GERMINATION OF ASYSTASIA GANGETICA L.

SEED WITH
SPECIAL REFERENCE TO THE EFFECT OF AGE ON
THE TEMPERATURE REQUIREMENT FOR
GERMINATION1
ERNEST K. AKAMIN E
Received January 30, 1947

Introduction
Freshly harvested seed of Asystasia gangetica L., an ornamental shrub,
does not germinate well under ordinary conditions. This seed normally
requires a storage period of several months at room temperature before it
germinates. The purpose of this investigation is to determine the require-
ments for germination of the seed at different ages.
The literature on the use of alternating temperatures to break the dor-
mancy of seeds is voluminous, and no attempt will be made here to refer to
all of the reported cases. HARRINGTON (2) extensively studied the use of
alternating temperatures to increase the germination of seeds of grasses
and garden crops. MORINAGA (3) increased the germination of Bermuda
grass seed with alternating temperatures. TOOLE (4) also used alternating
temperatures to improve the germination of other grass seeds. It has been
the experience of the author that the germination of seeds of some of the
grass species grown on Hawaiian ranges is materially improved by the use
of alternating temperatures (1).
Methods /
Seeds used in the investigation reported herein were harvested at
maturity from Asystasia plants grown on the Hawaii Agricultural Experi-
ment Station grounds. Curing was effected by storing the seeds at room
temperature conditions until they were used for germination tests.
Germination tests were conducted in Petri dishes using filter paper
moistened with distilled water. Ovens and cold storage chambers were
used for temperature variations. Temperatures employed consisted of:
6.60-10.20, 15.00, 20.8°-28.0° (room temperature), 29.90-33.2o, and 36.00 C.
and combinations of these for the alternating temperatures. For tempera-
ture alternations, the Petri dishes with the seeds therein were transferred
from one temperature to another at regular intervals. The schedule of
daily alternating temperatures listed in table I was followed, and except
in some unavoidable cases, the daily 8-hour period extended from 8 A.M. to
4 P.M. and the 16-hour period from 4 P.M. to 8 A.M. the following day. The
number of seeds used for the germination tests varied from 25 to 100 seeds
per treatment. In a few cases where the seed supply was sufficient, re-
plicated tests were conducted.
1 Published with the approval of the Director as Technical Paper no. 155 of the
Hawaii Agricultural Experiment Station.
603
604 PLANT PHYSIOLOGY

Results
Results of many tests to increase the germination of freshly harvested
seed of Asystasia indicated that temperature has a marked influence on
germination and that the alternating temperature treatment is the most
effective. At continuous temperatures the dormant seed does not germinate
as well as at alternating temperatures. Whatever temperature treatment
is necessary to break the dormancy, a minimum temperature of approxi-
mately 200 C. must be provided for actual germination.
Removal of the seedcoat of normally dormant seed produces a germina-
tion of approximately 75%c at room temperature, whereas the intact seed
produces no germination or at best a very low germination. The maximum
germination produced by this treatment, however, is not so high as the
TABLE I
SCHEDULE OF DAILY ALTERNATING TEMPERATURE TREATMENTS

PERIOD AT PERIOD AT
TEMPERATURE ALTERNATION LOWER HIGHER
TEMPERATURE TEMPERATURE
°C. hours hours
6.6-10.2 to 15.0 8 16
6.6-10.2 to 20.8-28.0
(room temperature) 8 16
6.6-10.2 to 29.9-33.2 8 16
6.6-10.2 to 36.0 8 16
15.0 to 20.8-28.0 8 16
15.0 to 29.9-33.2 8 16
15.0 to 36.0 8 16
20.8-28.0 tb 29.9-33.2 16 8
20.8-28.0 to 36.0 16 8
29.9-33.2 to 36.0 16 8

germination (about 90% and over) obtained by the use of optimum alter-
nating temperatures.
After performing a number of germination tests with seeds harvested
at different times, it became apparent that seeds of different ages respond
differently to the various temperature treatments. Attempts were then
made to determine the temperature requirement for maximum germination
at different ages (since harvest). A complete tabulation of all of the data
is not included in this report, but a compilation of some of the results ob-
tained in these trials is recorded in table II and a resumse of all tests con-
ducted on different lots of seed harvested at various times is presented in
table III which shows the temperature requirement for germination of seed
of different age groups.
From tables II and III it is seen that for germination after one week of
curing, the fresh seed requires a severe temperature treatment, and as the
seed ages, this treatment becomes less and less severe'until firally the seed
requires no special treatment and germinates at room temperature (20.8°-
28.00 C.). When the seed is only two to seven days old, it requires an
 AKAMINE: GERMINATION OF ASYSTASIA 605

TABLE II
EFFECT OF TEMPERATURE VARIABLES ON THE GERMINATION OF Asystasia gangetica L.
SEED OF DIFFERENT AGES

GE
OIP
SEED SINCE TEMPERATURE TREATMENT ~~~~~~~~No.oFp
~
SEEDS USED NATION IN
GERM

HARVEST* ~~~~~~~~~~~TWO
WEEKS
days °C. %
Alternating 6.6-10.2 to 20.8-28.0
(room temperature) 100 51.7
it
6.6-10.2 to 29.9-33.2 100 67.2
it
6.6-10.2 to 36.0 100 51.7
It
15.0 to 20.8-28.0 100 86.0
7 " 15.0 to 29.9-33.2 100 88.0
15.0 to 36.0 100 63.0
Continuous 20.8-28.0 100 1.7
Alternating 20.8-28.0 to 29.9-33.2 100 84.7
"i 20.8-28.0 to 36.0 100 93.2
Alternating 6.6-10.2 to 29.9-33.2 75 91.9
6.6-10.2 to 36.0 75 100.0
Continuous 20.8-28.0 75
14 Alternating 20.8-28.0 to 29.9-33.2 75 30.0
"i 20.8-28.0 to 36.0 75 60.0
Continuous 29.9-33.2 75 0.0
"i 36.0 75 26.1t
Alternating 6.6-10.2 to 29.9-33.2 100 97.4
6.6-10.2 to 36.0 100 68.4
15.0 to 20.8-28.0 100 93.0
20 15.0 to 29.9-33.2 100 94.0
15.0 to 36.0 100 76.0
20.8-28.0-to 29.9-33.2 100 44.7
20.8-28.0 to 36.0 100 50.0
Alternating 6.6-10.2 to 20.8-28.0 100 94.7
6.6-10.2 to 29.9-33.2 100 90.0
6.6-10.2 to 36.0 100 84.2
28 Continuous 20.8-28.0 100 5.2
Alternating 20.8-28.0 to 29.9-33.2 100 79.7
"i 20.8-28.0 to 36.0 100 87.5
Continuous 29.9-33.2 100 12.1
" 36.0 100 29.3t
Alternating 15.0 to 20.8-28.0 100 99.0
58 " 20.8-28.0 to 29.9-33.2 lOO 56.0
20.8-28.0 to 36.0 100 35.0
Alternating 15.0 to 20.8-28.0 50 87.0
84 Continuous 20.8-28.0 50 8.0
Alternating 20.8-28.0 to 29.9-33.2 50 84.0
"i 20.8-28.0 to 36.0 50 92.0
Continuous 20.8-28.0 25 16.7
Alternating 20.8-28.0 to 29.9-33.2 25 91.7
112 " 20.8-28.0 to 36.0 25 89.0
Continuous 29.9-33.2 25 0.0
36.0 25 41.7t
Alternating 15.0 to 29.9-33.2 25 91.7
Continuous 20.8-28.0 25 96.8
140 Alternating 20.8-28.0 to 29.9-33.2 25 100.0
Continuous 29.9-33.2 25 41.7
"i 36.0 25 50.0 t
* Seed cured and stored at room temperature conditions.
t Even though germination at 36.00 C. was superior to that at 29.90-33.20 C., it is
probable that the higher temperature is excessive for germination of the seeds continu-
ously exposed to it because injury to the seeds was apparent at this temperature.
606 PLANT PHYSIOLOGY
alternation between room temperature and 36.00 C. for germination. When
the seed is 8 to 14 days old, it requires a more drastic treatment-an alterna-
tion between the lowest and the highest temperatures used. This differ-
ence in the germination requirement in the two age groups is probably due
to the fact that in one case the seed-especially the seedcoat-is not fully
cured, thus requiring a less severe treatment, whereas in the other, the seed
is fully cured, thus requiring more extreme temperatures in the alternation.
When the seed is older than 14 days, it requires less and less drastic treat-
ment for germination as the seed ages. Finally when the seed is 135 days
old and older it germinates normally at room temperature.
TABLE III
THE OPTIMUM TEMPERATURE CONDITIONS FOR MAXIMUM GERMINATION AT THE VARIOUS
AGES OF THE SEED OF Asystasia gangetica L.

AGE OF
SEED SINCE TEMPERATURE REQUIRED FOR MAX]'IMUM GERMINATION
HARVEST*
days OC.
2 to 7 Alternating 20.8-28.0 (room temperrature) to 36.0
8 to 14 " 6.6-10.2 to 36.0
15 to 26 " 6.6-10.2 to 29.9-33.2
27 to 36 " 6.6-10.2 to 20.8-28.0
37 to 82 " 15.0 to 20.8-28.0
83 to 88 " 20.8-28.0 to 36.0
89 to 134 " 20.8-28.0 to 29.9-33.2
135 to 168 Continuous 20.8-28.0 or alternatingg 20.8-28.0 to 29.9-33.2
* Seed cured and stored at room temperature conditions.

Discussion
Since removal of the seedcoat promotes germination at room tempera-
ture, though not to a degree of alternating temperatures, it seems that the
cause of dormancy in the freshly harvested seed of Asystasia mainly lies
in the nature of the seedcoat. The difference in germination percentage
resulting from the removal of seedcoat and the use of alternating tempera-
tures is probably due to the more favorable influence of the alternating
temperatures on the respiration process of the seed. The seedcoat readily
allows passage of water, but it probably restricts the passage of gases in-
volved in respiration. Removal of the seedcoat might eliminate this ob-
struction to gas movement.
A higher germination with alternating temperatures than with con-
tinuous temperatures was obtained in dormant Asystasia seed (table II).
HARRINGTON (2) states that the beneficial effect of alternating temperatures
on germination is due not to the specific effect of the extreme temperatures
of the alternation or to the mean temperature of the alternation but to the
changes in the temperatures. Siniee removing the seedcoat allows for
germination, it seems probable that the alternating temperatures render
the intact seedcoat more permeable to essential materials required for
germiiination.
 AKAMINE: GERMINATION OF ASYSTASIA 607
Since the severity in the temperature alternation necessary for germi-
nation decr_a$eis with the increase in age of thq seed (tables II and- 111), it
seems that normally the change in permeability comes about through the
aging (after 7 days) of th, .seedeoat. -Perhaps simultaneously the after-
ripening process proceeds as the seed ages. Thus with the aging of the seed,
the severity df the temperature treatment required for germination de-
creases until finally when the seedcoat is no longer an obstruction to essential
materials the seed germinates under ordinary conditions.
Summary
1. Freshly harvested seed of Asystasia gangetica L. does not germinate
well at room temperature.
2. After a period of approximately 135 days in ordinary storage, it
germinates readily at room temperature.
3. Seeds between the ages of 8 and 134 days require an alternation of
temperatures for germination, the severity of which is related to the age of
the seed-the younger the seed the more severe the required treatment and
vice versa.
The author is gratefully indebted to DR. HARRY F. CLEMENTS, Plant
Physiologist, Hawaii Agricultural Experiment Station, for his aid in the
preparation of the manuscript and to H. KAMEMOTO of the Horticulture
Department for making the seed available.
DIVISION OF PLANT PHYSIOLOGY
HAWAII AGRICULTURAL EXPERIMENT STATION
UNIVERSITY OF HAWAII
HONOLULU 10, T. H.

LITERATURE CITED
1. AKAMINE, E. K. Germination of Hawaiian range grass seeds. Hawaii
Agr. Expt. Sta. Tech. Bul. No. 2. 1944.
2. HARRINGTON, G. T. Use of alternating temperature in the germination
of seeds. Jour. Agr. Res. 23: 295-333. 1923.
3. MORINAGA, T. Effect.of alternating temperatures upon the germination
of seeds. Amer. Jour. Bot. 13: 141-158. 1926.
4. TOOLE, V. K. Germination of seed of vine-mesquite, Panicum obtusum,
and plains bristle-grass, Setaria macrostachya.. Jour. Amer. Soc.
Agron. 32: 503-512. 1940.