Arm. Bot.

51, 661-668, 1983 661

Viability and Storage of Pollen of the Oil Palm,

Elaeis guineensis Jacq.
S. N. R. EKARATNE* and the late S. SENATHIRAJAH
Department of Botany, University of Colombo, Colombo, Sri Lanka

Accepted: 28 January 1982

ABSTRACT
The effects of drying pollen of the oil palm, Elaeis guineensis, and storage under various conditions, on its
viability have been examined. Viability was based on the ability of pollen to germinate on a sucrose medium
with and without added boric acid, and on its capacity to set fruit when applied to receptive female flowers.
Oven-drying at 37 °C for 2-8 h followed by storage in a deep freezeT proved to be the best method of storing the
pollen. Pollen treated in this way and stored for 12 months was capable of producing fruits which were equal
in weight and appearance to those produced by fresh pollen.

Key words: Elaeis guineensis, oil palm, pollen, storage, viability.

INTRODUCTION
The oil palm, Elaeis guineensis Jacq., is tropical in distribution, being found in the wild
and cultivated in the equatorial regions of Africa, Asia and South America. Its fruit
provides two oils of commercial importance, palm oil and palm kernel oil, from the
mesocarp and kernel respecitvely, and it is by far the highest yielding/oil crop in the
world (Hartley, 1970). '
The oil palm is cross pollinated, the pollinating agent usually being the wind.
Inflorescences on adult oil palms appear as males, followed by females, or vice versa.
It is unusual to find ripe male and receptive female inflorescences on a palm at the same
time. It has been shown that pollen falling on female inflorescences as much as 6 days
before they become receptive is capable of effecting a good fruit set, but that pollen
remaining on an inflorescence for more than 6 days rapidly loses its viability (Hardon
and Turner, 1967). Thus, if fresh pollen can be stored in a viable state, it can be applied
to the female flowers at the correct time and an increased yield may be obtained. This
artificial application of pollen is referred to as assisted pollination. Assisted pollination
is known to reduce loss of crop due to inadequate pollination which may cause bunch
abortion. It is also known to increase bunch weight although it may decrease the number
of bunches (Hartley, 1970). Storage of pollen in a viable state is also important for
breeding programmes (Hartley, 1970).
In Sri Lanka, at present, the demand for palm oil far outstrips its supply. The major
determining factors for this state of low supply is the limited extent of land presently under
oil palm cultivation, and the need for assisted pollination. It is this last mentioned factor
that led us to study the most suitable and practical methods of pollen storage.

MATERIALS AND METHODS

Site of sample collection
Samples were collected from the State Oil Palm Plantation situated at Nakiadeniya, Sri
Lanka. Male inflorescences with flowers in which the anthers had just begun to dehisce
were selected.
• Present address: The Library, Postgraduate Institute of Medicine, Kynsey Road, Colombo 8, Sri Lanka.
0305-7364/83/050661 +08 $03.00/0 © 1983 Annals of Botany Company
662 Ekaratne and Senathirajah—Viability and Storage of Pollen

Treatment of sample material

Pollen from the male inflorescences was shaken on to a clean paper, sieved through
a 0-5 mm sieve to free the pollen grains from foreign matter and other debris. It was then
divided into portions and subjected to the following treatments:
(1) undried,
(2) oven-dried at 37 °C for 2, 4, 6, 8, 10 and 12 h,
(3) sun-dried for 4 h (sun-drying was performed only for 4 h due to the frequent
absence of continuous sunlight for longer periods).
Each sample after treatment was placed in a dry McCartney bottle. Representative
samples from each of the treatments were stored as follows:
(1) deep freezer (-15 °C),
(2) desiccator with silica gel as desiccant (29-75 °C),
(3) desiccator with 50 per cent aqueous glycerol as desiccant (29-5 °C).
McCartney bottles containing pollen samples to be stored in the deep freezer were
sealed in double polythene bags to ensure that the pollen grains were not exposed to high
humidity. A control sample of undried pollen contained in McCartney bottles was left
at room temperature (29-5-30 °C).

Testing for percentage germination

In this study, percentage germination, i.e. emergence of the pollen tube, was considered
to indicate percentage viability.
In testing percentage germination, the pollen grains were allowed to germinate in
drops of 10 per cent sucrose in 0- 5 per cent agar placed on a microscope slide and enclosed
in a covered Petri dish with moist cotton wool to maintain a high humidity. Germination
counts were taken 2 h from the time of transference of the pollent to the germination
medium. Each pollen sample was tested in duplicate and approximately 250 pollen grains
from each sample counted on each occasion.
Percentage germination of each set of pollen stored under the conditions described
above was tested once a fortnight except in the case of undried pollen stored at room
temperature which was tested every 2-3 days.

In vivo pollination with dried stored pollen

Mature female inflorescences which had been bagged 6 days before receptivity were
pollinated with pollen which had been oven-dried for 2-8 h and stored in a deep freezer
for a period of 12 months. After pollination the inflorescences were bagged again for
3 days to prevent any contamination with fresh pollen and also the possible loss of test
pollen by wind and rain. Thereafter the bags wereremovedand the test bunches allowed
to develop.
The extent of fruit set was observed periodically and the bunch weights determined
at maturity.

Effect of boric acid on pollen germination

An attempt was made to enhance the process of germination by supplementing the
medium with boric acid. Sucrose/agar media containing 0005, 001, 002 and 005 per
cent boric acid were prepared (Vasil, 1964). In these studies selected pollen samples were
allowed to germinate as described above. The percentage germination was evaluated in
each case.
 Ekaratne and Senathirajah—Viability and Storage of Pollen 663

RESULTS
Viability of pollen with storage
Figure 1 shows the time course of decrease in the percentage viability of undried pollen
stored under natural conditions at room temperature. It is evident from these results that
after the fifth day, reduction in percentage viability is pronounced and the pollen falls
below 50 per cent viability on about the seventh day. Pollen stored under these conditions
loses its viability fully by the 12th day.

100
\

80

£ 60

40

20

5 10 15
Doy of storoge

Fio. 1. Percentage viability of undried pollen during storage at room temperature (29-5 to 30 °C).

Figure 2 shows the decrease in percentage viability with time for the undried
and dried pollen samples stored under the various conditions. It is seen that the decrease
in percentage viability was lowest when the pollen was oven-dried for 6 h.
Under all storage conditions the undried pollen lost viability very rapidly when
compared with the dried samples. Figure 3 shows that the viability of undried pollen was
prolonged when it was stored under humid conditions. Among the three different storage
conditions, it is evident that the deep freezer is the best means of pollen storage
(Figs 3, 4).
Pollen oven-dried for 2, 4, 6, 8 or 10 h and sun-dried for 4 h showed similar losses in
viability and in all these cases the fall in percentage viability with time was very slow,
particularly with pollen stored in the deep freezer. When stored in the desiccator with
silica gel as desiccant, the dried pollen (oven- and sun-dried) maintained a greater than
50 per cent viability up to about 50 days, after which there was a rapid decrease in
percentage viability with complete loss of viability by about the 115th day. Dried pollen
grains stored in the desiccator with 50 per cent glycerol as desiccant were dead by
approximately the 30th day. The pollen grains when dead were wrinkled and their
diameter was reduced to about 2-9 pm in comparison with normal pollen which has a
diameter of about 4-3 /on. Undried pollen stored in the desiccator with 50 per cent
glycerol was very moist and was infected with a fungus by the 18th day.
664 Ekaratne and Senathirajah—Viability and Storage of Pollen
Undried
100
80
60
40
20
0 R
Oven dried at 37°C for 2 h
100
80
60
40
20
0
Oven dried at 37°C for 4 h
100
80
60
40
20
0 SL.
Oven dried at 37°C for 6 h
too

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Oven dried at 37°C for 8 h

100
80
60
40
20
0
Oven dried at 37°C for 10 h
100
80
60
40
20

Oven dried at 37°C for 12 h

100
80
60
40
20
0 01
Sun dried for 4 h
lOOi
80
60
40
20

5 5 5 10 10 10 30 30 30 52 52 52 75 75 75 87 87 87 125 125 125

Day of storage

FIG. 2. Changes in percentage viability of undried, oven-dried (2, 4, 6, 8, 10 and 12 h) and sun-dried
(4 h) pollen samples, under different storage conditions. D , Deep freezer; £3, desiccator with silica
gel; • , desiccator with SO per cent glycerol.
 Ekaratne and Senathirajah—Viability and Storage of Pollen 665

100

2 60-

20-

40 60
Doy of storoge
FIG. 3. Changes in percentage viability of undried polkn stored in the deep freezer (#), desiccator
with silica gel ( • ) and desiccator with SO per cent glycerol (A)-

100

20 40 60 80 140
Doy of storoge

FIG. 4. Changes in percentage viability of pollen oven-dried at 37 °C for 6 h and stored in the deep
freezer (#), desiccator with silica gel ( • ) and desiccator with 50 per cent glycerol (A).

In vivo pollination
Mature femaleflowerswithreceptivestigmas when pollinated with pollen oven-dried
for 2-8 h and stored in the deep freezer for 12 months gave bunches having a weight of
8-6±0-4 kg. Control bunches had a comparable weight of 80±0-6 kg. Appearance and
texture of the fruit was similar to the control in all respects.
666 Ekaratne and Senathirajah—Viability and Storage of Pollen

Effect of boric acid on pollen germination

Results obtained from the experiment are summarized in Table 1. These results indicate
that boric acid causes an increase in percentage germination of oil palm pollen at
concentrations of 001 and 002 per cent. At these two concentrations the time for
initiation of germination of pollen was accentuated by 2 and 4 min respectively. At a
concentration of 004 per cent boric acid proved to be inhibitory to the germination of
viable oil palm pollen.

TABLE 1. Effect of boric acid on pollen germination

Pollen Sample
Percentage Percentage
Treatment and Time of germination germination
Percentage of method of Day of observation in the control in the medium
boric acid storage storage (h) medium with boric acid

0005 Oven-dried for 2 h and 9 0-5 30-2±0-76 28-5±0-63

in freezer
0005 Oven-dried for 6 h and 9 0-5 61-5±0-45 62-1 ±0-32
in freezer
001 Oven-dried for 6 h and 10 0-5 56-3 ±0-49 6O-5±O-21*
in freezer
001 Oven-dried for 6 h and 10 0-5 4-2±015 9-5±0-34*
in desiccator with
silica gel
001 Undried and in 14 0-5,2 00 00
desiccator with
50% glycerol
002 Oven-dried for 6 h and 18 0-5 50-7±0-32 6-4 ±0-6*
in freezer
002 Oven dried for 6 h and 18 0-5 2-8±0-22 9-3±0-19*
in desiccator with
silica gel
004 Oven dried for 6 h and 18 0-5 52-5 ±0-38 00
in freezer
004 Undried and in 18 0-5,2 00 00
desiccator with
silica gel

Significant at 5 per cent significance level.

DISCUSSION

The fact that dried pollen (i.e. oven-dried at 37 °C for 2-12 h and sun-dried for 4 h)
maintained viability for longer periods than the undried pollen indicates that removal
of a certain amount of moisture helps the pollen to remain viable during storage. Also,
it is evident from the results that drying for longer periods such as 10 and 12 h is less
satisfactory because the pollen grains tend to lose viability at a faster rate when compared
with those that are dried for less than 10 h. This may be attributed to deleterious effects
of desiccation or the effects of the accumulated temperature sum. The existence of an
optimal drying time may be due to the fact that removal of moisture beyond a certain
critical level may be unfavourable as the protoplasm becomes dehydrated.
The results of this study indicate that the best method of treatment before storage of
oil palm pollen appears to be oven-drying for 2-8 h. The sun-dried samples (4 h),
however, gave results very close to that of the samples oven-dried at 37 °C for 10 h. Thus
 Ekaratne and Senathirajah—Viability and Storage of Pollen 667
sun-drying for 4 h is not as satisfactory as oven-drying for 2-8 h and may be advisable
only in the absence of oven-drying facilities. Drying in an oven may give better results
because it ensures uniform drying over the required length of time.
Of the three methods of storage used in this study, deep freezer conditions proved to
be very efficient in maintaining the viability of pollen during storage as compared with
the desiccator with silica gel and desiccator with 50 per cent glycerol. This is in
accordance with data recorded by Henry (1959), Devreux and Malingraux (1960),
Kernick (1961) and Purseglove (1972), that viability of oil palm pollen can be maintained
for prolonged periods if it is stored at low temperatures such as 5 °C. When undried pollen
was stored in the desiccator with silica gel it remained viable for 2-3 weeks while the
dried pollen was viable for periods of up to 3 months. It is apparent that both temperature
and moisture content affect pollen viability in the oil palm.
Although Darlington and La Cour (1942) mention that pollen can be stored in
desiccators with humidity controlled at 50 per cent (glycerol in water) it was not found
to be successful here. Even dried pollen lost its viability very rapidly under these
conditions, ending in loss of viability by about the fourth week. Further, the presence
of excess moisture and fungal attack in the undried pollen stored under these conditions
reveal that the moisture level in this environment is unsuitable for pollen storage.
It has been recorded that pollen grains of many plants are deficient in boron (O'Kelley,
1957), but in nature this deficiency is often met by the high levels of boron in the stigmatic
and stylar tissues (Bertrand and Silberstein, 1938; Bobko and Zerling, 1938a,6; Gartel,
1952). Pollen grains of some plants, such as Capsicum annuum, Corchorus capsularis and
Crotalariajuncea, yield optimum germination and optimum pollen tube length in sucrose
solutions alone, and the addition of boric acid either inhibits germination and tube length
or has no effect at all. These plants may therefore have sufficiently high natural levels of
boron and any additional supply proves ineffective or inhibitory (Vasil, 1964). In certain
other plants, however, specific concentrations of boric acid have a stimulatory effect on
pollen germination. Boric acid (001 per cent) stimulates pollen germination in Dolichos
lablab (Vasil, 1964), Lilium bulbiferum L., Cucumis sativus, Solanum citrullifolium (Bobko
and Zerling, 1938a), while 0015 per cent boric acid is stimulatory in Brassica nigra
T. 257 and Pennisetum typhoideum. Higher concentrations of boric acid (002 per cent)
proved stimulatory in Cucumis melo but toxic in Pennisetum typhoideum (Vasil, 1964).
Oil palm pollen too shows a significant response to boric acid at concentrations of 001
and 002 per cent. However, germination was inhibited at a concentration of 004 per
cent boric acid.
In this study viability of dried stored pollen wasfirstdetermined through its ability to
germinate in an artificial medium. In vitro elongation of the pollen tube is not the perfect
criterion for viability; good fruit set would be the ideal. According to Broekmans (1957)
good fruit set may be obtained when pollen viability as shown by laboratory tests is as
low as 10 per cent. He is also of the view that actual germination of pollen is higher on
stigmas than on artificial media. This may be a contributory factor to the results of our
field tests with stored pollen (12 months old). Bunch size and appearance were
comparable in all respects to that of the control. The results of this in vivo test confirm
that oil palm pollen can be stored for a period of 12 months at least, provided the pollen
is dried and then kept dry, and especially if it is also stored at —15 °C.

ACKNOWLEDGEMENTS
Our thanks are due to Professor R. N. de Fonseka of the Department of Botany,
University of Colombo for providing us with all facilities to conduct this study, and to
Mr Rex H. Perera and the staff of the State Oil Palm Plantation for supplying the material,
and assisting in the pollination of female inflorescences.
668 Ekaratne and Senathirajah—Viability and Storage of Pollen

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19386. Vliianie bora na reproduktivnoe razvitie rasentenii. Zh. Bot. USSR, 23, 3-11.
BROEKMANS, A. F. M., 1957. Studies of the factors influencing the success of controlled pollination of the oil
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DARLINGTON, C. D. and LA COUR, L. F., 1960. The Handling of Chromosomes, 248 pp., 3rd edn. George Allen
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HENRY, P., 1959. Prolongation de la viabilite du pollen chez Elaeis guineensis Jacq. C.r. hebd Seanc. Acad.
Sci., Paris 248, 772-4.
KERNICK, M. D., 1961. Agronomy. In Agricultural and Horticultural Seeds. FAO Agricultural Studies, no. 55,
531 pp. Food and Agricultural Organization of the United Nations, Rome.
O'KELLY, J. C , 1957. Boron effects on growth, oxygen uptake and sugar absorption by germinating pollen.
Am. J. Bot. 44, 239-44.
PURSBGLOVE, J. W., 1972. Tropical Crops - Monocotyledons, vol. 2, 607 pp. Longman, London.
VASL, I. K., 1964. Effect of boron on pollen germination and pollen tube growth. Int. Symp. Pollen Physiol.
Fert. Univ. Nijmegen, The Netherlands, August 1963, pp. 107-19.