WO1991015110A1 - Procedes et appareil de micropropagation - Google Patents

Procedes et appareil de micropropagation Download PDF

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Publication number
WO1991015110A1
WO1991015110A1 PCT/GB1990/001001 GB9001001W WO9115110A1 WO 1991015110 A1 WO1991015110 A1 WO 1991015110A1 GB 9001001 W GB9001001 W GB 9001001W WO 9115110 A1 WO9115110 A1 WO 9115110A1
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WO
WIPO (PCT)
Prior art keywords
plantlets
nutrient medium
cut
growing
comb
Prior art date
Application number
PCT/GB1990/001001
Other languages
English (en)
Inventor
Jane Mary Allard
Jennet Blake
David Peter Griffin
Matthew Hutchinson
Original Assignee
British Technology Group Plc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by British Technology Group Plc filed Critical British Technology Group Plc
Publication of WO1991015110A1 publication Critical patent/WO1991015110A1/fr

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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
    • A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/003—Cutting apparatus specially adapted for tissue culture

Definitions

  • the present invention relates to methods and apparatus for use in micropropagation.
  • the invention is concerned in particular, but not exclusively, with micropropagation of seed potatoes, and harvesting of potato microtubers from plantlets which have been grown by micropropagation.
  • Micropropagation of plants involves the use of the techniques of plant tissue culture and the application of these to the propagation of plants.
  • micropropagation consists initially of surface sterilising and excising small pieces of actively growing tissue, normally shoot tips or nodes cut from the stems of plants. Then, under aseptic conditions, the pieces of tissue are transferred to a nutrient medium which supports plant growth. The plant material will finally develop into entire plantlets. These plantlets must then be weaned from the axenic conditions in which they have existed within the laboratory into viable, rooted plants capable of survival in conventional horticultural or agricultural environments.
  • the step of cutting a plantlet into small pieces, referred to as explants, for regrowth is repeated several times before a batch of plantlets is grown to viable plants. Where the plants propagate by propagules such as tubers, a further stage is introduced. The plantlets at the end of the multiplication process are grown until small tubers, called microtubers, are formed, which are then removed for planting to produce viable plants.
  • the growth of plants from tissue culture is a technique which can produce large numbers of genetically identical plants, perhaps possessing a desirable quality such as disease resistance, in a short time.
  • the cuts are required to cut from the donor a shoot tip, or a node, from which a side shoot will develop from an axillary bud.
  • the cut portion is then transferred by the forceps to the soft nutrient medium, which is in the nature of a gel, and the cutting is then placed upright with the stem part of the cutting in the soft nutrient medium.
  • the positioning is normally carried out with the use of forceps. It is an object of the invention, in a number of different aspects, to provide techniques for automating the micropropagation of plantlets, particularly of the kind normally propagating by tubers, in which very simple methods of cutting, transfer, planting and harvesting can be used which do not depend upon robotic selection of particular plantlets for processing, nor individual planting of cut portions of plant material.
  • the techniques of the invention provide for easy automation, and are therefore particularly suitable for bulk micropropagation of relatively low value plants such as seed potatoes, thus leading to lower labour costs and reduction in the cost of the final product.
  • a method of micropropagation including the steps of growing a plurality of plantlets in a nutrient medium, the plantlets having stems bearing tips and/or nodes, cutting through a plurality of stems of the plantlets at a predetermined distance from the nutrient medium to provide a plurality of plant portions containing tips and/or nodes, repeating the cutting step at a further, smaller, predetermined distance from the nutrient medium, to produce from the remaining cut stems before regrowth a further plurality of cut plant portions, and transporting the cut plant portions under aseptic conditions in bulk to a fresh nutrient medium.
  • the method includes repeating the cutting step at a series of different distances from the nutrient medium to produce a series of pluralities of cut plant portions.
  • the step of transporting cut plant portions includes depositing the cut plant portions at random on the surface of the fresh nutrient medium, where it is found such explants will grow satisfactorily without individual siting and positioning.
  • the step of transporting the cut plant portions to the fresh nutrient medium can be achieved conveniently at least partly by gravity, and/or by a forced airflow carrying the cut plant portions in the airflow.
  • the method may include the step of harvesting propagules from the plantlets by combing through the plantlets with a comb and separating the propagules from the plantlets.
  • apparatus for use in micropropag ⁇ ation of plants comprising a first support means for supporting a nutrient medium for growing plantlets having stems bearing tips and/or nodes, cutting means adapted for cutting at least twice through a plurality of stems of the plantlets at decreasing predetermined distances from the nutrient medium to remove before regrowth at least two pluralities of plant portions containing tips and/or nodes, and a second support means for supporting a second nutrient medium in which plantlets may be grown, the second support means being positioned to receive cut plant portions in bulk under aseptic conditions, cut from the said plantlets.
  • the cutting means may comprise shear cutting means, for example by relative reciprocatory motion of sets of cutting teeth, generally in the manner of a clipper for pet animals.
  • the plantlets may be presented at a workstation for cutting, in a conventional configuration with the stems growing substantially vertically from a horizontal nutrient medium.
  • the nutrient medium may conveniently be presented with its normal upper growing surface oriented away from the normal horizontal position.
  • the supporting container may be inverted so that the plantlets are directed downwardly from a horizontal surface of the nutrient medium, or the surface may be vertical with a cutter moving upwardly to release material sequentially onto a second moving nutrient medium in a generally regular manner.
  • the orientation of the nutrient medium may provide a further independent aspect of the present invention.
  • a method of micropropagation including the steps of presenting at a workstation a plurality of plantlets held in a nutrient medium, the medium being presented in an orientation such that a portion of a plantlet which is cut from the plantlet falls free from the plantlets held in the nutrient medium, cutting through a plurality of the growing plantlets to release in bulk cut-away plant portions large enough to contain tips and/or nodes, and receiving the released plant portions on a second nutrient medium.
  • apparatus for use in micropropagation of plants comprising a first support means for supporting a nutrient medium with plantlets growing in the medium, means for presenting the nutrient medium and plantlets in an orientation such that a portion of a plantlet which is cut from the plantlet falls free of the plantlets held in the nutrient medium, cutting means for cutting through a plurality of growing plantlets to release cut-away plant portions large enough to contain tips and/or nodes, and second support means for supporting a second nutrient medium, the second support means being arranged in a position such as to receive in bulk the released plant portions cut by the cutting means.
  • a later step in the method may comprise harvesting propagules from plantlets growing in a nutrient medium.
  • This step may be utilised independently of the other steps of the invention, and there is therefore provided in accordance with another independent aspect of the invention a method of harvesting propagules from plantlets growing in a nutrient medium comprising producing relative movement between the plantlets and a comb, the relative movement being such as to cause the teeth of the comb to enter into an array of the stems of the plantlets, and also being such as to cause the comb to move away from the bases of the stems, in such a manner that the teeth comb through the plantlets and remove propagules from the plantlets.
  • this aspect apparatus for harvesting propagules from plantlets growing in a nutrient medium comprising a comb, support means for supporting a nutrient medium having plantlets growing therein and means for producing relative movement between the support means and the comb, the relative movement being such as to cause, in operation, the teeth of the comb to enter into an array of stems of plantlets growing in a nutrient medium supported by the support means, and also being such as to cause the comb to move away from the bases of the stems, in such a manner that the teeth comb through the plantlets and remove propagules from the plantlets.
  • an apparatus for use in particular, but not exclusively, in the methods set out above concerned with moving cut plant portions by gravity and concerned with combing propagules comprising a main support structure, engaging means extending from the support structure and adapted to engage a container containing a nutrient medium with a plurality of plantlets growing therein, and a plurality of fingers extending from the support structure in a region spaced from the engaging means, the fingers being adapted to be positioned between plantlets and in contact with the surface of the nutrient medium to retain the nutrient medium in the container.
  • a method of operating on a plurality" of plantlets during micropropagation including the steps of inserting between plantlets growing in a nutrient medium a plurality of fingers, and contacting the surface of the medium by the fingers to retain the nutrient medium during an operating step being carried out on the plantlets during micropropagation.
  • the invention extends to icroplants and microtubers whenever produced by any method set out in the previous paragraphs in accordance with the invention.
  • microtuber refers to a small tuber grown by the process of micropropagation, and is to be distinguished from the term minituber which indicates merely the small size of the tuber rather than the method of production.
  • a microtuber is a small (1-lOmm, usually 3-8mm diameter) potato tuber produced on a potato microplant, or part thereof, in vitro, which may be maintained in an aseptic state after harvest until planting. It is to be differentiated from a minituber which is a small (5-45mm usually 20-30mm diameter) potato tuber produced on a potato microplant, or part thereof, in vivo, usually under protected, clean, though not aseptic conditions, e.g. in sterilized peat in an aphid proof glasshouse.
  • plantlet is meant a small plant grown in aseptic conditions, in a nutrient medium.
  • explant is meant a portion of plant material cut from a plant or plantlet and suitable to be transferred to a sterile nutrient medium for growth into a plantlet.
  • An example of a propagule is a microtuber.
  • a microtuber is meant an enlarged part of a stem of a plantlet, which can be removed from the plantlet and used for subsequent growth of a plant.
  • the microtubers of potatoes are very small, for example approximately 5mm in diameter.
  • Figures la and lb illustrate diagrammatically steps in a method of micropropagation embodying invention
  • Figures 2a, 2b and 2c illustrate diagrammatically further steps in a method of micropropagation embodying the invention
  • Figures 2d and 2e illustrate diagrammatically a tool for holding a container in the step shown in Figure 2c during a method of micropropagation embodying the invention
  • Figure 3 is a diagrammatic perspective view of a reciprocatory cutter for use in embodiments of the invention.
  • Figure 4 is a diagrammatic perspective view of apparatus embodying the invention for use in micropropagation
  • - 10 - Figure 5" is a diagrammatic schematic view of a comb for use in a method of harvesting propagules for use in micropropagation embodying the invention.
  • Figure 6 is a diagrammatic side view of apparatus for harvesting propagules for use in micropropagation, embodying the invention, Figure 6a showing a diagrammatic plan view of a comb shown in Figure 6.
  • Embodiments of the invention will now be described by way of example with reference to micropropagation of seed potatoes.
  • the gelling agent such as agar
  • the gelling agent is mixed with specific nutrient solution which may consist of Murashige and Skoog medium without Agar, IAA, Kinetin or Sucrose used at 4.71g 1 ⁇ 1, with the addition of extra iron to double that in the Murashige and Skoog medium and sucrose at 4% w/v.
  • the agar may consist of Oxoid Technical Agar No. 3, used at 0.7 - 0.8% w/v.
  • the plants should be grown in growth rooms at a temperature of between 17 * C and 21 * C, preferably 19"C.
  • the surface of the nutrient medium should be between 30cm and 40cm from the light source, which may be provided by, for example, Philips 84HF tubes. After one month of sixteen hour “days" in a temperature- growth room the stems of the plants need to be cut up into nodes and tips, and the cut plant portions transferred to new containers for regrowth. These nodes and tips may now be regrown as before to produce more nodes, or used to produce a harvest of microtubers.
  • This latter stage uses a slightly different nutrient solution (as above but with 8% sucrose instead of 4%), and shorter, eight hour light periods are given for two months following one month of sixteen hour days again with the same type of light source and similar temperature regime; thus this part of the process takes three months.
  • the microtubers, 3 to 10mm in diameter, are harvested and replanted conventionally after a three month dormancy period, to produce conventional seed potatoes.
  • the plantlets At the end of the first growth stage, the plantlets would, in accordance with known techniques, be cut up using a scalpel and forceps, (frequently sterilised), and the whole operation would occur within a laminar air-flow cabinet in order to maintain aseptic conditions.
  • the process of plant transfer is time consuming, and the labour costs form a substantial proportion of the overall cost of a microtuber. A reduction in labour costs of this stage would lead to a lower production cost for potato microtubers.
  • the cutting step at the end of growth stage is carried out as shown for example in Figures la and lb.
  • a plurality of plantlets 11 are grown in a gelled nutrient medium 12 placed in a container 13.
  • the first concept of the invention is that, instead of cutting each individual stem of a plantlet 11 at a position dictated by the required nodes 14, a single cut, indicated at Cl is made at a predetermined distance LI from the growing surface 16 of the gel 12.
  • This distance Ll is set to be such as to be likely to give in portions 17 cut from the plantlets, on average, one tip or one node in each cut plant portion.
  • the cut plant portions are then removed, as will be described hereinafter, and a second cut is made at the line indicated at C2.
  • the cut C2 is set at a level L2 above the growing surface 16 of the agar, and is again set to be such that the cut portions contain on average one node.
  • the levels Ll and L2 may be varied according to the characteristics of the plant species or cultivar being handled.
  • cut Cl contains principally the tips of the shoots.
  • FIGS. 2a, b and c are shown three alternative methods of transporting cut plant portions to fresh gelled medium, whilst minimising contact with operational elements and working in sterile conditions.
  • the cut portions 17 of the plantlets are removed by suction and transported along a conduit 18 and deposited on the growing surface of fresh gelled medium 12A in a second container 13A.
  • the container 13 is suspended upside down so that the plantlets 11 protrude downwardly from the gelled medium.
  • the container 13 is positioned above a second container 13A containing gelled medium 12A, and both containers 13 and 13A are moved in unison (to the right in Figure 2c) to meet a stationary reciprocatory cutter 20.
  • the cut portions 17 fall onto the growing surface of the gelled medium 12a.
  • FIG. 2(d) there may be a difficulty with a large container that the gelled nutrient medium may not stay in the container when it is inverted.
  • a tool for overcoming this difficulty In Figure 2(d) there is shown a hand tool 60 comprising a handle 61, a transverse arm 62, a set of fingers 63 extending from the arm 62 and a backing plate 64 with sides 65.
  • the fingers 63 and transverse arm 62 are arranged generally in the shape of a garden hand fork, but the fingers 63 are set at the top of the transverse arm 62, so as to be spaced from the backing plate 64 by approximately the depth of the gelled medium 12 plus the base of the container 13.
  • the tool 60 In use the tool 60 is slid over the container 13, as shown in Figure 2(e), so"that the fingers 63 pass between plants 11 and backing plate 64 passes under the container 13. The assembly is then inverted and the fingers 63 hold the gelled medium in place.
  • the tool 60 will of course also be useable in an automatic system.
  • FIG. 3 shows a suitable cutting means 20 for cutting plantlets.
  • the cutting means has an upper set of cutting teeth 21 and a lower set of cutting teeth 22.
  • the set 22 is maintained stationary and the other set 21 is reciprocated, although in other arrangements both sets may be reciprocated.
  • the teeth cut by shear between the upper and lower teeth 21 and 22.
  • the arrow C indicates the direction of cut of the cutting means. Shear cutting is preferred so as to avoid the cutter imparting a translational motion to the cut plant portions.
  • the cutter itself will need frequent cleaning and sterilisation, and must have height adjustment to perform the different cuts. It may therefore have detachable, sterilisable blades (or the whole motor unit and blades may be removed from position for cleaning) and preset positions which are easy to select.
  • Figure 4 shows schematically apparatus for carrying out the cutting and transfer steps shown diagrammatically in Figures la and 2.
  • a main frame 23 has upper ledges 24 and 25 on which can rest an inverted container 13 containing a gelled nutrient medium 12 in which are growing plantlets 11.
  • the container is preferably held by the tool 60, which is omitted for simplicity.
  • the nature of the gelled nutrient medium is such that the plantlets are held in the gel and do not fall when the container is inverted.
  • the frame 23 has second ledges 26 and 27 on which may be rested and held, by means not shown, a second container 13A containing fresh gelled nutrient medium 12A, this container 13A being positioned in the normal upright position.
  • the frame 23 is moveable on wheels 28 resting on rails 30.
  • a cutter driver assembly 31 from which protrudes a cutter means 20, for example as shown in Figure 3.
  • the cutter 20 is mounted on an support 32 which may be raised and lowered by passing along a vertical slot 33 in the side of the cutter driver assembly 31.
  • a manual adjustment of the height of the arm 32 is conveniently provided by a cranked handle 34.
  • the reciprocatory cutter 20 is driven, by an electric motor within the cutter driver assembly 31.
  • the container 13 and container 13A are placed on the frame 23 and the cutter 20 is set at a height equivalent to the line Cl in Figure la.
  • the frame 23 is then moved either manually or for example by electrical drive means not shown, from left to right in Figure 4, so that the plantlets 11 are cut by the cutter 20.
  • the cut portions then fall by gravity onto the fresh gelled medium 12A.
  • the entire operation takes place within a sterilised aseptic environment.
  • the tool of Figure 5 is a manual tool, and has a handle 35 supporting a tray-like structure 34 having side walls 36, 37 and 38, and being open at the end remote from the handle 35.
  • the base 39 of the tray-like structure 34 is divided to form a comb having teeth 40 projecting away from the handle 35.
  • Each tooth has a rounded distal end 41, tapered sides for part of its length, and a main portion 42 with parallel sides. The end of each slot formed by the parallel sides is rounded at the base at the region 43. It is a feature of the system of harvesting by combing that the tubers are in effect being graded by the combing action, since tubers smaller than the spacing of the parallel sided parts of the teeth, are not removed by the combing action.
  • the gap between the parallel sided parts 42 of the teeth 40 indicated at G is set to be smaller than the smallest usable microtuber, but large enough to allow passage of stems, leaves, etc. through between the teeth.
  • the tool is combed through the plantlets growing in the agar, and lifted gently away from the agar to pluck the microtubers.
  • the tool illustrated in Figures 2d and 2e may be employed also to prevent removal of plants from the agar whilst combing is carried out.
  • Figure 6 illustrates an automated version of the combing tool of Figure 5.
  • the teeth 40 of the comb 39 are shown in Figure 6a, the region indicated at Rl being for entrainment of tubers, and the region indicated at R2 being for plucking.
  • containers 13 of plantlets ready for harvesting are fed as indicated by arrow F onto a conveyor system 50 which carries the containers 13 downwardly and to the right in Figure 6.
  • the conveyor system 50 consists of a conveyor belt 51 entrained about three pulleys 52, 53 and 54, the conveyor belt 51 having, for example, apertures into which the container 13 can fit.
  • two conveyor belts can be positioned side by side, with the container 13 resting between the two conveyor belts.
  • the comb 39 is positioned without net translational movement relative to the pulleys 52, 53 and 54 but is vibrated by a vibration drive system 55.
  • the comb 39 is set at an angle to the surface of the conveyor belt 51, such that the teeth 40 of the comb 39 diverge from the surface of the conveyor belt 51 in a direction away from the distal ends of the teeth 40.
  • the effect of this is that as the container 13 is carried downwardly on the conveyor belt 51, the distal tips of the teeth 40 intrude into the stems of the plantlets 11, and progressively comb upwardly through the plantlets 11.
  • tubers 56 are plucked from the plantlets 11 and then roll down the comb 39 onto a conveying chute 57 contiguous with the comb 39.
  • the stripped plantlets 11 in the waste container 13 are then ejected from the conveyor system 50 over the pulley 53 as indicated at the arrow W.

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  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Developmental Biology & Embryology (AREA)
  • Cell Biology (AREA)
  • Botany (AREA)
  • Environmental Sciences (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

Abstract

Dans un procédé de micropropagation, on effectue une série de coupes horizontales à travers une masse de plantules (11) à tiges pratiquement droites et croissant dans un milieu (12) nutritif gélifié, afin d'obtenir des parties coupées (17) contenant chacune en moyenne une terminaison ou un n÷ud au moins. Après chaque coupe, les parties coupées (17) sont transférées en vrac, au moyen d'un courant d'air ou par gravité, vers un nouveau milieu nutritif gélifié (12A), dans lequel on les plante en les éparpillant. Dans l'étape finale, on récolte les propagules (56) des plantules (11) en passant un peigne (39) dans le semis et en séparant les propagules par mouvement relatif du peigne (39) effectué à partir du milieu gélifié (12).
PCT/GB1990/001001 1990-04-05 1990-06-29 Procedes et appareil de micropropagation WO1991015110A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB909007685A GB9007685D0 (en) 1990-04-05 1990-04-05 Methods and apparatus relating to micropropagation
GB9007685.2 1990-04-05

Publications (1)

Publication Number Publication Date
WO1991015110A1 true WO1991015110A1 (fr) 1991-10-17

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PCT/GB1990/001001 WO1991015110A1 (fr) 1990-04-05 1990-06-29 Procedes et appareil de micropropagation

Country Status (5)

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EP (1) EP0523040A1 (fr)
AU (1) AU5839890A (fr)
CA (1) CA2079773A1 (fr)
GB (1) GB9007685D0 (fr)
WO (1) WO1991015110A1 (fr)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993019587A1 (fr) * 1992-03-27 1993-10-14 Microcrop (Ireland) Limited Systeme de micro-reproduction amelioree
WO1993019586A1 (fr) * 1992-03-27 1993-10-14 Microcrop (Ireland) Limited Procede et appareil de micropropagation ameliores
US5584140A (en) * 1995-04-04 1996-12-17 Byrne; Michael Rooting method for vegetative plant propagation of hard-to-root plants
WO2011085446A1 (fr) * 2010-01-13 2011-07-21 Bses Limited Procédés de régénération de plante et appareil associé
WO2020010412A1 (fr) * 2018-07-13 2020-01-16 Lowes TC Pty Ltd Systèmes, dispositifs et procédés de propagation de plantes

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US1354283A (en) * 1918-11-29 1920-09-28 William B Clapp Cranberry-picker
US2459471A (en) * 1946-12-19 1949-01-18 Harold E Cole Cranberry picking machine
WO1988006618A1 (fr) * 1987-03-06 1988-09-07 KONTAKTA Villamosszerelési Vállalat Agencement pour la micropropagation de plantes
EP0298722A1 (fr) * 1987-07-07 1989-01-11 Mitsui Petrochemical Industries, Ltd. Appareil de culture
CA1249130A (fr) * 1988-03-14 1989-01-24 Rene L. Girard Chariot de cueillette de baies
EP0320239A2 (fr) * 1987-12-08 1989-06-14 Mitsui Petrochemical Industries, Ltd. Procédé de préparation de bandes de tissu végétal pour la culture de tissu de plantes

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US1354283A (en) * 1918-11-29 1920-09-28 William B Clapp Cranberry-picker
US2459471A (en) * 1946-12-19 1949-01-18 Harold E Cole Cranberry picking machine
WO1988006618A1 (fr) * 1987-03-06 1988-09-07 KONTAKTA Villamosszerelési Vállalat Agencement pour la micropropagation de plantes
EP0298722A1 (fr) * 1987-07-07 1989-01-11 Mitsui Petrochemical Industries, Ltd. Appareil de culture
EP0320239A2 (fr) * 1987-12-08 1989-06-14 Mitsui Petrochemical Industries, Ltd. Procédé de préparation de bandes de tissu végétal pour la culture de tissu de plantes
CA1249130A (fr) * 1988-03-14 1989-01-24 Rene L. Girard Chariot de cueillette de baies

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993019587A1 (fr) * 1992-03-27 1993-10-14 Microcrop (Ireland) Limited Systeme de micro-reproduction amelioree
WO1993019586A1 (fr) * 1992-03-27 1993-10-14 Microcrop (Ireland) Limited Procede et appareil de micropropagation ameliores
GB2272824A (en) * 1992-03-27 1994-06-01 Microcrop Improved micropropagation apparatus and method
GB2272824B (en) * 1992-03-27 1996-11-13 Microcrop Improved micropropagation apparatus and method
US5584140A (en) * 1995-04-04 1996-12-17 Byrne; Michael Rooting method for vegetative plant propagation of hard-to-root plants
WO2011085446A1 (fr) * 2010-01-13 2011-07-21 Bses Limited Procédés de régénération de plante et appareil associé
WO2020010412A1 (fr) * 2018-07-13 2020-01-16 Lowes TC Pty Ltd Systèmes, dispositifs et procédés de propagation de plantes
CN112804873A (zh) * 2018-07-13 2021-05-14 洛斯Tc 私人有限公司 植物繁殖系统、装置和方法

Also Published As

Publication number Publication date
AU5839890A (en) 1991-10-30
GB9007685D0 (en) 1990-06-06
CA2079773A1 (fr) 1991-10-06
EP0523040A1 (fr) 1993-01-20

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