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Cool germination test can be an alternative vigour test for maize

Article  in  Seed Science and Technology · July 2009

DOI: 10.15258/sst.2009.37.2.29

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 H. ILBI, S. KAVAK AND B. ESER

Ilbi, H., Kavak, S. and Eser, B. (2009), Seed Sci. & Technol., 37, 516-519

Research Note

Cool germination test can be an alternative vigour test for maize

H. ILBI1, S. KAVAK2 AND B. ESER1

1
Ege University, Faculty of Agriculture, Department of Horticulture, 35100, Bornova, Izmir, Turkey
(E-mail: hulya.ilbi@ege.edu.tr)
2
Süleyman Demirel University, Faculty of Agriculture, Department of Horticulture, 32260, Isparta, Turkey

(Accepted December 2008)

Summary
An alternative to the cold test to evaluate maize seed vigor was investigated. The standard germination test,
cool germination tests at 12, 15 and 18°C, the cold test were performed for seven maize seed lots and field
emergence at one site determined. The standard germination of all lots was >90% and was not correlated with
field emergence, but the results of the cool germination test at 18°C and the cold test were significantly correlated
with field emergence (r=0.890** and 0.974** respectively). The results of the cool germination test at 18oC were
also significantly correlated with the cold test (r=0.828**). These results suggest that a cool germination test at
18°C can be considered as an alternative to the cold test to evaluate vigour differences among maize seed lots.

Experimental and discussion

In many countries maize is sown in early spring when soil temperatures are low. At low
temperatures, the seeds imbibe, but often can not germinate and may be attacked by fungi.
Therefore, the germination test, as carried out in accordance with the International Rules
(ISTA, 2006) under optimal conditions of temperature and moisture, does not provide a
reliable prediction of field emergence. In these circumstances, it is necessary to carry out
a test under sub-optimal conditions and the cold test is widely used for this purpose, to
evaluate the vigour of maize seeds (Fiala, 1987). However the test procedure for the cold
test is not standardized. For example, while one procedure uses soil and takes 12 days (7
days at 10°C, then 5 days at 25°C), another method, the tray method, uses wetted paper
and takes 10 days at 10°C. Therefore, much research had been done on alternative vigour
tests for maize, including the accelerated aging test (Woltz and TeKrony, 2001; Lovato
et al., 2005), low temperature germination test and the date of primary root emergence
(Van de Venter and Lock, 1991; Toledo et al., 1999), the Complex Stressing Vigour Test
(Lovato and Balboni, 1997), and the rate of germination at lower temperature (Matthews
and Hosseini, 2006).
The present research was conducted to compare the ability of a cool germination at
different temperatures to evaluate vigour in maize seed lots, and to propose an alternative
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COOL GERMINATION TEST CAN BE AN ALTERNATIVE VIGOUR TEST FOR MAIZE

vigour test which is more straight forward and easily standardized than the cold test. The
laboratory tests and the field emergence test were conducted to determine the appropriate
method to evaluate vigour in 7 hybrid maize seed lots (from Toros Tarım seed company,
Turkey).
The standard germination test (SG) was carried out using 8 × 50 seed samples per seed
lot sown between moistened rolled papers and kept at 25°C for seven days. The normal
seedlings were counted according to the evaluation criteria established by ISTA (2006)
and the data were expressed as germination percentage.
The cold test (CT) used 8 × 50 seed samples, placed between moistened rolled papers
with soil (70% of water holding capacity) at 10°C for 7 days, after which they were
transferred to 25°C for 6 days. Thirteen days after the start of the cold test, emerged
seedlings were classified into five groups as described by Fiala (1987). All the seedlings of
group 1 (strong seedlings with no damage) and group 2 (strong seedlings but development
slightly delayed, or with slight injuries) were counted and expressed as a percentage.
The cool germination test was performed by placing seeds between rolled paper (4 ×
50 seeds) as in the standard germination test but at each of three different temperatures
(12, 15 and 18°C) for twelve days. The normal seedlings at 12°C (CG-12), at 15°C (CG-
15) and at 18°C (CG-18) according to ISTA (2006) were evaluated and expressed as a
percentage.
The field emergence of the seed lots was assessed for 4 × 50 seeds which were hand
sown on 12th April 2006 in 4.5m long rows spaced 0.15m apart. The lots were distributed
completely at random within each experimental block. The soil was a sandy-clay-loam
with a moisture content of 16% at sowing. During field emergence (FE), the lowest air
temperature was 6°C and highest air temperature was 16°C. Emerged seedlings were
evaluated as a percentage.
All of the data from the various tests were analyzed using ANOVA and the mean
comparisons among the seed lots were separated by the Duncan’s test. Similarities in seed
quality tests were determined by correlation analysis on SPSS version 10.0 for Windows.
There were no significant differences in SG values of the 7 seed lots, all of which had
≥90% germination (table 1). In contrast, the field emergence data indicated that the
lots showed differences in vigour. Lots 2, 1, 3 and 4 had the highest FE ( 90-96%) and
therefore, were grouped as high vigour, while the significantly lower field emergence of
lots 5, 6 and 7 (62-75%) indicated that they had lower vigour.
Both CT and CG at different temperatures also revealed significant differences among
lots. Thus, as seen in the field emergence data, lots 1, 2 and 3 consistently gave higher
germinations, suggesting high vigour in all tests, while lots 6 and 7 were consistently
lowest.
There was no significant correlation (r=0.316) between SG and FE (table 1) and
the coefficient of determination showed that the SG test contributed only 9.9% of the
variation in the FE (figure 1A). Thus, it can be said that the SG test could not be used
satisfactorily to predict field emergence of maize seed lots at this site. A similar poor
correlation between standard germination and field emergence in maize was also reported
by Lovato et al. (2005). Therefore, although standard germination indicated seed lot
viability, it failed to detect differences in seed vigour.
517
 H. ILBI, S. KAVAK AND B. ESER

In contrast, both CT and CG at 18°C were highly correlated with FE (r=0.974** and
0.890** respectively, table 1). In addition, there was a significant correlation between CG-
18 and CT (r=0.828, p<0.01)) (table 1, figure 1D) indicating that both tests identified the
same lots as having high or low vigour.
Table 1. Results of the standard germination test (SG), cold test (CT), cool germination test carried out at 12°C
(CG-12), 15°C (CG-15), and 18°C (CG-18) and the field emergence of 7 seed lots of maize.

Lots SG (%) FE (%) CT (%) CG-12 (%) CG-15 (%) CG-18 (%)

1 99 95 a 78 a 57 ab 85 ab 96 a
2 98 96 a 76 a 66 a 88 a 97 a
3 92 94 ab 74 a 51 bc 82 b 92 b
4 94 90 b 67 b 34 d 52 d 82 c
5 91 75 c 60 b 39 cd 54 d 65 f
6 92 74 c 50 c 36 d 61 c 77 d
7 96 62 d 41 d 40 cd 54 d 70 e
Mean 95 n.s. 84 64 46 68 83
Correlation
with FE 0.316 - 0.974** 0.673 0.757* 0.890**
Significance levels: * p< 0.05, ** p< 0.01

(A) (B)
y = 1.2795x - 37.227
100 100 y = 0.9286x + 24.599
R2 = 0.0998
R2 = 0.9481
90 90
FE (%)

FE (%)

80 80

70 70

60 60

50 50
80 90 100 30 50 70 90
SG (%) CT (%)
(C) (D)
100 y = 0.932x + 6.7363 100 y = 0.9096x - 11.439
R2 = 0.7922 R2 = 0.6862
90 80
CT (%)
FE (%)

80 60

70 40

60 20

50 0
50 60 70 80 90 100 0 20 40 60 80 100
CG-18 (%) CG-18 (%)
Figure 1. Regression analysis of various tests for 7 maize seed lots: (A) Field Emergence (FE) and Standard
Germination (SG); (B) Field Emergence (FE) and Cold Test (CT); (C) Field Emergence (FE) and Cool
Germination at 18°C (CG-18); (D) Cold Test (CT) and Cool Germination at 18°C (CG-18).

518
 COOL GERMINATION TEST CAN BE AN ALTERNATIVE VIGOUR TEST FOR MAIZE

The cool germination test at 18°C was therefore equally as good as the cold test in
identifying differences in vigour in maize, which were reflected in the field emergence
of the seed lots. Van de Venter and Lock (1991) also reported that a low temperature test
at 13°C had the greatest overall predictive value for maize field emergence compared
to both a soil cold test and the accelerated aging test. Matthews and Hosseini (2006)
found that the rate of germination at a low temperature (13°C) was a good indicator of
vigour in maize, where seed lots that germinated slowly had poor soil emergence and
produced small and variable seedlings. Thus, the reduced germination seen in the present
work for low vigour seed lots may reflect the slow germination of these lots at the low
temperature.
In conclusion, this paper reveals that germination at 18°C may provide an alternative
vigour test for maize which, unlike the cold test, may be easily standardized.

References

Fiala, F. (1987). Cold test. In: Hand Book of Vigour Test Methods. 2nd Edition, Published by The International
Seed Testing Association, 28-36.
ISTA. (2006). International Rules for Seed Testing. International Seed Testing Association, Bassersdorf,
Switzerland.
Lovato, A. and Balboni, N. (1997). Seed vigour in maize (Zea mays L.): two year laboratory and field test
compared. Italian Journal of Agronomy, 1, 1-6.
Lovato, A., Noli, E. and Lovato, A.F.S. (2005). The relationship between three cold temperatures, accelerated
ageing test and field emergence of maize seed. Seed Science and Technology, 33, 249-253.
Matthews, S. and Khajeh Hosseini, M.K. (2006). Mean germination time as an indicator of emergence
performance in soil of seed lots of maize (Zea mays). Seed Science and Technology, 34, 339-347.
Toledo, F.F.De., Novembre, A.D.da L.C., Chamma, H.M.C.P. and Maschietto, R.W. (1999). Evaluation of maize
(Zea mays L.) seed vigour through the date of primary root emergence. Scientia Agricola, 56, 191-196.
Venter, H.A. van de. and Lock, H.W. (1991). A comparison of seed vigour tests for maize (Zea mays L.). South
African Journal of Plant and Soil, 8, 1-5.
Woltz, J.M. and TeKrony, D.M. (2001). Accelerated aging test for corn seed. Seed Technology, 23, 21-34.

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