CA1309588C - Method for cutting plant material - Google Patents

Method for cutting plant material

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Publication number
CA1309588C
CA1309588C CA 562325 CA562325A CA1309588C CA 1309588 C CA1309588 C CA 1309588C CA 562325 CA562325 CA 562325 CA 562325 A CA562325 A CA 562325A CA 1309588 C CA1309588 C CA 1309588C
Authority
CA
Canada
Prior art keywords
cutting
laser
cut
power
plant material
Prior art date
Legal status (The legal status 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 status listed.)
Expired
Application number
CA 562325
Other languages
French (fr)
Inventor
Reetta Puska
Esa Manneri
Donald Jonasson
Kari Jokinen
Timo Tormala
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Stora Enso Oyj
Kemira Oyj
Original Assignee
Enso Gutzeit Oy
Kemira Oyj
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 Enso Gutzeit Oy, Kemira Oyj filed Critical Enso Gutzeit Oy
Application granted granted Critical
Publication of CA1309588C publication Critical patent/CA1309588C/en
Expired legal-status Critical Current

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Classifications

    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01HNEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/003Cutting apparatus specially adapted for tissue culture

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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)
  • Laser Beam Processing (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Sampling And Sample Adjustment (AREA)
  • Laser Surgery Devices (AREA)

Abstract

(57) Abstract The invention relates to a method for cutting a plant material for the purpose of plant propagation, especially micropropagation, in which method the cutting is carried out by using a laser beam.

Description

~309~8 A method for cutting plant material It was observed in the 1960s that it was possible to produce plants from plant parts or from undifferentiated callus tissue. This technique is called micropropagation, which is thus asexual reproduction of plants in the labora-tory. The objective of micropropagation is to produce genetically identical valuable individual elite plants.
Thus the choice of the mother plant is an essentially im-portant step, since the plants to be produced will be its clones.

Micropropagation can be started from the growing point, bud or, for example, petiole, cut from the mother plant. The cultivation is carried out aseptically on a substrate which contains the principal and trace nutrients needed by the plants, and vitamins and hormones by means of which the growth is regulated. The substrate is usually made more solid by means of agar.

At the start the plant part forms a shoot, which is trans-planted from the test tube into a larger glass vessel to propagate. By using hormones, mainly cytokinins, new shoots are induced from axillary buds or, for example, from adven-titious buds forming in a leaf of the plant. After approxi-mately four weeks of growth, the propagated shoots are cut off and transplanted to new substrates to propagate. The propagation is continued until the desired number of plants has been produced.

For root formation, the shoots are transplanted usually to a substrate which contains auxin. After the development of roots, the plants are transplanted to soil in a greenhouse having a high air humidity. By gradually increasing light 130~88 the photosynthesis of the plant is induced.

The most important cost factor in micropropagation is the large amount of work requiring professional skill, which mainly consists of manual cutting of plants and their transplanting from one substrate to another.

In connection with the cutting, the sensitive plant mate-rial may also easily be damaged. Cutting with a knife is slow, especially when working with an easily contaminated material, in which case special attention must be paid to asepsis.

It has now been observed surprisingly that the above-mentioned problems can be reduced by using a laser beam for the cutting of plant material.

Live tissue has been cut by means of a laser beam only in surgery. In surgical operations, the cauterization of the blood vessels in the tissue can be regarded as an advan-tage, the cutting being facilitated as bleeding is pre-vented, especially bleeding from small blood vessels.
Instead, in connection with the present invention it was observed surprisingly that the vascular bundles of plant material are not damaged detrimentally in connection with laser cutting, but the tissue retains its capacity for taking in water and nutrients through the cut surface and, furthermore, the tissue close to the cut surface retains its totipotency.

The advantages of the use of a laser beam in cutting plant material also include the ease of use and rapidness of the method. Because of the high requirement of asepsis, when plant material is being worked on conventionally using a knife, the knife has to be sterilized between the cuttings by dipping it in ethanol and by passing it through a flame~

~30~8 This slows down the cutting, and ethanol may pass into the plant material; even in small amounts ethanol will cause a delay in the starting of growth or may cause the plant to die. The sterilization may also be carried out by heating the instrument. On the other hand, the laser beam is natu-rally sterile, whereupon asepsis is considerably improved.
Likewise, the speed of the cutting increases as the steri-lization step is eliminated.

Automation can also be connected with laser cutting. In this case the share of manual work is minor and the cutting is sped up considerably.

In laser cutting the damage to the plant material is mainly caused by heating. Inert shield gases such as nitrogen, carbon dioxide or argon are used for improving the result of the cutting. The shield gas is directed to the object to be cut in open space by means of a nozzle, or alternatively the cutting is carried out in a chamber filled with shield gas. The amount of shield gas is selected so that charring is as insignificant as possible.

In connection with the developing of the present invention, the e~uipment used for cutting plant material was laser equipment the most important parameters of which are de-scribed below.

The mode of the laser instrument indicates the form of distribution of its beam. In the cutting experiments the operating mode was continuously the TEM 00 mode, in which case the intensity of the beam is distributed according to the Gaussian bell curve.

The cutting power of the laser instrument indicates how much energy the instrument is capable of transferring per time unit to the object to be cut. Often all of the energy ~309~88 is not absorbed into the object to be cut but is reflected from the surface of the object and/or absorbed into the vapors and gases emanating from the object. The cutting power can also be expressed in terms of intensity, which indicates the cutting power per surface area of the beam.
Since the focussing lens focusses the beam in the focal point over a very small area, it is possible even with a low cutting power to achieve high intensity values. Experi-ments have shown that it is possible to cut plants using even a CO2 laser of a power of only 20 W, but in this case the cutting speed is not sufficient. It has been proposed in the literature that a power of approximately 40 W is sufficient for cutting live animal tissue. On the other hand, the price of a laser instrument increases almost in direct proportion to the power squared, and so the highest economically justifiable power for a laser instrument is approximately 100 W. On the basis of the above, the power of a laser instrument suitable for use in connection with the present invention should be within the range 30-100 W.

When the laser beam is pulsed, the values for the length of the pulse and the interval between pulses must be selected so that the material to be cut is heated as little as pos-sible. Values between 0.1 and 10 ms are suitable values.

The diameter of the beam at the focal point also affects the result of the cutting, but in general this parameter is constant and in an order of magnitude of 0.2 mm or less.

In the following examples, the cutting laser used was a longitudinal-flow CO2 laser (Coherent) the beam mode of which was TEM 00 and the resonator of which was provided with an ECQ module enabling the beam to be pulsed. The continuous power of the laser instrument was in principle adjustable within the range 90-350 W, but for the cutting experiment it was desired to lower the power by means of a 1309~88 special gas mixture, in which case the constant continuous power was 61 W. The pulse frequency was selectable within the approximate range 10-2500 Hz, and the length of an individual laser pulse was selectable within the range 0.1 ms - 10 s. The diameter of the beam at the focal point was approximately 0.2 mm. The shield gas used was nitrogen having a purity of 99.998 %. The work station was made up of an xy table the size of which was 600 x 600 mm.

Example The experiments were performed on birch. The effects of cutting were observed by means of a propagation experiment and subsequent cultivation in a greenhouse.

Since a laser beam heats the object being cut, plant tissue may dry or become charred. The damage to the cut surface was monitored by means of a microscope. If the cut surface was subjected to too high a power, it was often visible as charring and as total constriction of the vascular bundles, i.e. "fusion" of the tissue. By changing the parameters of the instrument, attempts were made to improve the trace of the cutting.

For purposes of cutting the plant parts were fixed in place by using small sterile Petri dishes filled with agar ~9 g/l).

Birch, Betula pendula Birch shoots (from an in vitro population) were cut into lengths containing one axillary bud each, by using the laser instrument. The control experiment was carried out by cutting shoots with a knife. The buds were placed on a propagation substrate and cultivated in an incubator (23 C, humidity 50 %, light 2000 lux, 16/8 h). Parallel ~3~ 3~8 experiments were performed in duplicate (3-7 shoot pieces per experiment).

The starting of the growth of the transplants was observed and the propagation coefficient was calculated twice, at 4 and at 6 weeks from the time of the transplanting. The parameters shown in Table 1 were experimented with in the laser cutting.

Table 1. Parameter alternatives experimented with in laser cutting of birch.

Cutting Pulsing parameters Maximum speed Mean method No. Tp/ms Te/ms T/ms f/Hz % 1 m/s power/W
1 0.1 0.4 0.5 2000 0.5-0.6 15.5 2 0.1 0.7 0.8 1250 0.4-0.5 10.5 3 0.1 1.0 1.1 909 0.2 8.5 1.0 1.0 2.0 500 2.0 33 6 1.0 10 11 91 0.2 12 7 10 10 20 50 0.6 32 8 continuous power 2.4 61 The laser-cut transplants propagated well. The propagation of the birches cut in different ways at 4 weeks and at 6 weeks of growth is shown in Figure 1.

The length of the pulse (Tp) was maintained at 0.1 ms in cutting methods 1-3, but the recovery phase (Te) was varied. When the cutting was by method No. 1, the resting phase between the pulses was 0.4 ms, and when method No. 2 was used it was 0.7 ms. It was possible to maintain the cutting speed almost at the same value in both methods. The lengthened recovery phase seems to improve the vitality of the tissue, which was manifested as an increased propaga-tion coefficient. The propagation seemed most effective when cutting methods 5, 6, and 7 were used. In experiment ``` 1309588 No. 5 the long pulse (1.0 ms) and rather long interval (1.0 ms) during a rapid operation (~ %/ms~l) produced a good result. The lengthening of the interval to 10 ms at the expense of the operation speed yielded the same propagation result in experiment No. 6. Experiment No. 7 surprisingly produced the best growth result: in it a very long pulse (10 ms) and a very long pulse interval (10 ms) were com-bined with a moderate operation speed (0.6 %/ms~l).

A tissue cut using a continuous power of 61 W (experiment No. 8) seemed to propagate on the average more weakly than tissue cut using a pulsed beam.

On the basis of the experiment it can be concluded that a laser beam is well suited for cutting birch. The propaga-tion was at least as effective as when the cutting was done with a knife. The birches took root in a normal manner after propagation following laser cutting. The best propa-gation was achieved with the pulsing of experiment No. 7, in which the long recovery time is assumed to have pre-vented the tissue from heating and burning too much. It can also be concluded on the basis of the experiment that a laser beam oscillating at a low frequency damages plants least (method 7).

Claims (8)

1. A method for cutting plant material for the purpose of propagation of plants, wherein the cutting is carried out using a laser beam and a shield gas is used around the object to be cut.
2. A method according to claim 1, wherein the propagation of plants is micropropagation.
3. A method according to claim 1, wherein the laser used is a CO2 laser.
4. A method according to one of claims 1, 2 or 3, wherein the cutting is carried out using continuous power or by pulsing the beam.
5. A method according to claim 4, wherein the continuous power used is a power within the range 30 - 100 w.
6. A method according to claim 4, wherein during pulsing, the mean power varies within the range 5 - 40 w.
7. A method according to claim 1, wherein in that the shield gas used is an inert gas.
8. The method of claim 7, wherein the inert gas is nitrogen or carbon dioxide.
CA 562325 1987-03-26 1988-03-24 Method for cutting plant material Expired CA1309588C (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
FI871333 1987-03-26
FI871333A FI80185C (en) 1987-03-26 1987-03-26 Procedure for cutting plant material

Publications (1)

Publication Number Publication Date
CA1309588C true CA1309588C (en) 1992-11-03

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ID=8524203

Family Applications (1)

Application Number Title Priority Date Filing Date
CA 562325 Expired CA1309588C (en) 1987-03-26 1988-03-24 Method for cutting plant material

Country Status (9)

Country Link
JP (1) JPS63291581A (en)
AU (1) AU598919B2 (en)
CA (1) CA1309588C (en)
DE (1) DE3809002A1 (en)
FI (1) FI80185C (en)
FR (1) FR2612732B1 (en)
GB (2) GB8805784D0 (en)
NL (1) NL190800C (en)
SE (1) SE469206B (en)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB8921403D0 (en) * 1989-09-21 1989-11-08 British Res Agricult Eng Method of and apparatus for cutting plant tissue
WO1992013443A1 (en) * 1991-02-01 1992-08-20 Plant Production Systems B.V. A method for use in a multiplication process of plants and a device for carrying out said method
US6172328B1 (en) * 1998-02-17 2001-01-09 Advanced Foliar Technologies, Inc. Laser marking of plant material
US6180914B1 (en) * 1998-02-17 2001-01-30 Advanced Foliar Technologies, Inc. Laser marking of foliage and cigars
WO2002017705A1 (en) * 2000-08-31 2002-03-07 Wolf-Garten Gmbh & Co. Kg Method and device for gardening and landscape conservation

Family Cites Families (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3483934D1 (en) * 1983-10-13 1991-02-21 Rikagaku Kenkyusho METHOD AND APPARATUS FOR PLANTING A FOREIGN SUBSTANCE INTO LIVING CELL.
JPS6083583A (en) * 1983-10-13 1985-05-11 Rikagaku Kenkyusho Perforation apparatus of live cell with laser
JPS60118473A (en) * 1983-11-29 1985-06-25 オリンパス光学工業株式会社 Micromanipulator
JPS60251872A (en) * 1984-05-25 1985-12-12 Hitachi Ltd Microoperation apparatus of biological cell
JPS60251875A (en) * 1984-05-30 1985-12-12 Hitachi Ltd Apparatus for microoperation of cell
DE3572612D1 (en) * 1984-11-23 1989-10-05 Basf Ag Process for producing cuts in biological material
JPH0644867B2 (en) * 1986-02-19 1994-06-15 株式会社日立製作所 Laser processing method for raw samples
DD262787A1 (en) * 1987-08-10 1988-12-14 Inst Ruebenforschung Kleinwanz METHOD AND DEVICE FOR STERILE CUTTING OF PLANT MATERIAL FOR IN VITRO REPRODUCTION

Also Published As

Publication number Publication date
GB2202723A (en) 1988-10-05
FR2612732B1 (en) 1992-04-24
AU1328388A (en) 1988-09-29
DE3809002A1 (en) 1988-10-06
DE3809002C2 (en) 1992-01-30
FI871333A0 (en) 1987-03-26
SE469206B (en) 1993-06-07
GB8806483D0 (en) 1988-04-20
FR2612732A1 (en) 1988-09-30
SE8801124L (en) 1988-09-27
NL8800760A (en) 1988-10-17
NL190800C (en) 1994-09-01
FI871333A (en) 1988-09-27
GB2202723B (en) 1990-09-05
JPH0446089B2 (en) 1992-07-28
NL190800B (en) 1994-04-05
GB8805784D0 (en) 1988-04-13
JPS63291581A (en) 1988-11-29
AU598919B2 (en) 1990-07-05
SE8801124D0 (en) 1988-03-25
FI80185C (en) 1990-05-10
FI80185B (en) 1990-01-31

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