Sciemia Horticulturae, 48 ( 199 i ) 17 I - 176 171

Elsevier Science Publishers B.V., A m s t e r d a m

Effects o f air temperature and time on rosette

formation in seedlings of Eustoma grandiflorum
(Raf.) Shinn.

K. Ohkawa, A. Kano, K. Kanematsu and M. Korenaga

Department of Biological Science. Faculty of Agriculture, Shizuoka University, 836 Ohya, Shizuoka
422, Japan
(Accepted 23 January 1991 )

ABSTRACT

Ohkawa, K., Kano, A., Kanematsu, K. and Korenaga, M., 1991. Effects o f air temperature and time
on rosette formation in seedlings ofEustoma grandiflorum (Raf.) Shinn. Scientia Hortic., 48:17 !-
176.

Seedlings of Eustoma grandiflorum (Raf.) Shinn. were induced by temperatures above 2 5 C to

grow as rosettes. The season had an additional u n k n o w n effect, especially on plants grown in the
intermediate temperature range. Exposing germinating seedlings to high temperatures for as little as
3 days reduced stem elongation, and treatment for more than 14 days completely inhibited elongation
even when planls were subsequently grown at lower temperatures.

Keywords: bolting; Eustoma grandijTorum; rosette; temperature.

INTRODUCTION

Eustoma grandiflorum (Raf.) Shinn. is a North American wild flower, na-

tive to the American states of Wyoming, South Dakota, Nebraska, Colorado,
Kansas, Oklahoma and Texas as well as northern Mexico (Shinner, 1957;
Wood and Weaver~ 1982 ). Eustoma was introduced into Japan more than 60
years ago for ornamental use, and since then many floral colors have been
developed, including its native color, purple, as well as pink, white, and pink-
or purple-edged white (Halevy and Kofranek, 1984). More than 100
E. grandiflorum cultivars were listed in Japanese seed catalogues in 1990. In
Japan, over 70 million cut flowers were sold in 1989. However, about 80% of
the total Japanese production was concentrated from June to September with
very little in the autumn or winter season. To harvest the flowers during the
winter months, the seeds must be sown from June to August. However, ro-
sette growth occurs and flowering stem elongation delays until the following
spring.

0304-4238/91/$03.50 1991 Elsevier Science Publishers B.V. All rights reserved.

172 K. OHKAWA ET AL.

Although no data are shown, R o h ( 1988 ) suggested that air temperatures

of 2 I. l / 15.5 C ( d a y / n i g h t ) and a soil temperature o f 24 C for 30 days after
sowing, followed by 15.5C at night for the following 60 days could avoid
such rosette formation.
We conducted the present study to investigate the relationship between ro-
sette formation o f young seedlings a n d high t e m p e r a t u r e regimes as well as
the age or stage o f seedling development.

MATERIALS AND METHODS

All experiments were conducted using the cultivar 'Fukushihai" in 1988 and
1989 under natural day length ( m a x i m u m and m i n i m u m day length were
15.5 and I 1.0 h d a y - i ) conditions. E x p e r i m e n t I h a d 128 plants, a n d Exper-
iments II, III a n d IV had 32 plants, all with four replications. Each experi-
mental plot was fertilized with 25 g o f coated long life fertilizer ( 1 4 N - 1 2 P -
14K) and irrigated by mist for 8 weeks after sowing and by h a n d afterwards.
Seven weeks after sowing, the seedlings were transplanted, after which shoot
elongation was checked every week. Plants h a v i n g internodes longer than 3
m m were j u d g e d as 'bolted plants'.
The data were recorded for 18 or 19 weeks after sowing. Unless otherwise
noted, all experiments were carried out inside natural-light phytotrons where
the m a x i m u m and m i n i m u m day lengths were 15.5 a n d I 1.0 h d a y - ~.

Experiment I. m To test at what temperature rosette plant growth could be

induced, seeds were sown on J u n e 6 and October 17, 1988 and February 4,
1989 at 2 3 / 1 8 C , 2 8 / 2 3 C or 3 3 / 2 8 C .

Experiment H . ~ To d e t e r m i n e at what age plants could still be induced to

form rosettes by high temperature exposure, seeds were sown at 2 3 / 1 8 C on
June 9, 1988. After 2, 4, 6 or 8 weeks, seedlings were transferred to a 3 3 / 2 8 C
phytotron.

E~periment I I I . m To d e t e r m i n e how long plants had to be exposed to high

temperature in order to induce rosette growth, seeds were sown on October
17, 1988 at 3 3 / 2 8 C . After 3, 5, 7, 9, 11 or 14 days, plants from each experi-
mental plot were m o v e d to another phytotron r o o m at 2 3 / 1 8 C .

Experiment I V . ~ Seeds were sown on March 16, 1989 and g e r m i n a t e d at 25,

30 or 35 C with continuous i l l u m i n a t i o n (35.0 lLmol m -2 s - ~ photosynthet-
ically active radiation ) by fluorescent lamps. They were then transferred to a
greenhouse, 3, 5, 7, 9 or l I days later. The greenhouse was m a i n t a i n e d at a
d a y / n i g h t temperature o f 25 _+3 / 2 0 _ 3 C, respectively. This e x p e r i m e n t was
EFFECTS O F AIR T E M P E R A T U R E A N D T I M E ON E U S T O M A ROSETTE FORMATION 173

designed to determine the effects of different temperatures and durations of

exposure to the temperatures on induction of rosette growth.

RESULTS

Experiment L Effect o f growth temperature on bolting, m More than 90% of

the seedlings at 2 3 / 1 8 C bolted regardless o f the time of sowing. At 2 8 / 2 3 C
the bolting rate varied between 30 and 90%, depending on the season. Some
90 to 95% of seedlings grown at 3 3 / 2 8 C formed rosettes (Fig. 1 ).

Experiment IL Effect o f duration o f low temperature exposure on bolting. - -

The bolting percentage increased as the duration of exposure to 2 3 / 1 8 C in-

creased. Phnts exposed to 6 weeks at 2 3 / 1 8 C after sowing had a similar
bolting percentage to those never exposed to 3 3 / 2 8 C . Seedlings grown for 6
weeks at 2 3 / 1 8 C had four completely developed leaves at the end of the
period, and the fifth and sixth leaves were visually identified (Fig. 2 ).

Experiment IH. Effect o f high temperature duration on bolting. - - The bolting

percentage for 18 weeks after sowing is shown in Fig. 3. More than 90% of the

I00 100

8O ~ 80
.9
60 -~ 6o
o'l
c

20 2O

0 0 ~
0 8 9 10 Ii 12 13 14 15 16 17 18 08 9 io i 12 13 14 15 16 17 18

Weeks after sowing Weeks after sowing

100
B
80 A:Sown on June 9
6
B:So%~ on October 17
60 C:Sown cn Vebruary 4
c
40 9---- 2 3 / 1 8 o c
m 28/23C
20 o 33/28Oc

0
0 8 9 i0 Ii 12 13 14 15 16 17 18
Weeks after sowing
Fig. 1. Percent o f bolting plants as affected by different temperatures.
174 K. OHKAWA ET AL.

100

80
dO
60

40
r-~
O
m 20

0 v

0 2 4 6 8 conti-
~UOUS
Weeks after sowing

Fig. 2. Effect o f initial duration of 2 3 / 1 8 C before being transferred to 3 3 / 2 8 C on bolting.

Standard error bars smaller than the symbols are not shown.

100

80

"~ 4o
~ 2o
0 -
0 3 5 7 9 ii 14

Time in high temperature (days)

Fig. 3. Effects o f d u r a t i o n s o f 3 3 / 2 8 C temperatures on bolting. Standard error bars smaller

than the symbols are not shown.

plants bolted when continuously exposed to 2 3 / 1 8 C conditions. For the

seedlings which were exposed to the high temperatures ( 3 3 / 2 8 C), the bolt-
ing percentage of the plants was lower, depending on the duration at the high
temperature. As an example, when the plants were kept for 14 days at
3 3 / 2 8 C, no shoot elongation was observed.

Experiment IV. Effect o f temperature and duration on bolting. - - Scant rosette

formation was observed at 25C and most of the seedlings bolted. Only 5
days at 30C decreased the bolting percentage. Similar results were obtained
for the 35C treatment (Fig. 4).
Early germination was observed at 25 and 30C with a final germination
rate of about 80%. At 35C it was about 70% (data not shown).
EFFECTS OF AIR TEMPERATURE A N D T I M E O N EUSTOMA R O S E T T E F O R M A T I O N 175
I00

do 80

60

j_}
40
O
2O

0 I i I ~ I I
0 3 5 7 9 II

T i m e in h i g h t~nDerature (days)
25'C zL 30oc ~ 35oc

Fig. 4. Effects of different temperatures and their durations on bolting. Standard error bars
smaller than the symbols are not shown.

DISCUSSION

Very young seedlings of E. grandiflorum generally form rosettes. They nor-

mally start shoot elongation after three leaf pairs (three nodes) have ex-
panded. However, when seedlings are exposed to high temperatures, the ro-
sette growth habit continues to develop without shoot elongation. When
seedlings were maintained in a 3 3 / 2 8 C phytotron, they continued to de-
velop leaves on a compressed stem, forming a rosette growth habit for more
than 21 months (K. Ohkawa, unpublished data, 1988).
Rosette formation of the young seedlings was induced by high tempera-
tures, such as 33/28 C, and few plants bolted when they were continuously
grown under such conditions after sowing. On the other hand, at 2 8 / 2 3 C
induction of bolting depended on the date of sowing. This is possibly because
of higher leaf temperature in the summer than in the winter due to the differ-
ent irradiance levels. The bolting rate of seedlings at 2 3 / 1 8 C was always
higher than 90%, regardless of the sowing time. From this, we concluded that
the main cause of the rosette induction was the high temperature, and a small
change in temperature influenced the bolting percentage when the tempera-
ture was moderately high, such as 28/23C. It was also possible that some
other factors, such as the irradiance level, day length, and stress caused by
transplanting, could have a minor influence on inducing the rosette state, al-
though none of them had been proven to affect rosette induction. It also seems
that the difference in bolting rates among the different cultivars could be sig-
nificant at this temperature regime (Ohkawa, 1988 ).
The seeds of E. grandiflorum were sensitive to high temperature immedi-
ately after they were hydrated, and, during the following two weeks plants
were sensitive to high temperature. Similar results have been reported using
Limonium sinuatum Mill plants, which induced rosette growth at 2 7 / 2 2 C
176 K. OHKAWAET AL.

immediately after sowing and continued such growth for 6 months, regardless
ofthe day length (Shillo and Zamski, 1985).
High temperature induced rosette growth until seedlings produced an av-
erage of two leaf pairs (two nodes). After this stage, the reaction of seedlings
to high temperature depends on their temperature histories. In other words,
high temperature enhances rosette leaf formation of seedlings already in the
rosette state, whereas it stimulates shoot elongation of plants which have never
been exposed to the high temperature and, therefore, are not in the rosette
state.
Seedlings must be grown under conditions with an average temperature of
about 25 C and with a minimum temperature of 20C or lower from the time
of sowing until two true leaf pairs expand to prevent the rosette induction.
The first 2 weeks after seed hydration are critical, and temperature control
during this period is important.

ACKNOWLEDGEMENT

This study was supported by a Grant-in Aid for Scientific Research from
the Japanese Ministry of Education (No. 01560030).

REFERENCES

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Ohkawa, K., 1988. Effect of temperature and light intensity on the rosette formation uf
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Soc. Holt. Sci., pp. 578-579 (in Japanese).
Roh, M.S., 1988. Growing alternatives. Suitable as a potted plant, this new Eustoma variety is
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Shillo, R. and Zamski, E., 1985. Limonium sinuatum. In: A.H. Halevy (Editor), H a n d b o o k of
Flowering 3. C R C Press, Boca Raton, FL, pp. 292-301.
Shinner, L.H., 1957. Synopsis of the genus Eustoma. The Southwestern Naturalist, 2: 38- 43.
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