International Journal of Agriculture: Research and Review. Vol.

, 3 (1), 172-175, 2013

Available online at http://www.ecisi.com
ISSN 2228-7973 ©2013 ECISI Journals

SEED GERMINATION AND DORMANCY BREAKING METHODS FOR

PHEASANT'S EYE (Adonis vernalis L.)

HOSSEIN REZA ROUHI1*, MOHAMMAD ALI ABOUTALEBIAN1, MARYAM SAMAN2, FATEMEH KARIMI1
ROYA MAHMOUDIEH CHAMPIRI3

1- Department of Agronomy and Plant Breeding, Faculty of Agriculture, Bu-Ali Sina University,
Hamedan, I. R. Iran
2- Department of Agricultural Sciences, Payame Noor University, Tehran, I. R. Iran
3- Agriculture Jahad Institute of Technical and Vocational higher Education, Isfahan Education Center, I.
R. Iran

*Corresponding Author Email: hosseinroohi@ut.ac.ir

ABSTRACT: Dormancy and germination requirements were investigated in seeds of Adonis

vernalis (Ranunculaceae). Seeds were subjected to different treatments (i) concentrated
sulphuric acid for 30, 60 and 90 seconds,(ii) hot water at 70 and 90°C for 5 and 10 minutes
(iii) three levels of KNO3 (0.1, 0.2 and 0.3% v/v) and (iv) 3 different stratification periods (1,
2 and 3 weeks). The highest percentage of germination was observed in the case of
stratification applied for 3 weeks (90%), but 2 and 3 weeks treatments resulted in positive
effect not stronger than 3 weeks. Same as stratification, KNO3 treatment had a positive effect
on seed germination in all levels especially at 0.3% v/v but this was lower than stratification
treatment. Germination percentage, day of first emergence and 50% emergence and mean
germination time were affected by both stratification and KNO3 treatments. In contrast to
stratification and KNO3 treatments, seeds showed weak response to periods of sulfuric acid
98% (v/v) and hot water treatments.

Key words: Dormancy; germination; seed; stratification.

INTRODUCTION
Pheasant's eye (Adonis vernalis L.) has a long
history of medicinal use and is still retained in
the Pharmacopoeias of several European
countries (Stuart, 1990). The plant contains
cardiac glycosides similar to those found in the
foxglove (Digitalis purpurea). These substances
improve the heart's efficiency, increasing its
output at the same time as slowing its rate
(Chevallier, 1996). It also has a sedative action
and so is generally prescribed for patients whose
hearts are beating too fast or irregularly
(Chevallier, 1996). Pheasant's eye has seed
dormancy. Seed dormancy is a temporary
suspension of visible growth of any plant
structure containing a meristem does not refer to
commencement of temporary suspensions of
growth (Lang et al., 1987). Seed dormancy has
been further negatively categorized in terms of
the requirements for release from this block,
such as disruption of the seed coat
(scarification), a period of dry storage (after-
ripening) or moist chilling (stratification), or
exposure to light. The situation is further
complicated by the fact that, although
germination is an all or nothing event for each
seed, populations display variable degrees of
dormancy that are reflected in the rate or
percentage of germination under specific
conditions. Presumably, each seed is in a state
somewhere along the continuum from deeply
dormant to nondormant, but it remains unclear
how the tipping point between nonpermissive
and permissive for germination is sensed for
each seed (Finkelstein et al., 2008).
Stratification plays an important role in
providing the stimulus required to overcome
dormancy. Stratification has been reported to
induce an increase in GA3 concentration
(Yamauchi et al., 2004). Potassium nitrate is
well documented as a compound, which
increases the germination of photo-dormant
seeds (Yamauchi et al., 2004). Many gardeners
choose potassium nitrate to break seed
Intl. J. Agric: Res & Rev. Vol., 3 (1), 172-175, 2013
dormancy and increase the health of plants.
According to Bewley and Black (1994), KNO3
raises the ambient oxygen levels by making less
oxygen available for citric acid cycle. Aliero
(2004) reported that use of hot water, sulfuric
acid and sand paper scarification affected
Parkia biglobosa seed dormancy-breaking. The
objective of this research was to determine the
effects of different seed dormancy breaking
treatments which are able to stimulate and
enhance germination of this medicinal plant.
MATERIALS AND METHODS
This research was done at the Department of
Agronomy and Plant Breeding, Faculty of
Agriculture, Bu-Ali Sina University, Islamic
Republic of Iran. Pheasant's eye seeds were
received from Hamedan. Measured traits were:
final germination percentage, mean germination
time, E1st, E50 and abnormal seedling.
Seed treatments
For stratification treatment, Pheasant's eye seeds
were mixed in perlite medium and distilled
water in vessels, then transferred to a
refrigerator for 1, 2 and 3 weeks at 5±1◦C.
These vessels put into sealed plastic bags to
avoid moisture loss. After this time the seeds
were rinsed with tap water completely. For
KNO3 treatment, seeds were treated by 0.1, 0.2
and 0.3% (v/v) KNO3 for 24 hours. In this case,
flasks were shaken on orbital shaker in light
condition. Scarification with sulfuric acid 98%
(v/v) was done in 3 periods (30, 60 and 90
seconds). For hot water treatment, seeds were
treated at 70°C and 90°C in a hot water bath, for
5 and 10 minutes, and then taken away from the
heat source. The seeds were left in the water
overnight (ca.12 hr) while it gradually cooled
down to room temperature.
Germination tests
Four replicates of 100 seeds were germinated
between double layered papers [7] with 5 ml of
water in 8cm Petri dishes. These Petri dishes
were put into sealed plastic bags to avoid
moisture loss. Seeds were allowed to germinate
at 15±1◦C in the dark condition for 14 days.
Germination was considered to have occurred
when the radicles were 2mm long. Germination
percentage was recorded every 24h for 14 days.
Mean germination time (MGT) was calculated
by following equation (Schelin et al., 2003).
Mean Germination Time (MGT) :
 fini
N expression of the GA catabolic gene GA2ox2
173
fi: Day during germination period (between 0
and 14 day).
ni: Number of germinated seeds per day
N: Sum of germinated seeds
Statistical analysis
The statistical design was a completely
randomized design. Four replications and 100
seeds per replicate were used. Data for
germination and abnormal germination
percentage were subjected to arcsine
transformation before analysis of variance.
Statistical analysis was carried out using SAS
program. Mean comparison was performed with
Duncan’s test at the 5% level of significance.
RESULTS AND DISCUSSION
In our experiment, neither sulfuric acid (98%)
nor treating with hot water had any strong effect
on germination indexes compared to control
group (Table 1). El Siddig et al. (2001) found
that scarification with 90% sulfuric acid
decreased fresh weight, MGT and E1st of
Tamaridus indica seedling. Levels of KNO3 did
stimulate the germination of Pheasant's eye
(Table 1). Shanmugavalli et al. (2007) showed
that seeds of sorghum soaked in 0.5% and 1%
potassium nitrate (KNO3) improved germination
up to 44%. Potassium nitrate has been used for
many years, with positive studies beginning in
the 1980's and it often increased the germination
of photo-dormant. According to Bewley and
Black (1994), KNO3 raises the ambient oxygen
levels by making less oxygen available for citric
acid cycle. In addition to KNO3 treatment, seed
treatment with stratification for 1, 2 and 3 weeks
were significant and dormancy of Pheasant's eye
seeds broken by these treatments. Stratification
for 3 weeks was more effective than 1 and 2
weeks (Table 1). Stratification plays an
important role in improving sensitivity to other
treatment to overcome dormancy (Bretzloff and
Pellett,1979; Schelin et al., 2003; Yamauchi et
al., 2004). Stratification has been reported to
induce an increase in sensitivity to GA3
concentration [9]. In this study stratification
treatments and KNO3 levels were the most
effective treatment respectively. Similar to our
results, Oh et al. (2006) suggest that
stratification promotes germination by
increasing the potential for bioactive GA
accumulation. Stratification led to increased
expression of the GA biosynthesis genes
GA20ox1 (GIBBERELLIN 20 OXIDASE ),
GA20ox2, and GA3ox1 and decreased
(Yamauchi et al., 2004). In some reports
 Intl. J. Agric: Res & Rev. Vol., 3 (1), 172-175, 2013

stratification was introduced as the best method
for dormancy breaking. Beginning of the
embryo dormancy is associated with
accumulation of growth inhibitors such as ABA
and breaking of dormancy with a shift in the
balance of growth regulators towards growth
promoters such as GA3, that overcome the effect
of growth inhibitors (Rouhi et al., 2012).
Conclusions
The dormancy of Pheasant's eye seed was
broken by stratification and KNO3 treatments,
but scarification with hot water and sulfuric acid
had no strong effect. Our results showed that
applying stratification for 3 weeks could be the
best treatment for Pheasant's eye dormant seeds
(Table 1). Finally, we suggest that Pheasant's
eye has morphophysiological dormancy because
it has underdeveloped embryo, but in addition it
has a physiological component to their
dormancy. Therefore seeds require a dormancy-
breaking treatment, for example a defined cold
stratification which in some cases can be
replaced by GA application.

Table 1: influence of seed dormancy breaking treatments on germination traits of Pheasant's eye, In each column
means followed by the same letter are not significantly different at the P < 0.05 level
FGP MGT E1ts E50 Abnormal
Dormancy breaking treatments
(%) (seed/day) (days) (days) seedling (%)
Control 5d 23.38 d 26.6 d 0d 0a
Acid scarification 98% (v/v)
Sulfuric acid (30 sec) 3d 28.1 e 26.5 d 0d 0a
Sulfuric acid (60 sec) 3d 27.7 e 27.1 d 0d 0a
Sulfuric acid (90 sec) 4d 28.12 e 29.1 e 0d 5b
Scarification by hot water
70°C + 5 min 8d 23.4 d 25.5 d 0d 7b
70°C + 10 min 8d 23.22 d 25.2 d 0d 7b
90°C + 5 min 5d 24.1 d 26 d 0d 5b
90°C + 10 min 7d 24.5 d 26 d 0d 7b
Potassium nitrate (KNO3 v/v)
0.1 23 c 18 c 20.2 c 18.3 b 20 c
0.2 78 b 18.2 c 14.1 b 17.9 b 18 c
0.3 75 b 8.85b 16.2 b 19.7 b 7b
Stratification
1 week 81 a 9.11 b 19.4 c 18.2 b 5b
2 weeks 82 a 9.7 a 14.2 b 19.2 b 6b
3 weeks 90 a 9.9 a 13.4 a 15.1 a 3a
FGP: final germination percentage, MGT: mean germination time, E1st: day of first observation of seedling
emergence, E50: days to 50% emergence

REFERENCES El siddig K, Ebert, G and Ludders P (2001) A

comparison of pretreatment methods for
Aliero, BL (2004) Effects of sulfuric acid, scarification and germination of
mechanical scarification and wet heat Tamarindus indica seeds. Seed Sci
treatments on germination of seeds of Technol. 29: 271–274.
Parkia biglobosa. African journal of Finkelstein R, Reeves W, Ariizumi T, and
biotech. 3: 179–181. Steber C (2008) Molecular Aspects of
Bewley JD, and Black M (1994) Seeds: Seed Dormancy. Annu Rev Plant Biol.
Physiology of Development and 59:387-415.
Germination. Plenum, New York. ISTA (1996) Rules for Seed Testing.
Bretzloff LV, Pellett NW (1979) Effect of International Seed Testing Association.
stratification and gibberllic acid on the Seed Sci Technol.
germination of Carpinus caroliniana Lang GA, Early JD, Martin GC, Darnell RL
Walt. Hort Sci. 14: 621–622. (1987) Endo-, para-, and eco-dormancy:
Chevallier, A (1996) The Encyclopedia of physiological terminology and
Medicinal Plants. DK ADULT.

174
Intl. J. Agric: Res & Rev. Vol., 3 (1), 172-175, 2013
classification for dormancy research.
Horticulture Sci.22: 371–377.
Oh E, Yamaguchi S, Kamiya Y, Bae G, Chung
WI, Choi G (2006) Light activates the
degradation of PIL5 protein to promote
seed germination through gibberellin in
Arabidopsis. Plant J. 47:124–39.
Rehman S, Loescher RN, and Harris PJC
(1999). Dormancy breaking and
germination of Acacia saliciina seeds.
Seed Sci Technol. 27: 553–557.
Rouhi HR, Sepehri A, Karimi F (2012) Study
of dormancy-breaking of Black cumin
seeds (Nigella sativa L.) Annals of
Biological Res.3 (6):2651-2655.
Schelin M, Tigabu M, Eriksson I, Swadago L,
and Oden PC (2003) Effect of
scarification, gibberllic acid and dry heat
175
treatments on the germination of
Balanties Egyptian seed from the
Sudanian savanna in Burkina Faso. Seed
Sci Technol. 31: 605–617.
Shanmugavalli M, Renganayaki PR, and
Menaka C (2007) Seed dormancy and
germination improvement treatments in
fodder sorghum. Int Crops Res Inst Semi-
Arid Tropics. 3:1-3.
Stuart, M (1990) (Editor) The Encyclopedia of
Herbs and Herbalism. Black Cat, United
Kingdom.
Yamauchi Y, Ogawa M, Kuwahara A, Hanada
A, Kamiya Y, Yamaguchi S (2004)
Activation of gibberellin biosynthesis and
response pathways by low temperature
during imbibitions of Arabidopsis
thaliana seeds. Plant Cell. 16:367–78.