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DOI 10.1007/s10725-016-0165-z
ORIGINAL PAPER
significant difference between the four combination treatments and the control with regard to the mean germination
time and germination index. These results suggest that the
selected combinations strongly inhibited the seed germination speed and PHS of rice in the field. However, these
combinations had no inhibitory effects on the final germination percentage and showed no negative effect on seed
quality after short-term storage. The combinations of
inhibitors used in this study can be applied to decrease the
PHS damage caused by high temperatures and moist
weather during the production of hybrid rice.
Keywords Pre-harvest sprouting Inhibitor Hybrid
rice Seed Germination
Introduction
Rice is an important staple food that is planted worldwide
(Maclean et al. 2002; Valipour et al. 2014; Valipour 2014,
2015). Pre-harvest sprouting (PHS) occurs when grains
(e.g., in rice, wheat, barley and maize) germinate in ears
before field harvesting (Groos et al. 2002). Prior to harvest,
physiologically mature grains sprout easily within ears
under conditions of high humidity and temperature (Gubler
et al. 2005; Yang et al. 2007). PHS generally reduces the
cereal yield and degrades seed quality for planting,
resulting in severe economic losses of cultivars with high
rates of PHS (Chono et al. 2006; Depauw et al. 2009;
Ullrich et al. 2009). PHS also complicates seed separation
and processing because the sprouting seeds must be sorted
and discarded. Moreover, the lixivium of sprouting seeds
may affect other, normal seeds during seed soaking before
sowing (Wang et al. 2008). Rice hybrids showing a yield
advantage of 20 % over conventional varieties have been
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Uniconazole
Maleic hydrazide
1000a
1000
100
10
1200
750
75
12
750
1200
75
14
750
750
120
123
Statistical analysis
Statistical analyses of all data were performed using analysis of variance (ANOVA) with SAS software, version 8.0
(SAS Institute, Cary, NC). The percentage data were
transformed according to y = arcsin [sqr (x/100)] before
analysis. When a significant difference occurred in the
treatments, the least significant difference was calculated
(a = 0.05, LSD).
Results
Effects of combinations of inhibitors on expression
of a-amylase biosynthetic genes
MGT (day)
GE (%)
GP (%)
GI
Control
2.93a
94.3a
96.0ab
16.96a
3.59b
88.0bc
94.3ab
13.62b
10
12
3.55b
3.54b
85.3c
90.3b
92.7b
96.7a
13.65b
14.23b
14
3.52b
91.3ab
94.7ab
13.97b
Control: seeds soaked in water; 8, 10, 12 and 14: seeds soaked in the
combinations shown in Table 1. Each value represents the mean
(n = 3); within each column, different letters following the values
indicate a significant difference (a = 0.05, LSD) between the control
and the combined treatments
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Treatment
RL (cm)
SH (cm)
RL/SH
RDW/SDW
Control
6.20a
6.96a
0.89b
35.0a
53.5a
0.65b
3.56bc
3.34b
1.07a
26.0b
38.5b
0.67b
10
3.45bc
3.25b
1.06a
25.0b
39.0b
0.64b
12
3.07c
3.02b
1.02a
25.5b
41.0b
0.62b
14
3.85b
3.43b
1.12a
28.0b
38.5b
0.73a
Control: seeds soaked in water; 8, 10, 12 and 14: seeds soaked in the combinations shown in Table 1. Each
value represents the mean (n = 3); within each column, different letters following the values indicate a
significant difference (a = 0.05, LSD) between the control and the combined treatments
80
B
a
60
b
c
40
20
0
control
10
12
14
Treatment
Sprouting Index
0.8
0.6
0.4
0.2
0
control
10
12
14
Treatment
compared with the control (Table 5). There were no significant differences in the GP, shoot height or root/shoot
length ratio between seeds that were freshly harvested from
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0.6
OsAMY3A
OsAMY3B
OsAMY3E
OsAMY1A
0.2
0.2
-0.2
0
24
48
72
-0.2
-0.4
-0.4
-0.6
-0.6
-0.8
-0.8
Log
12
24
48
72
-1
-1
12
0.8
0.8
0.6
0.6
0.4
0.4
0.2
0.2
-0.2
12
24
48
72
0
-0.2
-0.4
-0.4
-0.6
-0.6
-0.8
-0.8
-1
Control
27.9a
1.34a
16.5b
0.76b
10
14.6b
0.72b
12
14
17.1b
17.2b
0.81b
0.83b
Control: sprayed with water; 8, 10, 12 and 14: sprayed with the
combinations shown in Table 1. Each value represents the mean
(n = 3); within each column, different letters following the values
indicate a significant difference (a = 0.05, LSD) between the control
and the combined treatments
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0.6
0.4
0.8
Control
12
10
0.4
Treatment
1
0.8
Log
-1
14
12
24
48
72
Discussion
In addition to genetic improvement, the application of
exogenous chemicals is an effective way to reduce PHS
damage to seed production (Wang et al. 2008). Maleic
hydrazide and uniconazole are well-known plant growth
regulators (Paterson et al. R1952; Tso et al. 1965; Izumi
et al. 1988). In our preliminary experiment, maleic
hydrazide, uniconazole and eugenol postponed or inhibited seed germination. The combinations of the three
chemicals used in the present study were designed based
on previous results. Because of its possible toxicity, MH
was limited to a lower dosage of 75120 mg/L, which was
approximately one-tenth the dosage of each of the other
two chemicals.
Treatment
MGT (day)
GE (%)
Control
3.02b
3.20ab
99.3ab
10
3.32a
95.3c
12
3.22a
96.0c
14
3.29a
96.7bc
100a
GP (%)
99.3a
GI
RL (cm)
SH (cm)
R/S (length)
0.75a
17.0a
5.36a
7.27a
100a
15.9b
4.61ab
6.98a
0.65a
100a
15.5b
3.73b
6.12a
0.61a
98.7a
15.6b
4.61ab
6.55a
0.70a
99.3a
15.6b
4.36ab
6.92a
0.63a
Control: seeds sprayed with water; 8, 10, 12 and 14: seeds sprayed with the combinations shown in Table 1.
Each value represents the mean (n = 3); within each column, different letters following the values indicate
a significant difference (a = 0.05, LSD) between the control and the combined treatments
Treatment
MGT (day)
GE (%)
Control
3.05b
98.0a
3.12ab
99.3a
10
3.26ab
96.0a
12
3.16ab
99.0a
14
3.33a
97.3a
GP (%)
GI
RL (cm)
SH (cm)
R/S (length)
16.50a
4.03a
6.79a
0.60a
100a
16.43ab
2.79a
5.25b
0.53a
100a
15.79ab
3.28a
5.08b
0.65a
100a
16.18ab
2.91a
4.78b
0.60a
15.19b
3.04a
4.80b
0.63a
98.0a
98.7a
Control: seeds sprayed with water; 8, 10, 12 and 14: seeds sprayed with the combinations shown in Table 1.
Each value represents the mean (n = 3); within each column, different letters following the values indicate
a significant difference (a = 0.05, LSD) between the control and the combined treatments
Treatments with the combinations of chemicals significantly slowed seed germination speed compared with the
control but resulted in no significant difference in the final
germination percentage, indicating that these combination
treatments significantly delayed, but did not completely
inhibit, the germination of rice seeds. After the seed germination test, shorter root length and shoot height were
observed in the four combination treatments compared
with the control. This phenomenon might be related to the
inhibitory effect of uniconazole on GA biosynthesis, which
leads to slower growth and stocky seedlings (Saito et al.
2006). In this study, no abnormal seedlings were observed,
suggesting that the selected range of MH concentrations
might be acceptable for safe application. In the PHS laboratory simulation, the four combination treatments produced a lower sprouting rate and sprouting index than the
control.
During grain seed germination, a-amylase plays a
prominent role in hydrolyzing endosperm starch into
metabolizable sugars and providing energy for radicle
protrusion (Sithichoke et al. 2005; Akazawa and HaraNishimura 1985; Beck and Ziegler 1989). Unlike other
amylolytic enzymes, it is responsible for activating carbohydrate reserves by breaking a-1,4-linked polyglucans
and initiating starch breakdown (Janecek et al. 2014). In
the present study, a-amylase activity was almost undetectable when seeds were imbibed in water for 12 h;
however, it increased markedly with increasing imbibition
times. The application of exogenous inhibitors significantly
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References
Akazawa T, Hara-Nishimura I (1985) Topographic aspects of
biosynthesis, extracellular secretion, and intracellular storage
of proteins in plant cells. Annu Rev Plant Physiol Plant Mol Biol
36(1):441472
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