HORTSCIENCE 43(3):845–852. 2008. peratures (Cantliffe et al., 1981; Valdés et al.

,
1985). Still, these enhancement treatments
Maternal Light Environment During represent a cost and additional manipulation
of the harvested seeds. A superior approach
would be to produce more vigorous or less
Seed Development Affects Lettuce Seed dormant seeds in the field.
There are several reports about the effects
Weight, Germinability, and Storability of the maternal environment on different
aspects of seed quality, including germina-
Samuel Contreras1 bility, dormancy, size, and composition (Bas-
Departamento de Ciencias Vegetales, Pontificia Universidad Católica de kin and Baskin, 1998; Fenner, 1991, 1992;
Chile, Casilla 306-22, Santiago, Chile Gutterman, 2000; Hilhorst and Toorop,
1997). Some of the frequently studied envi-
Mark A. Bennett and James D. Metzger ronmental factors are temperature, water
Department of Horticulture and Crop Science, Ohio State University, availability, light (quality and photoperiod),
altitude, and mineral nutrition. In most stud-
Columbus, OH 43210-1086 ies where photoperiod effects on seed pro-
David Tay2 duction were addressed, seeds produced
under shorter days had higher germinability,
Ornamental Plant Germplasm Center, Ohio State University, Columbus, OH e.g., Ononis sicula Guss. and lettuce (Gutter-
43210-1086 man, 1973), Beta vulgaris L. var. crassa
Additional index words. Lactuca sativa, photodormancy, photoperiod, thermoinhibition, seed Mansf. (Heide et al., 1976), Portulaca oler-
acea L. (Gutterman, 1974), Amaranthus retro-
storability, seed development, red to far-red ratio
flexus L. (Kigel et al., 1977), and Chenopodium
Abstract. Seed germinability and storability are important aspects of seed quality album L. (Karssen, 1970). In fewer cases,
determined by the genotype and environment of seed development. Lettuce (Lactuca shorter days resulted in the production of
sativa L.) is produced commercially in most temperate and subtropical areas of the seeds with lower germinability, e.g., lettuce
world. The objective of this study was to determine how photoperiod and light quality of (Koller, 1962), Carrichtera annua L. (Gut-
the mother plant environment affects lettuce seed quality. Seeds of cv. Tango were terman, 1973), and Polypogon monspeliensis
produced in growth chambers under one of two treatments: a) short day (SD), consisting L. (Gutterman, 2000). Light quality, specif-
’310 mmolm–2s–1) plus 16 hours of darkness daily, and b)
of 8 hours of fluorescent light (’ ically the red to far-red (R:FR) ratio, during
long day (LD), consisting of 4 hours of incandescent light (’’21 mmolm–2s–1), 8 hours of seed development affected the light require-
fluorescent light, 4 hours of incandescent light, and 8 hours of darkness daily. The red to ments for seed germination in Arabidopsis
far-red ratio was ’6.8 and 1.0 for the fluorescent and incandescent light, respectively. In thaliana L. (Hayes and Klein, 1974; McCul-
both treatments, the temperature was 23 8C. The LD treatment produced significantly lough and Shropshire, 1970), Bidens pilosa
heavier seeds; however, germination at optimal conditions (20 8C-light) was similar for L. (Fenner, 1980), Cucumis sativus L. and
both treatments. Germinability (percentage and rates) at suboptimal conditions (30 8C, Cucumis prophetarum L. (Gutterman and
20 8C with different external ABA concentrations, negative osmotic potentials, or dark) Porath, 1975), and Piper auritum Kunth
was higher for seeds produced under the LD treatment. On the other hand, seeds (Orozco-Segovia et al., 1993). Cresswell
produced under the LD treatment presented better storability (evaluated by the and Grime (1981) studied light requirements
accelerated aging test and standard germination after storage at 30 8C and 74% RH). for seed germination of 21 species, and
The critical period for light environment effects was also studied. Seed weight patterns concluded that light quality during seed
were determined early in seed development, during the first 6 days after flowering. drying strongly affect light requirements for
Conversely, light environment effects on seed germinability and storability were germination.
determined at the end of seed development, after physiological maturity, which occurred In spite of the importance that high-
by 11 days after flowering. These results show that lettuce seed germinability and quality seed production has for agriculture
storability may be modified by management of light conditions during seed production in general and horticulture in particular, the
and provide useful information for seed producers, seed companies, and seed conserva- mechanisms operating during seed develop-
tion institutions. ment that control germinability in the mature
seed are still poorly understood (Fenner, 1991;
Gutterman, 2000; Hilhorst and Toorop, 1997),
Lettuce (Lactuca sativa L.) is one of the year with an annual crop value of 2 billion and the management of particular environ-
most important vegetables in the world. In dollars, which makes it the most important mental conditions, such as photoperiod or
the United States, between 2001 and 2006, fresh vegetable in the country (USDA, 2007). light quality, for specific improvement of
lettuce was cultivated on over 121,000 ha per Lettuce seed quality is important because it some aspects of seed quality is not a frequent
affects seedling emergence and uniformity of practice in seed production for most species.
growth, which is fundamental for attaining Storability or longevity may be defined as
Received for publication 19 Nov. 2007. Accepted
high yield and quality in a single harvest the ability of the seed to survive long periods
for publication 21 Jan. 2008. (Smith et al., 1973b; Wien, 1997; Wurr and of time until the initiation of germination. In
Salaries and research support were provided by Fellows, 1985). Thermoinhibition (sensitiv- contrast to dormancy, storability represents a
state and federal funds appropriated to The Ohio ity to high temperatures) and photodormancy desirable seed trait for agronomic, vegetable,
State University, Ohio Agricultural Research and (lack of germination in dark) are two char- and ornamental crops and is commonly
Development Center, as well as a Fulbright Schol- acteristics frequently found in some lettuce included as an attribute of seed quality. Al-
arship to S. Contreras. cultivars that present reduced speed and though dormancy and storability often occur
We thank Hank Hill from Seed Dynamics, Inc., for uniformity in seed germination and seedling coincidently in the same seed, it is not clear if
providing lettuce seeds and technical advice used emergence in the field (Ryder, 1999; Wien, a cause–effect relationship exists between
in this study, and Miller McDonald for critical
review of the manuscript.
1997). A common approach to overcome ger- them. This knowledge is important for man-
1
To whom reprint requests should be addressed; mination problems in lettuce has been to agement of seed stocks by seed companies
e-mail [email protected]. treat the seeds before sowing. For instance, and producers, preservation of target geno-
2
Current address: International Potato Center, seed priming improves germination and types in gene banks, and management of
Apartado 1558, Lima 12, Peru. emergence of lettuce seeds under high tem- natural seed banks of weeds and wild species.

HORTSCIENCE VOL. 43(3) JUNE 2008 845

 The main objectives of this study were: a) parent plastic boxes (11 · 11 · 4 cm). These 10 min. ABA content was determined from
to determine how photoperiod and light boxes were placed in a germination chamber this supernatant by using anti-ABA mono-
quality of the mother plant environment af- at 20 C and constant light. After 4 and 7 d, clonal specific antibodies and competitive
fects lettuce seed quality, and b) to assess the only normal seedlings were counted as ger- ELISA test according to instructions by Phy-
relationship between lettuce seed germina- minated (ISTA, 1999). todetekÒ ABA Test Kit (Agdia, Elkhart, IN).
bility and storability. Other germination tests were conducted Vigor index and average radicle length
using two groups of 50 seeds per replication, measurements were determined on 80 seeds
Materials and Methods planted over two layers of blotters saturated per replication (two groups of 40) using the
in 10 mL of distilled water, a solution of Seedling Vigor Imaging System (SVIS, Ohio
Two experiments were performed to (±) abscisic acid (ABA; Sigma-Aldrich, St. State University, Columbus, OH) according
determine: a) effects of maternal light envi- Louis, MO) or polyethylene glycol (PEG to methodology described by Sako et al.
ronment on lettuce seed quality, and b) the 8000, Sigma-Aldrich) and placed in 9-cm (2001). Before being placed in germination
critical period during lettuce seed develop- petri dishes. The PEG concentrations were boxes, seeds were imbibed 8 h in light to
ment for light environment effects. calculated to obtain water potentials of –0.15, alleviate photodormancy. Seedlings were
–0.30, –0.45, and –0.60 MPa according to scanned 72 h after initiating imbibition.
Expt. 1: Maternal light environment Michel (1983). Germination tests at different The effects of brief interruptions with far-
effects on lettuce seed quality ABA and PEG concentrations were per- red (FR) light on dark germination at 20 C
‘Tango’ lettuce plants were produced in formed at 20 C and constant light, with daily were investigated on two subsamples of 50
the greenhouse in 1.75-L plastic pots filled counts of germinated seeds (radicle emer- seeds per replication. FR breaks for 4 min at
with a soilless growing medium (Metromix gence) to 14 d. Germination at 30 C and 2, 4, 6, 8, and 24 h after sowing were provided
360, Scotts, Marysville, OH). Plants were constant light was evaluated daily to 7 d. The by light-emitting diodes (Quantum Devices,
irrigated daily and each pot was fertilized germination index (GI) was calculated Barneveld, WI) with a wavelength peak at
weekly with 50 mL of a solution containing according with the following equation (adap- 732 nm, a photon flux of 160 mmolm–2s–1,
35 mg N, 15 mg P, and 29 mg K (Peters ted from Maguire, 1962): and a R:FR ratio of 0.01. Seed germination
Professional, Scotts, Marysville, OH). After was evaluated 4 d after sowing.
GI = ðratio of germinated seeds day 1Þ=1
bolting and before flowering, plants were The data were analyzed by the ANOVA
transferred into growth chambers represent- +   +ðratio of germinated seeds day‘‘XÞ’’=‘‘X’’ procedure. Before the analysis, germination
ing one of two treatments: a) short day (SD), +    + ðratio of germinated seeds last countÞ= percentages and GI values were transformed
consisting of 8 h fluorescent light plus 16 h of days to final count: to the arcsin of the square root of the fraction
darkness daily, or b) long day (LD), consist- value. Correlation coefficients between dif-
ing of 4 h incandescent light, 8 h fluorescent Germination in dark was performed using ferent parameters of germinability and stor-
light, 4 h of incandescent light, and 8 h of black-painted petri dishes placed on a ther- ability were calculated.
darkness daily. The photosynthetic photon mogradient table (Series #16065, Seed
flux (PPF), measured by using a portable Processing Holland B.V., Enkhuizen, Expt. 2: Critical period for light
light meter (LI-189, LI-COR Biosciences, Netherlands) at 14, 19, 24, or 29 C; germi- environment effects
Lincoln, NE), was 310 and 21 mmolm–2s–1 nation was evaluated 4 d after sowing. Four lettuce plants (cv. Tango) with 25
for the fluorescent and incandescent light, For the accelerated aging (AA) test, let- flower heads each labeled by flowering day
respectively. The R:FR ratio, calculated as tuce seeds were aged at 41 C and 100% were assigned to one of each of the following
the sum of wavelengths between 656 and 664 RH for 72 h and then germinated following six treatments: L (LD throughout seed devel-
nm divided by the sum of wavelengths the SG protocol. Normal seedlings (ISTA, opment), S (SD throughout seed develop-
between 726 and 734 nm, was 6.8 and 1.0 1999) were evaluated 10 d after planting. ment), L6 (6 d in LD, then SD), S6 (6 d in SD,
for the fluorescent and incandescent light, Seed storage. Seeds were stored in square then LD), L12 (12 d in LD, then SD), and S12
respectively. Light spectral irradiance was mea- plastic boxes (11 · 11 · 4 cm) containing 100 (12 d in SD, then LD). From 3 d after
sured by using a portable spectroradiometer mL of a saturated NaCl solution; the seeds flowering (DAF) and each alternating day,
(LI-1800, LI-COR Biosciences). In both treat- were placed inside aluminum pots over a five flower heads from plants in L and S
ments, the temperature was a constant 23 C. mesh tray so there was no direct contact treatments were sampled to determine seed
The experiment was repeated three times between seeds and the salt solution. The wet and dry weight accumulation. For all
using plants from different sowing dates. boxes, containing the seeds, were placed treatments, management of the plants was
Each replication was considered a block inside plastic bags and put in a dark chamber performed as described in Expt. 1. Seed
and consisted of 10 plants randomly assigned at 30 C. The relative humidity inside the harvest was performed manually by extract-
to each chamber (randomized complete block boxes, measured with a data logger (HOBO ing only labeled and fully matured flower
design). Seeds (achenes) were harvested by U12-012, Onset, Bourne, MA), was 74% heads of each plant. The experiment was
manually extracting only fully matured flower and the seed water content under these repeated two times.
heads (dry and open, with visible seeds of storage conditions was 7.2% ± 0.8% (wet Seed evaluation and data analysis. Seed
8.5% water content on a wet basis) of each basis). Seed samples were extracted after 2, 4, dry weight, germination at 30 C, germina-
plant. Seeds were cleaned and stored in paper 6, 8, and 10 months of storage, and SG was tion in 50 mM ABA solution, dark germina-
envelopes inside a storage room at 4 C and evaluated. tion at 20 C, and germination of normal
25% RH until evaluation. The equilibrium Abscisic acid extraction and determi- seedlings after AA were evaluated as de-
seed water content during storage was 4.7% ± nations. ABA extraction and determination scribed for Expt. 1. Differences among treat-
0.2% (wet basis). from mature lettuce seeds were performed as ments were analyzed using ANOVA and
Seed evaluation. For seed fresh and dry described by Roth-Bejerano et al. (1999) with significantly different means separated using
weight determination, three groups of 100 some modifications. Sixty seeds were frozen least significant difference (LSD, a = 0.05).
seeds each were extracted from each replica- in liquid nitrogen and stored at –80 C. After Specific groups of treatments were compared
tion and weighed before and after drying in freeze-drying (lyophilization), the seeds were by contrast analysis. Statistical analysis of
an oven at 103 C for 48 h. ground to powder in liquid nitrogen and then Expts. 1 and 2 were conducted using SAS
The standard germination (SG) test weighed. Methanol containing 0.5 gL–1 citric (SAS Institute, Cary, NC).
(ISTA, 1999) was conducted in two groups acid monohydrate and 100 mgL–1 butylated
of 50 seeds for each replication. Seeds were hydroxytoluene was added at a ratio of Results and Discussion
planted over two layers of blotter paper 1.0 mL for each 10 mg of dry tissue. The
(Anchor Paper Co., St. Paul, MN) saturated suspension was stirred at 4 C in dark for at Heavier seeds are commonly believed to
in distilled water and placed in square trans- least 20 h and then centrifuged at 1500 g for perform better in most seedling establishment

846 HORTSCIENCE VOL. 43(3) JUNE 2008

environments (Fenner, 1992; Wulff, 1995), types and cultivars, differences among see- cultivars, treatment application, or evaluation
although exceptions have been reported dlots within cultivars have also been methodologies.
(Bennett, 2004). In lettuce, seed weight has observed (Wurr et al., 1986). Several reports Lettuce seed germinability was also
been positively correlated with seed vigor have documented the effect of producing assessed by germination in dark at 14, 19,
(Smith et al., 1973a) and seedling growth lettuce seed at higher temperatures (e.g., 24, and 29 C. Germination of seeds from
after emergence (Smith et al., 1973b). In 30/20 C compared with 20/10 C, day/ both treatments was affected by the absence
Expt. 1, seed performance was evaluated by night) in reducing seed thermoinhibition of light; however, seed from LD had signif-
a) SG test or the ability to produce normal (Drew and Brocklehurst, 1990; Gray et al., icantly higher germination percentage than
seedlings under optimal (20 C-light) con- 1988; Koller, 1962; Kozarewa et al., 2006; seed from SD at 14 (P = 0.018), 19 (P =
ditions, b) germinability (radicle emergence) Sung et al., 1998); however, the effect of 0.019), and 24 C (P = 0.039) (Fig. 1). Along
at different optimal and suboptimal condi- maternal light environment on lettuce seed with thermoinhibition, photodormancy is a
tions, c) seedling growth and uniformity thermoinhibition has rarely been studied. common problem affecting lettuce seed
(vigor index and radicle length from SVIS), Koller (1962) compared germination at emergence and crop establishment (Wien,
and d) the AA test. Seeds produced under LD 20, 23, and 26 C of lettuce seed produced 1997) and has been extensively studied
were significantly heavier than seeds from under 8 or 24 h of light and found that (Ikuma and Thimann, 1964; McArthur,
SD; however, no differences were observed lettuce plants had poor seed production 1978; Toyomasu et al., 1998; Van der Woude
in SG or germination at 20 C (Table 1). At under constant light. But the available data and Toole, 1980). The degree of light sensi-
20 C, germination rates, expressed as GI, indicate that germinability at any of the tivity in lettuce varies among cultivars, and
were slightly (P = 0.041) higher for seed from three temperatures evaluated was better for Tango is described as a photosensitive geno-
SD (Table 1), which could be due to faster seed produced under longer days. Koller’s type (H.J. Hill, pers. comm.), coincident with
imbibition associated with smaller seed size. data contradict our results, where SD our results. Additionally, light requirements
Similar associations between smaller seeds during lettuce seed production improved for germination of photosensitive lettuce
and faster germination were reported for seed germination at higher temperatures genotypes increase with temperature (Ikuma
Triticum aestivum L. (Lafond and Baker, compared with LD (Table 1). Possible rea- and Thimann, 1964; Sung et al., 1998; Van
1986), Zea mays L. (Bennett et al., 1988; sons for this may be differences in lettuce der Woude and Toole, 1980), which explains
Muchena and Grogan, 1977; Shieh and
McDonald, 1982), Erodium brachycarpum
Godr. (Stamp, 1990), and Pastinaca sativa L. Table 1. Quality attributes for ‘Tango’ lettuce seed produced under one of two light treatments.z
(Hendrix, 1984). Parameter LD SD P valuey
Average seedling radicle length after 3 d Dry weight (mg/seed) 0.84 0.73 0.001
of germination has been used for vigor Normal seedlings at 20 C (%) 99.7 98.7 0.423
evaluation of lettuce seeds (Smith et al., Germination at 20 C (%) 100.0 100.0 —
1973a), and a positive correlation of this Germination index at 20 C 0.98 1.00 0.041
parameter with lettuce field emergence and Germination at 30 C (%) 21 60 0.034
Germination index at 30 C 0.05 0.35 0.066
yield has been observed (Contreras and Normal seedlings after AAx (%) 66 5 0.025
Barros, 2005; Smith et al., 1973b; Wurr and Vigor indexw 785 778 0.793
Fellows, 1985). The SVIS integrates param- Radicle length (pixels/seedling)w 357 387 0.085
eters of seedling growth (radicle and hypo- Seed ABA content (pg/mg dry weight) 84 37 0.038
cotyl length) and uniformity (standard z
Short day (SD; 8 h fluorescent light + 16 h darkness daily) or long day (LD; 4 h incandescent light + 8 h
deviation from seedling lengths) to produce fluorescent light + 4 h of incandescent light + 8 h of darkness daily).
a vigor index from 0 (minimum vigor) to y
Calculated from analysis of variance. In the case of germination percentage and germination index
1000 (maximum vigor) (Sako et al., 2001). In P values were calculated with transformed data (arcsin of the square root of the fraction value).
x
our experiments, no differences between AA, accelerated aging of the seeds at 41 C and 100% RH for 72 h.
w
seeds from SD and LD treatments were Values from SVIS (Seed Vigor Image System).
observed for the vigor index and average
radicle length (Table 1). Seeds from LD were
heavier than seeds from SD, thus production
of larger seedlings would be expected. On the
other hand, seeds from SD were smaller but
germinated faster, and this could explain why
at seedling evaluation (3 d after sowing) no
differences in seedling growth were observed
between treatments.
For both treatments, germination per-
centage and rate (expressed as GI) were
lower at 30 C; nevertheless, seeds from
the SD treatment were less affected and
had significantly higher germination per-
centage than seeds from the LD treatment
(Table 1). Thermoinhibition of lettuce seed
germination at high temperature ($25–30
C) is one of the most important problems
affecting lettuce seedling establishment
(Wien, 1997), and seeds from cv. Tango
are known to be sensitive to high temper-
atures (H.J. Hill, pers. comm.). Different
levels of thermoinhibition during seed ger-
mination have been observed among lettuce Fig. 1. Germination percentage after 4 d at different temperatures in dark of ‘Tango’ lettuce seed produced
types and cultivars (Gray, 1975; Kozarewa under one of two light treatments: short day (SD; 8 h fluorescent light + 16 h darkness daily) or long day
et al., 2006). In addition to differences in (LD; 4 h incandescent light + 8 h fluorescent light + 4 h of incandescent light + 8 h of darkness daily).
high temperature germination among lettuce Data are means ±SE of three replications.

HORTSCIENCE VOL. 43(3) JUNE 2008 847

the lack of germination observed at 29 C
(Fig. 1). Higher germinability in dark of seed
produced by lettuce plants under short days
was also observed by Gutterman (1973), who
reported that lettuce seeds from plants grown
under 8 h of light had higher germination than
seeds from plants under 16 h of light (germi-
nation was evaluated after 48 h at 26 C in
dark with one light break of 5 min of white
light 1.5 h after sowing). Improvement of
dark germination in seeds produced under
shorter days has also been reported in Che-
nopodium album L. (Karssen, 1970) and
Amaranthus retroflexus L. (Kigel et al.,
1977).
Seeds produced under SD had lower
sensitivity to increased exogenous ABA
(Fig. 2a) and decreased water potentials
(Fig. 2b) during germination. Seed dormancy
has been positively related with ABA pres-
ence or sensitivity of seeds to this phytohor-
mone (Benech-Arnold et al., 1991; Finch-
Savage and Leubner-Metzger, 2006; Ni and
Bradford, 1993; Yogeesha et al., 2006), and
also sensitivity of germination to water
potential (Ni and Bradford, 1993). Thus, it
may be said that seeds from the LD treatment
are more dormant than those from the SD
treatment. Additional evidence supporting
this concept is the significantly higher ABA
content in mature seeds from the LD treat-
ment (Table 1). Variations in seed ABA
concentration have been observed among
different genotypes of the same species
(Goldbach and Michael, 1976; Groot and
Karssen, 1992; Steinbach et al., 1995; Yogee-
sha et al., 2006) and for the same genotype
produced at different temperatures (Gold-
bach and Michael, 1976) or water treatments
(Benech-Arnold et al., 1991). However, no Fig. 2. Germination percentage (square) and germination index (triangle) at different external abscisic acid
report about variation in seed ABA content (ABA) concentrations (A) and water potential (B) of ‘Tango’ lettuce seed produced under one of two
associated with different maternal light envi- light treatments: short day (8 h fluorescent light + 16 h darkness daily; broken line) or long day (4 h
ronment was found. When variations in seed incandescent light + 8 h fluorescent light + 4 h of incandescent light + 8 h of darkness daily; solid line).
ABA accumulation and final content have Data are means ±SE of three replications.
been observed, higher ABA concentrations
usually have been associated with lower
germinability and higher dormancy (Ni and
Bradford, 1993; Steinbach et al., 1995;
Yogeesha et al., 2006). Our results suggest
that the higher germinability observed in
seeds from SD could be explained, in part,
by lower seed ABA sensitivity and content.
The AA test has been used for evaluation
of seed storability and vigor (Copeland and
McDonald, 2001). ‘Tango’ lettuce seeds
from LD performed better after AA, pro-
ducing a greater number of normal seedlings
than seed from SD conditions (Table 1).
These results suggest that seeds from LD,
despite their lower germinability, are more
vigorous and longer-lived than seeds from
SD. This assumption was corroborated by the
evaluation of SG after different periods of
storage at 30 C and 74% RH. Seeds from the
SD treatment deteriorated faster than seeds
from the LD treatment, which, after 4 months
of storage, produced an average of 91%
normal seedlings compared with 22% from Fig. 3. Lettuce seed germination percentage of normal seedlings after different storage periods at 30 C and
the SD seeds (Fig. 3). Based on these results, 74% RH of Tango’ lettuce seed produced under one of two light treatments: short day (SD; 8 h
the maternal light environment during lettuce fluorescent light + 16 h darkness daily) or long day (LD; 4 h incandescent light + 8 h fluorescent light +
seed development affected not only seed 4 h of incandescent light + 8 h of darkness daily). Data are means ±SE of three replications.

848 HORTSCIENCE VOL. 43(3) JUNE 2008

germinability but also seed storability. A Table 2. Correlation coefficients (r value) between ‘Tango’ lettuce seed germinability and storability
causal relationship between seed storability parameters.
and some forms of physiological dormancy Germinability parameter
has been suggested (Hilhorst and Toorop, Storability Germination Dark germination Germination in Germination
1997; Tesnier et al., 2002; Zarbakhsh et al., parameter after AAy at 19 C ABA, 50 mM at 30 C
1999). However, this hypothesis remains Normal seedlings after
controversial (Fueyo et al., 2003). The results 4 mo. of storagez 0.940 (0.005)x –0.983 (<0.001) –0.586 (0.221) –0.687 (0.132)
from the AA test had a strong correlation (r = Germination index after
0.94; P = 0.005) with SG after 4 months of 4 mo. of storage 0.821 (0.045) –0.865 (0.026) –0.262 (0.616) –0.401 (0.431)
storage (Table 2). The correlation coefficient Germination after AA — –0.984 (<0.001) –0.375 (0.463) –0.469 (0.348)
z
(r value) also was calculated between the Storage at 30 C and 74% RH.
y
values of lettuce seed performance after 4 AA = accelerated aging: 72 h at 41 C and 100% RH.
x
P value for the correlation.
months of storage (normal seedlings percent-
age and GI) and lettuce seed germinability
under different conditions; some of the r
values and their significance (P values) are
presented in Table 2. In general, seed germi-
nability and storability parameters were
inversely related (negative r values); how-
ever, the only parameter of germinability that
had a significant (P < 0.05) correlation with
the three parameters used for storability was
dark germination (Table 2). According to
these results, the type of physiological dor-
mancy exhibited by ‘Tango’ lettuce seeds is
significant and inversely related to seed stor-
ability. From an ecological perspective, this
correlation makes sense because seeds with
lower germinability (i.e., higher dormancy)
will remain in the soil for longer periods,
until optimal conditions permit germination.
Thus, the ability to survive is more important
for dormant seeds than for seeds with high
germinability that likely will germinate
shortly after being shed from the mother
plant. The significant correlation observed
in our study between lettuce seed photo-
dormancy and storability may assist in the Fig. 4. Dry weight accumulation (squares) and seed water content (diamonds) during development of
management of seed stocks by germplasm Tango’ lettuce seed produced under one of two light treatments: short day (8 h fluorescent light + 16 h
centers and seed companies. Careful evalua- darkness daily; broken line) or long day (4 h incandescent light + 8 h fluorescent light + 4 h of
tion of dark germination would permit the incandescent light + 8 h of darkness daily; solid line). Data are means ± SE of two replications.
identification of seed lots most suitable for
storage. Further research should be directed
to corroborate the existence of this correla- observed, indicating that the effect of the average from L, S6, and S12 by contrast
tion in other lettuce cultivars, as well as in light treatment on individual seed dry weight analysis, the difference was significant (P <
other species, especially those of the Aster- was produced early in seed development, 0.001). Differences were also detected when
aceae family. during the first 6 DAF or first half of the similar contrast analyses were performed for
Although LD and SD treatments differed PM process (Figs. 4 and 5a). It was pre- germination percentage at 30 C (P = 0.013),
in the amount of dry matter accumulated by viously observed that longer days affected GI at 30 C (P = 0.002), and GI in 50 mM ABA
lettuce seeds, the pattern of development was flowering and the number of seeds produced solution (P = 0.005). Based on these results,
similar (Fig. 4). Seed physiological maturity by lettuce plants (Koller, 1962). Addition- the effect of the day-length treatments on
(PM, defined as the moment of maximum ally, the number of seeds produced per lettuce seed germinability occurred in the last
seed dry weight accumulation), determined lettuce plant has an inverse relationship with portion of seed development, after PM. Sim-
by an iterative regression analysis procedure individual seed weight (Contreras, 2007; ilar to the results observed for germinability,
(Pieta-Filho and Ellis, 1991), occurred 10.4 ± Izzeldin et al., 1980), because of the higher maternal light environment effect on seed
0.4 and 10.7 ± 0.1 DAF for LD and SD plants, competition for resources among seeds dur- storability (assessed by the AA test) was
respectively (Fig. 4). Consequently, 6 DAF ing seed filling. Thus, it may be that the dif- produced during the last part of seed devel-
(time when the plants from L6 and S6 treat- ference in dry weight between ‘Tango’ seeds opment, after PM. The percentage of normal
ments where moved from one light treatment from SD and LD treatments was caused by lettuce seedlings after AA of seeds from L,
to another) represented about half of PM, the production of more flower heads and seeds S6, and S12 differed (P = 0.002) when
whereas 12 DAF (time of plant movement for in plants under SD. However, this hypothesis compared with those of seeds from S, L6,
the L12 and S12 treatments) represented cannot be supported or rejected by our results and L12 treatments (Fig. 5c). Maximum seed
about 1 d after PM. because the number of flower heads or seeds quality during seed development is believed
Individual seed dry weights increased in per plant were not evaluated. to coincide with PM, or the moment of
seeds from S6 and S12 in relation to S; In Expt. 2, seed germinability was maximum seed dry weight accumulation,
however, the greatest values were for seeds assessed as dark germination at 20 C (Fig. after which viability and vigor would de-
from the L, L6, and L12 treatments (Fig. 5a). 5b), germination in light at 30 C (data not crease (Abdul-Baki and Anderson, 1972;
When the average seed weight of L, L6 and shown), and germination with light in 50 mM Harrington, 1972). However, there is evi-
L12 was compared with the average value ABA solution (data not shown). When the dence that in some species seed quality may
of S, S6, and S12 by a contrast analysis, average dark germination at 20 C of seed increase after PM (Demir and Ellis, 1992a,
a significant difference (P < 0.001) was from S, L6, and L12 was compared with the 1992b; Sinniah et al., 1998; Welbaum, 1999).

HORTSCIENCE VOL. 43(3) JUNE 2008 849

 further accumulation of dry matter in the
seed, but seed water content is sufficient to
allow metabolic activity. In our experiments,
lettuce seeds were 40% water content (wet
basis) at PM, and above 35% for at least 4 d
following PM; 6 d after PM, seeds were
8.5% water content, which was maintained
until harvest (Fig. 4).
The light requirement for lettuce seed
germination is mediated by the action of
phytochrome, a soluble protein found in
two interconvertible forms: Pr (red light
absorbing, biologically inactive) and Pfr
(far-red light absorbing, biologically active)
(Shinomura, 1997). According to Vertucci
et al. (1987), phytochrome photoconversion
occurs when seed water contents are over 8%
in lettuce, so conversion is possible in the
seed during desiccation and until harvest.
Pfr (or some stable intermediate able to yield
Pfr in dark) may persist in the seed after
final maturation and dehydration (Taylorson,
1982). The amount of this pre-existent Pfr
will depend on the light quality and intensity
to which seeds were exposed at the end of
seed development and dehydration (Taylor-
son, 1982). In our experiments, the SD
treatment consisted only of fluorescent light,
which is relatively rich in red light (R:FR 
6.8). Conversely, incandescent light, which
was used to extend day-length in the LD
treatment, is relatively rich in far-red light
(R:FR  1.0). These differences in light
quality favor higher accumulation of pre-
existent Pfr in seeds from plants under SD
compared with seeds from LD treatment,
which would explain the higher dark germi-
nation of seeds from SD treatment and why
the effect was produced at the end of seed
development. The suppression of dark ger-
mination of seeds from SD treatments by
breaks of FR light (Table 3) supports the idea
that seeds from SD had a higher content of
pre-existent Pfr than seeds from LD treat-
ments. Thus, light quality, and not hours of
light, would be the critical factor explaining
differences in ‘Tango’ lettuce seed germina-
bility and storability for seeds produced
under SD vs. LD treatments. This hypothesis
is supported by results from other authors that
have reported reductions in photodormancy
caused by seed development under environ-
ments with higher R:FR ratios (Cresswell and
Grime,1981; Hayes and Klein, 1974; McCul-
lough and Shropshire, 1970).
There is evidence that Pfr would promote
seed germination by promoting gibberellin
biosynthesis and suppressing ABA formation
(Roth-Bejerano et al., 1999; Toyomasu et al.,
Fig. 5. Seed dry weight (A), dark germination at 20 C (B), and normal seedling after accelerated aging (C) 1998). Additional research should be con-
from ‘Tango’ lettuce seed produced under different combinations of long day (LD; 4 h incandescent ducted to determine if differences in ABA
light + 8 h fluorescent light + 4 h of incandescent light + 8 h of darkness daily) and short day (SD; 8 h concentration of mature lettuces seeds devel-
fluorescent light + 16 h darkness daily) light conditions: L (LD throughout seed development), S (SD
throughout seed development), L6 (6 d in LD, then SD), S6 (6 d in SD, then LD), L12 (12 d in LD, then
oped under different light environments is
SD), and S12 (12 d in SD, then LD). Data are means ± SE. In the same graphic, treatments with different mediated by Pfr action. Possible cause–effect
letters are significantly different (LSD, a = 0.05). relationships between pre-existent Pfr and
lettuce seed germinability or storability
should be also studied.
Based on our results, ‘Tango’ lettuce seed development is known as maturation drying In conclusion, the light treatments applied
germinability and storability may increase or (Bewley and Black, 1994) and is character- in these experiments affected the weight,
decrease after PM depending on the maternal ized by fresh weight loss and a decline in seed germinability and storability of lettuce seeds
light environment (Fig. 5). This phase of seed water content. During this period, there is no cv. Tango. Effects on seed weight were

850 HORTSCIENCE VOL. 43(3) JUNE 2008

Table 3. Germination percentage after 4 d at 20 C in continuous dark and dark plus far-red light breaks fruits storage on the phytochrome and on the
(FR; 4 min. at 2, 4, 6, 8, and 24 h after sowing) for ‘Tango’ lettuce seed produced under one of two light germination of Cucumis prophetarum L. and
treatments.z Cucumis sativus L. seeds. Oecologia 18:37–45.
Day-length treatment Hayes, R.G. and W.H. Klein. 1974. Spectral
quality influence of light during development
Germination condition LD SD P valuey of Arabidopsis thaliana plants in regulating
Dark 2.3 ± 1.5 69.3 ± 12.4 0.010 seed germination. Plant Cell Physiol. 15:643–
Dark + FR 1.0 ± 0.6 10.3 ± 7.33 0.147 653.
z
Short day (SD; 8 h fluorescent light + 16 h darkness daily) or long day (LD; 4 h incandescent light + 8 h Harrington, J.F. 1972. Seed storage and longevity,
fluorescent light + 4 h of incandescent light + 8 h of darkness daily). p. 145–245. In: T.T. Kozlowski (ed.). Seed
y
Calculated from analysis of variance with transformed data (arcsin of the square root of the fraction biology, Vol. 3. Academic Press, New York.
value). Heide, O.M., O. Juntilla, and R.T. Samuelsen.
1976. Seed germination and bolting in red beet
as affected by parent plant environment. Phys-
determined during the first 6 d of seed Copeland, L.O. and M.B. McDonald. 2001. Prin- iol. Plant. 36:343–349.
ciples of seed science and technology. Kluwer Hendrix, S.D. 1984. Variation in seed weight and
development (first half of the time needed its effects on germination in Pastinaca sativa L.
to reach PM), while maternal light environ- Academic Publishers, Boston.
Cresswell, E.G. and J.P. Grime. 1981. Induction of (Umbelliferae). Amer. J. Bot. 71:795–802.
ment effects on seed germinability and stor- Hilhorst, H.W. and P.E. Toorop. 1997. Review on
a light requirement during seed development
ability occurred after PM, during the phase dormancy, germinability, and germination in
and its ecological consequences. Nature
of seed maturation and drying. Lettuce seed 291:583–585. crop and weed seeds. Adv. Agron. 61:111–165.
storability had an inverse and significant Demir, I. and R.H. Ellis. 1992a. Changes in seed Ikuma, H. and K.V. Thimann. 1964. Analysis of
correlation with dark germination. These re- quality during seed development and matura- germination processes of lettuce seed by means
sults are of practical interest for seed pro- tion in tomato. Seed Sci. Res. 2:81–87. of temperature and anaerobiosis. Plant Physiol.
ducers, and contribute to our knowledge Demir, I. and R.H. Ellis. 1992b. Development of 39:756–767.
about the factors affecting quality during pepper (Capsicum annuum) seed quality. Ann. International Seed Testing Association. 1999.
Appl. Biol. 121:385–399. International rules for seed testing. Seed Sci.
seed production and the relationship between
Drew, R.L.K. and P.A. Brocklehurst. 1990. Effects Technol. 27 (Suppl.), 333 pp.
different aspects of seed quality. Based on the Izzeldin, H., L.F. Lippert, and F.H. Takatori. 1980.
results of these experiments, we hypothesize of temperature of mother-plant environment on
yield and germination of seeds of lettuce An influence of water stress at different growth
that light quality (R:FR ratio) during lettuce (Lactuca sativa). Ann. Bot. (Lond.) 66:63–71. stages on yield and quality of lettuce seed. J.
seed production plays a critical role in seed Fenner, M. 1980. The induction of a light require- Amer. Soc. Hort. Sci. 105:68–71.
germinability and storability. ment in Bidens pilosa seeds by leaf canopy Karssen, C.M. 1970. The light promoted germina-
shade. New Phytol. 84:103–106. tion of the seeds of Chenopodium album L. III.
Fenner, M. 1991. The effects of the parent envi- Effects of the photoperiod during growth and
Literature Cited development of the plants on the dormancy
ronment on seed germinability. Seed Sci. Res.
Abdul-Baki, A.A. and J.D. Anderson. 1972. Phys- 1:75–84. of the produced seeds. Acta Bot. Neerl. 19:81–
iological and biochemical deterioration of Fenner, M. 1992. Environmental influences on 94.
seeds, p. 283–315. In: T.T. Kozlowski (ed.). seed size and composition. Hort. Rev. (Amer. Kigel, J., M. Ofir, and D. Koller. 1977. Control of
Seed biology, Vol. 2. Academic Press, New Soc. Hort. Sci.) 13:183–213. the germination responses of Amaranthus ret-
York. Finch-Savage, W.E. and G. Leubner-Metzger. roflexus L. seeds by their parental photothermal
Baskin, C.C. and J.M. Baskin. 1998. Seeds: ecol- 2006. Seed dormancy and the control of ger- environment. J. Expt. Bot. 28:1125–1136.
ogy, biogeography, and evolution of seed mination. New Phytol. 171:501–523. Koller, D. 1962. Preconditioning of germination in
dormancy and germination. Academic Press, Fueyo, P.A., R.A. Sánchez, and R.L. Benech- lettuce at time of fruit ripening. Amer. J. Bot.
San Diego, CA. Arnold. 2003. Seed longevity in two sorghum 49:841–844.
Benech-Arnold, R.L., M. Fenner, and P.J. varieties with contrasting dormancy level prior Kozarewa, I., D.J. Cantliffe, R.T. Nagata, and P.J.
Edwards. 1991. Changes in germinability, to harvest. Seed Sci. Technol. 31:639–650. Stoffella. 2006. High maturation temperature
ABA content and ABA embryonic sensitivity Goldbach, H. and G. Michael. 1976. Abscisic acid of lettuce seeds during development increased
in developing seeds of Sorghum bicolor (L.) content of barley grains during ripening as ethylene production and germination at ele-
Moench. induced by water stress during grain affected by temperature and variety. Crop Sci. vated temperatures. J. Amer. Soc. Hort. Sci.
filling. New Phytol. 118:339–347. 16:797–799. 131:564–570.
Bennett, M.A., L. Waters, Jr., and J.H. Curme. Gray, D., D.C.E. Wurr, J.A. Ward, and J.R. Lafond, G.P. and R.J. Baker. 1986. Effects of
1988. Kernel maturity, seed size, and seed hy- Fellows. 1988. Influence of post-flowering genotype and seed size on speed of emergence
dration effects on seed quality of a sweet temperature on seed development, and sub- and seedling vigor in nine spring wheat culti-
corn inbred. J. Amer. Soc. Hort. Sci. 113:348– sequent performance of crisp lettuce. Ann. vars. Crop Sci. 26:341–346.
353. Appl. Biol. 113:391–402. Maguire, J.D. 1962. Speed of germination- aid in
Bennett, M.A. 2004. Seed and agronomic factors Gray, D. 1975. Effects of temperature on germina- selection and evaluation for seedling emer-
associated with germination under temperature tion and emergence of lettuce (Lactuca sativa gence and vigor. Crop Sci. 2:176–177.
and water stress, p. 97–123. In: R.L. Benech- L.) varieties. J. Hort. Sci. 50:349–361. McArthur, J.A. 1978. Light effects upon dry lettuce
Arnold and R.A. Sánchez (eds.). Handbook of Groot, S.P.C. and C.M. Karssen. 1992. Dormancy seeds. Planta 144:1–5.
seed physiology: applications to agriculture. and germination of abscisic acid-deficient McCullough, J.M. and W. Shropshire, Jr. 1970.
The Haworth Reference Press, Binghamton, tomato seeds. Plant Physiol. 99:952–958. Physiological predetermination of germination
NY. Gutterman, Y. 1973. Differences in the progeny responses in Arabidopsis thaliana (L.). Heynh.
Bewley, J.D. and M. Black. 1994. Seeds: Physiol- due to daylength and hormone treatment of Plant Cell Physiol. 11:139–148.
ogy of development and germination. 2nd ed. the mother plant, p. 59–80. In: W. Heydecker Michel, B.E. 1983. Evaluation of the water poten-
Plenum Press, New York. (ed.). Seed ecology. Butterworth & Co., tials of solutions of polyethylene glycol 8000
Cantliffe, D.J., K.D. Shuler, and A.C. Guedes. London. both in the absence and presence of other
1981. Overcoming seed thermodormancy in a Gutterman, Y. 1974. The influence of the photo- solutes. Plant Physiol. 72:66–70.
heat sensitive romaine lettuce by seed priming. periodic regime and red-far red light treatments Muchena, S.C. and C.O. Grogan. 1977. Effect of
HortScience 16:196–198. of Portulaca oleracea L. plants on the germi- seed size on germination of corn (Zea mays L.)
Contreras, S. and M. Barros. 2005. Pruebas de nability of their seeds. Oecologia 17:27–38. under simulated water stress conditions. Can. J.
vigor en semillas de lechuga y su correlación Gutterman, Y. 2000. Maternal effects on seeds Plant Sci. 57:921–923.
con emergencia. Cienc. Invest. Agr. 32:3– during development, p. 59–84. In: M. Fenner Ni, B.R. and K.J. Bradford. 1993. Germination and
11. (ed.). Seeds: the ecology of regeneration in dormancy of abscisic acid- and gibberellin-
Contreras, S. 2007. Effects of maternal plant plant communities. 2nd ed. CABI Publishing, deficient mutant tomato (Lycopersicon escu-
environment on lettuce (Lactuca sativa L.) seed Wallingford, England. lentum) seeds: sensitivity of germination to
dormancy, germinability, and storability. Ohio Gutterman, Y. and D. Porath. 1975. Influences of abscisic acid, gibberellin, and water potential.
State Univ., Columbus, PhD Diss. photoperiodism and light treatments during Plant Physiol. 101:607–617.

HORTSCIENCE VOL. 43(3) JUNE 2008 851

Orozco-Segovia, A., M.E. Sánchez-Coronado, and carpum, Geraniaceae). Amer. J. Bot. 77:874– lettuce seeds by seed priming. HortScience
C. Vázquez-Yánez. 1993. Effect of maternal 882. 20:1112–1114.
light environment on seed germination in Piper Steinbach, H.S., R.L. Benech-Arnold, G. Kristof, Van der Woude, W.J. and V.K. Toole. 1980.
auritum. Funct. Ecol. 7:395–402. R.A. Sánchez, and S. Marcucci-Poltri. 1995. Studies of the mechanism of enhancement of
Pieta-Filho, C. and R.H. Ellis. 1991. The develop- Physiological basis of pre-harvest sprout- phytochrome-dependent lettuce seed germina-
ment of seed quality in spring barley in four ing resistance in Sorghum bicolor (L.) Moench. tion by prechilling. Plant Physiol. 66:220–224.
environments. I. Germination and longevity. ABA levels and sensitivity in devel- Vertucci, C.W., F.A. Vertucci, and A.C. Leopold.
Seed Sci. Res. 1:163–177. oping embryos of sprouting-resistance and 1987. Water content and the conversion of
Roth-Bejerano, N., N.J.A. Sedee, R.M. VanDer- susceptible varieties. J. Expt. Bot. 46:701– phytochrome regulation of lettuce dormancy.
Meulen, and M. Wang. 1999. The role of 709. Plant Physiol. 84:887–890.
abscisic acid in germination of light-insensitive Sung, Y., D.J. Cantliffe, and R.T. Nagata. 1998. Welbaum, G.E. 1999. Cucurbit seed development
lettuce seeds. Seed Sci. Res. 9:129–134. Seed development temperature regulation of and production. HortTechnology 9:341–348.
Ryder, E.J. 1999. Lettuce, endive, and chicory. thermotolerance in lettuce. J. Amer. Soc. Hort. Wien, H.C. 1997. Lettuce, p. 479–509. In: H.C.
CABI Publishing, Wallingford, England. Sci. 123:700–705. Wien (ed.). The physiology of vegetable crops.
Sako, Y., M.B. McDonald, K. Fujimura, A.F. Taylorson, R.B. 1982. Interaction of phytochrome CABI Publishing, Wallingford, England.
Evans, and M.A. Bennett. 2001. A system for and other factors in seed germination, p. 323– Wulff, R.D. 1995. Environmental maternal effects
automated seed vigour assessment. Seed Sci. 346. In: A.A. Khan (ed.). The physiology and on seed quality and germination, p. 491–505.
Technol. 29:625–636. biochemistry of seed development, dormancy In: J. Kigel and G. Galili (eds.). Seed develop-
Shieh, W.J. and M.B. McDonald. 1982. The and germination. Elsevier Biomedical Press, ment and germination. Marcel Dekker, New
influence of seed size, shape and treatment on New York. York.
inbred seed corn quality. Seed Sci. Technol. Tesnier, K., H.M. Strookman-Donkers, J.G. Van Wurr, D.C.E. and J.R. Fellows. 1985. A determi-
10:307–313. Pijlen, A.H.M. Van der Geest, R.J. Bino, and nation of the seed vigour and field performance
Shinomura, T. 1997. Phytochrome regulation of S.P.C. Groot. 2002. A controlled deterioration of crisp lettuce seedstocks. Seed Sci. Technol.
seed germination. J. Plant Res. 110:151–161. test for Arabidopsis thaliana reveals genetic 13:11–17.
Sinniah, U.R., R.H. Ellis, and P. John. 1998. variation in seed quality. Seed Sci. Technol. Wurr, D.C.E., J.R. Fellows, D. Gray, and J.R.A.
Irrigation and seed quality development in 30:149–165. Steckel. 1986. The effects of seed production
rapid-cycling brassica: seed germination and Toyomasu, T., H. Kawaide, W. Mitsuhashi, Y. techniques on seed characteristics, seedling
longevity. Ann. Bot. (Lond.) 82:309–314. Inoue, and Y. Kamiya. 1998. Phytochrome growth and crop performance of crisp lettuce.
Smith, O.E., N.C. Welch, and T.M. Little. 1973a. regulates gibberellin biosynthesis during ger- Ann. Appl. Biol. 108:135–144.
Studies on lettuce seed quality: I. Effect of seed mination of photoblastic lettuce seeds. Plant Yogeesha, H.S., K.K. Upreti, K. Padmini, K.
size and weight on vigor. J. Amer. Soc. Hort. Physiol. 118:1517–1523. Bhanuprakash, and G.S.R. Murti. 2006. Mech-
Sci. 98:529–533. United States Department of Agriculture. 2007. anism of seed dormancy in eggplant (Solanum
Smith, O.E., N.C. Welch, and O.D. McCoy. 1973b. Vegetables and melons outlook. VGS-321/21 melongena L.). Seed Sci. Technol. 34:319–325.
Studies on lettuce seed quality: II. Relationship June 2007. USDA Economic Research Ser- Zarbakhsh, A.J., H.S. Prakash, and H.S. Shetty.
of seed vigor to emergence, seedling weight, vice. 18 July 2007. <http://www.ers.usda.gov/ 1999. Effect of storage on chemical composi-
and yield. J. Amer. Soc. Hort. Sci. 98:552–556. publications/vgs/tables/fresh.pdf>. tion, germination and vigour of dormant and
Stamp, N.E. 1990. Production and effect of seed Valdés, V.M., K.J. Bradford, and K.S. Mayberry. non-dormant seeds of sunflower genotypes.
size in a grassland annual (Erodium brachy- 1985. Alleviation of thermodormancy in coated Seed Sci. Technol. 27:771–778.

852 HORTSCIENCE VOL. 43(3) JUNE 2008