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HORTSCIENCE 36(5):979–981. 2001.

materials required at the time of sowing (Tay-


lor et al., 1998). Seed enhancements for spin-
Seed Enhancements to Improve ach include removal of the pericarp, hydra-
tion, or chemical treatment of imbibing seeds.
Decoating improves germination and extends
Spinach Germination the range of temperatures at which spinach
seeds can germinate (Atherton and Farooque,
Leslie S. Katzman1 1983; Leskovar and Esensee, 1999; Suganuma
Department of Horticulture, Cornell University, Ithaca, NY 14853-5908 and Ohno, 1984). Soaking in NaOCl and H2O2
can also stimulate germination by weakening
Alan G. Taylor2 the seed covering tissues, enhancing gas ex-
Department of Horticultural Science, New York State Agricultural Experiment change, or a combination of both (Katzman,
1999; Ku et al., 1996).
Station, Cornell University, Geneva, NY 14456-0462 Chemical inhibitors in the seed coverings
Robert W. Langhans3 may also reduce germination. Germination of
spinach seeds was improved at 30 °C by leach-
Department of Horticulture, Cornell University, Ithaca, NY 14853-5908 ing the seeds to remove water-soluble inhibi-
Additional index words. Spinacia oleracea, decoating, dormancy, germination inhibitors, tors (Atherton and Farooque, 1983). Seeds
hydrogen peroxide, sodium hypochlorite, hydroponics, temperature imbibed with this leachate germinated poorly
in comparison with those imbibed in water.
Abstract. Rapid, synchronized, and high percentage of germination is required for Oxidants such as H2O2 and NaOCl may oxi-
commercial spinach (Spinacia oleracea L.) production using hydroponic techniques. Seed dize inhibitors within the seed coat (Bewley
treatments examined to improve seed germination were: 1) decoating; 2) leaching in and Black, 1982). In addition, sodium hy-
water; and 3) soaking seeds for 4 hours in 0.5% NaOCl, leaching for 15 hours in water, and pochlorite can kill fungi and bacteria on the
sowing in 0.3% H2O2 (this treatment will be referred to as NaOCl/H2O2). Germination seed surface and thereby enhance seedling
studies were conducted on four cultivars at a constant 18 °C (optimal) or 30 °C (inhibitory). root and shoot growth (Chun et al., 1997).
At 18 °C, germination rate (T50) was maximized by both hydration treatments, but Heydecker et al. (1969) suggested that
uniformity of germination (Tsd) was greatest for decoated seeds; final germination was high temperatures might inhibit spinach seed
≥89% for all treatments. At 30 °C, decoating resulted in greatest uniformity of germina- germination by reducing the amount of dis-
tion. The NaOCl/H2O2 treatment resulted in highest germination (94%) at the high solved oxygen in water, while increasing
temperature, whereas decoating was least effective (69%). Reduced germination of embryo respiration rate and metabolic de-
decoated seeds was attributed to atypical germinants. Cultivars differed in response to the mand. Therefore, another method of enhanc-
treatments at both temperatures. Component analysis of the NaOCl/H2O2 treatment was ing germination is exposure of imbibed seeds
studied with two slow-to-germinate cultivars. Treatment with H2O2, with or without to oxygen levels higher than that of air, e.g.,
NaOCl, improved the rate, uniformity, and percentage of germination of seeds of both exposure to pure oxygen or to oxygen-
cultivars, but NaOCl alone did not. Pericarp removal or pericarp removal plus NaOCl/ liberating compounds such as H2O2 (Heydecker
H2O2 treatments reduced variability in germination time and enhanced speed of germina- et al., 1969). Catalase, which is present in
tion at 30 °C, but decoating produced a higher percentage of atypical seedlings than did seeds, converts H2O2 to oxygen and water
other treatments. Therefore, the NaOCl/H2O2 treatment is recommended for growers who (Bewley and Black, 1982). Collectively, fa-
are unable to maintain cool germination temperatures and/or cannot afford the costs vorable conditions for spinach seed germina-
associated with cooling. If growers can maintain a germination temperature of ≈18 °C, tion involve cool temperature (18 to 20 °C)
decoated seeds are preferable, based on the high uniformity of germination. and high availability of oxygen.
The main goal of this study was to improve
Spinach is a challenging crop to germinate may be useful for enhancing germination. the percentage, rate, and uniformity of germi-
and grow in a hydroponic system. Seed dor- The spinach “seed” is botanically a fruit nation of spinach cultivars selected for a com-
mancy may cause poor germination and vari- with a spirally coiled embryo surrounding a mercial hydroponic production facility. To
able plant stands. Nongerminating seeds waste pocket of nonliving tissue called the perisperm, achieve this goal, the effects of selected seed
expensive space and resources in the growth but the term “seed” will be used throughout treatments (decoating, leaching, and a NaOCl/
chamber or greenhouse. Speed and uniformity this paper (Taylor, 1997). Orientation of the H2O2 sequence of chemical and hydration treat-
of germination are also important, since slow- embryo is peripheral, and it is covered by the ments) were evaluated at both 18 and 30 °C.
to-germinate seeds and poor uniformity of pericarp. The NaOCl/H2O2 presowing seed treatment
germination result in uneven growth. Exogenous and endogenous factors may was applied to slow-to-germinate cultivars,
Presowing hydration or physical treatments be responsible for germination inhibition or and selected steps were examined indepen-
dormancy in spinach (Katzman, 1999). Ger- dently to determine their effects.
Received for publication 22 May 2000. Accepted for mination declines progressively at tempera-
publication: 6 Nov. 2000. This paper is a portion of a tures >20 °C. Dormancy has been attributed to
thesis submitted by Leslie S. Katzman in fulfilling a
Materials and Methods
graduate degree requirement. We thank the Dept. of
chemical inhibitors and/or physical properties
Horticulture for their financial support, David Paine of the pericarp (outer fruit coat), as pericarp Seed sources. Smooth leaf Japanese culti-
for statistical assistance, Young-Hoe Woo for assis- removal permits germination at high tempera- vars Okame, Megaton, Alrite, and Dash are
tance in developing the NaOCl/H2O2 treatment, Mike tures. The pericarp may reduce germination used in commercial hydroponic production in
Freimer for editing the manuscript, and Takii and Alf by physically blocking radicle emergence and/ Japan. Seeds of these cultivars were provided
Christianson Seed Companies for donating seeds. or limiting the amount of oxygen available to by Takii Seed Co. (Kyoto, Japan) and com-
The cost of publishing this paper was defrayed in part the embryo for respiration. mercially prepared as both decoated and intact
by the payment of page charges. Under postal regula- Seed treatments may overcome dormancy seeds. Semi-savoy leaf American cultivars
tions, this paper therefore must be hereby marked and enhance germination by altering the physi- Rushmore and Whitney were obtained from
advertisement solely to indicate this fact. Alf Christianson Seed Co. (Mount Vernon,
1
Graduate Student.
cal integrity of seed coverings, allowing the
2
Professor of Seed Science and Technology. To seed to complete early phases of germination Wash.). These cultivars were selected based
whom reprint requests should be addressed. E-mail under more favorable conditions, or both. Seed upon growth characteristics, such as high yields
address: agt1@cornell.edu enhancements are postharvest treatments that and tolerance to bolting.
3
Emeritus Professor of Controlled Environment improve germination and seedling growth, or Experimental procedures. Seeds were
Agriculture. facilitate the delivery of seeds and other placed on blue blotter papers in acrylic germi-
nation boxes (11.2 × 11.2 × 3.5 cm high) ordinary logistic model of GENSTAT ≥95% for all four cultivars. However, the
(Hoffman Manufacturing Co., Albany, Ore.). (Hacisalihoglu et al., 1999). Data were ana- nontreated ‘Okame’ and decoated ‘Dash’ had
Each box contained 50 seeds evenly spaced on lyzed as a 2-way (cultivar × seed treatment) slightly lower germination (91% and 89%,
two layers of blotter paper, which was moist- analysis of variance for each dependent vari- respectively).
ened with 15 mL of reverse osmosis (RO) able (T50, Tsd, and percentage of germination The effects of presowing seed treatments
water or 0.3% H2O2 solution, depending on on day 5) for all experiments, and Fisher’s LSD on germination at 30 °C. NaOCl/H2O2-treated
treatment. No additional solution was sup- was used to compare treatment means. Means seeds had the fastest germination at 30 °C
plied to the seeds during the experiments. A with standard errors were calculated for the followed by increasingly higher T50 values for
seed was considered germinated if 1 mm of germination of normal versus atypical seed- decoated, leached, and control seeds (Table
radicle had emerged through the pericarp. lings at 30 °C. 1). Decoating generally resulted in the lowest
Atypical germination occurred when the coty- Tsd values and final percentage of germination
ledons emerged before the radicle; these atypi- Results and Discussion (average 69%). Atypical germinants ranged
cal seedlings were counted as germinated if from 15% to 35% for the four seed lots, and
and when the radicle emerged. The number of The effects of presowing seed treatments were responsible for the reduced normal ger-
germinated seeds was counted at 24-h inter- on germination at 18 °C. In each cultivar mination (Table 2). ‘Megaton’ had the lowest
vals for a period of 5 d after initial sowing. Day studied, T50 was highest for the control treat- percentage of atypical germinants, and there-
zero was designated as the day on which the ment, followed by the decoated treatment, fore this seed lot was least sensitive to high
seeds were sown. Each 50-seed germination while the hydration treatments resulted in the temperature. Percentage of germination was
box was replicated four times for a total of 200 fastest germination (Table 1). However, ≥94% when the data for normal and atypical
seeds per treatment. cultivar differences in response resulted in seedlings were combined.
The effects of presowing seed treatments significant interaction. This acceleration was The NaOCl/H2O2 treatment consistently
on germination at 18 °C and 30 °C. Seeds, attributed to the prehydration phase, since the induced the highest final percentage of germi-
provided by Takii Seed Co. (which utilizes a leached and NaOCl/H2O2 treated seeds were nation of all the presowing seed treatments at
proprietary technology for removing the peri- imbibed for 19 h, whereas control and 30 °C, and may have counteracted the effects
carp), were treated and then placed on two decoated seeds were not hydrated prior to of inhibitors in the pericarp that are respon-
blotters in germination boxes with 15 mL germination. sible for thermosensitivity. In support of this
reverse RO or 0.3% H2O2 solution depending Decoated seeds had the lowest mean Tsd hypothesis, Hsiao and Quick (1984) showed
upon presowing treatment. Treatments were value, while ‘Megaton’ showed little response that NaOCl and H2O2 break dormancy in wild
as follows: 1) intact seeds were sown in RO to any treatment (Table 1). This suggests that oat (Avena fatua L.) seeds by modification of
water (control); 2) decoated seeds were sown alteration or removal of the pericarp may be an the seed coverings. In addition to counteract-
in RO water; 3) intact seeds were soaked in 2 important source of seed-to-seed variation in ing chemical inhibitors, H2O2 increases avail-
L of vigorously aerated RO water for 19 h and time required for germination. The final per- ability of oxygen to seeds at high temperatures
sown in RO water (leached); and 4) intact centage germination on day 5 was generally by providing supplemental oxygen for respi-
seeds were soaked in 2 L of 0.5% NaOCl for
4 h, then in 2 L of vigorously aerated RO
Table 1. Seed treatment effects on germination of spinach cultivars incubated at 18 °C and at 30 °C.
water for 15 h, then sown in 0.3% H2O2. Comparison of means using Fisher’s LSD test for rate (T50) and uniformity (Tsd) of germination, and
Treatment 4) will be referred to as NaOCl/ percentage germination after 5 d.
H2O2. Bleach with a labeled concentration of
5.25% NaOCl was diluted 1:9 with water. 18 °C 30 °C
The 0.3% H2O2 solution was obtained by Cultivar Treatment T50 (d) Tsd (d) Germ % T50 (d) Tsd (d) Germ %
diluting a labeled 3% U.S.P. solution of H2O2 Dash Control 2.2 1.4 96 2.0 1.4 76
to 1:9 with water. Decoatedz 1.8 0.7 89 1.1 0.8 60
Component analysis of the NaOCl/H2O2 Leached 1.2 1.6 95 1.7 1.8 77
treatment at 18 °C. Selected steps of the NaOCl/ NaOCl/H2O2 1.3 1.0 97 0.8 1.4 93
Alrite Control 2.2 1.0 99 2.0 1.1 96
H2O2 treatment were examined independently Decoated 1.7 0.5 98 1.2 1.3 70
to determine their effects on germination of Leached 1.1 1.4 98 2.1 1.7 85
seeds of the slow-to-germinate cultivars, NaOCl/H2O2 1.2 0.9 98 0.8 1.0 96
Rushmore and Whitney. The solutions of Okame Control 2.2 1.6 91 2.1 1.3 78
NaOCl and H2O2 were prepared as previously Decoated 1.8 0.6 97 1.2 0.8 65
described. Seeds were treated prior to germi- Leached 1.2 1.5 99 1.4 1.6 79
nation at 18 °C as follows: 1) leaching (as NaOCl/H2O2 1.2 1.4 99 1.0 1.3 92
described above); 2) NaOCl (same as NaOCl/ Megaton Control 2.8 0.9 96 2.8 2.5 71
H2O2 above, except that seeds were sown in Decoated 2.0 0.8 96 1.5 0.7 83
Leached 1.6 1.0 97 2.5 2.1 81
RO water); 3) H2O2 (same as leached, except NaOCl/H2O2 1.5 0.7 98 1.1 0.8 94
that seeds were sown in 0.3% H202; and 4) Interaction LSD0.05 0.2 0.3 5.1 0.3 0.5 7.5
NaOCl/H2O2 (as described above).
Treatment means
Data analysis. Germination counts were Control 2.4 1.3 95 2.2 1.6 79
recorded daily for the first 5 d after sowing. Decoated 1.8 0.6 95 1.3 0.9 69
From these counts, two other measures of Leached 1.3 1.4 97 1.9 1.8 80
germination were derived: speed of germina- NaOCl/H2O2 1.3 1.0 98 0.9 1.1 94
tion and uniformity of germination. Germina- Cultivar means
tion curves from the daily germination counts Dash 1.6 1.2 94 1.4 1.4 77
were plotted. Speed of germination, T50, was Alrite 1.6 1.0 98 1.5 1.3 87
the number of days required for half the seeds Okame 1.6 1.3 97 1.4 1.3 79
to germinate; uniformity of germination, Tsd, Megaton 2.0 0.9 97 2.0 1.5 82
was the number of days required for one stan- Significance
dard deviation of the total number of germinants Cultivar ** ** * ** NS **
to germinate (as described by Hacisalihoglu et Treatment ** ** NS ** ** **
al., 1999). The T50 and Tsd were obtained for Cultivar × treatment * ** * ** ** **
each replication of each treatment by perform- z
Av. for three replicates for T50 and Tsd data at 30 °C.
NS, *, **
ing a standard nonlinear curve fit using the Nonsignificant, significant at 0.05 and 0.01, respectively.
Table 2. Effects of decoating seeds on percentage germination of nation at specific temperatures. When germi-
four spinach cultivars after 5 d at 30 °C. nating seeds at higher temperatures, we rec-
Germination (%) ommend the NaOCl/H2O2 presowing seed
Cultivar Normal germinants Atypical germinants Total
treatment, given the significant number of
Dash 60 ± 3z 35 ± 3 95
atypical seedlings that resulted from decoating.
Alrite 70 ± 1 27 ± 2 97 However, if germination temperature can be
Okame 65 ± 6 29 ± 1 94 maintained at 18 °C, decoated seeds are rec-
Megaton 83 ± 1 15 ± 1 98 ommended. They consistently had the most
z
Mean ± SE. uniform germination and were convenient to
use, since no additional time or effort was
required to apply the treatment. Also, decoated
Table 3. Component analysis of the effects of the NaOCl/H2O2 pre-sowing seed treatment with NaOCl and
seeds can be sown dry with an automatic
H2O2, alone and together, on germination of two spinach cultivars at 18 °C. seeder, and no posttreatment drying procedure
is required. Thus, for the most uniform germi-
Cultivar Treatment T50 (d) Tsd (d) Germination (%) nation in a hydroponic system, sowing decoated
Rushmore Water 3.4 2.6 62 seeds is recommended.
NaOCl 2.6 2.2 70
H2O2 2.0 1.4 90
NaOCl/H2O2 1.8 1.2 92 Literature Cited
Whitney Water 4.4 2.7 86 Atherton, J.G. and A.M. Farooque. 1983. High
NaOCl 3.6 2.6 74 temperature and germination in spinach. I. The
H2O2 2.4 2.0 94 role of the pericarp. Scientia Hort. 19:25–32.
NaOCl/H2O2 1.9 0.9 100 Bewley, J.D. and M. Black. 1982. Physiology and
Interaction LSD0.05 8.0 biochemistry of seeds in relation to germination.
Means Vol. 2. Springer-Verlag, Berlin.
Treatment Water 3.9 az 2.7 a 74 Chun, S.-C., R.W. Schneider, and M.A. Cohn. 1997.
NaOCl 3.1 b 2.4 a 72 Sodium hypochlorite: Effect of solution pH on
H2O2 2.2 c 1.7 b 91 b rice seed disinfestation and the direct effect on
NaOCl + H2O2 1.9 c 1.0 c 96 b seedling growth. Plant Dis. 81:821–824.
Cultivar Rushmore 2.4 b 1.8 a 79 Hacisalihoglu, G., A.G. Taylor, D.H. Paine, M.B.
Whitney 3.1 a 2.0 a 88 Hilderbrand, and A.A. Khan. 1999. Embryo
Significance Cultivar * NS * elongation and germination rates as sensitive
Treatment * * * indicators of lettuce seed quality: Priming and
Cultivar × treatment NS NS * aging studies. HortScience 34:1240–1243.
z
Mean separation within columns and sets by Fisher’s LSD test, P ≤ 0.05. Heydecker, W., P.I. Orphanos, and R.S. Chetram.
NS, *
Nonsignificant or significant at 0.05. 1969. The importance of air supply during seed
germination. Proc. Intl. Seed Testing Assn.
34:297–303.
ration and metabolic activities. Heydecker et NaOCl/H2O2 treatment were examined for Hsiao, A.I. and W.A. Quick. 1984. Actions of so-
dium hypochlorite and hydrogen peroxide on
al. (1969) found that soaking spinach seeds in their effects on germination in a 2 × 4 (two seed dormancy of wild oats, Avena fatua L.
H2O2 stimulated germination at high tempera- cultivars × four treatments) factorial experi- Weed Sci. 24:411–419.
tures and in excessively wet conditions by ment. The main effects of cultivar and treat- Katzman, L.S. 1999. Developing a system to germi-
increasing the partial pressure of oxygen. ment were significant for T50, while only nate and grow hydroponic spinach, Spinacia
High temperature (30 °C) reduced final treatment affected Tsd. Interaction was sig- oleracea. MS Thesis, Dept. of Horticulture,
percentage of germination for all except the nificant for final percentage of germination, Cornell Univ., Ithaca, N.Y.
NaOCl/H2O2 treated seeds, especially for but not for T50 and Tsd (Table 3). Ku, J.H., T.H. Kim, and D.W. Jun. 1996. Effect of
decoated seeds. The lower germination of This experiment was performed at 18 °C, sodium hypochlorite treatment on germination
decoated seeds was attributed to the large since most hydroponic growth facilities are of spinach seeds. J. Korean Hort. Sci. 37:357–
361.
number of atypical seedlings at 30 °C (Table able to maintain low temperatures during Leskovar, D.I. and V. Esensee. 1999. Pericarp,
2); however, the total percentage of germinants germination/seedling establishment. Seeds leachate, and carbohydrate involvement in
(atypical plus normal seedlings) was similar to soaked in water (leached) or NaOCl alone thermoinhibition of germinating spinach seeds.
the percentage of normal germinants at 18 °C had a slower rate and lower uniformity of HortScience 34:301–306.
(cf. Tables 1 and 2). Therefore, the elevated germination (highest T50 and Tsd) than did those Suganuma, N. and H. Ohno. 1984. Role of pericarp
temperature did not induce dormancy of the treated with H2O2. The final germination of in reducing spinach (Spinacia oleracea L.) seed
decoated seeds, but rather affected seedling leached ‘Rushmore’ seeds was lower than that germination at supra-optimal temperatures. J.
growth. In a parallel study, when sown in peat of ‘Whitney’ seeds. Finally, both H2O2 and a Jpn. Soc. Hort. Sci. 53:38–44.
medium at 30 °C and then transferred at daily combination of H2O2 and NaOCl improved the Taylor, A.G. 1997. Seed storage, germination, and
quality, p. 1–36. In: H.C. Wien (ed.). The
intervals to 20 °C, decoated seeds germinated rate, uniformity, and percentage of germina- physiology of vegetable crops. CAB Intl.,
well provided that exposure to high tempera- tion, whereas NaOCl alone did not. Wallingford, U.K.
ture did not exceed 3 d. Practical implications for commercial sys- Taylor, A.G., P.S. Allen, M.A. Bennett, K.J.
Component analysis of the NaOCl/H2O2 tems. Selected seed enhancements increased Bradford, J.S. Burris, and M.K. Misra. 1998.
treatment at 18 °C. Selected steps of the the rate, uniformity and percentage of germi- Seed enhancements. Seed Sci. Res. 8:245–256.