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PHYSIOLOGIAE
PLANTARUM
Vol. 22. No. 2. 2000:129-134
Institute of Forest Ecology, Slovak Academy of Sciences, Vieska nad Zitavou, 951 52 Slepcany, Slovak Republic
Key words: Magnolia x soulangiana Soul.-Bod., al. 1995). Shoot induction, proliferation and bio-
axillary shoots, media composition, media concen- mass production directly and indirectly are influ-
tration, type of vessel closure enced by the type of culture vessels and their clo-
sures as well, (Monette 1986, Mackay, Kitto 1988,
McClelland, Smith 1990). The type of culture ves-
Abstract sel closures stimulated growth of the explant cul-
Proliferationof axillaryshoots from nodal segmentsof saucer ture (Tricoli 1983, Kamenicka 1996). The culture
magnolia (Magnolia x soulangiana Soul.-Bod.)was achieved closures can have not only a stimulative effect on
on modifiedStandardiand Catalano(S medium)and Lloydand
the growth of explant culture (Fitter, Krikorian
McCown (WPM) medxa containing 1.33 gmol.dm- BA and
0.54 ~mol.dm " NAA. The greatestnumber of axlllaryshoots 1985), but they can induce necroses of shoot tips
was producedon S-mediumwith full strengthmacronutrients. and branching of shoots (Sha et al. 1985).
Statisticallysignificantwere the differencesin biomassof axil-
lary shoots cultured in vessels sealed with plastic closures. Tissues of magnolias are difficult to disinfect prior
Rootingof the shoots was achievedon half strengthS medium to attempting to place them in sterile culture
supplemented with 4.9, 9.8, 14.7 and 19.6 gmol-drn- IBA.
Rootedplantletswere able to resume independentgrowthafter (Tubesing 1998). For the establishment of magno-
a short period of acclimatization. lia culture juvenile shoots, axillary buts, shoot tips
(direct regeneration) or unripe embryos (somatic
List of abbreviations: 6-benzylaminopurine (BA), embryogenesis) are used as primary explant in mi-
(x-naphtaleneacetic acid (NAA), 4-(3-indolyl) bu- cropropagation. Biederman (1987) concluded that
tyric acid (IBA) the best time for primary explants is the period after
dormancy. Success in regeneration depends on the
choice of the culture media and growth conditions.
Introduction
Some species and cultivars of magnolia have differ-
Environmental factors are very important for the ent culture media requirements, modification of
growth, development and morphogenesis of tissue culture medium is often inevitable for magnolia re-
cultures. Plant growth in vitro is affected by many generation in vitro (Biedennan 1987, Kamenicka,
factors, such as photoperiod, temperature, exoge- Valka 1997).
nous growth regulators, relative humidity in the
culture vessels and medium composition (Kozai et
129
A. KAMENICKA & M. LANAKOVA
The objective of this research was to study the in- After 8 weeks number, length, fresh and dry weight
fluence of media composition, media concentration of axillary shoots were recorded. In each experi-
and type of vessel closure on propagation of mag- ment, 70 explants were used in three repetition.
nolia in vitro. Shoots 3.0 to 4.0 cm long were separated from the
culture and rooted on S1/2 medium suplemented
with 4.9, 9.8, 14.7 and 19.6 ~amol.dm-3 IBA, 0.3 %
Materials a n d M e t h o d s
activated charcoal, 20.0 g.dm -3 sucrose and 7.0
Apical shoots (7-10 cm long) of saucer magnolia g.dm -3 agar (Phyto Agar from Duchefa Biochemie
(Magnolia x soulangiana Soul.-Bod.) consisting of B.V. Haarlem, The Netherlands). Rooting medium
3 to 4 internodes were taken in spring (April) from without IBA was marked as control. All chemicals
100-year old trees growing at the Arboretum were purchased from SIGMA (St. Louis, MO,
Mlyfiany. After removal of leaves the shoots were USA). Five shoots were placed into 250 ml culture
sterilized in solution of HgC12 (0.1-0.3 %) with jar. Growth conditions were the same as above.
three drops of Tween 20 (0.03-0.05 %) for 5-7 min.
After 5 weeks, number of rooted shoots, number of
Then the shoots were soaked in 70 % ethanole (20 s)
roots, total root length, fresh and dry weight of roots
and rinsed (three times) with sterile water. Shoots
were recorded. Results were evaluated using
were cut for nodal segments (3-5 mm long) and cul-
SAS/Statistical program package for personal com-
tured on S medium. The primary culture was trans-
puters (SAS Institute Inc., USA, 1995). Signifi-
ferred after 6-8 weeks on fresh medium (Fig. 1).
cance of the treatment effects was determined by
Shoots were cultured on S medium o r S1/2 concen-
analysis of variance (Statgraphics, Version 6, SAS
tration medium (Standardi, Catalano 1985) or
Institute Inc.USA).
WPM or WPM1/2 m e d i u m (Lloyd, M c C o w n
1980). Shoots were cultured in 100 ml culture ves-
sels containing 25 ml of medium. The culture ves- Results and Discussion
sels were closed with metal (SMet) or plastic (Spvc)
light permeable closures with vent membranes Culture e s t a b l i s h m e n t a n d multiplication
(FORING-LOCK M 82.5 x 24/6). The culture me-
The effect of different culture media on the forma-
dia were supplemented with 20 g-dm -3 of sucrose, 7
tion of axillary shoots was examined (Table 1).
g.dm -3 of agar, 1.33 gmol.dm -3 BA and 0.54
Contamination (caused by fungi) of primary cul-
~tmol.dm-3 NAA. All cultures were kept at a 16 h
ture was generally low. A comparison of nodal ex-
light (35-40 ~tmol.m-2.s -1) and 8 h dark photocycle
plants cultured on WPM medium with WPM1/2 in-
at 20-24 °C.
dicated that differences were similar. Shoots pro-
130
EFFECT OF MEDIUM COMPOSITION ...
Dcor~rot
Jcu~e
0 10 20 30 56
days
131
A. KAMENICKA & M. LANAKOVA
Table 3. Effect of vessel closures on growth of magnolia cul- Table 4. Effects of IBA concentration on rooting of magnolia
ture microcuttings
Media Number of Fresh weight_+SE IBA Number Length of Rooting %
shoots+-SE (lamol.dm3) roots/shoot roots (mm)
SMct 4.42-+0.225 0.79-+0.033 0.0 0.9 12.0 28.7
Spvc 5.78+-0.237 1.21-+0.042 4.9 1.3 30.4 45.0
9.8 3.9 26.8 65.7
Analysis of variance 14.7 5.3 24.5 83.3
19.6 6.8 23.4 96.2
Source of df Mean squares
variation Analysis of variance
No shoots/explant Productionoffmshweight
Vessels 1 0.7 7.1"
closure Source of df Mean squares
variation
Error 98 0.1 0.3 No roots/shoot Length of roots
Auxins 4 54.4* 8.3*
*Significant at P = 0.01 Error 55 7.2 0.4
Culture vessels with metal (SMct)and plastic (Spvc) closure
*Significant at P = 0.01
was not significant. McClelland and Smith (1990)
described that the effect of types vessel closure also
depends on the requirements of individual plant R o o t i n g o f microcuttings
species. Our results confirmed that multiplication
S h o o t s with 2-3 c m length were exiced from cul-
of axillary shoots was more intensive in culture ves-
tures and placed on rooting m e d i u m (Table 4). In
sels sealed with light permeable closures. The sig-
the absence of plant growth regulators, shoots grew
nificance o f differences in fresh weight was meas-
normally, but root production was sporadic (0.9).
ured by the analysis of variance.
Increasing level of I B A stimulated root production,
but length of roots decreased. Vitrification and cal-
132
EFFECT OF MEDIUM COMPOSITION ...
2: @: 7:¸¸¸¸ i £ ¸¸ :: /
133
A. KAMEN1CKA & M. LANAKOVA
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134