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Journal of Horticultural Science
ISSN: 0022-1589 (Print) (Online) Journal homepage:
The production of strawberry plants by in vitro
P.h. Boxus
To cite this article: P.h. Boxus (1974) The production of strawberry plants by
in vitro micro-propagation, Journal of Horticultural Science, 49:3, 209-210, DOI:
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Published online: 27 Nov 2015.
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Date: 26 October 2017, At: 07:50
J. hort. Sci. (1974) 49, 209-210
Research Note
The production of strawberry plants by in vitro micro-propagation
Fruit and Vegetable Research Station, Gembloux, Belgium
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In the presence of 6-benzylaminopurine strawberry plants produce many axillary
buds which grow into shoots bearing more axillary buds. Rooted plantlets, however, are only formed in the absence of added cytokinins. A technique that can be
used to multiply large numbers of strawberry plants very quickly is described.
STRAWBERRY plants growing in vitro readily form axillary buds, and the presence of the
cytokinin 6-benzlyaminopurine in the medium stimulates this growth. This paper describes
how this stimulus can be regulated to permit the large-scale production of strawberry plants.
Young strawberry plantlets, obtained from meristems, 'are initially maintained in a
medium containing undiluted Knop solution, the micro-elements used by Murashige and
Skoog (1962), nicotinic acid 0.5 mg/1, pyridoxine HCl 0.5 mg/1, glycine 2.0 mg/1, thiamine
HCl 0.1 mg/1, meso-inositol 100.0 mg/1, indolybutyric acid 1.0 mg/1, glucose 40.0 g/1,
agar 8.0 g/1, adjusted pH 5.6. This medium is based on the mixture of two media recommended by Vine (1968) for strawberry meristem culture.
Under our experimental conditions, at a constant temperature of 24 oc, under 16 h
light (using 40-watt fluorescent lamps, Gro-Lux W.S. : 6 lamps per area of 1.2 X 1 m) the
young plants grow very well. Tl1;ey are put in jam jars with 60 ml of the medium (Fig. 1).
A few leaves and roots are quickly formeq, and then growth slows down and almost stops
after 2 or 3 months of culture. The plants can remain at this stage of growth for several
months without any attention. After one to one and half years, they must be aseptically
transplated into a fresh medium. At this time, the plants are rejuvenated by removing all the
formed roots and all except the 2 or 3 newest leaves. In the fresh basic medium these
rejuvenated plants develop like plants which have been grown from meristems.
If plants, whether grown from meristems or from rejuvenated plants, are cultivated on
basic medium enriched with 1 mg/1 of 6 benzylaminopurine they develop differently. These
plants stop producing roots and trifoliate leaves. After 3 or 4 weeks, 2 or 3 axillary buds
appear at the base of the petiole of the oldest leaves. These axillary buds grow very quickly
and in their turn produce new axillary buds· in a few days. Within 2 months, the initial
plantlet is transformed into a mass of more or less developed buds. Externally, these appear
as a tuft of very small unifoliate leaves, with short petioles, closely pressed against each
other, and at the base of these petioles new buds are formed. There are no roots. These
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Production of strawberry plants by in vitro micro-propagation
tufts can reach several centimetres in diameter, comprising I 5 to 25 buds each of which has
2 or 3 leaves {Fig. 2).
These buds can be divided and separately transplanted into a fresh medium. If they are
planted in a medium containing cytokinin, the axillary buds will continue to proliferate, but
if they are planted on the basic medium without cytokinin, the development of the axillary
bud ceases immediately, and these buds produce normal young plants with some roots and
trifoliate leaves in about 6 weeks (Fig. 3).
This evolution can be easily controlled by the addition or omission of 6-benzylaminopurine (Fig. 4). All the varieties tested, Domanil, Fanil, Gorella, Goupil, Senga gigana,
Senga precosana, Surprise des Hailes, have reacted essentially in the same way. Over
a 2-year period, a few Domanil plantlets have been submitted to alternating cycles in which
cytokinins were present or absent. These repeated treatments never modified the type of
reaction. The plantlets obtained from the various treatments are now under observation in
the field to determine whether varietal characteristics and productivity have been maintained.
It seems probable that our observations apply to many other strawberry varieties.
Adams (1972) also recorded the appearance of shootlets around a small basal callus on
plantlets of two strawberry hybrids grown from meristems in a medium containing benzylaminopurine. The concentration of cytokinin .he used, however, was only one-tenth that
used by us, which may explain why only 22 plants were formed from 4 meristems .
.The adaptation of in vitro strawberry plants to normal cultural conditions is very simple.
The young plants are transplanted into a non-disinfected leaf-mould and maintained for a
few days under a plastic cover or under mist. Transplantation is 100% successful.
By this process one can obtain within a year several million plants (from 156 to 25 6)
from a single mother plant. The rapidity of multiplication, and its ease of regulation in all
seasons, makes it possible to industrialize the clonal multiplication of strawberry plants
in vitro on lines similar to those already used for carnations and orchids.
Provided that our subsequent studies do not reveal the appearance of mutants, we
believe that the micro-propagation of strawberry plants in vitro will replace the expensive
but not wholly effective traditional methods of multiplying healthy plants (Boxus, 1973).
ADAMS, A. N. (1972). An improved medium for strawberry meristem culture. J. hort. Sci.,
47, 263-4.
Boxus, PH. (1973). La production de plants sains de fraisiers. Acta Hort., 30, 187-91.
MURASHIGE, T., and SKOOG, F. (1962). A revised medium for rapid growth and bioassays
with tobacco tissue cultures. Physiologia Pl., 15, 474-97.
VINE, S. J. (1968). Improved culture of apical tissues for production of virus-free strawberries. J. hort. Sci., 43, 293-7.
(Received 13th Ju(v 1973.)
in l'itro
Bv PH . Boxus
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FIG . 2
Tuft of strawberry buds cv. Domanil, in a 25 mm diameter test tube. after 6 weeks with
[face p. 110
in vitro
BY PH . Boxus
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FIG. 3
Fifteen strawberry plantlets, cv. Domanil, in 900 ml jar after 5 weeks' growth without cytokinin.
FIG . 4
Strawberry plantlets, cv. Domanil , after 5 weeks without cytokinin (left) and with
purine/1 (right).
mg 6-benzylamino[face p. 211
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