全 文 ：Journal of Forestry Research (2008) 19(2):125–130
DOI 10.1007/s11676-008-0021-8





Beneficial effects of cold-moist stratification on seed germination be-
haviors of Abies pindrow and Picea smithiana

Balbir Singh Rawat 1, Vinod Prasad Khanduri 2*, Chandra Mohan Sharma 1
1 Department of Forestry, HNB Garhwal University, Post Box-76, Srinagar Garhwal-246174 (Uttaranchal), India
2 Department of Forestry, Mizoram University, Aizawl-796001, Mizoram, India

Abstract: A study was conducted to evaluate the effect of GA3, moist–chilling and temperature on seed germination of Abies pindrow and
Picea smithiana from five different provenances. Seeds were soaked in GA3 (10 mg⋅L-1) for 24 h, then chilled at 3−5°C for 15 days. Four
temperature regimes viz. 10°C, 15°C, 20°C and 25°C were used for stimulating seed germination. Results showed that soaking and chill-
ing significantly increased germination percentage. The germination percentage was highest at 10°C. Overall results showed that soaking
seeds in GA3 (10 mg⋅L-1) for 24 h, moist chilling for 15 days, and germinating at 10°C produced an effective germination in both the spe-
cies studied. The statistical analysis of the data proclaimed significant effect of treatment, temperature, provenance and treatment with
temperature interactions on seed germination.
Keywords: stratification; provenance; seed germination; GA3; silver fir; spruce



Introduction

The Western Himalayan temperate Forests are distinguished as
Moru Oak Forests, Moist Deodar Forests, Western Mixed Conif-
erous Forests, Low Level Blue pine forests, Kharsu Oak Forests
and West Himalayan Upper Oak and fir forests (Champion et al.
1968). The most common coniferous species in these forests are
Blue Pine (Pinus wallichiana A. B. Jacks), Himalayan cedar
(Cedrus deodara Royal ex D. Don), Himalayan cypress (Cu-
pressus torulosa Don), Spruce (Picea smithiana wall. Boiss),
Silver fir (Abies pindrow spach.) and Himalayan Yew (Taxus
baccata Linn.). The natural regeneration of silver fir, spruce and
Himalayan yew is generally poor and first attention to this prob-
lem was paid by Redcliffe (1906). Since then, the problem of
natural regeneration of these species has been constantly engag-
ing the attention of the forest scientists. A number of factors are
considered responsible for the absence of natural regeneration of
these species, such as lack of adequate light on the forest floor,
dense weed growth (Troup 1921), thick layer of humus (Troup
1921; Taylor et.al. 1934; Glover 1936; Kaul 1970), accumulation
of debris (Hafizullah 1970), and continuous grazing (Redcliffe

Received: 2007-10-15; Accepted: 2008-01-19
© Northeast Forestry University and Springer-Verlag 2008
The online version is available at http://www.springerlink.com
Biography: Balbir Singh Rawat (1975-), male, Project Associate in De-
partment of Forestry, HNB Garhwal University, Post Box- 76, Srinagar
Garhwal-246174 (Uttaranchal), India. (E-mail: bsrawat74@gmail.com)
∗Corresponding author: (E-mail: vpkhanduri@yahoo.co.in)
Responsible editor: Zhu Hong

1906; Flewett 1930; Sufi 1970). Infrequent seed years and low
germinative capacity of the seeds could be also considered being
important factors contributing poor natural regeneration.
The reproductive system of conifers is exclusively sexual and
the natural regeneration, in turn, depends on the production, dis-
persal and germination capacity of seeds and successful estab-
lishment of seedlings. Seed germination of most temperate co-
niferous species is inhibited by evolved trait seed dormancy
(Leadem 1986; West et al. 1970; Singh 1989). Cold-moist strati-
fication is a commonly used practice to break dormancy in seeds
and to attain vigorous, speedy, maximum, and uniform germina-
tion for laboratory testing, green house and nursery sowing
(Fowler et al. 1964), which has been reported in many studies on
conifers (Mergen 1963; Fowler et al. 1964; Roberts et al. 1982;
Wang et al. 2000;) and several hardwood seeds (Schopmeyer
1974; Villiers 1971; Nikolaeva 1977; Bevington 1986; Barnett et
al. 1978; Farmer et al. 1972; Farmer 1974; Farmer et al. 1981).
The plant hormone, GA3, plays an important role in control of
the various physiological processes in plant growth and devel-
opment including seed germination, shoot growth, cell division,
internode elongation and the formation of flower buds. Gibberel-
lin is well known to break dormancy of seeds and buds in many
plants (Brian et al. 1959; Stuart et al. 1961; Weaver 1959). The
Gibberellic acid (GA3) has been reported to promote germi-
nation of seeds (Vogt 1970; Krishnamurthy 1973; Chandra et
al. 1976). However, the germination percentage increased in
the seeds of Nothofagus obliqua, when they were pre-chilled
after soaking in GA3 solutions for 24 hours (Shafiq 1980).
Moist chilling and gibberellin treatments have been reported very
effective for seed germination in some woody species, viz. Aes-
culus hippocastanum (Tompsett et al. 1998), Pinus taeda (Wu et
al. 2001), Ginkgo biloba (Wilson et al. 2004), Acer pensylvani-
RESEARCH PAPER
Journal of Forestry Research (2008) 19(2):125–130

126
cum (Bourgoin et al. 2004), Aribotrya japonica (El-Dengawy,
2005), Larix deciduas (Gorian et al. 2007) and Prunus cam-
panulata (Chen et al. 2007). Variation in germination can be
manifested through provenance tests designed to assess the de-
gree and pattern of variation across species ranges. Such tests are
actually based on the phenotypic variations among seed lots from
provenances, and not on genetic variation. Rowe (1964), and
Baskin and Baskin (1973) have noted that the difference in seed
characteristics of ecologically important provenances may also
be due to genetic variability. Considering all the aforesaid facts,
the present study aims at understanding the effect of pre-chilling,
after soaking in GA3 solutions for 24 hours, on the seed ger-
mination of different provenances of Abies pindrow, and Picea
smithiana.

Materials and methods

Selected seed provenances

The seeds of A. pindrow and P. smithiana were collected from 5
different provenances from Garhwal Himalaya, India. The details
of the provenances selected within the study area were presented
in Table 1 and Fig. 1.

Table 1. Geographical and meteorological description of the selected provenances of A. pindrow and P. smithiana
Temperature (°C) Provenances Species occurred Altitude
(m)
Latitude
(N)
Longitude
(E) Min. Max.
Mean annual rainfall
(mm)
Bharsar A. pindrow 2697 30º24 79º31 -0.70 27.5 1084.00
Dudhatoli A. pindrow 3122 30º5 79º12 -0.65 25.8 1935.00
Ransolikhal A. pindrow 2750 30º25 79º17 -0.56 26.9 1475.00
Surkanda A. pindrow 3030 30º27 79º18 -0.75 25.3 1595.00
Tapovan A. pindrow & P. smithiana 3798 30º31 79º36 -0.84 24.5 1892.00
Banjbagad P. smithiana 2775 30º15 79º34 1.13 30.8 1276.00
Hanumanchatti P. smithiana 2880 30º41 79º30 -0.10 25.3 2098.00
Helang P. smithiana 2595 30º33 79º37 1.34 28.9 1860.00
Pandukeshwar P. smithiana 2657 30º31 79º32 -0.91 27.0 1932.00


Fig. 1 Location map of the study area

The seed germination tests were conducted under laboratory
conditions at various constant temperatures viz., 10°C, 15 °C,
20°C and 25°C inside a seed germinator (Model No. 8LT-SGL
CALTAN). The seeds of all the provenances of both species
were germinated at all the aforesaid temperatures to obtain the
best temperature range for seed germination after applying fol-
lowing treatments: Treatment 1 (soaking of the seeds in distilled
water at room temperature (25°C) for 24 hours) was as control
treatment. Treatment 2 (seeds in Gibberellic acid (GA3 10
mg⋅L-1) were soaked at room temperature for 24 hour and then
chilled for 15 days (at 3−5°C).
For germination, five replicates of 20 pre-treated seeds were
placed in Petri dishes containing two filter papers, kept in the
germinator, and maintained at desired temperatures in light con-
dition. Observation data were recorded daily upto 21 days.
Radical emergence was taken as the criteria for germinability.
The recorded data on seed germination were quantified in terms
of germination percentage and germination value. The germina-
tion percentage was the value of seeds germinated at the comple-
tion of the germination period, whereas, germination value is an
index, combining speed and completeness of germination. The
germination value according to Czabator (1962) can be ex-
pressed as:
GV= PV × MDG (1)
where, GV is the germination value, PV the peak value of germi-
nation, and MDG is the mean daily germination.

Statistical analysis

The statistical analysis was conducted on mean values and the
analysis of variance (ANOVA) was performed using SPSS
package. The critical difference (Cd) was calculated as:
Cd = SEd × t0.01 (2)
where, SEd is the standard error of difference calculated as
SEd = √2Me / r (3)
where, Me is the mean sum of square due to error, and r is the
number of replicates.

Results and discussion

Germination of seeds of various provenances of A. pindrow and
P. smithiana after pre-soaking and pre-chilling treatments under
different temperature regimes, (10°C, 15°C, 20°C and 25 °C) has
Journal of Forestry Research (2008) 19(2):125–130

127
yielded significant differences in seed germination. The detailed
treatment-temperature interactions are as following. Five differ-
ent provenances, viz. Bharsar, Dudhatoli, Ransolikhal, Surkanda
and Tapovan of Abies pindrow manifested maximum germina-
tion percentage at 10 °C, for both control and GA3 treated chilled
seeds. The seeds of different provenances which had given dis-
tilled water treatment proclaimed a germination percentage range
of 21%−32 % at 10°C, 14%−18% at 15°C, 15%−22% at 20°C
and 10%−19% at 25°C, whereas, pre-chilled seeds manifested
34%−56%, 29%−47%, 28%−33% and 25%−30% germination at
10°C, 15°C, 20°C, and 25°C, respectively (Table 2). Similarly,
the seed germination of five provenances, i.e, Banjbagad, Hanu-
manchatti, Helang, Pandukeshwar and Tapovan, of P. smithiana
also revealed the maximum germination percentage at 10°C. The
range of germination percentage at 10°C, 15°C, 20°C and 25°C
for both control and prechilled seeds was from 31%−37% and
52%−72%, respectively; at 10°C, from 22%−30% and 32%−55%,
respectively; at 15 °C, from 25%−30% and 39%−48%, respec-
tively; at 20°C and 15%−26% and 32%−47%, respectively at
25°C (Table 3).

Table 2. Effect of different treatments and temperatures on seed germination percentage and germination value in different provenances of
Abies pindrow (Italics represent germination value)
Provenances Treatments
Bharsar Dudhatoli Ransolikhal Surkanda Tapovan
10°C
Control 22±3.75
0.34±0.08
26±4.86
0.26±0.05
26±4.01
0.54±0.18
21±2.92
0.41±0.08
32±2.00
0.67±0.15
Chilling 34±1.87
0.53±0.10
56±5.80
0.80±0.21
45±4.19
0.63±0.11
45±5.49
0.58±0.18
52±5.84
0.56±0.07
GA3 (10 mg⋅L-1) 28±2.55
0.39±0.04
41±4.86
0.64±0.14
45±4.19
0.69±0.12
33±3.40
0.48±0.11
34±4.86
0.47±0.14
15°C
Control 14±4.01
0.30±0.10
17±2.55
0.25±0.07
17±1.22
0.22±0.06
17±2.55
0.39±0.09
18±4.37
0.25±0.14
Chilling 29±1.87
0.29±0.02
43±3.40
0.54±0.09
34±3.68
0.51±0.11
37±2.00
0.45±0.09
47±5.39
0.64±0.11
GA3 (10 mg⋅L-1) 27±5.16
0.47±0.15
25±1.58
0.35±0.03
21±2.92
0.38±0.13
21±4.31
0.29±0.07
31±2.92
0.44±0.05
20°C
Control 15±3.17
0.15±0.04
15±3.54
0.29±0.11
22±9.59
0.49±0.38
21±7.50
0.12±0.02
17±2.00
0.11±0.01
Chilling 31±4.90
0.26±0.06
33±2.55
0.39±0.08
28±3.00
0.41±0.07
33±4.64
0.30±0.06
30±2.74
0.33±0.03
GA3 (10 mg⋅L-1) 23±6.26
0.44±0.08
28±3.40
0.47±0.17
25±2.74
0.27±0.07
18±4.07
0.20±0.06
25±1.58
0.26±0.03
25°C
Control 10±8.28
0.30±0.28
12±2.00
0.19±0.05
19±1.00
0.26±0.05
10±3.17
0.09±0.03
10±2.74
0.10±0.05
Chilling 27±3.00
0.36±0.05
30±5.71
0.43±0.13
27±4.64
0.43±0.13
30±6.53
0.39±0.11
25±3.17
0.27±0.05
GA3 (10 mg⋅L-1) 20±6.53
0.20±0.09
16±4.31
0.16±0.05
23±2.00
0.29±0.05
26±4.86
0.46±0.17
21±2.45
0.28±0.05

A critical review of the data presented in Tables 2 & 3 reveals
that among all the selected temperature regimes, 10°C was the
best temperature for the seed germination of both species, as the
highest germination percentage and germination value was ob-
served in this constant temperature, whereas, the least percentage
of germination and germination value was recorded at 25°C.
However, the seeds which were followed chilling treatment after
soaking in GA3 for 24 h manifested highest percentage of ger-
mination in all the selected provenances of both species. On the
other hand, seeds treated with distilled water (as control) exhib-
ited poor germination percentage and germination value in all the
provenances of A. pindrow, and P. smithiana. In nature, dormant
seeds of most temperate conifer’s species are exposed to cold
wet conditions during winter, which germinate when temperature
rise in early spring. The dormant seeds of such species would
germinate at controlled, low temperature in the laboratory (Ed-
wards 2004). Among different provenances of both species, the
Dudhatoli provenance of A. pindrow and Tabovan provenence of
P. smithiana were the most successful in respect of germination
percentage and germination value. The statistical analysis of the
data revealed significant effect of treatment, temperature, prove-
nance and treatment with temperature interactions on seed ger-
mination in both the studied taxa (Table 4).
Moist chilling has long been recognized as a useful method of
treating seeds to improve the rate and percentage of germinabil-
ity (Outcall 1991) in addition to other pre-sowing treatments that
increase germination (Heydecker et al. 1977). The treatment may
also facilitate germination at sub-optimal temperatures
(10−20°C), which is particularly important for spring sowing in
nurseries in temperate climates. Allen (1960) was of the opinion
that the longer the chilling period, the better was the germination
of the coniferous seeds. Improved germination by stratification at
Journal of Forestry Research (2008) 19(2):125–130

128
different time period has been reported in Ginkgo biloba for 12
weeks stratification period (West et al. 1970), Ceanothus san-
guinus for 4 months at 2−5 °C (Radwan et al. 1977), Carpinus
caroliana for 18 weeks at 4−5 °C (Bretzloff et al. 1979), Cedrus
deodara for one week (Thapliyal et al. 1980) and Picea smithi-
ana for two months at 2°C (Singh 1989). In the present study,
moist chilling after GA3 treatment has resulted in better germina-
tion for seeds, which was duly supported by many other studies
(Roos et al. 1971; Willemsen et al. 1972). Nevertheless, the
stratification-redry method has been shown to improve germina-
tion in Pacific silver fir (Abies amabilis), (Edwards 1982; 1997;
Leadem 1986), subalpine fir (A. lasiocarpa), (Leadem 1988,
1989), and Nordmann fir (A. nordmanniana), (Jensen 1997).
However, controlling fir seed moisture content during stratifica-
tion is not a new idea, having been recommended for prolonged
pretreatment of seeds of hybrid firs (Wright 1950). Additionally,
reduced seed moisture content was reported for pretreatment of
Guatemalan fir (A. guatemalensis), (Donahue et al. 1985).

Table 3. Effect of different treatment and temperature on germination percentage and germination value in different provenances of Picea
smithiana (Italics represent germination value)
Provenances Treatments
Banjbagad Hanumanchatti Helang Pandukeshwar Tapovan
10°C
Control 31±1.86
0.54±0.11
31±6.61
0.83±0.29
37±2.25
0.69±0.09
36±3.68
1.26±0.30
27±3.75
0.68±0.09
Chilling 52±3.40
0.73±0.18
63±6.05
1.17±0.21
54±8.59
0.82±0.27
59±5.35
1.48±0.29
72±7.53
1.26±0.31
GA3 (10 mg⋅L-1) 42±5.62
0.83±0.29
53±4.07
0.93±0.18
45±5.01
1.05±0.24
52±3.75
1.07±0.25
56±6.01
1.29±0.07
15oC
Control 29±2.92
0.49±0.09
22±2.55
0.41±0.10
29±1.12
0.59±0.05
30±1.58
0.61±0.05
22±2.00
0.82±0.17
Chilling 32±5.16
0.31±0.04
41±5.80
0.50±0.09
35±5.25
0.38±0.10
50±3.17
0.99±0.32
55±4.19
0.84±0.13
GA3 (10 mg⋅L-1) 38±3.00
1.07±0.22
30±4.48
0.64±0.13
38±6.65
0.59±0.12
38±2.00
0.78±0.13
36±5.58
0.61±0.05
20°C
Control 29±6.22
0.57±0.13
26±4.31
0.55±0.09
26±4.31
0.46±0.08
25±1.58
0.42±0.06
30±2.74
0.71±0.21
Chilling 39±4.31
0.49±0.07
48±6.05
0.91±0.18
48±6.05
0.62±0.16
45±4.75
0.41±0.15
45±5.25
0.57±0.11
GA3 (10 mg⋅L-1) 36±3.68
0.85±0.18
43±3.75
1.02±0.13
43±3.75
1.16±0.39
54±11.90
1.57±0.53
42±2.00
1.22±0.13
25°C
Control 26±4.01
0.42±0.10
21±1.87
0.35±0.15
21±1.87
0.30±0.09
25±3.54
0.41±0.15
15±5.71
0.22±0.10
Chilling 32±3.00
0.30±0.03
42±5.84
0.49±0.12
42±5.84
0.48±0.10
57±5.16
1.08±0.34
31±3.32
0.36±0.06
GA3 (10 mg⋅L-1) 30±2.74
0.41±0.07
28±4.37
0.70±0.24
28±4.37
0.45±0.12
35±4.48
0.84±0.23
24±2.45
0.46±0.11

Table 4. ANOVA effects of provenance, temperature, treatment and treatment with temperature on seed germination of Abies pindrow and Picea
smithiana
Abies pindrow
F-critical CD Source of variation
d. f SS MSS F-ratio
5% 1% 5% 1%
Provenances 4 0.79 0.1975 5.64** 3.26 5.41 0.26 0.36
Temperature 3 1.89 0.63 18** 3.49 5.95 0.28 0.40
Error[a] 12 0.42 0.035
Treatment 3 1.56 0.52 4.7** 2.79 4.26 0.47 0.63
Treatment X Temperature 9 2.16 0.24 2.16* 2.08 2.81 0.29 0.39
Split plot Error[b] 48 5.31 0.1106
Picea smithiana
F-critical CD Source of variation
d. f SS MSS F-ratio
5% 1% 5% 1%
Provenances 4 1.46 0.365 6.08** 3.26 5.41 0.34 0.47
Temperature 3 3.14 1.04667 17.43** 3.49 5.95 0.37 0.53
Error[a] 12 0.72 0.06
Treatment 3 1.46 0.48667 1.87 2.79 4.26 0.72 0.95
Treatment X Temperature 9 7.45 0.82778 3.19** 2.08 2.81 0.46 0.61
Split plot Error[b] 48 12.45 0.25938
Notes: *----Significant at 5% and **----significant at 1%; MSS---- mean sum of squares SS---- Sum of squares.
Journal of Forestry Research (2008) 19(2):125–130

129
For shallow-dormant seeds of Douglas-fir (Pseudotsuga men-
ziesii), lodgepole pine (Pinus contorta) and Stika spruce (Picea
sitchensis), moist chilling is not only a requirement to alleviate
dormancy, but a prolonged period of chilling is necessary to
achieve rapid and uniform germination under the low tempera-
ture in early spring (Jones et al. 1994; Jinks et al. 1996). Moist
chilling for 15 days did improve the rate and percentage of ger-
mination of A. pindrow and P. smithiana seeds at 10−15°C over
21 days. This effect of moist-chilling on the activation of germi-
nation may be utilized for short-term maintenance of seeds in a
moistened condition at 3−5°C, which could be considered im-
portant for forestry practices, and afforestation programmes in
temperate regions. Further, the results of the present study sug-
gest that the seeds of A. pindrow and P. smithiana are dormant.
The level of dormancy may vary from one provenance to another
even for seeds from the same parent trees, among parent trees in
the same stand in any one crop, among cones on the same parent
tree, and from seed to seed in the same cone. Thus, in any seed
lot, some seeds may be non-dormant, slightly dormant, some-
what dormant, while others are very dormant. Standard stratifi-
cation/prechilling treatments to promote germination could ac-
commodate such wide variations.
Exogenous application of GA3 has been reported to be effec-
tive in breaking dormancy and in substituting for a cold stratifi-
cation requirement in many seeds (Chien et al. 2002; Hidayati et
al. 2000; Chen et al. 2005). Results of our study showed that the
application of soaking seeds in 10-mg⋅L-1 GA3 solution for 24 h
followed by 2-week moist chilling at 3−5°C was very effective in
enhancing germination, however, soaking seeds in GA3 (10mg⋅L-1)
for 24 h produced average germination for all five seed sources
of both species studied. Gibberellic Acid-3 (GA3) is a naturally
occurring plant growth regulator which may cause a variety of
effects including the stimulation of seed germination in some
cases. GA3 occurs naturally in the seeds of many species. Studies
on woody plants indicated that the GA3 content of seeds in-
creases during cold stratification. For example, cold stratification
induced an increase in GA3 and in GA7 in peach (P. persica)
seeds (Mathur et al. 1971), in GA1 in Corylus avellana embryos
(Williams et al. 1974), in GA4b in apple seeds (Halin´ ska et al.
1987) and in GA1, GA3 and GA4 in P. buergeriana seeds (Chen
et al. 2005). Cold stratification induced an increase of GA3 levels
in embryos of European hazel (Corylus avellana), suggesting
that gibberellins synthesized during cold treatment were respon-
sible for dormancy break (Williams et al. 1974). Yamauchi et al.
(2004) demonstrated that a gene involved in GA3 biosynthesis in
seeds of Arabidopsis thaliana was activated by cold stratification
at 48°C. Increase in tissue sensitivity to gibberellins during cold
stratification is another factor that may be involved in controlling
seed germination (Derkx et al. 1993; Koornneef et al. 2002).

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Chinese Abstracts 2
现指数性增长。在一定温度范围内（1−20ºC），利用温度系数
函数（Q10）拟合通量率来描述森林地被物的温度敏感性。各
试验点的森林地被物的温度敏感性拟合曲线与净碳矿化和
N2O 氮释放率都显正相关。各试验林的全部数据表明，每个
单位的净碳矿化和 N2O氮释放的温度系数函数值（Q10）分别
为 1.73−2.10和 2.81−3.58，可用以描述试验地净碳矿化和N2O
氮释放率对温度的依赖性。在三毛举和云杉的单一树种和混
合种中，净碳矿化率 和 N2O 氮释放率的温度依赖性没有明
显的差异，表明净净碳矿化率和 N2O 氮释放量不受不同树种
凋落物质量的影响。图 4表 8参 31。
关键词：三毛举；挪威云杉；净碳矿化；氮氧化释放；温度；
温度敏感性指数（Q10）
CLC number: S153.61 Document code: A
Article ID: 1007−662X (2008)02-0107-06
DOI: 10.1007/s11676-008-0018-3

08−02−004
科尔沁沙地南缘樟子松人工林主要外生菌根真菌的最适培养
条件 /许美玲（中国科学院沈阳应用生态研究所，沈阳
110016；中华人民共和国临沂出入境检验检疫局，临沂
276000）, 朱教君（中国科学院沈阳应用生态研究所，沈阳
110016）, 康宏樟（中国科学院沈阳应用生态研究所，沈阳
110016）, 许爱华（山东省计量科学研究院，济南 250014）,
张金鑫（中国科学院沈阳应用生态研究所，沈阳 110016）
//Journal of Forestry Research.−2008, 19 (2): 113–118.
在实验室条件下模拟环境因子进行沙地樟子松林下常见
3种外生菌根真菌的纯培养试验，探讨培养基、pH值、水分
和温度对外生菌根真菌生长的影响，以期得到 3 种外生菌根
真菌的最适生长条件。结果表明：外生菌根真菌在含有松针
汁、马铃薯汁、酵母粉和维生素的培养基里生长较好；pH值
对外生菌根真菌的生长影响较小，L. deliciosus 和 B. sp. 的最
适生长 pH分别为 6.0和 5.0，L. sp. 在较广的 pH范围内均可
生长得良好，且在中性和弱碱性的培养基中长势均好于另外
两种；水分对实验菌株的影响较大，在较低的 PEG浓度(100 g
PEG·kg-1 H2O)下，三种菌株生长良好，最适得生长 PEG浓度
为 10%；高温容易导致外生菌根真菌死亡，3 种外生菌根真
菌的最适生长温度范围是 25−28℃。图 4表 3参 35。
关键词：樟子松（Pinus sylvestris var. mongolica）；外生菌根
真菌；培养基；pH值；水势；温度
CLC number: S154.38; S791.253 Document code: A
Article ID: 1007−662X (2008)02-0113-06
DOI: 10.1007/s11676-008-0019-2

08−02−005
利用核糖体小亚基 RNA 序列探讨四种腹毛目纤毛虫的系统进
化/李艺松，牛延宁，刘兰侠 (华东师范大学，生命科学学院，
上海 200062) // Journal of Forestry Research .−2008, 19 (2):
119−124.
以四个在环境监测中比较有代表性的纤毛虫属－游仆虫
属、尾柱虫属、棘尾虫属和伪角毛虫属纤毛虫（Euplotes,
Urostyla, Stylonychia, Pseudokeronopsis）为材料，对其 16s小
亚基核糖体 RNA（16s－like Small Subunit rRNA，SSrRNA）
基因进行序列测定，并与其他种腹毛目纤毛虫相比较，以美
国赭虫（Blepharisma americanum）为外群构建系统进化树。
结果显示：游仆虫属最先从腹毛目纤毛虫中分化出来，尾柱
虫属（Urostyla）与全列虫属（Holosticha）纤毛虫亲缘关系
较近。贻贝棘尾虫（Stylonychia mytilus）和浮萍棘尾虫
（Stylonychia lenmae）是姊妹种，在外观上及其相似，但可
以用构建进化树的方法从分子角度加以区分。从构建的进化
树拓扑结构上可知，红色伪角毛虫与全列虫科种类的亲缘关
系较近，与尾柱虫科种类的亲缘关系较远。图 2表 1参 50。
关键词：包囊游仆虫（Euplotes encysticus）；系统进化；红色
伪角毛虫（Pseudokeronopsis rubra）；SSrRNA基因；贻贝棘
尾虫（Stylonychia mytilus）；大尾柱虫（Urostyla grandis）
CLC number: Q344.11 Document code: A
Article ID: 1007−662X (2008)02-0119-06
DOI: 10.1007/s11676-008-0020-9

08−02−006
冷湿层积法对印度冷杉（Abies pindrow）和长叶云杉（Picea
smithiana）种子萌发的影响/Balbir Singh Rawat (Department of
Forestry, HNB Garhwal University, Post Box- 76, Srinagar
Garhwal), Vinod Prasad Khanduri (Department of Forestry,
Mizoram University, Aizawl 796001, Mizoram), Chandra Mohan
Sharma (Department of Forestry, HNB Garhwal University, Post
Box- 76, Srinagar Garhwal) // Journal of Forestry Research.
−2008, 19 (2): 125−130.
研究赤霉素（GA3），冷湿和温度 对五个种源的印度冷
杉（Abies pindrow）和长叶云杉（Picea smithiana）种子萌发
的影响。种子被浸泡在 GA3 (10 mg⋅L-1)中 24 小时，然后在
3−5°C 温度的条件下冷藏 15 天。设计 4 个温度(10°C, 15°C,
20°C 和 25°C)条件来促进种子的萌发。结果表明，浸泡和冷
湿处理明显增加种子的萌发率。在 10 °C 时种子的萌发率最
高。总体结果表明，种子被浸泡在 GA3 (10 mg⋅L-1)中 24小时，
冷湿藏 15天，可以有效地促进印度冷杉和长叶云杉的种子萌
发。统计数据表明，浸泡处理、温度和种源以及与温度的相
互作用都对种子的萌发有明显的影响。图 1表 4参 67。
关键词：层积；种源；种子萌发；赤霉素；冷杉；云杉
CLC number: Q945.34 Document code: A
Article ID: 1007−662X (2008)02-0125-06
DOI: 10.1007/s11676-008-0021-8

08−02−007
杉木纯林、混交林土壤微生物特性和土壤养分的比较研究/王
清奎（中国科学院沈阳应用生态研究所会同森林生态实验站，
沈阳 110016）， 汪思龙（中国科学院沈阳应用生态研究所会