Production of Male Flowers on Rambutan (Nephelium lappaceum L.

) Trees in
Hawai'i
Andrea M. Kawabata, Mike A. Nagao and Laura K. Awong
University of Hawaii at Manoa, Beaumont Agricultural Research Center, 875 Komohana
St. Hilo, HI 96720
ABSTRACT
Production of deformed rambutan fruits has been attributed to poor pollination
and an insufficient amount of male flowers in Hawai'i orchards. The objectives of this
study were to monitor the development of hermaphroditic functionally male (HFM)
flowers on trees of the Silengkeng cultivar, determine if naphthaleneacetic acid (NAA)
could induce development of HFM flowers on cultivars grown in Hawai'i, identify the
stage at which floral buds are responsive to NAA, and document the viability of pollen
from HFM flowers. Results showed that with the exception of Silengkeng, cultivars
planted in Hawai'i produced few or no HFM flowers for pollination. Silengkeng
panicles produced large numbers of HFM flowers throughout the anthesis period which
made it a suitable pollinator cultivar. NAA applied to the R7, R9, R134, R162,
R167, R156 Red, R156 Yellow, Binjai, Jitlee and Rongrien panicles composed
of predominantly hermaphroditic functionally female (HFF) flowers stimulated
development of HFM flowers within 6 days after treatment. By 12 days after treatment,
production of HFM flowers was decreased. Only floral buds at a stage in development
where the apex of the pistil began protruding through the unopened sepals were
responsive to NAA. Pollen obtained from naturally produced HFM flowers on
Silengkeng panicles and from HFM flowers produced after NAA treatment was viable
and germinated within 24 hours after incubation in a culture medium. Incorporating
Silengkeng and male trees in the orchard or treating HFF panicles at the appropriate
stage of development can be strategies for increasing male flowers and fruit set and
reducing the development of fruit without arils.
KEYWORDS: flowering, fruit set, NAA, parthenocarpic fruit.
INTRODUCTION
Poor fruit set and development of fruits that lack an aril has been a recurring
problem in many rambutan orchards in Hawai'i. Studies in affected orchards showed that
exclusion of insect pollinators during anthesis resulted in development of a high
percentage of these deformed fruits and indicated that insufficient pollination was the
cause of the problem (Nagao et al., 2002). Production of deformed fruits was similar to
reports from Thailand where parthenocarpic type fruits fail to develop fully and contain
little aril (Kosiyachinda and Salma, 1987). During rambutan flowering, 3 types of
flowers are produced, and trees are classified into 3 groups based upon flower
morphology (Tindall et al., 1994); Valmayor et al., 1970). One group produces only
staminate male flowers, which lack a functional ovary and possess 5 to 7 well-developed
stamens with anthers that dehisce and shed pollen at anthesis. A second group produces

only hermaphroditic functionally female (HFF) flowers which set fruit but do not shed
pollen. At anthesis HFF flowers have a well-developed ovary with a prominent bifid
stigma extending beyond stamens that possess anthers which do not dehisce. A third
group of trees will produce panicles with both HFF and hermaphroditic functionally male
(HFM) flowers. The HFM flowers closely resemble HFF flowers, but have well
developed stamens often extending beyond an under-developed pistil. Anthers on these
flowers dehisce and shed pollen.
A shortage of male flowers results in poor fruit set and development of
parthenocarpic type fruits that contain little aril (Kosiyachinda and Salma, 1987;
Salakpetch, 2005). Establishing male trees into the orchard, suspending male panicles
from branches during flowering and using napthaleneacetic acid (NAA) to increase male
flower development have been used to aid pollination in Thailand (Salakpetch, 2005).
Application of NAA to panicles with HFF flowers during early anthesis has been shown
to induce development of HFM flowers, and is essential for fruit set and production in
some Thailand growing locations (Kosiyachina and Salma, 1987; Tindall et al., 1994).
Experiments with Jitlee and Binjai in Hawai'i have also shown that 90 mg/L NAA
stimulated production of HFM flowers (Nagao et al., 2002), however, the response of
other cultivars planted in Hawai'i and the stage of development at which individual
flowers are responsive to NAA have not been determined. Also the viability of pollen
from NAA induced HFF flowers has not been documented.
Most cultivars produce a high percentage of HFF flowers and a low percentage of
HFM flowers which may amount to only 0.05 to 0.9% of the flowers found on a panicle
(Nakasone and Paull, 1998; Tindall et al., 1994). Some orchards in Hawai'i have
established the Silengkeng cultivar to aid in pollination, but its ability to produce HFM
flowers under Hawai'i growing conditions has not been documented. The objectives of
this study were to monitor development of HFM flowers on Silengkeng trees, to
determine if NAA could induce development of HFM flowers on cultivars grown in
Hawai'i, to identify the stage at which floral buds are responsive to NAA treatments and
to document the viability of pollen produced on HFM flowers.
MATERIALS AND METHODS
HFM flower development on Silengkeng panicles. To observe occurrence of
HFM flowers, 22 panicles distributed among four Silengkeng trees were randomly
selected in an orchard in Onomea, HI (115 m elevation) in 2003 July. Trees were at least
12 years-old, and each panicle was monitored from the onset to the termination of
anthesis by observing the number of newly opened HFM flowers at 3 to 4 day intervals.
Response of cultivars to NAA. Individual panicles were sprayed to runoff with a
hand-held sprayer containing distilled water or an aqueous solution of 90 mg/L potassium
salt of NAA (K+NAA). Treatments were applied when approximately 10% of the
flowers on the panicle were at anthesis based on a visual assessment of each panicle. The
experiment was conducted between 2004 June to August in orchards at Onomea,
Kurtistown, (250 m elevation) and Keaau, HI (75 m elevation). A minimum of 15
replicate panicles distributed on at least 3 trees of each cultivar were treated. Because
flowering among rambutan trees in Hawai'i is non-synchronous (Kawabata et al., 2007),
for an individual cultivar an equal number of treated and control panicles was sprayed on

each tree, but the number of replicate panicles/tree was variable. Table 1 shows the
cultivars, location of the experiments, number of replicate panicles treated, number of
replicate panicles/tree, and number of trees treated. The number of newly-opened HFM
flowers was recorded at 0, 4, 6, 8, 10, and 12 days after treatment.
Stage of flower development responsive to NAA. In 2004 August, 18 replicate
panicles equally distributed on 3 Jitlee trees in Onomea, HI were sprayed with 0 or 90
mg/L K+NAA to run-off with a hand-sprayer. Individual flowers on each panicle were
monitored by marking a section of the panicle with indelible ink and photographing the
flowers at 0, 3 and 6 days after treatment to identify the stage of flower development
responsive to NAA treatment.
Pollen viability of Silengkeng and NAA induced HFM flowers. Binjai, R156
Red, Jitlee, Rongrien, R7, R9, R134, R167, R162, and R156 Yellow
panicles were treated with 90 ml/L K+NAA between 2004 August and September. At 7
days after treatment, panicles were removed from the tree and transported to the
laboratory in enclosed polyethylene bags containing moistened paper towels. Pollen
from 5 HFM flowers produced on untreated Silengkeng panicles and from treated
panicles of each cultivar was tested for germination. Flowers with dehisced anthers were
removed from each panicle. A drop of germination medium was placed on a microscope
slide and pollen grains were removed from 2 anthers per flower and placed in the drop of
germination medium. The medium consisted of a modified Brewbaker and Kwack
(1963) solution containing 50 ppm H3BO3, 150 ppm Ca(NO3)2.4H2O, 100 ppm
MgSO4.7H2O, 50 ppm KNO3, and 5% sucrose. The bottom of 100 x 10 mm glass Petri
dishes was fitted with filter paper moistened with distilled water. The slide was placed
on the moistened filter paper, the dish covered, and incubated for 24 to 48 hours at
22.8C. Pollen was considered viable, if pollen germination occurred and pollen tubes
were visible within 48 hours.
RESULTS AND DISCUSSION
Anthesis on individual Silengkeng panicles occurred over a 17 to 161 day
period, and the mean number of days for duration of anthesis for panicles was 81 days.
Figure 1 shows that production of HFM flowers occurred at highest frequency during the
initial third of the anthesis period, with smaller numbers produced during the remaining
anthesis period. At the peak of HFM flower production, approximately 84 HFM flowers
were observed on each panicle interspersed with HFF flowers. The duration of flowering
for Silengkeng panicles was much longer than observed on other cultivars planted in
Hawai'i (Kawabata et al., 2007) and in Thailand (Tongumpai et al., 1980), and production
of HFM flowers on Silengkeng panicles was more numerous than reported for other
cultivars. Studies in the Philippines and Malaysia (Tindall et al., 1994) showed that the
Maharlika and Seejonja cultivars also produced both HFM and HFF on the same
panicle. Approximately 99.94% of the flowers on Maharlika panicles were functionally
female, while the remaining flowers were functionally male. The percentage of HFF
flowers on Seejonja panicles was 99.55% at bloom. Tindall et al. (1994) also reported
that generally between 200 to 800 HFF flowers are borne on each panicle and that HFM
flowers make up about 0.05 to 0.9% of the flowers. The high frequency and the

continued production of HFM flowers on Silengkeng panicles over the entire anthesis
period confirm that Silengkeng may serve as a suitable source of pollen for fruit set.
All cultivars treated with 90 mg/L NAA produced HFM flowers by 4 to 6 days
after treatment (Table 2). HFM flower production was greatest between 6 and 8 days
after treatment and progressively decreased at 10 and 12 days. Since the appearance of
HFM flowers decreased at 12 days after NAA treatment, re-application to adjacent
panicles will be necessary to sustain HFM flower production throughout the anthesis
period which can extend over several weeks (Kawabata et al., 2007). Within each
location some cultivars tended to have a greater response to NAA. At Onomea,
Rongrien produced the greatest number HFM flowers (124.7 flowers per panicle) while
R156 Red produced the fewest HFM flowers. Elevation also may have had an effect
since the R9 cultivar at Kurtistown was not as responsive as at Onomea.
Since male trees do not produce fruits, some growers in Thailand tend to remove
these trees, and the result is an insufficient amount of pollen available to fertilize the HFF
flowers. NAA concentrations between 40 and 160 mg/L are then used to insure fruit set
and are essential for production in some areas (Kosiyachina and Salma, 1987; Lam and
Tongumpai, 1987). Sprays are applied to panicles that are 1.0 meter apart and when onethird of the flowers on the panicle are open (Tindall et al., 1994). HFM flowers are
visible 5-7 days later. After the pollen is shed from NAA treated panicles, panicles are
pruned off and newly emerging panicles will produce normal HFF flowers (Salakpetch,
2005).
HFM flowers were not produced on the control panicles except for R156 Yellow
and R134 at Onomea, and in both cultivars natural production of HFM flowers was
very low (Table 2). The small number of HFM flowers produced by these cultivars
without NAA is probably too few to adequately pollinate the large number of HFF
flowers.
Figure 2 shows Jitlee flower buds on the day of treatment and at 3 and 6 days
after NAA application. Buds on which the apex of the pistil began protruding through
the unopened sepals responded to NAA and developed into HFM flowers that were at
anthesis by 6 days after treatment. HFF flowers that were at partial or full anthesis and
buds that were tightly closed at the time of treatment did not respond to NAA. These
buds later developed into HFF flowers or abscised from the panicles. Untreated HFF
flowers at anthesis possess a pistil topped with a bifid stigma which is surrounded by 6
short stamens arising from the base of the bilocular ovary. Stamens do not extend
beyond the stigma, and anthers do not dehisce and brown by the second day after
anthesis. In HFM flowers, NAA stimulated elongation of the stamens and promoted
anther dehiscence while suppressing pistil development. NAA application to individual
panicles should be timed to coincide with a period when there is an abundance of floral
buds that are at a stage responsive to NAA. At 10% anthesis, panicles possess numerous
floral buds at this stage of development.
Germination was observed with pollen obtained from naturally occurring HFM
flowers on Silengkeng panicles and from HFM flowers produced after NAA treatment
for all cultivars tested. Germination of pollen grains and growth of pollen tubes could be
seen 16 to 48 hours after pollen was incubated in the germination medium (Figure 3).
There appeared to be a population effect during pollen germination, because germination
percentage tended to be lower when fewer pollen grains were incubated in the medium.

Germination tests confirmed that viable pollen was produced by naturally occurring HFM
flowers from Silengkeng panicles and from NAA induced flowers. Our attempts to
germinate pollen taken from anthers on HFF flowers were unsuccessful and support the
conclusion that HFF flowers only function as female flowers (Tindall et al., 1994).
Bagging experiments in Malaysia and Hawai'i to exclude pollinators reinforce the
importance of pollination for rambutan production since poor pollination will result in
reduced fruit set and production of deformed fruit (Lim, 1984; Nagao et al., 2002). With
the exception of Silengkeng, most cultivars in Hawai'i produce very few or no HFM
flowers to adequately supply pollen in a large orchard. Treating panicles with NAA will
increase production of viable pollen when applied to flower buds at the appropriate stage
of development. Based on our observations, smaller, more compact panicles seemed to
be less responsive to NAA treatment compared to large, more elongated panicles since
some smaller panicles did not produce HFM flowers after NAA treatment. Rambutan
flowering is initiated following a 2-4 week period of drought stress, and flowering
intensity is thought to be closely related to intensity and duration of the water stress
(Nakasone and Paull, 1998; Whitehead, 1959). Our observations indicate that during the
cooler flowering season (April to May), smaller compact panicles are more common than
during the warmer flowering season (July to August) in Hawai'i. Therefore selection of
suitable panicles for treatment as well as the timing of NAA application will be important
for an effective NAA response.
ACKNOWLEDGEMENTS
We gratefully acknowledge the support of Onomea Orchards and Plant-It Hawaii
for the use of their orchards. This research was supported with grants from the Hawaii
Department of Agriculture and the Hawaii Farm Bureau Federation.

LITERATURE CITED
Brewbaker, J.L. and B.H. Kwack. 1963. The essential role of calcium in pollen
germination and pollen tube growth. Am. J. Bot. 50(9):859-865.
Kawabata, A.M., M.A. Nagao, T. Tsumura, D.F. Aoki, K.Y. Hara and L.K. Pena. 2007.
Phenology and fruit development of rambutan (Nephelium lappaceum L.) grown
in Hawai'i. J. Hawaiian Pacific. Agric. 14:31-39.
Kosiyachinda, S. and I. Salma. 1987. Changes in rambutan during growth and
development. In: Rambutan: Fruit Development, Postharvest Physiology, and
Marketing in ASEAN. Kuala Lumpur, Malaysia, 16-27.
Lam, P.F. and P. Tongumpai. 1987. Preharvest factors affecting postharvest quality of
rambutan. In: Rambutan: Fruit Development, Postharvest Physiology, and
Marketing in ASEAN. Kuala Lumpur, Malaysia, 27-32.
Lim, A.L. 1984. The reproductive biology of rambutan, Nephelium lappaceum L.
(Sapindaceae). Gard. Bull. Sing. 37(2):181-192.
Nagao, M.A., H.M. Leite, A.M. Kawabata, and A.Y. Terada. 2002. Abstract: Update of
tropical fruit research in Hawaii. In: Proceedings of the 12th Annual International
Tropical Fruit Conference. Keauhou Kona, Hawaii, 8.
Nakasone, H.Y. and R.E Paull. 1998. Tropical fruits. CAB International. Wallingford,
UK.
Salakpetch, S. 2005. Rambutan production in Thailand. Acta Horticulturae 665:67-72.
Tindall, H.D., U.G. Menini and A.J. Hodder. 1994. Rambutan cultivation. Food and
Agriculture Organization of the United Nations. Rome, Italy.
Tongumpai, P., R. Sethpakdee, and B. Silayoi. 1980. Effect of some growth regulators on
sex expression of Seechompoo rambutan. J. of Hort. 15:31-38.
Valmayor, R.V., C.O. Palencia, H.B. Aycardo, and D.B. Mendoza. 1970. Growth and
flowering habits, floral biology and yield of rambutan (Nephelium lappaceum
Linn.). Philippine Agriculturalist 54(7/8):359-374.
Whitehead, C. 1959. The rambutan, a description of the characteristics and potential of
the more important varieties. Malayan Agr. J. 42:53-75.

Table 1. Location and cultivars treated with K+NAA.

Cultivar

Location

Replicate Replicate
panicles panicles/tree

R167
R9
Binjai
Jitlee
R162
R134
R9
R156 Red
Rongrien
R7
R156 Yellow

Kurtistown
Kurtistown
Keaau
Keaau
Keaau
Onomea
Onomea
Onomea
Onomea
Onomea
Onomea

15
15
15
17
22
16
15
18
17
18
16

No. trees
treated

5
5
5
4-8
5-10
2-6
2-5
3-5
5-7
2-8
2-8

3
3
3
3
3
5
4
5
3
4
3

Table 2. Production of HFM flowers on panicles treated with 0 (control) and 90 ml/L
K+NAA at 0, 4, 6, 8, 10, and 12 days after treatment.
No. HFM flowers/panicle

Cultivar

Location

R167

Kurtistown

R9

Kurtistown

Binjai

Keaau

10

12

NAA

NAA

NAA

NAA

NAA

NAA

16.5 5.5a

28.9 9.0

17.6 5.8

7.8 2.7

1.2 0.4

13.7 2.3

5.1 2.1

23.3 10.6

28.5 8.0

19.4 3.7

6.4 1.9

1.3 0.4
15.2 3.1

Jitlee

Keaau

10.5 2.8

65.8 17.1

84.8 13.1

65.0 9.0

R162

Keaau

2.7 1.9

50.8 19.6

40.5 14.5

22.5 9.8

12 5.0

R134

Onomea

0.1 0.1

2.7 1.5

37.0 16.0

36.5 14.4

18.3 9.0

5.6 4.3
9.2 2.1

R9

Onomea

50.6 5.3

51.3 6.2

20.9 4.1

R156 Red

Onomea

4.5 1.7

9.6 2.7

6.7 1.8

4.4 1.6

Rongrien

Onomea

18.5 9.5

106.2 32.8

111.2 24.9

88.9 19.9

41.6 13.0

R7

Onomea

0.6 0.4

27.4 7.3

81.7 18.2

47.3 12.0

17.0 6.3

R156 Yellow

Onomea

0.8 0.3

13.6 5.1

37.3 7.8

67.9 13.5

52.1 11.5

19.8 6.6

Control

Control

Control

Control

Control

Control

R134

Onomea

0.1 0.1

0.3 0.3

0.4 0.3

0.3 0.2

0.1 0.1

R156 Yellow

Onomea

0.2 0.1

0.4 0.3

0.1 0.1

Each value represents the mean SE

Figure 1. Frequency of naturally produced HFM flowers on Silengkeng panicles

observed at 3 to 4 days intervals from the onset to the termination of anthesis. Error bars
represent SE. n=22.

80

60

40

20

0
0
3
7
10
14
17
21
24
28
31
35
38
42
45
49
52
56
59
63
66
70
73
77
80
84
87
91
94
98
101
105
108
112
115
119
122
126
129
133
136
140
143
147
150
154
157
161

No. of HFM Flowers per Panicle

Figure 1. Frequency of naturally produced HFM flowers on Silengkeng panicles

120
observed at 3 to 4 days intervals from the onset to the termination of anthesis. Error bars
represent
SE.
100

Days After Anthesis

Figure 2. Development of Jitlee rambutan flower buds at 0, 3 and 6 days after treatment
with 90 mg/L NAA. Arrows show progressive development of buds responsive to NAA.

0 day

3 days

O day

6 days

3 days

6 days

Figure 3. Germination of pollen from HFM flowers from untreated Silengkeng panicles
(a) and from HFM flowers developing after 90 mg/L K+NAA treatment of Rongrien
panicles (b).