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不同处理方法对顶冰花种子萌发的影响研究(英文)



全 文 :Effects of Different Treatments on Seed
Germination of Gagea nigra
Kun YANG, Weijun ZENG*, Changqing ZHU*, Qing XIE
School of Life Sciences, Xinjiang Normal University, Urumqi 830054, China
*Corresponding author. E-mail: 1456559627@qq.com; 1559177022@qq.com
Received: March 11, 2016 Accepted: April 22, 2016A
Agricultural Science & Technology, 2016, 17(5): 1129-1133
Copyright訫 2016, Information Institute of HAAS. All rights reserved Agronomy and Horticultrue
G agea nigra L.Z. Shue (Lili-aceae: Gagea Salisb.) is atypical ephemeral perennial
bulb monocot.Bulbs provide most of
the nutrients demanded by G. nigra.
While summer is coming on and snow
has not melted completely, G. nigra
begins to grow, bloom and fruit, and its
entire life cycle lasts for amount 2
months [1]. In order to adapt to harsh
drought and other adverse physiologi-
cal stresses, ephemeral plants shorten
their growth cycles by utilizing melted
snow water and other favorable condi-
tions [2]. They have attracted much at-
tention for their short growth cycle, fast
development and unique drought toler-
ance [3]. The physiological ecology and
cloning of G. nigra are all the focuses
of concern[4-6].
As one typical ephemeral plant,
G. nigra has received much attention
from researchers. However, most do-
mestic and foreign researches focus
on bulbs and taxonomy of G. nigra.
The experiments about G. nigra are
mostly conducted by field observation
or with collected bulbs [1, 7-11]. Although
seed is one of important ways for bio-
logical breeding, there are rare reports
on seed germination of G. nigra. To
this end, the optimum conditions for
seed germination of G. nigra were ex-
plored primarily in this study, thereby
providing certain theoretical basis for
future studies on G. nigra and practical
significance for obtaining experimental
materials from labs and maintaining
biodiversity.
Materials and Methods
Materials
The experimental seeds were col-
lected from the Carp Mountain in Ur-
umqi, Xinjiang in 2013. They were
dried at room temperature and stored
for use.
Methods
Measurement of 1 000-grain weight
Total 3 000 G. nigra seeds were
selected randomly. They were evenly
divided into three groups (three repli-
cates). The seeds in each group were
weighed, thus the 1 000-grain weight
Abstract According to different mechanisms of dormancy, seeds of Gagea nigra L.
Z. Shue were treated with different methods so as to investigate the effects of dif-
ferent treatments on seed germination of G. nigra. The results showed that at 4 ℃,
the seeds of G. nigra began to germinate on day 32 with germination period of 80
d and germination rate of 72%; room temperature (25 ℃) and acid and alkali treat-
ments were all unsuitable for the germination of G. nigra seeds; no significant dif-
ferences were found in germination rate of G. nigra seeds among warm water (50
℃), ultrasound and 200 mg/L GA3 treatments (P>0.05); seed coat-removing treat-
ment effectively promoted the germination of G. nigra seeds (P=0.002<0.01) with
germination rate of 88%; seed coat piercing + 200 mg/L GA3 and 500 mg/L GA3 +
15% H2O2treatments effectively promoted the germination of G. nigra seeds with
germination rates of 84% and 82%, respectively (P<0.05); the germination vigor of
G. nigra seeds in the seed coat piercing + 200 mg/L GA3 treatment was highest,
and it was significantly higher than those in the other treatments.
Key words Gagea nigra; Ephemeral plants; Different treatments; Seed germination
不同处理方法对顶冰花种子萌
发的影响研究
杨坤,曾卫军*,祝长青*,谢青 (新疆师范大
学生命科学学院,新疆乌鲁木齐 830054)
摘 要 以新疆黑鳞顶冰花种子为实验对象,
根据不同休眠机制选取 10 余种处理方法进行
处理, 以此比较其对顶冰花种子萌发的影响。
研究结果显示: 黑鳞顶冰花种子 4 ℃时 32 d
左右开始萌发,萌发周期 80 d 左右,萌发率为
72%;室温 25 ℃和酸碱处理不适宜萌发;50 ℃
温水、超声波和 200 mg/LGA3处理均无显著性
差异(P > 0.05);去除种皮极有效促进其萌发
(P = 0.002<0.01), 萌发率达到 88%; 刺破种
皮+200 mg/L GA3 和 500 mg/L GA3+15%H2O2
处理也能有效促进萌发(P<0.05),萌发率分别
为 84%、82%,前者发芽势最好,明显高于其他
处理。
关键词 顶冰花;早春短命植物;不同处理;种
子萌发
作者简介 杨坤 (1990-),男 ,新疆乌鲁木齐
人,在读硕士研究生,研究方向:分子生物学。
*通讯作者。 曾卫军,讲师,博士,主要从事分
子生物学研究,E-mail: 1456559627@qq.com;祝
长青,副教授,主要从事新疆特色植物分子资
源开发与利用研究 ,E-mail: 1559177022@qq.
com。
收稿日期 2016-03-11
修回日期 2016-04-22
DOI:10.16175/j.cnki.1009-4229.2016.05.022
Agricultural Science & Technology 2016
Fig.1 Determination of water absorption capacity of G. nigra seeds
Table 1 Description of different treatments
No. Treatment method Reference species
Control Moistening in distilled water, inoculating in petri dish, and culturing at 4℃ Ixiolirion tataricum[12]
T1 Removing seed coat Saponaria officinalis[13]
T2 Soaking in warm water (50℃) for 5 min Spinacia oleracea[13-14]
T3 Soaking in 98% concentrated H2SO4 for 3 min Kobresia capillfolia, Vicia unijuga[11]
T4 Soaking in 10% NaOH for 2 h Carex heterostachy[14]
T5 Soaking in 30% acetone for 20 min Zoysia japonica[14]
T6 Soaking in 14% KNO3 for 24 h Chinese wild rye[15]
T7 Ultrasound 20 min Cycas revoluta[14], Polygonatum sibiricum[16]
T8 Soaking in 200 mg/L GA3 for 48 h
Arabidopsis pumila [17], Carpinus cordata [18], Syringa
reticulata[19]
T9 Piercing seed coat+soaking in 200 mg/L GA3 for 48 h
Malcolmia africana (Linn.) R. Br, Tetracme quadri-
cornis[20]
T10 Soaking in 500 mg/L GA3 for 3 d + soaking in 15% H2O2 for 20 min Lepidium perfoliatum[21]
T11
Soaking in 70% ethanol for 3 min + soaking in 30% NaOH for 20 min +
soaking in 200 mg/L GA3 for 10 min
Zoysia sinica[14]
T12 Soaking in 50 g/L KMnO4 for 2 h + soaking in 0.2% SA for 20 h Larix principis-rupprechtii
of G. nigra was obtained. The final
data was expressed as the mean of
the three replicates.
Determination of water absorption
capacity
Total 300 G. nigra seeds were se-
lected randomly, and they were evenly
divided into three groups. The weight
of the seeds in each group was
weighed and recorded. The seeds in
each group were placed in a 500-μl EP
tube and soaked in distilled water.
They were weighed after removing the
extra water once an hour until their
weight remained unchanged for three
consecutive times.The water absorp-
tion curve was drawn with the means
of the three replicates.
Germination condition treatments
According to the germination
conditions of other species, several
germination-promoting treatments
were designed for G. nigra seeds
(Table 1). In addition, one control
group was arranged. The treatments
with pretreatment time exceeding 1 h
were all conducted at 4 ℃ . The pre-
treated seeds were then transferred
into petri dishes (6 cm) and cultured at
4 ℃ . There were three replicates for
each group. The seeds with 1-2 mm
buds were considered as germinated,
and the germinated seeds in each
group were counted. The germination
vigor was determined on day 20 since
the seeds began to germinate. When
the number of germinated seeds in the
control group remained unchanged for
7 consecutive days, the statistics of
total germination rate ended.
Data processing and analysis
The processing of data and sig-
nificance analyses of differences
(LSD, Duncana) were conducted using
Excel and SPSS 19.0.
Morphological observation during
germination process
The changes of seed embryo ofG.
nigra were observed under a micro-
scope during the germination process.
Results and Analysis
1 000-grain weight
The seed shape of G. nigra was
similar to that of garlic bulbs. However,
G. nigra seeds were extremely small
and light with 1 000-grain weight of
only 0.640 g.
Water absorption capacity
G. nigra seeds absorbed water
quickly. The coat of dried G. nigra
seeds turned wet and swelling quickly
after they were soaked in water. Within
the first one hour, the water absorption
of the G. nigra seeds increased con-
tinuously; after 2 hours, the water ab-
sorption capacity of the G. nigra seeds
reduced, and their weight became sta-
ble; the water absorption of the G. ni-
gra seeds reached the saturation state
on 6th hour (Table 2, Fig.1). Different
from other seeds with thicker coat, the
coat of G. nigra seeds did not inhibit
their water absorption capacity.
Effects of different treatments on
seed germination of G. nigra
At temperature of 4 ℃ , the G. ni-
gra seeds in the control and treatment
groups all began to germinatearound
day 32 with germination cycle of 45 d.
The germination rate of untreated
seeds reached 72%. At temperature of
25℃, the G. nigra seeds turned brown
before their germination.
Effects of physical treatments on
seed germination of G. nigra
The three different physical treat-
ments all showed certain effects on
the germination rate and germination
vigor of G. nigra seeds. As shown in
Table 3, the germination rate of G. ni-
gra seeds in the T1 treatment group
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Agricultural Science & Technology2016
Table 2 Determination of water absorption capacity of G. nigra seeds
Time//h 0 1 2 3 4 5 6 7 8
Mean 6.45 E-02 9.67 E-02 1.10 E-01 1.15 E-01 1.19 E-01 1.20 E-01 1.26 E-01 1.27 E-01 1.28 E-01
Standard
deviation 2.12 E-03 2.08 E-03 3.21 E-03 3.61 E-03 1.53 E-03 2.31 E-03 4.58 E-03 3.79 E-03 3.00 E-03
Table 3 Effects of different treatments on germination rate of G. nigra seeds
N Mean Standard deviation Standard error
95% confidence interval
Lower limit Upper limit
Control 3 0.720 0 0.040 00 0.023 09 0.620 6 0.819 4
T1 3 0.880 0 0.060 00 0.034 64 0.731 0 1.029 0
T2 3 0.700 0 0.080 00 0.046 19 0.501 3 0.898 7
T3 3 0 0 0 0 0
T4 3 0.040 0 0.040 00 0.023 09 -0.059 4 0.139 4
T5 3 0.560 0 0.060 00 0.034 64 0.411 0 0.709 0
T6 3 0.520 0 0.080 00 0.046 19 0.321 3 0.718 7
T7 3 0.700 0 0.020 00 0.011 55 0.650 3 0.749 7
T8 3 0.660 0 0.020 00 0.011 55 0.610 3 0.709 7
T9 3 0.840 0 0.080 00 0.046 19 0.641 3 1.038 7
T10 3 0.820 0 0.060 00 0.034 64 0.671 0 0.969 0
T11 3 0.120 0 0.040 00 0.023 09 0.020 6 0.219 4
T12 3 0.520 0 0.080 00 0.046 19 0.321 3 0.718 7
Sum 39 0.544 6 0.298 54 0.047 80 0.447 8 0.641 4
N represents Number of replicates.
Fig.2 Morphological observation of seed germination of G. nigra
reached 88%, and those in the T2 and
T7 treatment groups were all 70% . In
the T1, T2 and T7 treatment groups, the
germination vigors of G. nigra seeds
were 0.48, 0.22 and 0.44, respectively
(Table 4).
Table 5 and Table 6showed that
seed coat-removing treatment had
significant effect on germination rate of
G. nigra seeds (P<0.01), while warm
water soaking (50 ℃, 5 min) and ultra-
sound (20 min) treatments showed no
significant effects on germination rate
of G. nigra seeds (P>0.05).
Effects of chemical treatments on
seed germination of G. nigra
Among the five chemical treat-
ments, T3 (98% concentrated H2SO4)
showed the greatest effect on the ger-
mination of G. nigra seeds with germi-
nation rate of 0; T8 (soaking in 200 mg/
L GA3 for 48 h) showed no significant
effect on the germination of G. nigra
seeds; T4 (soaking in 10% NaOH for 2
h), T5 (soaking in 30% acetone for 90
min) and T6 (14% KNO3) all inhibited
the germination of G. nigra seeds (P<
0.01) with germination rates of 0.04,
0.56 and 0.52, respectively.
Effects of comprehensive treat-
ments on seed germination of G.
nigra
As shown in Table 3, the germina-
tion rates of G. nigra seeds in the T9
(seed coat -piercing + soaking in 200
mg/L GA3 for 48 h) and T10 (soaking in
500 mg/L GA3 for 3 d + soaking in 15%
H2O2 for 20 min) treatments were high-
est (0.84, 0.82), while that in the T11
(soaking in 70% ethanol for 3 min +
soaking in 30% NaOH for 20 min +
soaking in 200 mg/L GA3 for 10 min)
treatment was lowest (0.12). Although
the germination rate in the T9 treat-
ment group was lower than that in the
T1 treatment group, the germination u-
niformity in the T9 treatment group was
highest among all the treatment
groups with germination vigor of 0.56
(Table 4).
Changes of embryo during germi-
nation process
As shown in Fig.2 (in onesame
petri dish), the germination process of
G. nigra seed was a process of em-
bryo’s breaking out from the en-
dosperm. Under the same conditions,
the changes of the embryos of simulta-
neously-treated same-batch G. nigra
seeds were even different.
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Agricultural Science & Technology 2016
Table 4 Effects of different treatments on germination vigor of G. nigra seeds
N Mean Standard deviation Standard error
95% confidence interval
Lower limit Upper limit
Control 3 0.300 0 0.040 00 0.023 09 0.200 6 0.399 4
T1 3 0.480 0 0.020 00 0.011 55 0.430 3 0.529 7
T2 3 0.220 0 0.040 00 0.023 09 0.120 6 0.319 4
T3 3 0.000 0 0.000 00 0.000 00 0.000 0 0.000 0
T4 3 0.020 0 0.020 00 0.011 55 -0.029 7 0.069 7
T5 3 0.180 0 0.040 00 0.023 09 0.080 6 0.279 4
T6 3 0.140 0 0.020 00 0.011 55 0.090 3 0.189 7
T7 3 0.440 0 0.040 00 0.023 09 0.340 6 0.539 4
T8 3 0.300 0 0.020 00 0.011 55 0.250 3 0.349 7
T9 3 0.560 0 0.020 00 0.011 55 0.510 3 0.609 7
T10 3 0.340 0 0.020 00 0.011 55 0.290 3 0.389 7
T11 3 0.040 0 0.020 00 0.011 55 -0.009 7 0.089 7
T12 3 0.200 0 0.020 00 0.011 55 0.150 3 0.249 7
Sum 39 0.247 7 0.174 21 0.027 90 0.191 2 0.304 2
N represents Number of replicates.
Table 5 Significance analysis of germination rate differences among different treatments (LSD)
Treatment Mean difference (I-J) Standard error Significance
95% confidence interval
Lower limit Upper limit
Control
-0.160 00 0.046 41 0.020 0 -0.255 -0.064 6
0.020 00 0.046 41 0.670 0 -0.075 4 0.115 4
0.720 00 0.046 41 0.000 0 0.624 6 0.815 4
0.680 00 0.046 41 0.000 0 0.584 6 0.775 4
0.160 00 0.046 41 0.002 0 0.064 6 0.255 4
0.200 00 0.046 41 0.000 0 0.104 6 0.295 4
0.020 00 0.046 41 0.670 0 -0.075 4 0.115 4
0.060 00 0.046 41 0.207 0 -0.035 4 0.155 4
-0.120 00 0.046 41 0.016 0 -0.215 4 -0.024 6
-0.100 00 0.046 41 0.041 0 -0.195 4 -0.004 6
0.600 00 0.046 41 0.000 0 0.504 6 0.695 4
0.200 00 0.046 41 0.000 0 0.104 6 0.295 4
T1
T2
T3
T4
T5
T6
T7
T8
T9
T10
T11
T12
Discussion
Seeds of many species have
dormancy phenomena. Dormancy pat-
terns include mainly physiological dor-
mancy (PD), morphological dormancy
(MD), morphological physiological
dormancy (MPD), physical dormancy
(PY) and complex dormancy (PY +
PD)[23].For different dormancy patterns,
different treatments can be designed
to break seed dormancy. For example,
removing seed coat and other physical
treatments can break the physical
dormancy; GA treatment can break
the physiological dormancy.
Although researches on seed
dormancy mechanisms and relevant
research methods have made great
progresses, and researches on seed
germination of many species have
obtained great gains [24], the seed ger-
mination of Gagea salisb. is rarely re-
ported, and its germination mecha-
nism is still unclear.
Based on the obtained experi-
mental data, it could be presumed that
the dormancy pattern of G. nigra
seeds is complex dormancy. The de-
termination results of water absorption
capacity showed that the permeability
of seed coat of G. nigra seeds was not
poor, which could not induce seed dor-
mancy. In addition, single GA treat-
ment showed no significant effect on
the germination rate of G. nigra seeds.
However, removing seed coat in-
creased the germination rate of G. ni-
gra seeds. The seed coat piercing +
GA complex treatment improved both
germination rate and germination uni-
formity of G. nigra seeds. G. nigra
seeds did not germinate at the room
temperature, but germinated normally
at low temperature (4 ℃).There were
no significant differences in germina-
tion beginning time (around day 32) or
germination cycle (about 50 d) among
all the treatment groups. How to re-
duce the germination cycle of G. nigra
seeds and nursing healthy and robust
G. nigra seedlings still need further
study. In this study, corresponding
treatments were designed for different
dormancy mechanisms, thereby pro-
viding certain theoretical basis for re-
searches on dormancy mechanisms of
G. nigra seeds.
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Table 6 Significance analysis of germination rate differences among different treatments
(Duncana)
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Subset for α=0.05
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T11 3 0.120 0
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