Classifications

• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
  • A01H3/00—Processes for modifying phenotypes, e.g. symbiosis with bacteria
• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
  • A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
  • A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
  • A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
  • A01H4/008—Methods for regeneration to complete plants

Definitions

• the present application relates generally to the field of plant propagation.
• the present invention relates to a method for the propagation of vegetatively reproducing plants and plants and plant parts produced by such methods.
• the invention also provides encapsulated propagules.
• the invention also provides various end uses for the encapsulated propagules and for plants grown from the same.
• the invention also provides a method for the modification of the architecture of rhizomes and rhizomes having modified architecture and a method for the modification of the architecture of stem cuttings and stem cuttings having modified architecture.
• the invention also provides a coating for a propagule and a propagule coated therewith.
• bio-fuels which is fuel derived from crops (energy crops) which can be burned to produce heat and electricity or treated with enzymes to produce sugars that can be used to produce ethanol or hydrogen.
• Miscanthus is an example of a perennial grass that has been identified as an energy crop due to its high biomass yields.
• Miscanthus species There are a number of Miscanthus species, but currently the most productive clone is a sterile hybrid, Miscanthus x giganteus, resulting from a natural cross between M. sacchariflorus and M. Sinensis. Miscanthus is a seed-bearing grass but the hybrid M. x giganteus is a triploid which is therefore sterile and does not produce seed. It can therefore only be multiplied by vegetative means which has severely restricted its commercial introduction. The main concern with seeded varieties of Miscanthus is their potential to become invasive, however the alternative to seeded varieties is to rely on vegetative propagation which is largely inefficient.
• Rhizome cuttings are obtained from these "root complexes", which are usually about 12-15cm long and weigh about 50g including the surrounding soil. Although these rhizomes look robust, they are rather sensitive to dehydration and therefore require careful storage and handling. The aforementioned factors make the multiplication of Miscanthus relatively expensive compared to other crops. For example, under typical growing conditions in Europe, the multiplication ratio of Miscanthus rhizomes is around 3-5 fold in one growing season, meaning than one rhizome will produce a maximum of five rhizomes after one year's growth. This is in contrast to cereals with a multiplication ratio of up to 80 fold and oilseed rape which has a multiplication rate of over 5,000 fold. Similarly, the costs of planting conventional Miscanthus rhizomes are also high compared with the costs of cleaning, processing and planting seeded crops.
• the multiplication of sugarcane from seed cane then begins, a process which typically takes about 3 years before commercial production can begin. Typically, about 20% of the total growing area is reserved for multiplication, meaning that about 20% of the land is tied up for at least 3 years, if not continuously, for successive cycles of multiplication.
• the multiplication ratios over the 3 year period are also relatively inefficient showing around an 8-fold increase.
• the present invention aims to solve some of the aforementioned problems associated with the propagation of Miscanthus and other vegetatively
• the efficiency gains obtained by performing the methods of the invention give an improvement, depending on the plant in question, of between about 50-fold to about 200-fold compared to the prior art methods, which show an improvement of about seven-fold compared to conventional techniques.
• step (ii) contacting the plantlets or a part thereof from step (i) with at least one plant hormone and growing the plantlets and harvesting mini rhizomes or mini stem cuttings therefrom; (iii) substantially encapsulating the mini rhizomes or mini stem cuttings produced from step (ii) in a plant growth medium.
• the present invention also provides encapsulated mini rhizomes and
• encapsulated mini stem cuttings which are collectively referred to herein as "encapsulated propagules”. Also provided herein are various uses for the encapsulated propagules and various uses for the plants obtained therefrom.
• the present invention also provides a biodegradable polymer for the coating of a propagule and propagules coated in the same.
• step (i) micro-propagation of plant material from a vegetatively reproducing plant followed by multiplication to produce plantlets; (ii) contacting the plantlets or a part thereof from step (i) with at least one plant hormone and growing the plantlets and harvesting mini-rhizomes or mini stem cuttings therefrom;
• step (iii) substantially encapsulating the mini-rhizomes or mini stem cuttings produced from step (ii) in a plant growth medium.
• Micro-propagation is a standard, well known horticultural technique used for the rapid bulking up of large numbers of plantlets.
• the micro-propagation described in stage (i) of the method aims at producing large quantities of material suitable for entering stage (ii) of the method.
• the micro-propagation techniques used may be any conventional micro-propagation technique, such as tissue culture or rooted stem cuttings.
• the micro-propagation results in plantlets which are ready to be multiplied, typically in a greenhouse. Conventional means for plant multiplication are used for this stage of the method.
• the greenhouse multiplication of the plantlets typically results in a 20-30 fold increase every 2-3 months.
• the micropropagation and multiplication results in every one plant generating about 10 plants in about an 8 week cycle; those 10 plants can then in turn generate about 100 plants in a further cycle of about 8 weeks.
• micro-propagation followed by multiplication can result in every 1 (one) plant generating 10 million plantlets, plug plants or bare root cuttings (which refer to individual stems which can be separated from larger plants) in less than 6 months or in less than 8 months or in less than 10 months or in less than 12 months or in less than 14 months or in less than 16 months or in less than 18 months, depending on the plant in question.
• micro-propagation followed by multiplication results in a decreased rate of mutation compared to conventional micropropagation, which again contributes to the overall efficiency gains brought about by performing the methods of the invention.
• the resulting plantlets are then ready for the hormone treatment in stage (ii) of the process.
• plantlet is used interchangeably herein and are taken to mean the small plants produced following the micro- propagation and multiplication techniques of stage (i) of the method.
• the size of the plantlet will vary depending on the plant being propagated, and a skilled person will have at his or her disposal suitable means for generating plantlets from any given plant. In the case of Miscanthus, the plantlets are typically in the range of at least 2-5cm in length.
• rhizome as used herein is used in its conventional sense to refer to an underground stem of a plant, which typically produces roots and shoots.
• rhizomatous plant as used herein refers to any plant capable of producing a rhizome.
• mini-rhizome is a term known in the art and refers to a whole rhizome from any given plant species which is about 10% of the size of a typical whole rhizome for that plant species, preferably about 5% of the size of a typical whole rhizome for that plant species.
• a typical whole rhizome from Miscanthus is about 50g in weight and between about 12 to 15 cm in length
• a mini-rhizome from the same plant species is typically about 5g in weight, preferably about 4g or 3g or 2g in weight and between about 2 to 5 cm in length, preferably between about 1 to 2 cm in length.
• stem cutting as used herein is used in its conventional sense where a piece of any given parent plant is removed and encouraged to grow as an independent plant by placing the removed plant piece on a suitable growth medium, such as one or more of the following: soil, compost, potting mix, rock wool, perlite, vermiculite, coir, expanded clay pellets, hydrogel and water, which facilitates the growth of new roots and/or stems, which enable the stem cutting to become a plant independent of the source plant.
• a suitable growth medium such as one or more of the following: soil, compost, potting mix, rock wool, perlite, vermiculite, coir, expanded clay pellets, hydrogel and water, which facilitates the growth of new roots and/or stems, which enable the stem cutting to become a plant independent of the source plant.
• mini stem cutting refers to a stem cutting from any given plant species which is about 10% of the size of a typical stem cutting for that plant species, preferably about 5% of the size of a typical stem cutting for that plant species.
• a typical stem cutting from Arundo donax is about 100g in weight and between about 40 to 60cm in length, whereas a mini stem cutting from the same plant species is typically about 10g in weight, preferably about 4g or 3g or 2g in weight and between about 2 to 4cm in length.
• a conventional stem cutting is 30 to 40 cm in length, whereas a mini stem cutting from the same sugarcane species is typically between about 2 to 4cm in length.
• stage (ii) of the process which comprises hormonal treatment of the plantlets with cytokinin(s) and/or auxin(s).
• Any one given plant hormone is applied at a rate of less than about l OOOppm, less than about 900ppm, less than about 800ppm, less than about 700ppm, less than about 600ppm, less than about 500ppm, less than about 400ppm, less than about 300ppm, less than about 200ppm, less than about 100ppm, less than about 90ppm, less than about 80ppm, less than about 70ppm, less than about 60ppm, less than about 50ppm, less than about 40ppm, less than about 30ppm, less than about 20ppm or less than about 10ppm.
• the cytokinin may be selected from the group consisting of kinetin, zeatin, 6- benzylaminopurine, diphenyl urea and thidiazuron (TDZ).
• the auxin may be selected from the group consisting of indole-3-actetic acid (IAA), 4-chloroindole- 3-acetic acid (4-CI-IAA), 2-phenylacetic acid (PAA) and indole-3-butyric acid (IBA).
• IAA indole-3-actetic acid
• PAA 2-phenylacetic acid
• IBA indole-3-butyric acid
• the benzylaminopurine, TDA and IAA are each applied at a rate of less than about 1000ppm, less than about 900ppm, less than about 800ppm, less than about 700ppm, less than about 600ppm, less than about 500ppm, less than about 400ppm, less than about 300ppm, less than about 200ppm, less than about 100ppm, less than about 90ppm, less than about 80ppm, less than about 70ppm, less than about 60ppm, less than about 50ppm, less than about 40ppm, less than about 30ppm, less than about 20ppm or less than about 10ppm.
• concentrations of plant hormones may readily be optimized, if necessary, by one skilled in the art depending on the plant to be propagated.
• the plants after stage (i) of the process and before, after or during the hormone treatment of stage (ii), but before the harvesting of mini rhizomes or mini stem cuttings, the plants may be subjected to a temporary abiotic or mechanical stress.
• the abiotic stress may comprise subjecting the growing plant to any temporary environmental change compared to the normal growth conditions for the plant in question.
• the stress may be any one or more of: (i) an osmotic stress (which may be caused by limited or excess salt or water compared to the normal levels of salt or water); (ii) a temperature stress (which may be caused by exposure of the plant to excessive heat or cold compared to normal growth conditions for the plant in question); (iii) a nutrient stress (which may be caused by a lack of nitrogen, phosphorous, sulphur etc.); or (iv) an oxidative stress.
• the exposure of the plants to the temporary abiotic and/or mechanical stress serves to encourage more bud formation.
• the mechanical stress refers to any non-environmental stress resulting from a physical action to a plant, such as cutting parts of the plant.
• a plant such as cutting parts of the plant.
• the aboveground parts of the plant are cut back to a point just above a node.
• Reference herein to a "temporary" stress is taken to mean exposure to a non- continuous stress, which may involve exposure of the plant to any one or more given stresses at intermittent periods.
• the intermittent periods of stress may be 1 , 2, 3, 4 or 5 or more separate occasions of exposure of the plant to a stress in between non-stress periods.
• Stage (ii) of the process (contacting the plantlets with at least one plant hormone), which may or may not comprise exposure of the plants to an abiotic and/or mechanical stress, can last from between about 12 and 24 weeks depending on the plant in question and depending on whether sufficient buds have formed.
• the contacting of the plantlets to at least one plant hormone may occur before, after or during exposure of the plants to a temporary abiotic or mechanical stress.
• Preferred plant hormones include benzylaminopurine, TDA and IAA, one or more of which may be applied at a rate of ⁇ 1000ppm (less than one thousand parts per million).
• a preferred method for the propagation of a vegetatively reproducing plant comprises the steps of:
• step (ii) contacting the plantlets or a part thereof from step (i) with at least one plant hormone;
• step (iii) exposing the plantlets before, after or during step (ii) above to an
• step (v) substantially encapsulating the mini rhizomes or mini stem cuttings produced from step (iv) in a plant growth medium.
• the hormone-treated plants are then grown in a field or greenhouse setting under optimal growth conditions.
• this growth phase may span a period of about 12 months.
• Shoots emerging from the new buds are removed and cut (by machine or hand) into short lengths resulting in mini stem cuttings.
• this growth phase may span a period of about 12 months or until the size of the plant is about 20cm in length and the rhizomes are between about 2 to 5g in weight and/or until the bud capacity of the rhizomes is around 20 buds per rhizome.
• the rhizomatous plant material growing in the field or greenhouse may then be subjected to a light shredding or separation of the root complex in order to separate out the mini rhizomes therefrom.
• this shredding or separation process may be automated to directly take plant material from the field or greenhouse and to separate out the mini rhizomes from the root complex ready for the encapsulation stage.
• the mini rhizomes are around 2 to 5g in weight compared to the about 30 to 50g weight of rhizomes produced by conventional production methods.
• a further advantage is that the Miscanthus mini rhizomes are also more uniform in shape and size than the rhizomes of conventionally-produced Miscanthus plants, meaning that they can be planted with minimal adaptation (where necessary) of available agricultural machinery, thereby avoiding the expense associated with providing specially adapted or custom-made agricultural machinery.
• the small, uniformly-shaped mini rhizomes or mini stem cuttings resulting from stage (ii) of the method are then in a form ready for encapsulation.
• the encapsulation process may advantageously be automated.
• the mini rhizome or mini stem section or cutting may advantageously be encapsulated in any plant growth medium, such as compost, potting mix, peat, hydrogel, soil, rock wool, perlite, vermiculite, foam, syrofoam, pumice, coir, expanded clay pellets etc.
• the resulting encapsulated rhizome or encapsulated mini stem cutting propagule is a stable planting unit of less than about 10g, or less than about 15g for some crops, and is in a form ready for precision planting using minimally adapted conventional farming equipment or ready for storage until required.
• the encapsulation material may also comprise compounds, such as plant hormones (such as cytokinins or auxins), plant growth regulators, mycorrhiza, endophytic organisms, symbiotic organisms or other beneficial organisms, surfactants, gels, fungicides, nematicides, insecticides, organic and inorganic nutrients, water, polymer and organic-based super absorbents and stabilization compounds etc. to aid the storage of the mini rhizomes and mini stem cuttings so as to prevent any loss of material due to deterioration and to enhance the survival and performance of the propagules once they are planted in the field.
• plant hormones such as cytokinins or auxins
• plant growth regulators such as cytokinins or auxins
• mycorrhiza such as cytokinins or auxins
• endophytic organisms such as cytophytic organisms, symbiotic organisms or other beneficial organisms
• surfactants such as cytokinins or auxin
• the encapsulated material is then substantially coated in a biodegradable polymer, which preferably has a melting point of between 30 to 65°C.
• a biodegradable polymer which preferably has a melting point of between 30 to 65°C.
• the polymer is selected from one or more of wax, polyester, petroleum-based paraffin or plastic, polysaccharide or any plant-based plastic.
• the coating also comprises a fibre component comprising at least up to 20%, at least up to 30%, at least up to 40%, at least up to 50%, at least up to 60%, at least up to 70%, at least up to 80% or at least up to 90% of the coating.
• a fibre component comprising at least up to 20%, at least up to 30%, at least up to 40%, at least up to 50%, at least up to 60%, at least up to 70%, at least up to 80% or at least up to 90% of the coating.
• the fibre may be one or more of the following: (i) fibre from agricultural biomass residue (for example, cereal straw, cotton, peanut hulls, soy straw, corn fodder); (ii) dedicated fibres (for example, Miscanthus, Arundo, sugarcane, bagasse, hemp, Kenaf); (iii) processed fibres (for example, paper, recycled cardboard, wood flour, wood saw dust); and (iv) artificial or processed fibres (for example, nylon, polyester, cotton).
• agricultural biomass residue for example, cereal straw, cotton, peanut hulls, soy straw, corn fodder
• dedicated fibres for example, Miscanthus, Arundo, sugarcane, bagasse, hemp, Kenaf
• processed fibres for example, paper, recycled cardboard, wood flour, wood saw dust
• artificial or processed fibres for example, nylon, polyester, cotton
• the coating may also comprise fungicides, endophytic organisms, plant nutrients, hormones, dyes or other means for identification, such as barcodes or transponders or the like, to aid sorting.
• the coating is applied to the encapsulated propagule by dipping (at least once) or by co-extrusion or by thermally forming the coating around the propagule.
• the coating covering the propagule is less than 1 mm (millimetre) thick, preferably less than 0.5mm thick.
• step (i) micro-propagation of plant material from a vegetatively reproducing plant followed by multiplication to produce plantlets; (ii) contacting the plantlets or a part thereof from step (i) with at least one plant hormone and growing the plantlets and harvesting mini-rhizomes or mini stem cuttings therefrom;
• step (iii) substantially encapsulating the mini-rhizomes or mini stem cuttings produced from step (ii) in a plant growth medium
• step (iii) in a biodegradable polymer.
• a vegetatively reproducing plant comprising the steps of:
• step (ii) contacting the plantlets or a part thereof from step (i) with at least one plant hormone;
• step (iii) exposing the plantlets before, after or during step (ii) above to an
• step (v) substantially encapsulating the mini rhizomes or mini stem cuttings produced from step (iv) in a plant growth medium
• step (v) cutting of step (v) in a biodegradable polymer.
• a preferred method for the propagation of a rhizomatous plant comprises the steps of:
• step (i) micro-propagation of plant material from a rhizomatous plant followed by multiplication to produce plantlets; (ii) contacting the plantlets or a part thereof from step (i) with at least one plant hormone and growing the plantlets;
• step (iii) exposing the plantlets before, after or during step (ii) above to an abiotic or mechanical stress
• step (vi) substantially encapsulating the mini rhizomes produced in step (iv) or (v) in a plant growth medium; and optionally
• step (vii) substantially coating the encapsulated mini rhizome of step (vi) in a biodegradable polymer.
• steps (i) to (vii) above are as described hereinabove.
• a preferred method for the propagation of a plant from a stem cutting comprises the steps of:
• step (ii) contacting the plantlets or a part thereof from step (i) with at least one plant hormone and growing the plantlets;
• step (iii) exposing the plantlets before, after or during step (ii) above to an abiotic or mechanical stress
• step (v) substantially encapsulating the mini stems produced in step (iv) in a plant growth medium;
• step (vi) substantially coating the encapsulated mini stem of step (v) in a
• the methods of the invention may be applied to any plant capable of vegetative reproduction.
• the methods of the invention are particularly suited to rhizomatous and stem propagated plants.
• the methods of the invention are particularly suited to energy grasses, such as Miscanthus (elephant grass), Pennisetum purpureum (napier grass), Panicum virgatum (switch grass), energy cane (the Saccharum complex), Arundo donax (Giant Reed), Sugarcane, Bambusa (bamboo), Curcuma, Humulus (hop), asparagus, Zingiber (ginger), iris, genus Erianthus , Faiiopia sac aiinensis (Igniscum) , Ipomoea batatas (sweet potato) and wasabi.
• energy grasses such as Miscanthus (elephant grass), Pennisetum purpureum (napier grass), Panicum virgatum (switch grass), energy cane (the Sacchar
• Arundo donax (giant reed), Pennisetum purpureum (napier grass), energy cane, Saccharum officinarum (sugar cane), Hevea (rubber) and Manihot (cassava), for example, are more suited to multiplication using stem cuttings.
• stem cuttings When multiplying using stem cuttings, the plantlets, derived from micro-propagation are planted and grown to a suitable height and any aboveground growth is cut back to a point just above a node. This procedure encourages more bud formation which may be further stimulated by the application of cytokinins and auxins. The shoots emerging from the new buds are removed and cut up by machine into short lengths to form mini stem cuttings which are then ready to be encapsulated. Although not all of the stem cuttings that are produced will contain buds, the multiplication rate that is achieved is many times greater than would be achieved by conventional procedures.
• a substantially encapsulated mini rhizome or substantially encapsulated mini stem cutting produced by the method according to the invention.
• the encapsulated mini rhizome or encapsulated mini stem cutting may be substantially coated in a biodegradable polymer.
• the present invention also provides a planting unit weighing less than about 25g, or less than about 20g, or less than about 15g or less than about 10g, or less than about 5g, or less than about 2g, comprising a substantially uniformly-shaped mini rhizome or mini stem cutting substantially contained within a plant growth medium and optionally coated in a biodegradable polymer.
• the weight of the planting unit described above does not include the growth medium.
• the planting unit comprises a rhizome from a Miscanthus plant, energy cane, Arundo donax or sugarcane plant.
• the invention also provides a method for the production of plants, comprising the steps of growing an encapsulated mini rhizome or mini stem cutting produced by the methods according to the invention. Also provided are plants obtainable from such an encapsulated mini rhizome or mini stem cutting and progeny and ancestors thereof.
• plants produced from the mini stem cuttings or mini rhizomes produced by the methods according to the present invention have increased vigour. Taking Miscanthus as an example, this is manifested by an increase in the number of tillers (200 to 400% increase); increased height (up to a 30% increase); increased root volume (up to 300% increase); increased belowground buds (up to a 400% increase); increased canopy closure (up to a 200%
• the plants produced by the methods of the invention resemble conventionally produced plants that are one year or older.
• an encapsulated mini rhizome or mini stem cutting, or plants produced thereform in increasing plant vigour relative to conventionally grown plants of the same species.
• the mini stem cuttings and mini rhizomes produced by the methods of the present invention offer starting material for plant propagation having the same growth capacity but of greatly reduced weight compared to conventional starting material, thereby saving transportation costs, storage and handling costs etc.
• the weight of the starting material will be about 0.25 tonnes per hectare compared to 1.5 tonnes per hectare in the case of conventionally grown Miscanthus, 6 tonnes per hectare for conventionally grown Napier grass and 10 tonnes per hectare for conventionally grown sugarcane.
• the mini stem cuttings or mini rhizomes produced by the methods of the present invention allow the harvest cycle of the crop plants grown therefrom to be shortened compared to the harvest cycle of the same plant grown by conventional methods.
• the mini stem cuttings or mini rhizomes produced by the methods of the present invention offer multiple planting windows compared to the planting window for the same plant grown by conventional methods.
• the plants produced by growing an encapsulated mini rhizome or mini stem cutting may also find use in the production of bio-fuels or bio-ethanol.
• an encapsulated mini rhizome or mini stem cutting or plant produced therefrom in biofuel or bioethanol production there is provided use of an encapsulated mini rhizome or mini stem cutting or plant produced therefrom in biofuel or bioethanol production.
• the mini stem cuttings and mini rhizomes produced by the methods of the present invention may find use in bioremediation applications for cleaning up areas having contaminated water.
• an encapsulated mini rhizome or mini stem cutting or plant produced therefrom in bioremediation there is provided.
• step (ii) contacting the plantlets from step (i) with plant hormones selected from the group consisting of benzylaminopurine, TDA and IAA;
• the aforementioned method for altering rhizome architecture may also comprise the additional steps of shredding and/or encapsulation of the rhizomes as described hereinabove.
• a preferred method for altering rhizome architecture comprises the steps of:
• step (ii) contacting the plantlets or a part thereof from step (i) with a plant hormone selected from the group consisting of benzylaminopurine, TDA and IAA and growing the plantlets;
• step (iii) exposing the plantlets before, after or during step (ii) above to an
• step (ii) contacting the plantlets from step (i) with a plant hormone selected from the group consisting of benzylaminopurine, TDA and IAA;
• a preferred method for altering stem architecture comprises the steps of:
• step (ii) contacting the plantlets or a part thereof from step (i) with a plant
• hormone selected from the group consisting of benzylaminopurine, TDA and IAA and growing the plantlets;
• step (iii) exposing the plantlets before, after or during step (ii) above to an
• the present invention provides mini rhizomes and mini stem cuttings obtainable by the methods described herein.
• the method for altering rhizome or mini stem architecture results in mini rhizomes or mini stem cuttings weighing less than about 25g or weighing less than about 20g or weighing less than about 15g or weighing less than about 10g or weighing less than about 5g or weighing less than about 2g.
• the benzylaminopurine, TDA and IAA are each (individually, not in combination) applied at a rate of less than about l OOOppm, less than about 900ppm, less than about 800ppm, less than about 700ppm, less than about 600ppm, less than about 500ppm, less than about 400ppm, less than about 300ppm, less than about 200ppm, less than about 100ppm, less than about 90ppm, less than about 80ppm, less than about 70ppm, less than about 60ppm, less than about 50ppm, less than about 40ppm, less than about 30ppm, less than about 20ppm or less than about 10ppm.
• the method for altering rhizome architecture is particularly suited to the rhizomes of Miscanthus plants, but may also be used for other rhizomatous plants.
• the combinations and concentrations of plant hormones may readily be optimized, if necessary, by one skilled in the art depending on the plant to be propagated.
• the resulting mini rhizomes or mini stem cutting having modified architecture may then be used in a method for the production of a plant, comprising the steps of growing a rhizome or an encapsulated rhizome or a mini stem cutting or encapsulated mini stem cutting obtainable by the aforementioned methods for altering rhizome or mini stem architecture.
• benzylaminopurine, TDA and IAA to modify the architecture of rhizomes, particularly Miscanthus rhizomes, or to modify the architecture of mini stems.
• use of benzylaminopurine, TDA and IAA to increase the yield of rhizomatous plants or the yield of plants propagated using stem cuttings compared to the yields obtained using conventional production methods.
• the use of benzylaminopurine, TDA and IAA is as described above in the method for plant propagation.
• the benzylaminopurine, TDA and IAA are each (individually, not in combination) applied at a rate of less than about l OOOppm, less than about 900ppm, less than about 800ppm, less than about 700ppm, less than about 600ppm, less than about 500ppm, less than about 400ppm, less than about 300ppm, less than about 200ppm, less than about 100ppm, less than about 90ppm, less than about 80ppm, less than about 70ppm, less than about 60ppm, less than about 50ppm, less than about 40ppm, less than about 30ppm, less than about 20ppm or less than about 10ppm.
• the architecture of the rhizomes and mini stem cuttings is modified such that the resulting mini rhizomes or mini stem cuttings after application of the plant hormones are substantially uniform in shape and weight less than about 25g or weigh less than about 20g or weigh less than about 15g or weigh less than about 10g or weight less than about 5g or weigh less than about 2g.
• a coating for a propagule comprising a biodegradable polymer, which preferably has a melting point of between 30 to 65°C.
• the invention also provides propagules coated with such a biodegradable polymer.
• the propagule is encapsulated prior to being coated in the
• the propagule may advantageously be encapsulated in any plant growth medium, such as compost, potting mix, peat, hydrogel, soil, rock wool, perlite, vermiculite, foam, syrofoam, pumice, coir, expanded clay pellets etc.
• the encapsulation material may also comprise compounds, such as plant hormones (such as cytokinins or auxins), plant growth regulators, mycorrhiza, endophytic organisms, symbiotic organisms or other beneficial organisms, surfactants, gels, fungicides, nematicides, insecticides, organic and inorganic nutrients, water, polymer and organic based super absorbents and stabilization compounds etc. to aid the storage of the propagule so as to prevent any loss of material due to deterioration and to enhance the survival and performance of the propagules once they are planted in the field.
• plant hormones such as cytokinins or auxins
• plant growth regulators mycorrhiza, endophytic
• the polymer is one or more of wax, polyester, petroleum-based paraffin or plastic, polysaccharide or any plant-based plastic.
• the coating may comprise Preferably, the coating also comprises a fibre, which may comprise at least up to 20%, at least up to 30%, at least up to 40%, at least up to 50%, at least up to 60%, at least up to 70%, at least up to 80% or at least up to 90% of the coating.
• a fibre which may comprise at least up to 20%, at least up to 30%, at least up to 40%, at least up to 50%, at least up to 60%, at least up to 70%, at least up to 80% or at least up to 90% of the coating.
• the fibre may be one or more of the following (i) fibre from agricultural biomass residue (for example, cereal straw, cotton, peanut hulls, soy straw, corn fodder); (ii) dedicated fibres (for example, Miscanthus, Arundo, sugarcane, bagasse, hemp, Kenaf); (iii) processed fibres (for example, paper, recycled cardboard, wood flour, wood saw dust); and (iv) artificial or processed fibres (for example, nylon, polyester, cotton).
• agricultural biomass residue for example, cereal straw, cotton, peanut hulls, soy straw, corn fodder
• dedicated fibres for example, Miscanthus, Arundo, sugarcane, bagasse, hemp, Kenaf
• processed fibres for example, paper, recycled cardboard, wood flour, wood saw dust
• artificial or processed fibres for example, nylon, polyester, cotton
• the coating may also comprise fungicides, endophytic organisms, plant nutrients, hormones, dyes or other means for identification, such as barcodes or transponders or the like, to aid sorting.
• the coating is applied to the (encapsulated) propagule by, for example, dipping (at least once) or by co-extrusion or by thermally forming the coating around the propagule.
• the coating covering the (encapsulated) propagule is less than 1 mm (millimetre) thick, preferably less than 0.5mm thick.
• a biodegradable polymer which preferably has a melting point of between 30 °C to 65°C, for the coating of a propagule.
• a “propagule” as defined herein is any plant part which is capable of being grown or regenerated into a whole plant.
• a propagule may therefore comprise, rhizomes, mini rhizomes, stem cuttings, mini stem cuttings, tubers, seeds etc.
• any commercial or other seed variety may optionally first be encapsulated as described above and then coated in a biodegradable polymer as described.
• the seed may be a transgenic or non- transgenic seed.
• Preferred seeds include melon and tomato seeds.
• Figure 1a shows a rhizome from Miscanthus produced by conventional methods (weighing about 30g) and Figure 1 b shows a mini rhizome from Miscanthus produced by the methods of the present invention (weighing around 2g).
• Figure 2 shows Miscanthus plantlets undergoing micro-propagation according to stage (i) of the method of the invention.
• Figure 3 shows the crop establishment in the field of Miscanthus plants produced from the encapsulated mini rhizomes (right hand half of the picture) produced by the methods of the present invention compared to plant produced from
• Explant material was selected from disease-free Miscanthus plant material.
• the explant material was surface sterilized, rinsed in sterilized water and seeded onto an agar-based growth medium containing nutrients (sucrose and other standard nutrients) and plant hormones to encourage cell division.
• the resulting callus was then re-plated onto a growth medium infused with nutrients and plant hormones (cytokinins and auxins) to stimulate cell differentiation into roots and shoots and the production of plantlets. Repeated cycles of this process allow a single explant sample to be increased from one to thousands of plantlets.
• the plantlets were then planted in compost and grown in a greenhouse until 5 to 10 shoots were established.
• the plantlets were then subjected to a temporary drought by withholding watering of the plants for a period of 1 day.
• the shoots when about 10cm tall, were subjected to a separation procedure which involved separating, by hand, each shoot having a dedicated root system and potting this shoot into compost.
• the potted shoots were then treated with a combination of benzylaminopurine, TDA and IAA each being at a rate of ⁇ 1000ppm (less than one thousand parts per million) to enhance shoot and root initiation.
• Arundo donax (giant reed), Pennisetum purpureum (napier grass) and energy cane are more suited to multiplication using stem cuttings.
• the Arundo donax , Pennisetum purpureum , sugarcane and energy cane plantlets were subjected to micro-propagation and multiplication as described in Example 1. Plantlets derived from micro-propagation and multiplication were planted onto compost and grown to a height of about 10cm. The aboveground growth was cut back to a point just above a node to encourage more bud formation, which was further stimulated by the application of benzylaminopurine, TDA and IAA, each being at a rate of ⁇ 1000ppm (less than one thousand parts per million). Shoots emerging from the new buds were removed and cut by machine into short lengths to form mini stem cuttings. These mini stem cuttings were then encapsulated as described in Example 4.
• Encapsulation was carried out using machinery available in the horticultural industry for encapsulating seeds or cuttings but adapted so as to fully encase the mini-rhizome or mini stem cutting.
• the mini-rhizome or stem cutting was introduced into a flow of compost containing cytokinins, auxins, mycorrhiza, surfactants, gels, fungicides and insecticides.
• the compost surrounding the mini-rhizome or stem cutting was then compressed.
• the resulting unit of mini- rhizome or mini stem cutting in compressed compost was then wrapped in a paper binding to increase the stability of the encased propagule.
• the encased propagule was then stored until required for planting.

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