherein the controller is configured to control the fluid-delivery structure and the negative pressure source.

In yet another embodiment, the apparatus can include a fluid-conditioning system in fluid communication with the fluid-delivery structure and configured to at least one of filter the input liquid and sterilize the input liquid. The fluid-conditioning system can include a membrane filter and a UV sterilizer.

In yet another embodiment, the cleaning station can be configured to remove polyethylene glycol from the multiple plant embryos.

Another aspect of the present invention relates to a method of preparing multiple plant embryos for plant production. The method includes supplying multiple plant embryos in a cleaning station, washing the multiple plant embryos by delivering an input liquid to the plant embryos, and controlling with a controller a flow of input liquid delivered to the plant embryos.

In an embodiment, the impingement of the input liquid can be maintained within a range of 0.00506 to 0.027 pounds per square inch at a normalized standard distance of twelve inches.

In another embodiment, the method can further include supplying a negative pressure to the cleaning station for controlling flow of output liquid, and controlling with the controller the negative pressure supplied to the cleaning station.

In yet another embodiment, the method can further include at least one of filtering the input liquid and sterilizing the input liquid.

In yet another embodiment, the method can include removing polyethylene glycol from the multiple plant embryos in the washing step.

Yet another aspect of the present invention relates to a method of preparing multiple plant embryos for plant production. The method includes supplying multiple plant embryos in a wash unit, washing the multiple plant embryos by delivering a first input liquid into the wash unit, transporting the multiple plant embryos to a rinse unit, rinsing the multiple plant embryos by delivering a second input liquid into the rinse unit, and controlling with a controller at least one of the steps of washing, transporting, and rinsing.

In an embodiment, the method can further include applying a first negative pressure to the wash unit for controlling flow of first output liquid from the wash unit, and applying a second negative pressure to the rinse unit for controlling flow of second output liquid from the rinse unit.

In another embodiment, the method can further include at least one of filtering the first and second input liquids and sterilizing the first and second input liquids.

In yet another embodiment of the method, the first input liquid and the second input liquid can have the same composition. Alternatively, the first input liquid and second input liquid can have different compositions.

Yet another aspect of the present invention relates to a method of preparing multiple conifer somatic embryos for plant production. The method includes positioning the multiple conifer somatic embryos on a porous material having a pore size within a range of 400 microns to 900 microns, and delivering fluid to the multiple conifer somatic embryos on the porous material to clean the conifer somatic embryos. In a further refinement, the pore size of the porous material can be within a range of 560 microns to 710 microns or within a range of 600 microns to 670 microns.

In one embodiment of the present invention at least one of the steps of washing and transferring are automated. Indeed, any one of the methods disclosed herein may comprises steps that are fully or partly automated and/or are computer-operated by software programs that may or may not require human input, intervention, or interaction. In this respect, the present invention also contemplates a fully-automated and semi-automated apparatuses or machines for harvesting embryos. Such an apparatus according to the present invention performs various automated functions pertinent to embryo harvesting techniques of the present invention. Hence, a fully- or semi-automated apparatus of the present invention may perform functions comprising (1) loading of embryos onto a surface, (2) washing of the embryos, (3) rinsing of the embryos, and (4) unloading or transferring of the embryos from the surface to another surface or vessel or container for further manipulation. The apparatus may transfer the treated embryos, by means of a robotic arm or a movable surface, for instance, to a conditioning environment without human intervention. Hence, human intervention may only ever be required at the step of bringing embryos to the apparatus and placing them into or onto the appropriate apparatus surface. From that point onwards, no further human intervention may be necessary until the embryos have been conditioned for a desired period of time. At that point, a human may remove one or more embryos from that conditioning environment to assess whether it is germination competent and then move onwards to plant that germination-ready embryo for plant propagation. Even then, that step, the step of removing the germination-competent embryos can be automated. That is, the apparatus may be designed such that the embryos are automatically removed from the conditioning environment after a period of time that is known to produce germination-competent embryos, and placed onto an appropriate seeding and rooting surface so as to promote germination and shoot growth.

A semi-automated apparatus that performs such functions may be semi-automated in the sense that it may require human intervention at certain points in the process, such as bringing embryos to the apparatus, permitting human intervention to increase or decrease a wash or rinse step, or simply to initiate the computer software that controls the operation of the components of the apparatus. Hence, the present invention contemplates the apparatus that is described herein and which performs the functions outlined above. See also Example 23 below.

The present invention also recognizes and appreciates that certain features of this apparatus can be modified or altered in due course and in response to the embryo harvesting task desired. Hence, the apparatus may be modified so as to increase the total numbers of embryos that can be treated according to the harvesting and washing protocols disclosed herein. For instance, the apparatus disclosed in Example 23, may include more than three units within which to wash embryos. That is, the apparatus may be adapted to include more units or units of larger capacity. Furthermore, the present invention contemplates the manipulation of the computer software that drives and operates the apparatus. In this respect, the present invention contemplates that an automated apparatus of the present invention is controlled by computer software that follows and implements, in computer terms, the process flow diagram depicted in FIG. 11. For instance, the apparatus described herein may be operated by and under the control of computer software that implements the process of FIG. 11. The skilled person appreciates that any of these parameters are open to manipulation. Hence, the present invention contemplates software that controls sensors, which determine the approximate load of embryos that are placed onto a loading surface. Depending on that determination, the software may make and send appropriate computer commands to increase or decrease the length of time of the wash and rinse steps, for example. Hence, if a subsequent batch of embryos is twice that of what was previously loaded, the sensors will direct the duration of the ensuing wash step to be longer or more powerful, or may require the steps of washing and rinsing to be repeated any number of times. Accordingly, the automated apparatus of the present invention for implementing the disclosed and novel harvesting techniques is adaptable, convenient, and useful for simultaneously processing multiple embryos. By multiple embryos, the present invention contemplates that 5,000, 10,000, 20,000, 30,000, 40,000, 50,000, 60,000, 70,000, 80,000, 90,000, 100,000, 110,000, 120,000, 130,000, 140,000, 150,000, 160,000, 170,000, 180,000, 190,000, 200,000, 210,000, 220,000, 230,000, 240,000, 250,000, 260,000, 270,000, 280,000, 290,000, 300,000 or more, or any integer in between, of embryos can be processed, e.g., washed and rinsed, per day by use of the methods and apparatuses disclosed herein.

The present invention also contemplates embryos that are prepared by any of the methods disclosed herein. In another aspect, the present invention encompasses a plant that is grown from any of the treated embryos disclosed herein.

It is to be understood that both the foregoing general description and the following detailed descriptions are exemplary and explanatory only, and are not restrictive of the invention as claimed.

BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying drawings, which are incorporated in and constitute part of this specification, illustrate embodiments of the invention and together with the description, serve to explain the principles of the invention.

FIG. 1 is a schematic showing the steps from embryogenic initiation, liquid bulk-up, embryo development, Mass Harvesting, cold storage, pre-germination, and germination steps.

FIG. 2 is a schematic drawing of an embodiment of a plant embryo cleaning apparatus according to the present invention.

FIG. 3 is a schematic drawing of a cleaning fluid source, a fluid-conditioning system, and a spray mechanism of the plant embryo cleaning apparatus of FIG. 2.

FIG. 4 is a schematic drawing of a cleaning station of the plant embryo cleaning apparatus of FIG. 2.

FIG. 5 is a schematic drawing of an outlet mechanism and negative pressure source of the plant embryo cleaning apparatus of FIG. 2.

FIG. 6 is a perspective view of the plant embryo cleaning apparatus of FIG. 2.

FIGS. 7A to 7F are perspective views of the of the plant embryo cleaning apparatus of FIG. 2 in operation.

FIGS. 8A and 8B are a cross-sectional view and a side view, respectively, of a spray mechanism, a mounting bracket, and a pneumatic cylinder of the plant embryo cleaning apparatus of FIG. 2.

FIGS. 9A and 9B are a plan view and a cross-sectional view, respectively, of holding units, a mounting bracket, a rotational device, and a pneumatic cylinder of the plant embryo cleaning apparatus of FIG. 2.

FIG. 10A is a plan view of wash units, rinse units, two electronic vacuum valves, and a horizontal moving structure of the plant embryo cleaning apparatus of FIG. 2.

FIG. 10B is a side view of the rinse units, a vacuum manifold, and output funnels of the plant embryo cleaning apparatus of FIG. 2.

FIG. 10C is a cross-sectional view showing the wash units, the output funnels, the vacuum manifold, and a horizontal rail of the plant embryo cleaning apparatus of FIG. 2.

FIG. 10D is a cross-sectional view showing the rinse unit, the output funnels, the vacuum manifold, and the horizontal rail of the plant embryo cleaning apparatus of FIG. 2.

FIG. 11 is a process flow diagram of the intermediate Mass Harvesting machine.



Download full PDF for full patent description/claims.

Advertise on FreshPatents.com - Rates & Info


You can also Monitor Keywords and Search for tracking patents relating to this somatic embryogenesis and embryo harvesting and method and apparatus for preparing plant embryos for plant production patent application.
###
monitor keywords

Browse recent Arborgen Inc. patents

Keyword Monitor How KEYWORD MONITOR works... a FREE service from FreshPatents
1. Sign up (takes 30 seconds). 2. Fill in the keywords to be monitored.
3. Each week you receive an email with patent applications related to your keywords.  
Start now! - Receive info on patent apps like somatic embryogenesis and embryo harvesting and method and apparatus for preparing plant embryos for plant production or other areas of interest.
###


Previous Patent Application:
Supplying treated exhaust gases for effecting growth of phototrophic biomass
Next Patent Application:
Fluidic device
Industry Class:
Chemistry: molecular biology and microbiology
Thank you for viewing the somatic embryogenesis and embryo harvesting and method and apparatus for preparing plant embryos for plant production patent info.
- - - Apple patents, Boeing patents, Google patents, IBM patents, Jabil patents, Coca Cola patents, Motorola patents

Results in 1.56127 seconds


Other interesting Freshpatents.com categories:
Electronics: Semiconductor Audio Illumination Connectors Crypto

###

Data source: patent applications published in the public domain by the United States Patent and Trademark Office (USPTO). Information published here is for research/educational purposes only. FreshPatents is not affiliated with the USPTO, assignee companies, inventors, law firms or other assignees. Patent applications, documents and images may contain trademarks of the respective companies/authors. FreshPatents is not responsible for the accuracy, validity or otherwise contents of these public document patent application filings. When possible a complete PDF is provided, however, in some cases the presented document/images is an abstract or sampling of the full patent application for display purposes. FreshPatents.com Terms/Support
-g2--0.6465
Key IP Translations - Patent Translations

     SHARE
  
           

stats Patent Info
Application #
US 20120276634 A1
Publish Date
11/01/2012
Document #
13544843
File Date
07/09/2012
USPTO Class
435422
Other USPTO Classes
435431, 435420
International Class
12N5/04
Drawings
17


Your Message Here(14K)


Embryo
Somatic Embryogenesis


Follow us on Twitter
twitter icon@FreshPatents

Arborgen Inc.

Browse recent Arborgen Inc. patents

Chemistry: Molecular Biology And Microbiology   Plant Cell Or Cell Line, Per Se (e.g., Transgenic, Mutant, Etc.); Composition Thereof; Process Of Propagating, Maintaining, Or Preserving Plant Cell Or Cell Line; Process Of Isolating Or Separating A Plant Cell Or Cell Line; Process Of Regenerating Plant Cells Into Tissue, Plant Part, Or Plant, Per Se, Where No Genotypic Change Occurs; Medium Therefore   Culture, Maintenance, Or Preservation Techniques, Per Se   Involving Conifer Cell Or Tissue (e.g., Pine, Spruce, Fir, Cedar, Etc.)