New Zealand Journal of Botany, 1996, Vol.

34: 273-278 273

0028-825X/96/3402-0273 $2.50/0 © The Royal Society of New Zealand 1996

Comparison of frost tolerance of Nothofagus solandrivar.

cliffortioides (Hook.f.) Poole and Nothofagus menziesii (HooW.i.)
Oerst.

OSBERT J. SUN (Hook.f.) Oerst. (silver beech) occur naturally at

NZ Forest Research Institute Limited similar latitudes and altitudes in New Zealand.
Private Bag 4800 Nothofagus solandri commonly dominates the for-
ests at high altitudes in the drier inland mountain
Christchurch, New Zealand
regions of the South Island; Nothofagus menziesii
dominates or co-dominates with N. solandri at
GEOFFREY B. SWEET
timberlines with high rainfall and improved soil
School of Forestry
conditions (Wardle 1984). Sun et al. (1995) relate
University of Canterbury the difference in the present distribution of N.
Private Bag 4800 solandri and N. menziesii to differences between the
Christchurch, New Zealand two species in tolerance to water stress and water-
logging. It is not known, however, whether the at-
tributes of other environmental variables, e.g., low
Abstract Frost tolerance was investigated in both winter temperature and summer frosts, are important
cold-hardened and actively growing seedlings of in differentiating the geographical distribution of the
Nothofagus solandri var. cliffortioides and two species.
Nothofagus menziesii in New Zealand. Seedlings of
Frost causes freezing of plant tissues, resulting in
the two species from the same geographical origins
injury due to intracellular and/or extracellular ice
showed similar tolerances to simulated mid winter
formation. Frost tolerance of plants is considered to
and summer frosts when grown in a common envi-
be a genetically determined physiological trait but
ronment. The maximum frost hardiness (LT50)
its expression is determined by environmental con-
ranged from -9.0 ± 0.8 to -12.1 ± 0.6°C in N.
straints (Sakai & Larcher 1987). Most woody plants
solandri, and from-10.0 ± 0.7 to-12.4 ± 0.1°C in
native to New Zealand show well-defined geographi-
N. menziesii. Both species exhibited provenance
cal limits along various temperature-related gradi-
variations in frost tolerance that corresponded with
ents (Sakai & Wardle 1978). Frost hardiness for N.
variation in long-term means of annual minimum
solandri was reported to range from -3.5 to —13°C
temperature and frost days per annum of the seed
in mid winter, apparently related to the altitude of
origins. It is suggested that low temperature may not
origin (Sakai & Wardle 1978; Sakai et al. 1981;
be a key environmental factor in differentiating the
Greer et al. 1989). Greer et al. (1989) suggest that
presence of the two species at upper timberlines in
the altitudinal variations in timberlines formed by N.
New Zealand.
solandri mainly reflect the frost hardiness limits of
seedlings. To elucidate the role of low temperature
Keywords frost tolerance; geographical distribu- in differentiating the natural distribution of N.
tion; Nothofagus menziesii; Nothofagus solandri var. solandri and N. menziesii, we investigated the frost
cliffortioides; provenance; timberline tolerance of both cold-hardened and actively grow-
ing seedlings of the two species with the same seed
origins and grown at the same conditions.
INTRODUCTION
Nothofagus solandri var. cliffortioides (Hook.f.)
Poole (mountain beech) and Nothofagus menziesii MATERIALS AND METHODS
Cold-hardened seedlings
B95034 Seed ofN. solandri and N. menziesii were collected
Received 14 August 1995; accepted 11 December 1995 from Lake Hauroko, Eglinton Valley, and Cobb
274
River, in the South Island, and germinated in a glass-
house at the University of Canterbury, Christchurch,
in spring 1990. At each site, collection was made on
eight maternal trees with heavy seed crops. The
minimum distance between trees used to provide
seed was 50 m. Table 1 lists the geographical loca-
tions and long-term means of annual minimum tem-
perature and frost days per annum of the three seed
collection sites (provenances).
Two weeks after germination, seedlings were
transplanted into 1 1 pots (15 cm top diameter x
12 cm height) containing a mix of peat (60%), soils
of a Nothofagus-dominated forest (20%), ground
pine bark (10%), coarse river sand (10%), and slow-
release fertiliser (2 kg/m3 Osmocote®Plus, Grace-
Sierra, Heerlen, The Netherlands). All seedlings
were kept in the glasshouse until the end of March
before being transferred into an open shade house
to induce cold hardiness. The one-year-old seedlings
were transported to Palmerston North in July to
undergo the frost treatment using low temperature
controlled environment (C.E.) facilities (Robertham
et al. 1978) at the HortResearch National Climate
Laboratory, Palmerston North.
Ninety seedlings, 15 for each provenance in each
species, were divided into 3 groups matched in size
and appearance, and placed randomly in the low-
temperature C.E. rooms with frost temperature set
for—5, —10, or—13°C. The C.E. rooms were run on
a 6—6-4 programme (Greer & Warrington 1982), i.e.,
6 hours for cooling from 12°C, 6 hours being main-
tained at the minimum temperature, and 4 hours for
warming up to 12°C. The actual temperatures dur-
ing frosting were recorded as-5.0 ± 0.8, -10.2 ± 0.2,
and—13.9 ± 1.1°C, respectively. The frosts were
conducted in darkness, with the lights turned on one
hour before the warming was completed. During the
frost process, soil temperature was maintained at 5°C
Table 1 Geographical locations of provenance and long-term means of frost
days per annum and annual minimum temperature (Tmin) of the sites.
Altitude
Provenance Latitude (m a.s.l.) frost (days)
Cobb River 41°08'S 870
Eglinton Valley 45°06'S 270
LakeHauroko 46°06'S 150
Source: New Zealand Meteorological Service (1983).
New Zealand Journal of Botany, 1996, Vol. 34
by a 100 watt heating element in the base of each
frost trolley. The soil surface was also well insulated.
Following the completion of all frost treatments,
seedlings were transported back to Christchurch and
placed into a glasshouse with temperatures set for
15°C by day and 10°C at night. Frost damage was
scored visually 4 weeks after treatment on a scale
of 0, no damage; 1, approx. 10% leaves damaged;
2, approx. 30% leaves damaged; 3, approx. 50%
leaves damaged (LT50); 4, approx. 80% leaves dam-
aged; and 5, shoot tissue dead, as described by
Menzies et al. (1981). Frost hardiness (LT50) was
defined as the temperature which caused 50% foliar
damage (Ritchie 1991). This was determined by
drawing free-hand curves of damage rating versus
frost temperatures for previously-matched seedlings
based on size and appearance under the three frost
treatments. Analysis of variance (ANOVA) was
conducted on the estimates of LT 5 Q.
Actively growing seedlings
Seedlings of the two species were lifted from the
floor of a mixed Nothofagus forest in the Eglinton
Valley in early January 1992, and potted into 11 pots
containing the same potting mix as described above.
The size and appearance of these seedlings were
similar to the cold-hardened seedlings of the previ-
ous experiment but their ages were not known. All
seedlings were placed initially in an open shade
house at the University of Canterbury. More than
80% of the seedlings had recovered from transplant-
ing after 4 weeks, and these were transported to the
HortResearch National Climate Laboratory,
Palmerston North, at the end of February 1992. In-
sufficient seed was available to provide seedlings
raised from seed in the glasshouse for this experi-
ment.
Seedlings were divided into 4 groups, with each
Annual Tmin Map
(°C) number
79 -6.5 NZMS260 M26
58 -6.1 NZMS260 D43
26 -4.9 NZMS260 C43
Sun & Sweet—Frost tolerance in Nothofagus 275

Fig. 1 Frost damage in cold-

hardened seedlings subjected to L. Hauroko -o- Eglinton V. • Cobb R.
different temperatures. Vertical
bars show standard errors of
5
means, n = 5, LT50 is indicated by N. solandri
the broken line. 4H

2-\
c
• ~ -I _

CD 0-

N. menziesii

2-

1 -

0
0 -2 -4 -6 -8 -10 -12 -14

Frost temperature (°C)

group containing 12 plants of each species. Seedlings RESULTS

in each group were placed randomly in a low-tem-
perature C.E. room set for a specific frost tempera-
ture. The rooms were run on the 6-6-4 programme
with temperatures of-1.0 ± 0.1, -2.5 ± 0.1,-3.9 ±
0.1,and-5.4±0°C.
Upon completion of the frost treatments, all seed-
lings were brought back to Christchurch and placed
in a glasshouse at the University of Canterbury for
visual assessment of frost damage. The assessment
was conducted 4 weeks after the treatment, with the
number of plants showing production of new shoots
at the stem base after complete shoot damage being
recorded 6 weeks after treatment. Differences in frost
damage between the two species were compared by
Student's Mest at each frost temperature.
Cold-hardened seedlings
Frost damage of cold-hardened seedlings increased
with decreasing temperatures for seedlings from all
three provenances in both species (Fig. 1). Seedlings
subjected to -5°C frost showed only minor damage
in all provenances of both species, but considerable
damage occurred to those subjected to approximately
—14°C. In both species, seedlings from the three
provenances differed clearly in damage rating at
—10°C; the most severe damage occurred in seed-
lings of the Lake Hauroko provenance, and the least
in the Cobb River provenance.
Frost hardiness (LT50) was significantly (P <
0.001) affected by provenance. In N. solandri, LT50
276
—_____==-—®—
__-—-—
Damage rating
4-
/
3-
2-
1- I "*" N. solandri
n-
0 -1 -2 -3 -4
Frost temperature (°C)
ranged from -9.0 ± 0.8°C for the Lake Hauroko
provenance, through-10.2 ± 1.0°C for the Eglinton
Valley provenance, to -12.1 ± 0.6°C for the Cobb
River provenance; while in TV. menziesii, it was—10.0
± 0.7, —11.3 ± 0.4 and-12.4 ± 0.1°C, respectively.
There was no significant difference between the two
species in frost hardiness in all three provenances.
Actively growing seedlings
Frost at -1.0°C caused minor damage to actively
growing seedlings of both N. solandri and N.
menziesii, while nearly all plants subjected to frost
temperatures of-2.5°C and below suffered severe
frost injury (Fig. 2). There was no significant differ-
ence between TV. solandri and N. menziesii in the
damage rating for seedlings subjected to -1.0°C
frost. LT50 for actively growing seedlings in the two
species is estimated to be at a temperature of approxi-
mately -1.8°C.
Six weeks after the frost treatment, some seed-
lings subjected to apparent lethal temperatures
showed production of new shoots at the base of the
main stem, especially in TV. menziesii. The percent-
age of plants showing production of new shoots dif-
fered considerably between the two species; one-third
ofN. menziesii seedlings produced new shoots after
being frosted at-2.5 and-3.9°C, but only one-sixth
of N. solandri seedlings at -2.5°C frost or N.
menziesii seedlings at—5.4°C were observed to have
such new shoots. Nothofagus menziesii seedlings
usually produced several vigorous new shoots on a
single plant, whereas N. solandri seedlings normally
produced a small, single shoot only.
New Zealand Journal of Botany, 1996, Vol. 34
Fig. 2 Frost damage in actively
m growing seedlings. Vertical bars
show standard errors of means,
n = 12, LT50 is indicated by the
broken line.
"•" N. menziesii
-5 -6
DISCUSSION
Low temperature plays a prominent role in the dis-
tribution of woody plants. Altitudinal limits for tree
growth are largely determined by low temperature.
Although N. solandri forms the highest timberline
in New Zealand, its cold tolerance in both cold-hard-
ened and actively growing seedlings was found to
be similar to that in N. menziesii for one-year-old
plants when the two species were grown in the same
environments. Both species showed a level of mid
winter frost hardiness similar to that of Nothofagus
cunninghamii (Read & Hill 1988, 1989; Read &
Hope 1989), Nothofagus betuloides and Nothofagus
dombeyi (Alberdi et al. 1985), and Nothofagus nitida
(Sakai et al. 1981); but greater than Nothofagus
fusca, Nothofagus truncata, and Nothofagus moorei
(Sakai etal. 1981; Read & Hope 1989). Two decidu-
ous subalpine Nothofagus species, Nothofagus
antarctica which occurs naturally in Chile, and N.
gunnii which is endemic to Tasmania, were found
to be the most cold tolerant species in the genus
(Sakai et al. 1981; Alberdi et al. 1985). The frost
hardiness in cold-hardened N. solandri seedlings
obtained in this study is consistent with those re-
ported in other studies (Sakai & Wardle 1978; Sakai
et al. 1981; Greer et al. 1989). \nN. menziesii, simi-
lar winter frost hardiness was also reported for adult
trees (Neuner & Bannister 1995).
The tests of tolerance to summer frost were con-
ducted using seedlings collected from the floor of a
Nothofagus forest in the Eglinton Valley because of
the lack of a sufficient seed source. Consequently the
Sun & Sweet—Frost tolerance in Nothofagus
age of seedlings tested was unknown. Neverthless,
the actively growing seedlings of the two species
exhibited a frost hardiness of approximately—1.8°C.
This is consistent with the results obtained by Greer
et al. (1989) who found that the mid summer frost
hardiness in N. solandri varied from —1 °C for seed-
lings collected from 460 ma.s.l. atGlentui, to-3.5°C
for those collected from 1100 m a.s.l. on the lower
slopes of Mt Cockayne in the Craigieburn Range.
Provenance variation in frost tolerance was
clearly displayed in both TV. solandri and N.
menziesii, with the variation corresponding appar-
ently to variations in long-term means of annual
minimum temperature and frost days per annum
among provenances even when seedlings of differ-
ent seed origins were grown in a common environ-
ment. This suggests that frost hardiness in the two
species is more related to the environmental condi-
tions of their natural habitats than to the genetic dif-
ference between them. It may be postulated that low
temperature has not acted as a prominent selection
pressure to differentiate the present distribution of
N. solandri and N. menziesii.
Frost survival capacity depends primarily on the
specific frost tolerance of a plant. However, the
importance of recovery from frost injuries has also
been stressed by Sakai & Larcher (1987) who stated
that the chance of frost survival of a plant in a given
environment consisted of mitigation or exclusion of
the freezing risk, frost tolerance, and recovery after
frost damage. During a summer frost, the basal buds
of a plant could remain undamaged partly due to
insulation provided by litter on the forest floor, and
partly due to their relatively high resistance to low
temperature compared with leaves and shoots higher
above (Larcher & Bauer 1981). Regrowth is thus
possible in species which are capable of producing
new shoots at their bases after suffering from severe
dieback. Although N. solandri and N. menziesii
showed similar frost hardiness in seedlings originat-
ing from the same sites, Nothofagus menziesii may
be more capable of surviving a summer frost than
N. solandri, because of its greater ability to produce
new shoots from the base of the stem. Thus, in ar-
eas with a high frequency of summer frost, Notho-
fagus menziesii might have a more extended
distribution than N. solandri.
The present study suggests that frost tolerance
may not contribute directly to the differences be-
tween N. solandri and N. menziesii in forming up-
per timberlines in New Zealand. The differentiation
between the two species in dominance at timberlines
might be related, at least partially, to the effects of
277
water stress. Nothofagus solandri has been shown
to be much more tolerant to water stress than N.
menziesii (Sun et al. 1995). A combination of high
radiation, low soil temperature, and shallow soil may
cause severe water stress to plants growing in sub-
alpine environments (Schulze & Hall 1982).
ACKNOWLEDGMENTS
We are grateful to E. A. Halligan and I. Warrington for
their kind support in use of the low-temperature facili-
ties at the HortResearch National Climate Laboratory,
Palmerston North. We thank D. H. Greer for his techni-
cal support and review of the manuscript. This study was
partially supported by a research grant from the Robert
C. Bruce Trust. O. J. Sun was a recipient of a T. W. Adams
Scholarship and BNZ Scholarship for Forestry Research.
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