Scientia Horticulturae 103 (2005) 441–451

www.elsevier.com/locate/scihorti

Effect of photoperiod and light integral

on flowering and growth of Eustoma
grandiflorum (Raf.) Shinn.
Nazrul Islam, Grete Grindal Patil,
Hans Ragnar Gislerød*
Department of Horticulture and Crop Sciences, Agricultural University of Norway,
P.O. box 5022, N-1432, Ås, Norway.
Accepted 9 June 2004

Abstract

The aim of the study was to examine the effects of different photoperiod and light integral on floral
initiation, development and subsequent growth of Eustoma grandiflorum (Raf.) Shinn. Six-weeks-old
seedlings of ‘Echo Blue’ and ‘Fuji Deep Blue’ were placed under short day (SD, 10 h) and were
transferred to long days (LD, 20 h) at 2-week intervals from 6 to 14 weeks after seeding. Plants
initiated flower buds regardless of light regimes. Flower bud initiation was delayed by SD compared
to LD; plants transferred after 6 weeks from seeding initiated flower buds at least 21 and 10 days
earlier at LD at high (HL) and low (LL) daily light integral, respectively, compared to those at SD.
Light regimes had little or no effect on time to flower bud development after initiation. Thus, it seems
likely that LD and HL affected the initiation rather than development. Both the photoperiod and light
integral strongly influenced the subsequent growth after initiation. SD delayed the time to visible bud
(VB), increased the number of nodes to first open flower, number of branches, stem diameter and
shoot dry weight compared to LD. HL promoted flowering and increased several shoot characteristics
and flowering compared to LL.

Abbreviations: HL, long day and high light integral; HPS, high-pressure sodium lamps; LD, long day; LL,
long day and low light integral; SD, short day
* Corresponding author. Tel.: +47 6494 7800; fax: +47 6494 7802.
E–mail address: hans.gislerod@ipm.nlh.no (H.R. Gislerød).

0304-4238/$ – see front matter # 2004 Elsevier B.V. All rights reserved.
doi:10.1016/j.scienta.2004.06.018
442 N. Islam et al. / Scientia Horticulturae 103 (2005) 441–451

The results indicate that Eustoma is a quantitative long-day plant. LD, and more specifically HL,
enhanced flower bud initiation, development and subsequent growth. An initial SD period is preferred
to increase the number of branches, number of flowering buds and flowers, stem diameter and shoot
dry weight.
# 2004 Elsevier B.V. All rights reserved.

Keywords: Eustoma; Flower bud initiation; Growth; Photoperiod; Light integral

1. Introduction

Eustoma grandiflorum (Raf.) Shinn. (Syn. Lisianthus russellianus) is a

seed propagated ornamental flower, produced worldwide and introduced to Norway
about 10 years ago. It is a North American wild flower, native to central and southern
US and mainly inhabits the moist prairies from Nebraska to Colorado and Texas
(Shinners, 1957; Wood and Weaver, 1982; Halevy and Kofranek, 1984). Eustoma has
been a very popular cut flower because of its floral colours and long vase life and is a
leading cut flower in Japan (Takeda, 1994; Hisamatsu et al., 1998). Eustoma is now
available all year round by using selected cultivars grown in appropriate climatic
conditions. At present, however, pot culture of Eustoma is not as significant as cut flowers
(Bradley et al., 2000; Ohkawa and Sasaki, 1999). In Norway, a year round greenhouse cut
flower production of Eustoma is achieved by supplementing light with high-pressure
sodium lamps (HPS).
Due to the complex response to photoperiod of Eustoma cultivars, it has been classified
differently, both as day-neutral plant (Azrak, 1984; Halevy and Kofranek, 1984) and as a
quantitative long-day plant (Tsukada et al., 1982; Grueber et al., 1984; Roh et al., 1989).
Growth of Eustoma, crop time and flower quality are affected by light intensity and day
length (Corr and Katz, 1997). At seedling stage, growth rate of Eustoma is very slow and
requires 50–140 days from germination to transplanting (Matsuo and Shirasaki, 1990;
Harbaugh, 1995) and totally about 22–28 weeks from seeding to anthesis of the terminal
flower bud (Halevy and Kofranek, 1984; Roh and Lawson, 1984). Recently, Zaccai and
Edri (2002) have reported a direct effect of photoperiod on floral transition in Eustoma as
well as a promoting effect of high temperature.
As light is one of the most important climatic factors in floral development and growth
of Eustoma, it is important in commercial production to know how the flower initiation and
development are affected by photoperiod and light integrals. Traditionally, appearance of
visible flower buds and time to first open flower along with the number of nodes to first
flower are often used to indicate the effect of the environment on the flowering process
(Mastalerz, 1977).
The aim of the present study was to examine the effects of light integral on floral
initiation and development and subsequent growth of Eustoma. In addition, we included
different time periods in short day (SD) before transfer to long day (LD) to see how plant
age is related to the effect of day length on flowering. The present study describes an
experiment performed in 2001. The results of preliminary studies in January–May 2000
were similar to those in 2001 and therefore not included.
 N. Islam et al. / Scientia Horticulturae 103 (2005) 441–451 443

2. Materials and methods

2.1. Plant material and growing condition

The experiment was conducted in a computer-controlled greenhouse during the time

period from December 2000–May 2001 at the Agricultural University of Norway, Norway.
Four-weeks-old seedlings of Eustoma ‘Echo Blue’ (double-flowered) and ‘Fuji Deep Blue’
(single-flowered) were bought in plugs from Holland (Hammer flower seeds Ltd.) and were
placed under SD conditions (10 h light), 21  1 8C, with 85 mmol m 2 s 1 supplementary
light for 2 weeks.
The experiment was started with 6-weeks-old seedlings, all grown further at 21  1 8C,
75% RH, 800 mmol mol 1 CO2. The plants were gradually transferred to bigger pots and
final potting was done 17 weeks after seeding in 15 cm plastic pots with standard limed (pH
5.5) and fertilized peat (2 kg of 15–4–12 NPK). The plants were watered daily with a
complete nutrient solution (2.0 mS cm 1). Additional photosynthetic light was provided
with high-pressure sodium lamps (HPS) at a level of 122 mmol m 2 s 1 at pot level and
three light regimes were established: low daily light integral and short day (SD: 10 h with
HPS giving 4.4 mol m 2 day 1), low daily light integral and long day (LL: 10 h with HPS
plus 10 h with 1.5 mmol m 2 s 1 incandescent light giving 4.4 mol m 2 day 1) and high
daily light integral and long day (HL: Extending the first 10 h with 10 more hours with HPS
giving 8.8 mol m 2 day 1). The two long day conditions with high and low light integral,
were established in two greenhouse compartments. All plants were covered with darkening
curtains mounted along the sides and the ceiling of the greenhouse for 14 h per day and the
lamps used for artificial lighting mounted under the curtains. During the time period the
plants were not covered with the curtains the HPS lamps were turned off at a natural light
level exceeding 25 mol m 2 day 1. The SD condition was located in the same
compartment as LL, but separated with SD curtains in the 14 h dark period. Global
radiation, calculated on monthly basis, provided a photosynthetic active radiation of 1.0,
1.9, 7.6, 16.0, 21.4 and 39.7 mol m 2 day 1 in December 2000 and January through May
2001, respectively.
At the start of the experiments the plants were divided in 11 groups, each of 70 plants.
After 6, 8, 10, 12 and 14 weeks from seeding, one group of plants were transferred from
SD to LL and one group to HL. One group was kept in SD throughout the whole
experiment.

2.2. Flower bud initiation

To identify the developmental stages of the flowers, apical dissection under a binocular
microscope were done and photos were taken by digital camera (Leica DC 200) connected
to the microscope. Due to variation in the plant size in each treatment, plants were selected
on the basis of height, and the result for the flower development stages was based on the
homogenous tall plants in each treatment, avoiding the plants that were too small or too tall
from the average. In ‘Echo Blue’, it was difficult to distinguish between second and/or third
layer of petals and the stamens initiation, since they developed simultaneously after sepals
and first layer of petals. Therefore, only the data for ‘Fuji Deep Blue’ are presented.
 444
N. Islam et al. / Scientia Horticulturae 103 (2005) 441–451
Fig. 1. Micrographs of flower bud development stages (0–9) of E. grandiflorum ‘Fuji Deep Blue’. 0: vegetative stage, apical dome with youngest pair of leaf primordia (L)
on the opposite side of the apex; 1: initiation of five sepal primordia (S); 2: sepal (s) elongation; 3: initiation of five petal primordia (P); 4: petal elongation; 5: initiation of
five stamen primordia (St); 6: stamen elongation; 7: initiation of gynoecium with carpel primordia (C); 8: elongation of carpel primordia; 9: visible bud ( 2 mm diameter).
Stages 0, 1, 3, 5–8, Bar = 0.5 mm; Stages 2, 4, 9, Bar = 1 mm.
 N. Islam et al. / Scientia Horticulturae 103 (2005) 441–451 445

For dissection under the microscope, two plants from each of the 11 treatments were
sampled every second day, starting 91 days after seeding and continued until visible bud
(stage 9, Fig. 1). To determine the developmental stages, all the visible leaves were
detached from the plants manually, and other developing young leaves removed surgically
under a binocular microscope until the leaf or flower primordia were observed. To expose
the inner floral organs, the outer ones had to be removed. The developmental stages were
divided in nine stages and they are defined in Fig. 1.

2.3. Flower and shoot characteristics at anthesis

All plants in the experiments were monitored daily for appearance of visible
bud (2 mm in diameter observed by the naked eye) and first open flower. Fourteen plants
were harvested per treatment at anthesis of third flower and the following data were
collected: node to first flower, stem diameter (approximately 5 cm above the medium
surface), number of branches on the stem, number of flower buds and flowers, and shoot dry
weight.

2.4. Data analysis

The effect of the light conditions and time to transfer to long day were subjected to a
two-factorial analysis of variance (general linear model, GLM, in Minitab, version 13) with
single plants as replicates. The plants kept continuously in SD were not included in the
analysis. The two cultivars were analysed separately.

3. Results

Eustoma developed pentamerous flowers centripetally with the sepals appearing first
followed by the petals, stamens and pistils (Fig. 1). LD (LL) from 6 and 8 weeks after
seeding enhanced the onset of floral primordia in ‘Fuji Deep Blue’ (stage 1, Fig. 1) with 10
days compared to continuous SD conditions (Fig. 2). Extending the period in SD until 10 or
12 weeks after seeding gave a progressive delay and finally no difference was observed
between constant SD conditions and transfer 14 weeks after seeding. Compared to a low
light integral (LL), a high integral (HL) enhanced the onset of floral primordia by at least 8
days and the delay in relation to the SD control was therefore, at least 18 days. A transfer to
HL up to 10 weeks after seeding promoted the onset strongly, while SD conditions up to
transfer at 12 and 14 weeks clearly delayed the process. Continuous SD delayed the
formation of the floral organs slightly compared to LL and HL, while no significant
difference between the two LD conditions (Fig. 2).
For plants grown until flowering, time to visible bud was delayed by 21 and 15 days
under SD compared to those transferred to LL (LD) 6 weeks from seeding for ‘Echo Blue’
and ‘Fuji Deep Blue’, respectively (Table 1). Increasing the time period in SD gradually
decreased the difference, but a transfer at 14 weeks still caused a deviation from SD of 7–11
days. A HL compared with LL enhanced time to visible bud with 12 days in both cultivars
and the delay was of course in general reduced with an increasing time period in SD before
446 N. Islam et al. / Scientia Horticulturae 103 (2005) 441–451

Fig. 2. Effect of light regimes and length of SD period before LD on floral development of E. grandiflorum ‘Fuji
Deep Blue’. Error bars indicate S.E. No error bars means either that only one plant represent the data point (9 and
11 points for LL and HL, respectively) or S.E. = 0.

transfer to one of the two LD conditions. No significant difference was found in the time
period required after visible bud to further develop into an open flower.
Plants grown at constant SD developed more nodes below the first flower than those
grown under LL (LD), and the number of nodes was increased with increasing time period
in SD (Table 1). HL also decreased the number by 0.7–1.7 nodes compared to LL. The
strongest effect of day length was found in ‘Echo Blue’, while the effect of light integral
was almost the same in the two cultivars tested.
The stem diameter was higher under constant SD conditions than for plants transferred
to LD, 6 weeks after seeding (Fig. 3). Compared to those transferred to LD this early, an
increasing period in SD increased the diameter and transfer to HL gave thicker stems than a
transfer to LL. Continuous SD conditions delayed the time to anthesis, and therefore,
 N. Islam et al. / Scientia Horticulturae 103 (2005) 441–451
Table 1
Effect of different light regimes and time to transfer of seedlings from short day (SD) to long day combined with either low (LL) or high light integral (HL) on time from
seeding to visible bud (VB) and on number of nodes to first flower in E. grandiflorum ‘Echo Blue’ and ‘Fuji Deep Blue’
Light regimes Number of weeks in SD before transfer to different light regimes
‘Echo Blue’ ‘Fuji Deep Blue’
6 8 10 12 14 6 8 10 12 14
Days from seeding to VB SD 146 – – – – 142 – – – –
LL 125 122 128 133 135 127 125 127 129 135
HL 113 112 116 125 131 115 113 117 122 127
Mean of LL and HL 119 b 116 a 121 c 129 d 133 e 121 b 119 a 122 b 126 c 131 d
Number of nodes to first flower SD 14.8 – – – – 12.8 – – – –
LL 11.0 10.9 11.1 12.6 13.7 10.5 10.4 10.5 10.8 11.5
HL 10.3 10.1 10.4 10.9 12.5 9.4 9.6 9.6 9.7 11.0
Mean of LL and HL 10.6 a 10.5 a 10.8 a 11.8 b 13.1c 10.0 a 10.0 a 10.1 a 10.3 a 11.2 b
See Section 2 (Materials and methods) for specifications of the light conditions. Effect of light integral (LL and HL) and time to transfer and their interaction were all
statistically significant at P  0.001 except for the interacting effect on no. of nodes to first flower for both ‘Echo Blue’ (P = 0.032) and ‘Fuji Deep Blue’ (not significant).
Different letters indicate differences among means according to Tukey’s method (a = 0.05).

447
448 N. Islam et al. / Scientia Horticulturae 103 (2005) 441–451

Fig. 3. Effect of light regimes and length of a SD period before LD on stem diameter, shoot dry weight and
number of flowering buds and flowers of E. grandiflorum ‘Echo Blue’ and ‘Fuji Deep Blue’. For the light regimes
(see Section 2). Standard error bars smaller than the symbols are not shown. Effect of light integral (LL and HL)
and time to transfer and their interaction were all statistical significant at P  0.001 except for the interacting effect
on shoot length for both ‘Echo Blue’ (not significant) and ‘Fuji Deep Blue’ (P = 0.076).

clearly gave a higher shoot dry weight than those transferred to LD at 6 weeks (Fig. 3). A
prolonged period in SD before transfer to LD, especially up to 12 and 14 weeks, increased
the dry weight and the effect was most apparent under high light integrals. The shoot dry
weight was strongly correlated with the stem diameter in both cultivars (r = 0.90 and 0.81
for ‘Echo Blue’ and ‘Fuji Deep Blue’, respectively). There was no significant effect of light
regimes on percent dry matter of the shoots (data not shown).
The number of buds and flowers was positively correlated with the number of branches
(r = 0.72–0.80) for the two cultivars. Both characters are again linked to the shoot dry
weight and follow the same response pattern to the different growing conditions;
 N. Islam et al. / Scientia Horticulturae 103 (2005) 441–451 449

continuous SD or a prolonged period in SD before transfer to LD increased the number for

branches per plant and the total number of flowers and flower buds. A positive effect of an
increasing period in SD was most pronounced under HL.

4. Discussion

The onset of flower bud formation in Eustoma in our experiment was promoted by LD
(LL) compared to SD conditions, even though flowers eventually were initiated under SD.
Our results are in agreement with those of Zaccai and Edri (2002) that described the same
effect of day length on floral transition. The enhancing effect of LD was also indicated by a
smaller number of nodes to first flower. Grueber et al. (1984) have also found that an
increasing photoperiod hastened flowering and reduced the total number of nodes to first
flower in Eustoma. On the other hand, Halevy and Kofranek (1984) found that photoperiod
had no effect on flowering and the species was categorized as a day-neutral plant.
Further development of floral organs after formation of the sepal primordia was only
slightly enhanced by LD compared to SD, but when counting the number of days to visible
bud seen by the naked eye, a clear promoting effect of LD was observed. Street and Öpik
(1984) stated that the photoperiod required for flower bud initiation may differ from the
optimal photoperiod for flower development, or the specific photoperiod may be of less
importance for further flower development, as reported by Tsukada et al. (1982) in
Eustoma and Bredmose (1997) in rose. We used a light source high in far-red for day
extension in LL, which promotes flowering in several LD plants (Thomas and Vince-Prue,
1997). This may have increased the differences between SD and LL. Taken together, our
results indicate that Eustoma is a facultative long-day plant.
Corr and Katz (1997) have found that crop time of Eustoma is affected by light intensity
and this is in agreement with the enhancing effect of HL compared to LL we found on
flower bud initiation. Grueber et al. (1984) have also reported that increasing light intensity
hastened flowering. However, even though initiation of flower buds started earlier in HL
than LL, there was no difference in time until all organs were formed. The far-red rich light
source used for day extension LL may have masked a possible promoting effect of HL on
floral formation.
Ohkawa et al. (1991) have found that 6-weeks-old seedlings of the cultivar Fukushihai
were no longer sensitive to high temperatures when it comes to affecting rosette formation.
There was no or little promoting effect of transferring plants after 6 as compared to 10
weeks from seeding. Looking at the number of nodes to first flower and time to visible bud
it is also clear that there was no effect of transferring plants to LD before 8–10 weeks on
flowering time. Our results are also in agreement with the study in the LD plant Gypsophila
by Islam and Willumsen (2001), where a 3-week SD-treatment delayed the flowering time.
Constant SD, and increasing the length of the SD period before LD, increased the
number of flowering buds and flowers compared to those transferred to LD, 6 weeks after
seeding. These findings disagree with a study by Tsukada et al. (1982) who found that SD-
treatment retarded development of lateral shoots compared to LD in Eustoma. Higher
number of branches could be a direct effect of SD or an effect of day extension light high in
far-red under LL, as it is known to inhibit lateral branching in several plants (Cathy and
450 N. Islam et al. / Scientia Horticulturae 103 (2005) 441–451

Campbell, 1975; Grimstad, 1987). Similarly, fewer branches were observed under LL than
under HL conditions, which could also be influenced by the far-red day extension light.
However, an effect of an increased light integral is likely to have affected the flowering
responses through an increased energy and dry matter production. However, there observed
a thicker stem with increasing period in SD before transferring the plants to long days and
high light integral (HL).
In conclusion, the results indicate that Eustoma is a quantitative long-day plant and long
day conditions promotes flowering already 6–10 weeks after seeding even though a high
light integral compensate for an extended period in SD of 2 weeks. High light intensities
reduce the time to flower transition and enhance the vegetative development as a basis for a
rich flowering response.

Acknowledgements

We wish to thank the Norwegian Research Council and the Norwegian Education Loan
Fund for financial support. We are grateful to Dr. Peter Adams for his useful comments and
critical review of this paper.

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