Trichosanthes (Cucurbitaceae) In Malesia
TRICHOSANTHES (CUCURBITACEAE)
IN MALESIA
Oleh
RUGAYAH
Nrp: 945511BIO
PROGRAM PASCA SARJANA
INSTITUT PERTANLAN BOGOR
1999
RINGKASAN
R U G A Y A H. Trichosanthes (Cucurbitaceae) in Malesio @i bawah
bimbigan Prof. Dr. Ir. Edi Guhardja, M.Sr sebagai ketua, Dr. WJ.J.0. d e
Wilde, Prof. Dr. Mien A. Rifai, Dr. Dedy Darnaedi, Dr. Gayuh Rnhayu
sebagai anggota)
Trichosanthes L. merupakan marga terbesar yang tergolong dalam suku
Cucurbituceae. Marga tersebut beranggotakan sekitar 100 jenis, tersebar di Asia
bagian timur, Malesia, Australia tropik sampai ke Fiji. Beberapa jenis anggotanya
mempunyai nilai ekonomi sebagai bahan obat tradisional maupun sayuran, bahkan
Cina telah mengembangkaa T. kirilowii, guna mendapatkan zat bioaktif yang
dapat digunakan untuk menggugurkan kandungan bahkan sebagai bahan anti HIV.
Di Malesia, marga tersebut belum banyak diteliti baik dari segi potensi
ekonomi maupun taksonominya secara menyeluruh. Oleh karena i t - perlu dilakukan revisi secara menyelumh untuk mendapatkan data dasar yang nantinya dapat
digunakan untuk memantapkan batasan takson pada peringkat marga maupun
jenisnya, mengetahui daerah persebaran dan memahami hubungan kekerabatan
antajenisnya, mengevaluasi seksi yang dipertelakan untuk jenis-jenis yang ada di
Cina untuk dipakai sebagai wadah jenis-jenis yang ada di Malesia, serta mencari
data pendukung melalui beberapa pendekatan seperti anatomi dam, sitologi, dan
isozim di samping hasil pengamatan morfologi.
Hasil penelitian ini menunjukkan adanya 39 jenis Trichosanthes di Malesia, 17 di antarmya me~pdCanjenis baru di samping dua anak jenis, dua varietas
and tujuh fonna baru. Jenis-jenis tersebut dapat digolongkan dalam lima seksi
yaitu seksi Cucumeroides (dengan anak seksi Cucumeroides dan Tetragonosperma), Trichosanthes, Foliobracteola, Involucraria (anak seksi Involucraria
clan Pedatae) clan seksi baru Edulis. Data persebaran jenis-jenis Trichosanthes
dapat dikelompokkan menjadi dua daerah subsenter yaitu Bomeo sebagai daerah
subsenter bagian barat Malesia yang terwakili oleh 21 jenis dengan tujuh jenis di
antarmya endemik, dan P. Irian sebagai daerah subsenter lainnya di bagian timur
Malesia yang terwakili oleh 12 jenis, tujuh jenis di antaranya endemik.
Dari hasil pengamatan sitologi delapan jenis dan satu varietas yang ada di
Java, dietahui bahwa Trichosanfhesmempunyai jumlah kromosom somatik yang
sama yaitu, 2n = 22. Data matomi tidak dapat dipakai sebagai ciri penanda jenis,
kecuali pada ketiga jenis yang sangat berdekatan (T. tricuspidata, T pubera dan
T. quinquanguIata)yang memperlihatkan perbedaan pada penampang paradermal
daun pemukaan atas dan bawah. Data SEM kulit biji, mungkin dapat dipakai
sebagai ciri pendukung untuk penciri jenis bahkan untuk seksi. Untuk data isozim,
temyata sistem enzim AAT, ACP, dan PRX, memberikan pita spesifik untuk
jenis-jenis yang berdekatan.
SUMMARY
R U G A Y A H. Trichosanthes (Cucurbitaceae) in Malesia (Under the
supervision of Prof. Dr. Ir. Edi Guhardja, MSc as a chairman, and Dr.
W.J.J.O. de Wide, Prof. Dr. Mien A. Rifai, Dr. Dedy Darnaedi, Dr. Gayuh
Rahayu as members)
Trichosanthes is one of the largest genera belonging to the family
Cucurbitaceae with about 100 species distributed in Eastern Asia, Malesia and
Tropical Australia to Fiji and subtropic Eurasia. The genus includes a number of
species which have economic value because they are used as vegetable and
medicine. In China about 33 species have been used for medicinal herb to induce
aborsion and T. Ririlowii is the one species which have been intensively cultivated
to isolate the bioactive trichosanthin for its HIV-inhibiting property.
In Malesia, a complete monographic study of this genus has never been
undertaken for the whole regions. Therefore a new revision is needed to provide
better understanding of the diversity as well as the relationship of the species and
its distribution, to evaluate the suitability of the delimitation of sections based on
species from China for the Malesian representatives, and to find out other
supporting taxonomic characters derived from leaf anatomy, SEM of seed coat,
cytology, and isozyme.
The result of this study indicated that there are 39 species that can be
recognized in Malesia including 17 new species, two subspecies, hyo varieties and
seven forms. They can be divided into five sections namely, Trichosanthes,
Cueumeroides (with two subsect.: Cucumeroides, Tetragonosprma), Foliobracteola, lnvolucraria (with subsect.: Involucraria, PeJafae), and the new
section Edulis. The pattern of distribution of this genus in Malesia indicated that
there are two subcenter distributions which can be recognized, with Borneo (21
species, seven endemic) as one of the subcenter in West Malesia, and New Guinea
(12 species, seven endemic) as the other subcenter in East Malesia.
From the cytological study it can be concluded that, the somatic metaphase
chromosomes of eight species and one variety (T. borneensis, T. cucumerina var.
anpina, T. montana, T. ovigera, T puberq T. quinqumplaa T. tricuspidata, T
villom, T. wawrae) have the same chromosome numbers of 2n = 22. Leaf
anatomical data cannot be used to identify species, except in the three closely
related species (T. pubera, T. quinquangt~lata,I: tricuspidafa) which show
differentiy on the paradermal section of the upper and lower surfaces. Preliminary
observation on SEM of seed coat indicated that, the data obtained might be usehl
as supporting character for the proposed classification of this genus. Isozyme
analysis using AAT, ACP, and PRX isozyme system indicated that, they can be
used to identify the different species including the three closely related species.
TRICHOSANTHES (CUCURBITACEAE)
IN MALESIA
Oleh
RUGAYAH
Nrp: 945511 BIO
In partial fulfilment of the requirement for
the Doctor degree in the field of Plant Taxonomy
PROGRAM STUD1 BIOLOGI
PROGRAM PASCA SARJANA
INSTITUT PERTANIAN BOGOR
Judul Disertasi
:
Trichosanthes (Cucurbitaceae) in Malesia
Nama
:
RUGAYAH
NRP
:
94551
Program Studi
:
Biologi
Menyetujui
1. Komisi Pembimbing
(Prof. Dr. Ir. ~ dGuhardia.
i
MSc.1
Ketua
(Prof. Dr. Mien A. Rifai)
Anggota
(Dr. Dedv Darnaedi)
Anggota
2. Ketua Program Studi Biologi
(Dr. davuh Rahavu)
Awgota
gram Pascasarjana
a Manuwoto, MSc.)
Tanggal Lulus : 21 Agustus 1999
CURRICULUM VITAE
Rugayah was born on 30 August, 1956 in Sob, Central of Java She spent
her childhood with her parents in Solo until she graduated from Senior High
School in 1975. She continued her study at Gajah Mada University in Yogyakarta
and graduated in 1981.
Since 1982 she has been working at Herbarium Bogoriense, Puslitbang
Biologi-LIPI, Bogor as a researcher in plant taxonomy. In September 1990, she
was admitted to the degree of MSc of Birmingham University (England)
sponsored by the British Council. In September 1994, LIP1 gave her an
opportunity to continue study at IPB for her PhD. She is rnamed and has 2
children.
TABLE OF CONTENS
Page
Ringkasan
Summaty
............................................................................
ii
..............................................................................
iv
Table of Contents
...................................................................
Lists of Tables. Maps and Figures
................................................
...................................................................
Acknowledgement
ix
xi
xiv
Historical Introduction
.............................................................
1
Materials and Methods
.............................................................
5
Result and Discussion ...............................................................
10
General Part
Morphology
Leaf Anatomy
.................................................................
...............................................................
SEM Observation of Seeds Coat Surface ................................
Cytology .....................................................................
Isozyme Analysis
....................................:.....................
Economic Botany
...........................................................
Diversity
....................................................................
Geographical Distribution
Cladistic Analysis
................................................
..........................................................
Special Part
Taxonomic
..................................................................
1. Trichosanthes cucumerina
.............................................
61
66
var. cucumerina.......................................................
68
var. anguina...........................................................
69
2 . T. ovigera
..............................................................
3. T. beccariana
......................................................
78
.....................................................
79
......................................................
80
................................................................
83
4 . T. mucronata
.
5 T. pendula
77
...............................................
subsp.beccariana
subsp.pussila
72
6 . T. kinabaluensis
........................................................
90
7 . T. obscura
...............................................................
92
8 . T. rejf.acza
................................................................
94
9 . T. longespicata
.........................................................
96
10. T. celebica
..............................................................
99
11.T.floresana
............................................................
101
12. T. pqpuana
............................................................ 103
13 . T. elmeri
..............................................................
105
.............................................................
108
forma w m a e
....................................................
111
forma hirsuta
.....................................................
113
..........................................................
113
14. T. wmurae
15. T. ellipsoidea
16. T.philippinensis
...................................................
..............................................................
17.T.pubera
18. T. tricuspidata
.........................................................
115
117
120
forma iricuspidata
................................................
123
forma mperifolia
.................................................
125
forma seramensis
.................................................
125
forma sibemtensis
................................................
126
19. T. leuserensis
........................................................
20. T. quinquangulata
127
....................................................
130
.......................................................
133
..........................................................
.........................................................
135
24. T. globosa
.............................................................
140
25 . T. montana
............................................................
143
21. T.planigans
.
22 T. borneensis
23 . T. emarginata
26. T. sepilokensis
.........................................................
138
146
27 . T. valida
................................................................ 148
28. T.villosa
...............................................................
subsp. villosa
......................................................
subsp. midorensis
29. T. rotundifolia
30. T. auriculata
3 1. T. coriacea
................................................
154
155
......................................................... 156
...........................................................
.............................................................
32. T. schlechteri
152
..........................................................
159
162
166
33 . T. h t a t a
..............................................................
34. T. dieniensis
..........................................................
169
172
.............................................................
forma laeoica ...................................................
173
.................................................
176
...............................................
177
35 . T. laeoica
forma sicyocarpa
forma sorongensis
forma yapenensis
.................................................
. T.pulleana .........................................................
37. T.densiJora .......................................................
36
38. T. dentifera
39. T. edulis
........................................................
.............................................................
176
178
179
180
182
184
var. edulis
..........................................................
var.sativa
.......................................................... 188
var.septembola
.................................................
187
189
Conclusion
............................................................................
191
References
...........................................................................
193
Appendix: General Key to the Maiesian species of Trichosanthes ........... 198
Plates o f Species
.......................................................
213
LISTS OF TABLES, MAPS,
FIGURES AND PLATES
TABLES
1.
Occurrence of stomata and trichome types on leaves of
Trichosanthes spp.
2.
Anatomical characteristic from transverse section of leaves in
Trichosanthes spp.
3.
...........................................................
Somatic chromosomes and their karyotipe formulation of
Trichosanthes spp.
4.
............................................................
...........................................................
Matrix characteristic of isozyme band patterns from PRX ,AAT,
and ACP in Trichosanthes spp.
............................................
...................
5.
List and the distribution of Trichosanthes in Malesia
6.
Morphological characters used in the cladistic analysis of
Trichosanthes
.................................................................
..............
7.
Matrix of morphological characters for cladistic analysis
8.
Characters, number of character states, steps and consistancy index
from morphological characters ...............................................
MAPS
1.
Distribution of Trichosanthes (endemic and non endemic) in
Malesia
2.
........................................................................
Distribution of T. mucronata ................................................
3.
Distribution of T. pendula
4.
Distribution of T. kinabaluensis
5.
Distribution of T. obscura
....................................................
94
6.
Distribution of T. reji-acta
...................................................
96
7.
Distribution of T. longispicata ...............................................
98
8.
Distribution of T. celebica
9.
Distribution of T.floresana
....................................................
.............................................
84
92
................................................... 100
.................................................
102
10. Distribution of T. p a p u a ~.................................................. 104
1 1. Distribution of T. elmeri
....................................................
107
...................................................
110
12. Distribution of T. wawrae
13. Distribution of T. philippinensis
............................................
117
....................................................
119
15. Distribution of T. Ieuserensis
..............................................
128
16. Distribution of T. planiglans
................................................
133
14. Distribution of T. pubera
17. Distribution of T. borneensis ................................................ 137
...............................................
139
19. Distribution of T. globosa
.................................................
142
20 . Distribution of T. montana
.................................................. 145
18. Distribution of T. emarginata
21 . Distribution of T. valida
.....................................................
149
..............................................
158
23 . Distribution of T. auriculata .................................................
160
..................................................
163
22 . Distribution of T. rotundifolia
24 . Distribution of T. coriacea
25 . Distribution of T. schlechteri
...............................................
168
26 . Distribution of T. hastata ....................................................
171
27 . Distribution of T. laeoica ....................................................
175
28. Distribution of T. Jel~~~@'ora................................................. 182
29. Distribution of T. edulis ......................................................
186
FIGURES
1.
Rachis of male flower .......................................................
16
2.
Type of petal of male flower ................................................
19
3.
Fruit type of Trichosmthes spp.................................. .
.. . . . .
22
4.
Seed of Trichosanthes spp...................................................
23
5.
Paradermal section of leaves ..............................................
26
6.
Trichome types on leaves of Trichosanthes spp..........................
27
7.
Transverse section of leaves ................................................
28
8.
SEM of seed coat Trichosanthes spp.......................................
33
9.
Somatic chromosomes at metaphase on T. cucumerina var. anpina, 7:
borneensis, T. montana. T. vilosa ................................ ... . . . .
10.
36
.
Somatic chromosomes at metaphase on T. ovigera T. prrbera. 7:
.............
trinrspidala. 7: quinquangulafa ..................... .
.
.
.
37
11. Explanatory drawing of metaphase chromosomes ......................
38
12. AAT and PRX isozyme band patterns of T. cucumerina var.
anguina ........................................................................
13. Isozyme band patterns from AAT
43
........................................
43
14. Isozyme band patterns from ACP ..........................................
44
15. Isozyme band patterns from PRX
..........................................
16. Cladogram based on morphological character ...........................
44
59
PLATES
1.
T. mucronata ................................................................
213
2.
T. kinabaluensis ...............................................................
214
3.
T. obscura .....................................................................
215
4.
T. longespicata ................................................................ 216
5.
T. celebica ....................................................................
217
6.
Zfloresana ................................................................
218
7.
Kpcqouana
8.
T. ellipsoidea ..................................................................
9.
T. philippinensis .............................................................. 221
10.
7: tricuspidata ................................................................ 222
1 1.
T. tricuspidda forma aspergolia .......................................... 223
12.
T. leuserensis ....................... .
.
.
.
...................................
13 .
T. quinquangulata ..................... .
.
.
.............................. 225
14.
T. planigans .................................................................. 226
15 .
T. emarginata ................................................................
16.
T. gIobosa ..................................................................... 228
17.
.............................. 229
T. montana ...................................
.
18 .
T.vali& ....................................................................... 230
19.
1: villosa subsp. midorensis ...............................................
....................................................................
219
220
224
227
231
20. T. rohrndifolia
21 . T. auriculata
................................................................
..................................................................
22. T. coriacea
....................................................................
23. T. dieniensis
..................................................................
24. T. pulleana
25 . T. densifora
26. T. edulis
....................................................................
..................................................................
.......................................................................
27 . T. edulis var. septembola
....................................................
xvii
ACKNOWLEDGEMENT
I would like to record my thanks to my supervisors: Prof Dr. 11. Edi
Guhardja, (chairman of the advisory committee), Dr. W.J.J.O. de Wilde, Prof. Dr.
Mien A. Rifai, Dr. Dedy Darnaedi and Dr. Gayuh Rahayu (members of the
advisory committee), for their advices, guidance and encouragement throughout
this study.
My thanks are due to Prof. Dr. Ir. Syafiida Manuwoto, MSc (Director of
the Post Graduate Programme, IPB), Prof Dr. Reviani Widjajakusuma (Head of
Biological Study Programme) and Dr. Arie B u d m @ad of the Research and
Development Center for Biology-LIPI), for the opportunity given to me to
undertake this study. I am also grateful to Prof. Dr. Peter Baas (Director of
Rijskherbarium/ H o w Bonaticus, Leiden) and Dr. Alex Hartana
(in
charge of
Biochemical Laboratorium in PAU-IPB) for allowing me to use the excellent
laboratorium facilities. Many thanks have to be extended to Dr. Irawati (Head of
Herbarium Bogoriense) for giving me her support.
I am very gratefull to LIPI, GEF Small Grand Project and Leiden
University for providing financial support.
I highly appreciate the assistence of Dr. Veldkamp in making the Latin
diagnoses of the new taxa, Dr. Ding Hou in translating some Chinese articles into
English, Ms. Bertie Joan van Heuven in preparing the seed coat preparation and
its photograph, Mr. Iskak Syamsudin and Mr. Van 0 s in making the illustrations,
Ms. Titien Ngatinem in preparing the cytological preparation, Ms. Siti Rohajawati
and Mr. Budi Rahman in typing this manuscript.
xviii
I am indebted to Brigitta Dudes-de Wilde for assisting me during the
execution of the research, and Dr. Harry Wiriadinata for his advice and criticism. I
sincerely thank all staffs at Rijskherbarium, Leiden and Herbarium Bogoriense,
for sharing their experties in different fields, and d l post graduate students of
Botany-Taxonomy, IPB for their friendship.
Finally, to my parents (especially to my father who has passed away on
1 5 February
~
1999), my husband Agoes Sriyanto and my daughters Arnalia and
Vinandia, I am grateful for their deep understanding and great patient and for
giving me their moral support.
HISTORICAL INTRODUCTION
Trichosanthes is one of the largest genera belonging to the family of
Cucurbitaceae, with about 100 species distributed in subtropical and tropical
Eastern Asia, Malesia and Tropical Australia to Fiji (Jeffky, 1990); there are 39
species in Malesia (Rugayah & de Wilde, 1999). The genus includes a number of
species which have economic value because they are used as vegetable and
medicine.
Most species of this genus are dioecious, they are climber with delicate
fimbriate flowers, their anthesis is taking place at night until the morning in the
following day (nocturnal). They muently die off after fruiting. These characters
make it difficult to collect the plant in the right and complete condition.
Consequently, some collections may be mixed with fruit and the vegetative part
collected from different plants, or even different species. Some species have been
based on a few specimens only and in some cases they are only known from a
single specimen, either with male flowers, or female flowers, or fruit only.
Trichosanthes was described for the first time by Linnaeus in his Genera
Plantarum in 1737. He mentioned four species namely, T. anguina., T. nervifolia,
T. cucumerina and T. amara. For Java, Blume (1826) enumerated 13 species
under Trichosanthes, three of which belong to different genera. Trichosanthes
costata B1. is Gymnopetalum chineme (Lour.) Merr., whereas T. hexasperma B1.
and T. macrocarpa BI. are Hodgsonia macrocarpa (Bl.) Cogn. Roxburgh (1 832)
treated this genus for India and described six species, four of which were new
ones. Miquel (1856) recognized 23 species of Trichosanthes for the Malesian
area, proposing several new species, some of which, however, are later treated as
synonyms of other species. In Australia, Bentham & Mueller (1866) enumerated
four species. Clarke (1879) in the Flora British India proposed four new species,
recognized eight others.
Cogniaux (1881) was the first author who made an extensive study of this
genus. He treated the genera Poppya Rumph., Cucumeroides Gaertn.,
Involucraria Ser. as synonyms of Trichosanthes. He m t e a monographic
treatment of Trichosanthes in de Candolle's Monographiae Phanerogamarum in
which 40 species were enumerated for the Old World, 12 of which were proposed
as new species: six species from Malesia (Sumatra, Singapore, Borneo, Celebes
and New Guinea), one from Ceylon, four species from India and one from
Australia. He divided this genus into two subgenera based on the flower
characters:
Eutrichosanthes
(with
male
flower
in
a
raceme)
and
Pseudotrichosanthes (with male and female flowers solitary). For Formosa,
Hayata (1921) accepted nine new species of Trichosanthes in lcones Plantarum
Formosanarum. Ridley (1922) described six species in the Malay Peninsula, two
of which were misidentifications. Harms (1925) enumerated seven species for
Papua New Guinea. For the Philippines, Merrill (1923) recognized six species in
An Enumeration ofPhilippines Flowering Plants.
More recent treatments are those of Kitamura & Yoshida (1949) for Japan,
with 12 species (paying special attention to seed characters). For the Indian
subcontinent, Chakravarty (1959) recognized 24 species, two of which were new
species, and he also described two new varieties h m H i i a y a and Kheri. In
1963, Backer and Bakhuizen van den Brink, Jr. in Flora of Java revised
Trichosanthes and accepted eight species. For Cambodgia, Laos and Vietnam,
Keraudren-Aymonim (1975) recognized 10 species and she treated T. pubera and
T quinq1~1ngulaia
as synonyms of T trtcuspidaia. Jeffrey (1 980) recognized 40
species for Eastern Asia, Indomalesia, tropical Australia to Fiji and subtropical
Eurasia in Miocene and Pliocene.
Intensive studies of Trichosanthes were undertaken by the Chinese in
modem times: Yueh and Cheng (1974) distinguished 55 species in their
preliminary investigation for China based on morphological characters,
34
species of which have medicinal properties. They divided the genus into two
subgenera, Cucumeroides and Trichosanthes, which were further divided into two
and five sections respectively, based on leaf, bract, pulp and seed characters. Later
on, Yueh & Zhang (1986) and Huang et al. (1997) used pollen character to
support the delimitation of the sections. Recently, Huang et al. (1998) recognized
37 species and six varieties, based on morphological, cytological, palynological
and anatomical characters. They accepted the two subgenera described by Yueh &
Cheng (1974): subgenus Trichosanthes with three sections (sect. Trichosanthes,
sect. Foliobracteola with subsect. Foliobracteola and subsect. Villosae, and sect.
Znvolucraria),and subgenus Cucurneroides with two sections (sect. Cucumeroides
and sect. Tetragonosperma); they proposed a new series, ser. Ovijolia, under
subsect. Foliobracteola.
In a preliminary treatment of Cucurbitaceae for Flora Malesiana,
Rugayah and de Wilde (1997) revised the genus for Java and accepted 10 species
in this area. They found that materials from Java passed under the name of T.
trifoliata (L.) Merr. needed to be renamed into T. wawrae Cogn., and that T.
anguina L. should be sunk as a variety of T. cucumerina L. They also found that
materials from Java under the name i? bracteata (Lam.) Voigt represented a
mixture of three different species namely T. pubera Bl., T. quinquangulata A.
Gray, and T. tricuspidata Lour.
Therefore there is a need to undertake an indepth taxonomic study in this
genus for Malesia, especially because of taxonomical problems like
misidentifications of T. wallichiana Wight in the Malay Peninsula by Ridley
(1922) and T. bracreata Voigt for Papua by Hanns (1925), and also because many
new materials need to be described. The present study is to revise this genus in
Malesia, to evaluate the suitability and the delimitation of sections as based on
species from China for the Malesian representatives, and to provide better
understanding of the diversity and the relationship of the Malesian species, with
the available extended materials now at hand.
MATERIALS AND METHODS
PLACE OF STUDY
Studies of herbarium specimens were conducted in Herbarium Bogoriense
(BO) of Puslitbang Biologi-LIPI, Bogor; Herbarium of the Royal Botanical
Gardens Kew (K); British Museum, London (BM), and Rijksherbarium, Leiden
(L). Observation on living plants, anatomical and cytological investigations were
undertaken in Herbarium Bogoriense, while the isozymes was analyzed in PAU,
IPB, Bogor. The SEM analysis of the seed coats was undertaken in
Rijskherbarium Leiden.
PLANT MATERIAL
For the present study a total of 969 herbarium sheets, including liquid and
carpological collections of Trichosanthes from several herbaria (BM, BO, BR,
CANB, L, LAE, MEL, P, S, SAN, SING, TNS, USA, U, W) were used to
provide morphological, distribution and economic botanical data for this study.
Living materials of nine species and one variety collected from Java, Sumatra,
Borneo, and Irian were cultivated in the premises of Herbarium Bogoriense, and
used for anatomical, cytological and isozyrne analyses.
Details of the living plant collections used for this study was as follows:
T. borneensis: MR s.n. (Kalimantan); Fanani s.n. (Kalimantan).
T. cucumerina var. anguina: Rugayah 209 (Labuan), 210 (Jakarta), 21 1 (Bogor),
212 (Madiun), 213 (Mad-);
de Wilde & Duyfjes 21673 (Bogor).
T. globosa: Rugayah 115 (Gunung Bunder); de Wilde & Duyfjes 2 1703, 21778
(Gunung Bunder).
T. montana: de Wilde & Duyfjes 21888 (Situ Gunung).
T. ovigera: de Wiide 21B68 (Situ Gunung), 21896 (Gunmg Bunder), 21927
(Gunung H d i u n ) .
T.pubera: de Wilde & Duyfjes 21667 (Cianten), 21840 (Cianten).
T. quinquangulata:de Wilde & Duyfjes 21661 ( h u n g Bunder); Wiriadinata s.n.
(Gunung Halimun); T. Uji s.n. (Bengkulu); Widjaja 7091 (Yapen, Isl., Irian Jaya).
T. tricwpidata: de Wilde & DuyfJes21660 (Gunung Bunder), 21767 (Cibodas),
21773 (Gunung Bunder); Rugayah 214 (Sukabumi); Titin 525 (Banten);
Wiriadinata s.n. (Gunung Salak), 6874 (f. siberutensis, from Sibemt, Sumatra).
T. villosa: de Wilde & Duyfjes 21839 (Cianten). 21883 (Cianten); Wiriadinata
8284 (Batu, Malang).
T. wawrae: Fanani s.n. (Gunung Halimun).
METHODS OF INVESTIGATIONS
Morphology, Distribution and Economic Botany
- Data and information on
morphology, distribution and economic botany were collected from the specimens
and literatures. The procedure for morphological observation followed those
described by Leenhouts (1968), Rifai (1976) and de Vogel(1985).
Anatomy - Leaf anatomy was investigated by first fixing the leaves (small part
of the middle base) in FAA and then embeded them in par&.
Transverse
sections (20 pm thick) were made, stained with safranin and fast green, and then
mounted in canada balsam. Paradermal sections were taken from the upper and
lower surfaces of leaves, then stained with safi.anin 1% in water and then mounted
in glycerin. Descriptions of the trichome and stomata types were based on the
criteria used by Inamdar & Gangadhara (1975,1976).
SEM of Seed Coat - Seeds of some species (T. auriculata, T. beccariana, T.
borneensis, T. cucumerina. var. anguina, T. elmeri, T. emarginata, T. laeoica, T.
montana. I: ovigera, T. pendula, T qquiquangulata, T. rotund~olia, T.
schlechteri, T. tricuspidata,
T.valida,) were soaked in detergent (amaloco) 1% for
one night, then rinsed with aceton for 5-10 minutes. The seed then were cut at the
middle and put on the cylindrical stub using carbon glue and coated with gold
SCD 005 (BAL-TEC) for 4 minutes. The SEM photographs were made by low
pressure JEOL-SEM.
Cytology -Root tips h m the living materials were pretreated with 0.0002 M 8hydroxyquinoline solution and kept in the refrigerator for three hours. The root
tips separated from root caps were fixed in 45% acetic acid for 10 minutes,
macerated in 1 N HCl solution at 60° C for 2-3 minutes, and then squashed in 2%
aceto-orcein solution.
Isozyme
- Enzyme
electrophoresis was performed by horizontal starch gel
electrophoresis using 12% of potato starch h m SIGMA. The procedures
followed those of Wendel & Weeden (1990). About 50 mg pieces of fresh young
leaves were cmshed in approximately 0.5 ml extract buffer consisting of 250 mM
ascorbic acid Na-salt, Tris-HCI buffer (pH 7.9, 0.25 M pvp, 0.1%
Mercaptoethanol, 20 mh4 Diethyldithimbamate, 20 m M sodium metabisulfite,
and 200 m M sodium tetraborate. Filter paper wicks were dipped into the leaf
extract and then inserted into the starch gel. The gel buffer was 5mM L-histidin,
adjusted with NaOH (pH 7.0) or 0.076 M Tris, adjusted with citric acid (pH 8.6).
The electrode buffer was 0.410 M citric-acid, Na2 salt, adjusted with free citric
acid (pH 7.0), or 0.300 M boric acid, adjusted with NaOH (pH 8). Gel were run at
18-22 mA for 3-4 hours. The gel then was sliced horizontally and stained with
three staining enzymes, PRX (peroxidase), AAT (aspartic aminotransferase), ACP
(acid phosphatase). The PRX staining mixer was 0.05 M sodium acetat (pH 5.0),
3-amino-9-etil-carbosol, aceton, 3% HzOz, CaCI2. The AAT staining mixer was
alfa-ketogluteric-acid, L-aspartic-acid, pvp-40, EDTA, Naz salt, sodium
phosphate. The ACP staining mixer was 0.1 M sodium acetat (pH 5.0), MgCIz,
Na-napthyl acid phosphate, fast garnet GBG salt.
Cladistic analysis - Vegetative and generative characters were used for
phylogenetic analysis of Trichosanihes, with Gymnopethalum chinemis and G.
integrifolia used as the outgroups. A total of 46 morphological characters (Table
5) involving habit, indument, stem, tendril, leaves, probract, flowers, fruits and
seeds were accumulated and analyzed using Hennig86 computer programme. For
this analysis, six species (T.dentgera, T. dieniensis, T. ellipsoidea, T. planiglans,
T. pulleana, T. refiacta) were not included because of incomplete material
available.
RESULTS AND DISCUSSIONS
GENERAL PART:
MORPHOLOGY
Habit
The species of Trichosanthes are predominantly perennial climbers, rarely
annual (only T. cucumerinu, including usually the cultivated var. anguina). They
are scandent or creeping herbs, but their habit is often not readily apparent from
the herbarium specimen. The female plants frequently will die off after fruiting.
Some species have a tuber, capable of producing new shoots.
Sexual Conditions
Whether a plant or species is monoecious or dioecious in this genus is not
easy to determine without observing the living plants directly. In this study, the
sexual condition of species was determinated on the base of the presence of male
flowers, or female flowers or fruits on the specimens examined. All species have
unisexual flowers, wholly, partly or incidentally and most species appear to be
dioecious, and only a few are monoecious (T. cucumerina, T. leuserensis, T.
montana, T. quinquangulata).
Indumentum
Plants are generally hairy, with hairs villous, puberulent or strigose or
rarely subglabrous. Some species, however, are glabrescent, the hairs disappear
with age. The colour of the hairs may be grey, white, brown or rusty. Most of the
species have whitish, sometimes brown, or blackish punctate dots originating
from hairs or hair scars on the upper leaf surface. Moreover the upper leaf surface
of some species may be very scabrid. White cystoliths may also occur on stem,
petiole, and nerves of lower leaf surface of several species.
Stem
The Iianescent main stems and branches are slender, the stems of the leafy
shoots grooved or angularly-winged (in T. auriculata. T. kinabaluensis, T.
pendulu, T. pubera), glabrous or pubescent, pale yellow or brown in drymg, rarely
reddish (T. pubera, T. leuserensis).They are thin in some species (T. cucumerina,
T. ovigera). The stem of most species contains colourless sap, odourless except in
the T. cucumerina var. anguina, where it has a strong smell latex.
Leaves
Leaves are arranged alternately, have long petioles, and simple or
compound blades. Simple blades are entire or with 3-5(-7) shallow to deep lobes;
compound leaves with 3 or 5 or 7 leaflets are characteristic of a few species: T.
celebica, T. elmeri, T.floresana, T. papuana, and T. wawrae. Some (or probably
most or all) species are heterophyllous e.g. T. montam and T. globosa of which
leaves in the juvenile stage are f entire but becoming deeply 3-5 lobed in the
adult plant; T. tricuspidata and T. pubera has deeply 5-7 lobed leaves in the
juvenile stage, but only (2)-3 lobed when adult; heterophylly also occurs in T.
celebica and T. wawrae, with simple (but f lobed) leaves in the juvenile stage.
Leaf-blade
- The shape of the
leaves is varying from ovate to orbicular in
outline. Their base is generally deeply (often f cuneate at the transition to the
petiole) cordate, with broad or narrow sinus; T. auricdata has small auricles at
the transition to the petiole. Their apex is acute to acuminate, whereas in T.
mucronata it is Iongly mucronate, in T. obscura, T. villosa regularly (or
irregularly) shortly mucronate. The margin is entire or minutely to conspicuosly
dentate or serrate, the teeth usually with a minute dark mucro. The lamina is thin,
membranous, chartaceow or coriaceous, glabrous or pubescent, and most species
have whitish, blackish or brownish punctated-dots at the upper surface. The
terminal (central) leaflets of compound leaves in T. celebica, T. elmeri, T.
papuana, and T. wavrae are symmetrical, but the lateral ones are asymmetrical
or oblique. In the 5-foliolate leaved species, the base of the leaflets are acutely
symmetrical. The living leaves are green, yellow, brown or blackish. In T. pubera,
T. Ieuserensis, and T. kinabaluensis, they are reddish tinged. On drying the lower
surface of leaves are lighter than the upper one, rarely with the same colour.
Nerves - Most species have 3 or 5(or 7) palmately radiating nerves arising fiom
the blade base, of which 2 (or 4) lateral ones may closely follow the margin of the
cuneate base. The leaflets of compound leaved T. celebica, T. elmeri, T. papuana,
and T. wawrae are penninewed. The nerves are either prominent on both sides, or
sometimes only the main nerves of the upper surface are prominent, and in some
cases are furrowed The nerves are densely pubescent, glabrescent, rarely
glabrous, sometimes with whitish cystoliths below.
Blade glands - Glands are flat and varying in size from 0.5 to 5 mm
diameter. They may be present in small or liuge numbers, located at the basal part
of the leaves or scattered, usually near the main nerves. Size and location on the
blade is diagnostic for the species, as in T.planiganr (k 3-5 mm diameter, located
at base of leave).
Petioles and petiolules -The petioles of Trichosanthes are slender, pubescent or
glabrous and in some species have whitish cystoliths. Petiolules only occur in the
compound leaved T. celebica, T. elmeri, T. papuana, and T. wawrae, although
sometimes the leaflets are (sub)sessile.
Probract - The probract is an organ of a stipule-like appearance, membranous
or f fleshy or coriaceous, found on the node beside the axil of leaves, and can be
easily seen on the young shoot. In Trichosanthes, they are present in various
shapes according to the species, linear-lanceolate, ovate, flat or concave, or they
are absent. In some species (T. auriculata, T. beccariana, T. cucumerina, T.
mucronafa, T. ovigera, T. pendula, T. villosa) they are minute and early caduceus.
The morphological origin of the probract is unknown.
Tendril
Tendrils in Trichosanthes often provide usefid taxonomic characters to
distinguish the species. In most species the tendrils are forked (with the main
branch strongest); most commoniy they are 2 or 3 branched, but those of T.
sepilokensis have 4 very short branches. Trichosanthes villosa has 3-5-fid(-9 in
juvenile stage) tendrils, whereas in T. globosa the tendrils are always simple, stout
or delicate. The tendrils are either pubescent at the base (except T. edulis var.
septemloba which has densely hairy tendril from base to apex) or glabrous.
Inflorescences
Trichosanthes is mostly dioecious, which means that male and female
flowers are formed on different individuals. The male flowers generally are
produced in a bracteate raceme. The racemes are situated single (rarely paired) on
the nodes, except in T. pendula where there may be several in a bundle.
Occasionally an additional single male flower can be found beside the raceme. In
female plants, the female flowers are solitary on the nodes. Sometimes the male
raceme contains also one or few female flowers on the node, or mixed in between
the males (T. auriculata, T. beccariana, T. montana, T. leuserensis, T. pendula).
In monoecious species, the female flowers can be found either singly on the node
or singly at the node beside the male raceme (e.g. T. cucumerina); not rarely the
female flowers develop later than the males. Some species from E Malesia,
especially Papua New Guinea may have a conspicuous straw-like appendage at
the node beside the male raceme, which represents the remnant of pedicel of an
earlier developed basal single flower.
Peduncle and rachis (Fig. 1) -The
peduncle may be slender or stout, densely or
sparsely pubescent or rarely glabrous. The rachis may be slender (T. ovigera),
usually as thick as peduncle, but is sometimes to 2 cm thick (T. globosa, T.
rnontana, T. sepilohnsis, and T. valida) or only somewhat stouter than the
peduncle (0.3-)0.5-0.9 cm diameter (T. densiflora, T. edulis, T. longispicata, T.
pulleana). Some species have zig-zag rachis (T. rafiacta, T. schlechteri). A few
species have the rachis apparently branched, that is, with up to 5 mm long sidebranches formed by the persistent lower part of the pedicels, the part below the
bract (T. auriculata, T. coriacea, T.pendula).
Bracts -Bracts are persistent or caduceus, located on the rachis, in some species
(T. auriculata, T. coriacea) on the side branches of the rachis. The bracts have
various shapes, varying according to the species from lanceolate, ovate, to
circular or rhomboid, with characteristically entire, dentate, to variously deep
lacineate (upper) margin. They are glabrous, or sparsely to densely pubescent,
with scattered flat glands or not. The texture of the bracts varies fiom thin
chartaceous (T. cucurnerina)to subcoriaceous (T. montana).
Fig. 1. Rachis of male flower: a. T. valida (thickened), b. T. montana (thickened),
c. T. densijZora (stout), d . T. ovigera (slender).
Plowers
The flowers of most species are nocturnal, they open at night and close
before sunrise. Some species e.g. i? cucumerina are diurnal, with flowers also
open a large part of the day.
The flowers in Trichosanthes are unisexual, showy and large, with corolladiameter (excluding petal threads) about 2.5-7 cm diam., among which those of
T. auriculata, T. beccariana, T. cucumerina, T. ovigera, T. rotundifolia, T.
wawrae (from Java), are species with comparatively small flowers. In general, the
male and female flower (corollas) are about the same size.
In the present study, the characteristics of the flowers were often ignored
because of the incomplete material available, and also because the flowers are
very fragile, and not easy to be analysed on dry material.
Pedicel - The pedicel is slender, mostly pubescent. The pedicel of a solitary
male or female flower is much longer than that of flower in a raceme.
Receptacle tube - The receptacle is tubifonn, several centimeters long, narrow,
mostly widened towards the apex, usually with long white hairs inside. In some
species, the receptacle in male flowers is swollen at the base (pseudo-ovary), and
contains 3 cylindrical disks-parts which are possibly rudiments of style and
stigma.
Sepals -The sepals are 5 and free,usually densely (finely and inconspicuously)
pubescent, rarely glabrous, subulate, or narrowly tciangular, lanceolate, or
narrowly ovate, with long acute apex, with the margin entire, dentate or lacineate.
In some species the sepals of the female flowers are entire, but dentate or incised
in the male flower.
Corolla (Fig. 2) - All species have 5 free petals with conspicuous thread-like
(rarely short) long-filiform appendages on the margin (fimbriate), the entire part
(which the size is given in the description) is very thin and delicate, ovate-oblong
to obovate-rhomboid, usually pubescent rarely glabrous, white as greenish white,
rarely pinkish (T. rubriflos, not in Malesia), or with pinkish thread-tips (T.
pubera). The precise shape of the petals are not known in all species, because of
the scanty material available in the herbarium.
Male flowers (Stamen) - The three stamens of all species are inserted in the
hypanthium tube towards the throat. The filaments are free, (very) short, glabrous
or pubescent. The anthers have S-shaped anther cells, two anthers are 2-thewus,
one 1-thecous, and all are united into an elongated f truncate synandrium,
included or hardly exterted. The yellow pollen, together with the often yellow
bases of the petals forming a yellow "eye" in the centre of open corolla, which
possibly has a function with (nocturnal) pollination by insects.
Fig. 2. Type of petal of male and female flowers: a. T. cucumerina var. anguina
(elliptic-oblong), b. T. pubera (pinkish thread with obovat-rhomboidshape of petal), c. T. trimpidata (obovate-rhomboid, male flower), d. T.
tricaspidata (obovate-rhomboid, female flower).
Female flowers (Pistil)
- The
styles are long, mostly slender, and either
glabrous or pubescent. The stigma is 3. The ovary is wholly hairy or glabrescent,
globose or ellipsoid, one celled with three placentas, and numerous ovules.
Fruit
The fruits are indehiscent, various in size and shape, from narrowly
ellipsoid, mostly ovoid or ellipsoid, to subglobose or globose. Most species from
New Guinea have longish cucumber or lagenaria-like fruit (Fig. 3). In some
species the died off base of the receptacle tube remains as a short beak on the apex
of the fruit. The pericarp is thin or thick, with the exocarp smooth, leathery,
mostly red or orange-red, rarely yellow (only in T. villosa), with yellow or pale
longitudinal markings (flames) in some species; the mesocarp is yellow or whitish
and soft.
Fruit stalk
-This is the fruiting pedicel. It is striated or smooth, slender, or thick
(T. globosa, T. valida, T. sepilobnsis and T. montana, T. longspicata). Some
species (like in T. borneensis, T. ernarginata) it is of "two-toned" appearance
because of the presence of alternating smooth and striated parts (Fig. 3). The
smooth part in fact is the narrow basal portion of the fruit.
Seed
The seeds are numerous, imbedded in a mostly f a p , greenish-black, very
bitter pulp; in some species the pulp is orange-red, sweet testing (T. cucumerina
var. anguina and T. beccariana); the pulp of T. pendula and T. villosa is creamywhite and especially in T. villosa of a sweet taste, whereas in some species fiom
New Guinea (T. edulis, T. h t a t a , T. shlechteri) the pulp is whitish-red. The
seeds are mostly smooth, sometimes with a longitudinal groove at the middle part.
Their apex is rounded, obtuse or acute, or rarely emarginate; the edge of the seed
is entire, rarely undulate, or it may form a broad or narrow and sometimes
thickened band, called 'margin'; their base is broadly rounded, truncate or
cuneate, rarely f acuminate. The colour of the seed is black or brown, rarely pale
brown in drying (T. villosa).
The seed characters are important taxonomic evidences at sectional and
species levels. The seeds can be classified into several groups: 1) turgid with two
inflated lateral sides (i? ovigera, ?i pendula, T. beccariana, T. mucronata), 2) flat
with undulate margin (T. cucumerim), 3) flat (broadly) elliptic or subcircular,
with broad margin (the margin radiate in T. auriculata, not radiation in T.
rotundifolia, T. coriacea, 2: villosa), 4 ) flat with crenulate edge and narrow
margin, notched at one ends or both ends (most species from New Guinea), 5)
flat, with chisel-shaped pointed base, without margin (T. kinabaluensis, T.
planiglans, T. quinquangulata), 6 ) flat, narrowly oblong or lanceolate, without
margin (T. globosa and its related species), 7) flat, broadly ovate-oblong, without
margin (T. elmeri and its related species), 8) elliptic, without or with faint margin,
rounded
IN MALESIA
Oleh
RUGAYAH
Nrp: 945511BIO
PROGRAM PASCA SARJANA
INSTITUT PERTANLAN BOGOR
1999
RINGKASAN
R U G A Y A H. Trichosanthes (Cucurbitaceae) in Malesio @i bawah
bimbigan Prof. Dr. Ir. Edi Guhardja, M.Sr sebagai ketua, Dr. WJ.J.0. d e
Wilde, Prof. Dr. Mien A. Rifai, Dr. Dedy Darnaedi, Dr. Gayuh Rnhayu
sebagai anggota)
Trichosanthes L. merupakan marga terbesar yang tergolong dalam suku
Cucurbituceae. Marga tersebut beranggotakan sekitar 100 jenis, tersebar di Asia
bagian timur, Malesia, Australia tropik sampai ke Fiji. Beberapa jenis anggotanya
mempunyai nilai ekonomi sebagai bahan obat tradisional maupun sayuran, bahkan
Cina telah mengembangkaa T. kirilowii, guna mendapatkan zat bioaktif yang
dapat digunakan untuk menggugurkan kandungan bahkan sebagai bahan anti HIV.
Di Malesia, marga tersebut belum banyak diteliti baik dari segi potensi
ekonomi maupun taksonominya secara menyeluruh. Oleh karena i t - perlu dilakukan revisi secara menyelumh untuk mendapatkan data dasar yang nantinya dapat
digunakan untuk memantapkan batasan takson pada peringkat marga maupun
jenisnya, mengetahui daerah persebaran dan memahami hubungan kekerabatan
antajenisnya, mengevaluasi seksi yang dipertelakan untuk jenis-jenis yang ada di
Cina untuk dipakai sebagai wadah jenis-jenis yang ada di Malesia, serta mencari
data pendukung melalui beberapa pendekatan seperti anatomi dam, sitologi, dan
isozim di samping hasil pengamatan morfologi.
Hasil penelitian ini menunjukkan adanya 39 jenis Trichosanthes di Malesia, 17 di antarmya me~pdCanjenis baru di samping dua anak jenis, dua varietas
and tujuh fonna baru. Jenis-jenis tersebut dapat digolongkan dalam lima seksi
yaitu seksi Cucumeroides (dengan anak seksi Cucumeroides dan Tetragonosperma), Trichosanthes, Foliobracteola, Involucraria (anak seksi Involucraria
clan Pedatae) clan seksi baru Edulis. Data persebaran jenis-jenis Trichosanthes
dapat dikelompokkan menjadi dua daerah subsenter yaitu Bomeo sebagai daerah
subsenter bagian barat Malesia yang terwakili oleh 21 jenis dengan tujuh jenis di
antarmya endemik, dan P. Irian sebagai daerah subsenter lainnya di bagian timur
Malesia yang terwakili oleh 12 jenis, tujuh jenis di antaranya endemik.
Dari hasil pengamatan sitologi delapan jenis dan satu varietas yang ada di
Java, dietahui bahwa Trichosanfhesmempunyai jumlah kromosom somatik yang
sama yaitu, 2n = 22. Data matomi tidak dapat dipakai sebagai ciri penanda jenis,
kecuali pada ketiga jenis yang sangat berdekatan (T. tricuspidata, T pubera dan
T. quinquanguIata)yang memperlihatkan perbedaan pada penampang paradermal
daun pemukaan atas dan bawah. Data SEM kulit biji, mungkin dapat dipakai
sebagai ciri pendukung untuk penciri jenis bahkan untuk seksi. Untuk data isozim,
temyata sistem enzim AAT, ACP, dan PRX, memberikan pita spesifik untuk
jenis-jenis yang berdekatan.
SUMMARY
R U G A Y A H. Trichosanthes (Cucurbitaceae) in Malesia (Under the
supervision of Prof. Dr. Ir. Edi Guhardja, MSc as a chairman, and Dr.
W.J.J.O. de Wide, Prof. Dr. Mien A. Rifai, Dr. Dedy Darnaedi, Dr. Gayuh
Rahayu as members)
Trichosanthes is one of the largest genera belonging to the family
Cucurbitaceae with about 100 species distributed in Eastern Asia, Malesia and
Tropical Australia to Fiji and subtropic Eurasia. The genus includes a number of
species which have economic value because they are used as vegetable and
medicine. In China about 33 species have been used for medicinal herb to induce
aborsion and T. Ririlowii is the one species which have been intensively cultivated
to isolate the bioactive trichosanthin for its HIV-inhibiting property.
In Malesia, a complete monographic study of this genus has never been
undertaken for the whole regions. Therefore a new revision is needed to provide
better understanding of the diversity as well as the relationship of the species and
its distribution, to evaluate the suitability of the delimitation of sections based on
species from China for the Malesian representatives, and to find out other
supporting taxonomic characters derived from leaf anatomy, SEM of seed coat,
cytology, and isozyme.
The result of this study indicated that there are 39 species that can be
recognized in Malesia including 17 new species, two subspecies, hyo varieties and
seven forms. They can be divided into five sections namely, Trichosanthes,
Cueumeroides (with two subsect.: Cucumeroides, Tetragonosprma), Foliobracteola, lnvolucraria (with subsect.: Involucraria, PeJafae), and the new
section Edulis. The pattern of distribution of this genus in Malesia indicated that
there are two subcenter distributions which can be recognized, with Borneo (21
species, seven endemic) as one of the subcenter in West Malesia, and New Guinea
(12 species, seven endemic) as the other subcenter in East Malesia.
From the cytological study it can be concluded that, the somatic metaphase
chromosomes of eight species and one variety (T. borneensis, T. cucumerina var.
anpina, T. montana, T. ovigera, T puberq T. quinqumplaa T. tricuspidata, T
villom, T. wawrae) have the same chromosome numbers of 2n = 22. Leaf
anatomical data cannot be used to identify species, except in the three closely
related species (T. pubera, T. quinquangt~lata,I: tricuspidafa) which show
differentiy on the paradermal section of the upper and lower surfaces. Preliminary
observation on SEM of seed coat indicated that, the data obtained might be usehl
as supporting character for the proposed classification of this genus. Isozyme
analysis using AAT, ACP, and PRX isozyme system indicated that, they can be
used to identify the different species including the three closely related species.
TRICHOSANTHES (CUCURBITACEAE)
IN MALESIA
Oleh
RUGAYAH
Nrp: 945511 BIO
In partial fulfilment of the requirement for
the Doctor degree in the field of Plant Taxonomy
PROGRAM STUD1 BIOLOGI
PROGRAM PASCA SARJANA
INSTITUT PERTANIAN BOGOR
Judul Disertasi
:
Trichosanthes (Cucurbitaceae) in Malesia
Nama
:
RUGAYAH
NRP
:
94551
Program Studi
:
Biologi
Menyetujui
1. Komisi Pembimbing
(Prof. Dr. Ir. ~ dGuhardia.
i
MSc.1
Ketua
(Prof. Dr. Mien A. Rifai)
Anggota
(Dr. Dedv Darnaedi)
Anggota
2. Ketua Program Studi Biologi
(Dr. davuh Rahavu)
Awgota
gram Pascasarjana
a Manuwoto, MSc.)
Tanggal Lulus : 21 Agustus 1999
CURRICULUM VITAE
Rugayah was born on 30 August, 1956 in Sob, Central of Java She spent
her childhood with her parents in Solo until she graduated from Senior High
School in 1975. She continued her study at Gajah Mada University in Yogyakarta
and graduated in 1981.
Since 1982 she has been working at Herbarium Bogoriense, Puslitbang
Biologi-LIPI, Bogor as a researcher in plant taxonomy. In September 1990, she
was admitted to the degree of MSc of Birmingham University (England)
sponsored by the British Council. In September 1994, LIP1 gave her an
opportunity to continue study at IPB for her PhD. She is rnamed and has 2
children.
TABLE OF CONTENS
Page
Ringkasan
Summaty
............................................................................
ii
..............................................................................
iv
Table of Contents
...................................................................
Lists of Tables. Maps and Figures
................................................
...................................................................
Acknowledgement
ix
xi
xiv
Historical Introduction
.............................................................
1
Materials and Methods
.............................................................
5
Result and Discussion ...............................................................
10
General Part
Morphology
Leaf Anatomy
.................................................................
...............................................................
SEM Observation of Seeds Coat Surface ................................
Cytology .....................................................................
Isozyme Analysis
....................................:.....................
Economic Botany
...........................................................
Diversity
....................................................................
Geographical Distribution
Cladistic Analysis
................................................
..........................................................
Special Part
Taxonomic
..................................................................
1. Trichosanthes cucumerina
.............................................
61
66
var. cucumerina.......................................................
68
var. anguina...........................................................
69
2 . T. ovigera
..............................................................
3. T. beccariana
......................................................
78
.....................................................
79
......................................................
80
................................................................
83
4 . T. mucronata
.
5 T. pendula
77
...............................................
subsp.beccariana
subsp.pussila
72
6 . T. kinabaluensis
........................................................
90
7 . T. obscura
...............................................................
92
8 . T. rejf.acza
................................................................
94
9 . T. longespicata
.........................................................
96
10. T. celebica
..............................................................
99
11.T.floresana
............................................................
101
12. T. pqpuana
............................................................ 103
13 . T. elmeri
..............................................................
105
.............................................................
108
forma w m a e
....................................................
111
forma hirsuta
.....................................................
113
..........................................................
113
14. T. wmurae
15. T. ellipsoidea
16. T.philippinensis
...................................................
..............................................................
17.T.pubera
18. T. tricuspidata
.........................................................
115
117
120
forma iricuspidata
................................................
123
forma mperifolia
.................................................
125
forma seramensis
.................................................
125
forma sibemtensis
................................................
126
19. T. leuserensis
........................................................
20. T. quinquangulata
127
....................................................
130
.......................................................
133
..........................................................
.........................................................
135
24. T. globosa
.............................................................
140
25 . T. montana
............................................................
143
21. T.planigans
.
22 T. borneensis
23 . T. emarginata
26. T. sepilokensis
.........................................................
138
146
27 . T. valida
................................................................ 148
28. T.villosa
...............................................................
subsp. villosa
......................................................
subsp. midorensis
29. T. rotundifolia
30. T. auriculata
3 1. T. coriacea
................................................
154
155
......................................................... 156
...........................................................
.............................................................
32. T. schlechteri
152
..........................................................
159
162
166
33 . T. h t a t a
..............................................................
34. T. dieniensis
..........................................................
169
172
.............................................................
forma laeoica ...................................................
173
.................................................
176
...............................................
177
35 . T. laeoica
forma sicyocarpa
forma sorongensis
forma yapenensis
.................................................
. T.pulleana .........................................................
37. T.densiJora .......................................................
36
38. T. dentifera
39. T. edulis
........................................................
.............................................................
176
178
179
180
182
184
var. edulis
..........................................................
var.sativa
.......................................................... 188
var.septembola
.................................................
187
189
Conclusion
............................................................................
191
References
...........................................................................
193
Appendix: General Key to the Maiesian species of Trichosanthes ........... 198
Plates o f Species
.......................................................
213
LISTS OF TABLES, MAPS,
FIGURES AND PLATES
TABLES
1.
Occurrence of stomata and trichome types on leaves of
Trichosanthes spp.
2.
Anatomical characteristic from transverse section of leaves in
Trichosanthes spp.
3.
...........................................................
Somatic chromosomes and their karyotipe formulation of
Trichosanthes spp.
4.
............................................................
...........................................................
Matrix characteristic of isozyme band patterns from PRX ,AAT,
and ACP in Trichosanthes spp.
............................................
...................
5.
List and the distribution of Trichosanthes in Malesia
6.
Morphological characters used in the cladistic analysis of
Trichosanthes
.................................................................
..............
7.
Matrix of morphological characters for cladistic analysis
8.
Characters, number of character states, steps and consistancy index
from morphological characters ...............................................
MAPS
1.
Distribution of Trichosanthes (endemic and non endemic) in
Malesia
2.
........................................................................
Distribution of T. mucronata ................................................
3.
Distribution of T. pendula
4.
Distribution of T. kinabaluensis
5.
Distribution of T. obscura
....................................................
94
6.
Distribution of T. reji-acta
...................................................
96
7.
Distribution of T. longispicata ...............................................
98
8.
Distribution of T. celebica
9.
Distribution of T.floresana
....................................................
.............................................
84
92
................................................... 100
.................................................
102
10. Distribution of T. p a p u a ~.................................................. 104
1 1. Distribution of T. elmeri
....................................................
107
...................................................
110
12. Distribution of T. wawrae
13. Distribution of T. philippinensis
............................................
117
....................................................
119
15. Distribution of T. Ieuserensis
..............................................
128
16. Distribution of T. planiglans
................................................
133
14. Distribution of T. pubera
17. Distribution of T. borneensis ................................................ 137
...............................................
139
19. Distribution of T. globosa
.................................................
142
20 . Distribution of T. montana
.................................................. 145
18. Distribution of T. emarginata
21 . Distribution of T. valida
.....................................................
149
..............................................
158
23 . Distribution of T. auriculata .................................................
160
..................................................
163
22 . Distribution of T. rotundifolia
24 . Distribution of T. coriacea
25 . Distribution of T. schlechteri
...............................................
168
26 . Distribution of T. hastata ....................................................
171
27 . Distribution of T. laeoica ....................................................
175
28. Distribution of T. Jel~~~@'ora................................................. 182
29. Distribution of T. edulis ......................................................
186
FIGURES
1.
Rachis of male flower .......................................................
16
2.
Type of petal of male flower ................................................
19
3.
Fruit type of Trichosmthes spp.................................. .
.. . . . .
22
4.
Seed of Trichosanthes spp...................................................
23
5.
Paradermal section of leaves ..............................................
26
6.
Trichome types on leaves of Trichosanthes spp..........................
27
7.
Transverse section of leaves ................................................
28
8.
SEM of seed coat Trichosanthes spp.......................................
33
9.
Somatic chromosomes at metaphase on T. cucumerina var. anpina, 7:
borneensis, T. montana. T. vilosa ................................ ... . . . .
10.
36
.
Somatic chromosomes at metaphase on T. ovigera T. prrbera. 7:
.............
trinrspidala. 7: quinquangulafa ..................... .
.
.
.
37
11. Explanatory drawing of metaphase chromosomes ......................
38
12. AAT and PRX isozyme band patterns of T. cucumerina var.
anguina ........................................................................
13. Isozyme band patterns from AAT
43
........................................
43
14. Isozyme band patterns from ACP ..........................................
44
15. Isozyme band patterns from PRX
..........................................
16. Cladogram based on morphological character ...........................
44
59
PLATES
1.
T. mucronata ................................................................
213
2.
T. kinabaluensis ...............................................................
214
3.
T. obscura .....................................................................
215
4.
T. longespicata ................................................................ 216
5.
T. celebica ....................................................................
217
6.
Zfloresana ................................................................
218
7.
Kpcqouana
8.
T. ellipsoidea ..................................................................
9.
T. philippinensis .............................................................. 221
10.
7: tricuspidata ................................................................ 222
1 1.
T. tricuspidda forma aspergolia .......................................... 223
12.
T. leuserensis ....................... .
.
.
.
...................................
13 .
T. quinquangulata ..................... .
.
.
.............................. 225
14.
T. planigans .................................................................. 226
15 .
T. emarginata ................................................................
16.
T. gIobosa ..................................................................... 228
17.
.............................. 229
T. montana ...................................
.
18 .
T.vali& ....................................................................... 230
19.
1: villosa subsp. midorensis ...............................................
....................................................................
219
220
224
227
231
20. T. rohrndifolia
21 . T. auriculata
................................................................
..................................................................
22. T. coriacea
....................................................................
23. T. dieniensis
..................................................................
24. T. pulleana
25 . T. densifora
26. T. edulis
....................................................................
..................................................................
.......................................................................
27 . T. edulis var. septembola
....................................................
xvii
ACKNOWLEDGEMENT
I would like to record my thanks to my supervisors: Prof Dr. 11. Edi
Guhardja, (chairman of the advisory committee), Dr. W.J.J.O. de Wilde, Prof. Dr.
Mien A. Rifai, Dr. Dedy Darnaedi and Dr. Gayuh Rahayu (members of the
advisory committee), for their advices, guidance and encouragement throughout
this study.
My thanks are due to Prof. Dr. Ir. Syafiida Manuwoto, MSc (Director of
the Post Graduate Programme, IPB), Prof Dr. Reviani Widjajakusuma (Head of
Biological Study Programme) and Dr. Arie B u d m @ad of the Research and
Development Center for Biology-LIPI), for the opportunity given to me to
undertake this study. I am also grateful to Prof. Dr. Peter Baas (Director of
Rijskherbarium/ H o w Bonaticus, Leiden) and Dr. Alex Hartana
(in
charge of
Biochemical Laboratorium in PAU-IPB) for allowing me to use the excellent
laboratorium facilities. Many thanks have to be extended to Dr. Irawati (Head of
Herbarium Bogoriense) for giving me her support.
I am very gratefull to LIPI, GEF Small Grand Project and Leiden
University for providing financial support.
I highly appreciate the assistence of Dr. Veldkamp in making the Latin
diagnoses of the new taxa, Dr. Ding Hou in translating some Chinese articles into
English, Ms. Bertie Joan van Heuven in preparing the seed coat preparation and
its photograph, Mr. Iskak Syamsudin and Mr. Van 0 s in making the illustrations,
Ms. Titien Ngatinem in preparing the cytological preparation, Ms. Siti Rohajawati
and Mr. Budi Rahman in typing this manuscript.
xviii
I am indebted to Brigitta Dudes-de Wilde for assisting me during the
execution of the research, and Dr. Harry Wiriadinata for his advice and criticism. I
sincerely thank all staffs at Rijskherbarium, Leiden and Herbarium Bogoriense,
for sharing their experties in different fields, and d l post graduate students of
Botany-Taxonomy, IPB for their friendship.
Finally, to my parents (especially to my father who has passed away on
1 5 February
~
1999), my husband Agoes Sriyanto and my daughters Arnalia and
Vinandia, I am grateful for their deep understanding and great patient and for
giving me their moral support.
HISTORICAL INTRODUCTION
Trichosanthes is one of the largest genera belonging to the family of
Cucurbitaceae, with about 100 species distributed in subtropical and tropical
Eastern Asia, Malesia and Tropical Australia to Fiji (Jeffky, 1990); there are 39
species in Malesia (Rugayah & de Wilde, 1999). The genus includes a number of
species which have economic value because they are used as vegetable and
medicine.
Most species of this genus are dioecious, they are climber with delicate
fimbriate flowers, their anthesis is taking place at night until the morning in the
following day (nocturnal). They muently die off after fruiting. These characters
make it difficult to collect the plant in the right and complete condition.
Consequently, some collections may be mixed with fruit and the vegetative part
collected from different plants, or even different species. Some species have been
based on a few specimens only and in some cases they are only known from a
single specimen, either with male flowers, or female flowers, or fruit only.
Trichosanthes was described for the first time by Linnaeus in his Genera
Plantarum in 1737. He mentioned four species namely, T. anguina., T. nervifolia,
T. cucumerina and T. amara. For Java, Blume (1826) enumerated 13 species
under Trichosanthes, three of which belong to different genera. Trichosanthes
costata B1. is Gymnopetalum chineme (Lour.) Merr., whereas T. hexasperma B1.
and T. macrocarpa BI. are Hodgsonia macrocarpa (Bl.) Cogn. Roxburgh (1 832)
treated this genus for India and described six species, four of which were new
ones. Miquel (1856) recognized 23 species of Trichosanthes for the Malesian
area, proposing several new species, some of which, however, are later treated as
synonyms of other species. In Australia, Bentham & Mueller (1866) enumerated
four species. Clarke (1879) in the Flora British India proposed four new species,
recognized eight others.
Cogniaux (1881) was the first author who made an extensive study of this
genus. He treated the genera Poppya Rumph., Cucumeroides Gaertn.,
Involucraria Ser. as synonyms of Trichosanthes. He m t e a monographic
treatment of Trichosanthes in de Candolle's Monographiae Phanerogamarum in
which 40 species were enumerated for the Old World, 12 of which were proposed
as new species: six species from Malesia (Sumatra, Singapore, Borneo, Celebes
and New Guinea), one from Ceylon, four species from India and one from
Australia. He divided this genus into two subgenera based on the flower
characters:
Eutrichosanthes
(with
male
flower
in
a
raceme)
and
Pseudotrichosanthes (with male and female flowers solitary). For Formosa,
Hayata (1921) accepted nine new species of Trichosanthes in lcones Plantarum
Formosanarum. Ridley (1922) described six species in the Malay Peninsula, two
of which were misidentifications. Harms (1925) enumerated seven species for
Papua New Guinea. For the Philippines, Merrill (1923) recognized six species in
An Enumeration ofPhilippines Flowering Plants.
More recent treatments are those of Kitamura & Yoshida (1949) for Japan,
with 12 species (paying special attention to seed characters). For the Indian
subcontinent, Chakravarty (1959) recognized 24 species, two of which were new
species, and he also described two new varieties h m H i i a y a and Kheri. In
1963, Backer and Bakhuizen van den Brink, Jr. in Flora of Java revised
Trichosanthes and accepted eight species. For Cambodgia, Laos and Vietnam,
Keraudren-Aymonim (1975) recognized 10 species and she treated T. pubera and
T quinq1~1ngulaia
as synonyms of T trtcuspidaia. Jeffrey (1 980) recognized 40
species for Eastern Asia, Indomalesia, tropical Australia to Fiji and subtropical
Eurasia in Miocene and Pliocene.
Intensive studies of Trichosanthes were undertaken by the Chinese in
modem times: Yueh and Cheng (1974) distinguished 55 species in their
preliminary investigation for China based on morphological characters,
34
species of which have medicinal properties. They divided the genus into two
subgenera, Cucumeroides and Trichosanthes, which were further divided into two
and five sections respectively, based on leaf, bract, pulp and seed characters. Later
on, Yueh & Zhang (1986) and Huang et al. (1997) used pollen character to
support the delimitation of the sections. Recently, Huang et al. (1998) recognized
37 species and six varieties, based on morphological, cytological, palynological
and anatomical characters. They accepted the two subgenera described by Yueh &
Cheng (1974): subgenus Trichosanthes with three sections (sect. Trichosanthes,
sect. Foliobracteola with subsect. Foliobracteola and subsect. Villosae, and sect.
Znvolucraria),and subgenus Cucurneroides with two sections (sect. Cucumeroides
and sect. Tetragonosperma); they proposed a new series, ser. Ovijolia, under
subsect. Foliobracteola.
In a preliminary treatment of Cucurbitaceae for Flora Malesiana,
Rugayah and de Wilde (1997) revised the genus for Java and accepted 10 species
in this area. They found that materials from Java passed under the name of T.
trifoliata (L.) Merr. needed to be renamed into T. wawrae Cogn., and that T.
anguina L. should be sunk as a variety of T. cucumerina L. They also found that
materials from Java under the name i? bracteata (Lam.) Voigt represented a
mixture of three different species namely T. pubera Bl., T. quinquangulata A.
Gray, and T. tricuspidata Lour.
Therefore there is a need to undertake an indepth taxonomic study in this
genus for Malesia, especially because of taxonomical problems like
misidentifications of T. wallichiana Wight in the Malay Peninsula by Ridley
(1922) and T. bracreata Voigt for Papua by Hanns (1925), and also because many
new materials need to be described. The present study is to revise this genus in
Malesia, to evaluate the suitability and the delimitation of sections as based on
species from China for the Malesian representatives, and to provide better
understanding of the diversity and the relationship of the Malesian species, with
the available extended materials now at hand.
MATERIALS AND METHODS
PLACE OF STUDY
Studies of herbarium specimens were conducted in Herbarium Bogoriense
(BO) of Puslitbang Biologi-LIPI, Bogor; Herbarium of the Royal Botanical
Gardens Kew (K); British Museum, London (BM), and Rijksherbarium, Leiden
(L). Observation on living plants, anatomical and cytological investigations were
undertaken in Herbarium Bogoriense, while the isozymes was analyzed in PAU,
IPB, Bogor. The SEM analysis of the seed coats was undertaken in
Rijskherbarium Leiden.
PLANT MATERIAL
For the present study a total of 969 herbarium sheets, including liquid and
carpological collections of Trichosanthes from several herbaria (BM, BO, BR,
CANB, L, LAE, MEL, P, S, SAN, SING, TNS, USA, U, W) were used to
provide morphological, distribution and economic botanical data for this study.
Living materials of nine species and one variety collected from Java, Sumatra,
Borneo, and Irian were cultivated in the premises of Herbarium Bogoriense, and
used for anatomical, cytological and isozyrne analyses.
Details of the living plant collections used for this study was as follows:
T. borneensis: MR s.n. (Kalimantan); Fanani s.n. (Kalimantan).
T. cucumerina var. anguina: Rugayah 209 (Labuan), 210 (Jakarta), 21 1 (Bogor),
212 (Madiun), 213 (Mad-);
de Wilde & Duyfjes 21673 (Bogor).
T. globosa: Rugayah 115 (Gunung Bunder); de Wilde & Duyfjes 2 1703, 21778
(Gunung Bunder).
T. montana: de Wilde & Duyfjes 21888 (Situ Gunung).
T. ovigera: de Wiide 21B68 (Situ Gunung), 21896 (Gunmg Bunder), 21927
(Gunung H d i u n ) .
T.pubera: de Wilde & Duyfjes 21667 (Cianten), 21840 (Cianten).
T. quinquangulata:de Wilde & Duyfjes 21661 ( h u n g Bunder); Wiriadinata s.n.
(Gunung Halimun); T. Uji s.n. (Bengkulu); Widjaja 7091 (Yapen, Isl., Irian Jaya).
T. tricwpidata: de Wilde & DuyfJes21660 (Gunung Bunder), 21767 (Cibodas),
21773 (Gunung Bunder); Rugayah 214 (Sukabumi); Titin 525 (Banten);
Wiriadinata s.n. (Gunung Salak), 6874 (f. siberutensis, from Sibemt, Sumatra).
T. villosa: de Wilde & Duyfjes 21839 (Cianten). 21883 (Cianten); Wiriadinata
8284 (Batu, Malang).
T. wawrae: Fanani s.n. (Gunung Halimun).
METHODS OF INVESTIGATIONS
Morphology, Distribution and Economic Botany
- Data and information on
morphology, distribution and economic botany were collected from the specimens
and literatures. The procedure for morphological observation followed those
described by Leenhouts (1968), Rifai (1976) and de Vogel(1985).
Anatomy - Leaf anatomy was investigated by first fixing the leaves (small part
of the middle base) in FAA and then embeded them in par&.
Transverse
sections (20 pm thick) were made, stained with safranin and fast green, and then
mounted in canada balsam. Paradermal sections were taken from the upper and
lower surfaces of leaves, then stained with safi.anin 1% in water and then mounted
in glycerin. Descriptions of the trichome and stomata types were based on the
criteria used by Inamdar & Gangadhara (1975,1976).
SEM of Seed Coat - Seeds of some species (T. auriculata, T. beccariana, T.
borneensis, T. cucumerina. var. anguina, T. elmeri, T. emarginata, T. laeoica, T.
montana. I: ovigera, T. pendula, T qquiquangulata, T. rotund~olia, T.
schlechteri, T. tricuspidata,
T.valida,) were soaked in detergent (amaloco) 1% for
one night, then rinsed with aceton for 5-10 minutes. The seed then were cut at the
middle and put on the cylindrical stub using carbon glue and coated with gold
SCD 005 (BAL-TEC) for 4 minutes. The SEM photographs were made by low
pressure JEOL-SEM.
Cytology -Root tips h m the living materials were pretreated with 0.0002 M 8hydroxyquinoline solution and kept in the refrigerator for three hours. The root
tips separated from root caps were fixed in 45% acetic acid for 10 minutes,
macerated in 1 N HCl solution at 60° C for 2-3 minutes, and then squashed in 2%
aceto-orcein solution.
Isozyme
- Enzyme
electrophoresis was performed by horizontal starch gel
electrophoresis using 12% of potato starch h m SIGMA. The procedures
followed those of Wendel & Weeden (1990). About 50 mg pieces of fresh young
leaves were cmshed in approximately 0.5 ml extract buffer consisting of 250 mM
ascorbic acid Na-salt, Tris-HCI buffer (pH 7.9, 0.25 M pvp, 0.1%
Mercaptoethanol, 20 mh4 Diethyldithimbamate, 20 m M sodium metabisulfite,
and 200 m M sodium tetraborate. Filter paper wicks were dipped into the leaf
extract and then inserted into the starch gel. The gel buffer was 5mM L-histidin,
adjusted with NaOH (pH 7.0) or 0.076 M Tris, adjusted with citric acid (pH 8.6).
The electrode buffer was 0.410 M citric-acid, Na2 salt, adjusted with free citric
acid (pH 7.0), or 0.300 M boric acid, adjusted with NaOH (pH 8). Gel were run at
18-22 mA for 3-4 hours. The gel then was sliced horizontally and stained with
three staining enzymes, PRX (peroxidase), AAT (aspartic aminotransferase), ACP
(acid phosphatase). The PRX staining mixer was 0.05 M sodium acetat (pH 5.0),
3-amino-9-etil-carbosol, aceton, 3% HzOz, CaCI2. The AAT staining mixer was
alfa-ketogluteric-acid, L-aspartic-acid, pvp-40, EDTA, Naz salt, sodium
phosphate. The ACP staining mixer was 0.1 M sodium acetat (pH 5.0), MgCIz,
Na-napthyl acid phosphate, fast garnet GBG salt.
Cladistic analysis - Vegetative and generative characters were used for
phylogenetic analysis of Trichosanihes, with Gymnopethalum chinemis and G.
integrifolia used as the outgroups. A total of 46 morphological characters (Table
5) involving habit, indument, stem, tendril, leaves, probract, flowers, fruits and
seeds were accumulated and analyzed using Hennig86 computer programme. For
this analysis, six species (T.dentgera, T. dieniensis, T. ellipsoidea, T. planiglans,
T. pulleana, T. refiacta) were not included because of incomplete material
available.
RESULTS AND DISCUSSIONS
GENERAL PART:
MORPHOLOGY
Habit
The species of Trichosanthes are predominantly perennial climbers, rarely
annual (only T. cucumerinu, including usually the cultivated var. anguina). They
are scandent or creeping herbs, but their habit is often not readily apparent from
the herbarium specimen. The female plants frequently will die off after fruiting.
Some species have a tuber, capable of producing new shoots.
Sexual Conditions
Whether a plant or species is monoecious or dioecious in this genus is not
easy to determine without observing the living plants directly. In this study, the
sexual condition of species was determinated on the base of the presence of male
flowers, or female flowers or fruits on the specimens examined. All species have
unisexual flowers, wholly, partly or incidentally and most species appear to be
dioecious, and only a few are monoecious (T. cucumerina, T. leuserensis, T.
montana, T. quinquangulata).
Indumentum
Plants are generally hairy, with hairs villous, puberulent or strigose or
rarely subglabrous. Some species, however, are glabrescent, the hairs disappear
with age. The colour of the hairs may be grey, white, brown or rusty. Most of the
species have whitish, sometimes brown, or blackish punctate dots originating
from hairs or hair scars on the upper leaf surface. Moreover the upper leaf surface
of some species may be very scabrid. White cystoliths may also occur on stem,
petiole, and nerves of lower leaf surface of several species.
Stem
The Iianescent main stems and branches are slender, the stems of the leafy
shoots grooved or angularly-winged (in T. auriculata. T. kinabaluensis, T.
pendulu, T. pubera), glabrous or pubescent, pale yellow or brown in drymg, rarely
reddish (T. pubera, T. leuserensis).They are thin in some species (T. cucumerina,
T. ovigera). The stem of most species contains colourless sap, odourless except in
the T. cucumerina var. anguina, where it has a strong smell latex.
Leaves
Leaves are arranged alternately, have long petioles, and simple or
compound blades. Simple blades are entire or with 3-5(-7) shallow to deep lobes;
compound leaves with 3 or 5 or 7 leaflets are characteristic of a few species: T.
celebica, T. elmeri, T.floresana, T. papuana, and T. wawrae. Some (or probably
most or all) species are heterophyllous e.g. T. montam and T. globosa of which
leaves in the juvenile stage are f entire but becoming deeply 3-5 lobed in the
adult plant; T. tricuspidata and T. pubera has deeply 5-7 lobed leaves in the
juvenile stage, but only (2)-3 lobed when adult; heterophylly also occurs in T.
celebica and T. wawrae, with simple (but f lobed) leaves in the juvenile stage.
Leaf-blade
- The shape of the
leaves is varying from ovate to orbicular in
outline. Their base is generally deeply (often f cuneate at the transition to the
petiole) cordate, with broad or narrow sinus; T. auricdata has small auricles at
the transition to the petiole. Their apex is acute to acuminate, whereas in T.
mucronata it is Iongly mucronate, in T. obscura, T. villosa regularly (or
irregularly) shortly mucronate. The margin is entire or minutely to conspicuosly
dentate or serrate, the teeth usually with a minute dark mucro. The lamina is thin,
membranous, chartaceow or coriaceous, glabrous or pubescent, and most species
have whitish, blackish or brownish punctated-dots at the upper surface. The
terminal (central) leaflets of compound leaves in T. celebica, T. elmeri, T.
papuana, and T. wavrae are symmetrical, but the lateral ones are asymmetrical
or oblique. In the 5-foliolate leaved species, the base of the leaflets are acutely
symmetrical. The living leaves are green, yellow, brown or blackish. In T. pubera,
T. Ieuserensis, and T. kinabaluensis, they are reddish tinged. On drying the lower
surface of leaves are lighter than the upper one, rarely with the same colour.
Nerves - Most species have 3 or 5(or 7) palmately radiating nerves arising fiom
the blade base, of which 2 (or 4) lateral ones may closely follow the margin of the
cuneate base. The leaflets of compound leaved T. celebica, T. elmeri, T. papuana,
and T. wawrae are penninewed. The nerves are either prominent on both sides, or
sometimes only the main nerves of the upper surface are prominent, and in some
cases are furrowed The nerves are densely pubescent, glabrescent, rarely
glabrous, sometimes with whitish cystoliths below.
Blade glands - Glands are flat and varying in size from 0.5 to 5 mm
diameter. They may be present in small or liuge numbers, located at the basal part
of the leaves or scattered, usually near the main nerves. Size and location on the
blade is diagnostic for the species, as in T.planiganr (k 3-5 mm diameter, located
at base of leave).
Petioles and petiolules -The petioles of Trichosanthes are slender, pubescent or
glabrous and in some species have whitish cystoliths. Petiolules only occur in the
compound leaved T. celebica, T. elmeri, T. papuana, and T. wawrae, although
sometimes the leaflets are (sub)sessile.
Probract - The probract is an organ of a stipule-like appearance, membranous
or f fleshy or coriaceous, found on the node beside the axil of leaves, and can be
easily seen on the young shoot. In Trichosanthes, they are present in various
shapes according to the species, linear-lanceolate, ovate, flat or concave, or they
are absent. In some species (T. auriculata, T. beccariana, T. cucumerina, T.
mucronafa, T. ovigera, T. pendula, T. villosa) they are minute and early caduceus.
The morphological origin of the probract is unknown.
Tendril
Tendrils in Trichosanthes often provide usefid taxonomic characters to
distinguish the species. In most species the tendrils are forked (with the main
branch strongest); most commoniy they are 2 or 3 branched, but those of T.
sepilokensis have 4 very short branches. Trichosanthes villosa has 3-5-fid(-9 in
juvenile stage) tendrils, whereas in T. globosa the tendrils are always simple, stout
or delicate. The tendrils are either pubescent at the base (except T. edulis var.
septemloba which has densely hairy tendril from base to apex) or glabrous.
Inflorescences
Trichosanthes is mostly dioecious, which means that male and female
flowers are formed on different individuals. The male flowers generally are
produced in a bracteate raceme. The racemes are situated single (rarely paired) on
the nodes, except in T. pendula where there may be several in a bundle.
Occasionally an additional single male flower can be found beside the raceme. In
female plants, the female flowers are solitary on the nodes. Sometimes the male
raceme contains also one or few female flowers on the node, or mixed in between
the males (T. auriculata, T. beccariana, T. montana, T. leuserensis, T. pendula).
In monoecious species, the female flowers can be found either singly on the node
or singly at the node beside the male raceme (e.g. T. cucumerina); not rarely the
female flowers develop later than the males. Some species from E Malesia,
especially Papua New Guinea may have a conspicuous straw-like appendage at
the node beside the male raceme, which represents the remnant of pedicel of an
earlier developed basal single flower.
Peduncle and rachis (Fig. 1) -The
peduncle may be slender or stout, densely or
sparsely pubescent or rarely glabrous. The rachis may be slender (T. ovigera),
usually as thick as peduncle, but is sometimes to 2 cm thick (T. globosa, T.
rnontana, T. sepilohnsis, and T. valida) or only somewhat stouter than the
peduncle (0.3-)0.5-0.9 cm diameter (T. densiflora, T. edulis, T. longispicata, T.
pulleana). Some species have zig-zag rachis (T. rafiacta, T. schlechteri). A few
species have the rachis apparently branched, that is, with up to 5 mm long sidebranches formed by the persistent lower part of the pedicels, the part below the
bract (T. auriculata, T. coriacea, T.pendula).
Bracts -Bracts are persistent or caduceus, located on the rachis, in some species
(T. auriculata, T. coriacea) on the side branches of the rachis. The bracts have
various shapes, varying according to the species from lanceolate, ovate, to
circular or rhomboid, with characteristically entire, dentate, to variously deep
lacineate (upper) margin. They are glabrous, or sparsely to densely pubescent,
with scattered flat glands or not. The texture of the bracts varies fiom thin
chartaceous (T. cucurnerina)to subcoriaceous (T. montana).
Fig. 1. Rachis of male flower: a. T. valida (thickened), b. T. montana (thickened),
c. T. densijZora (stout), d . T. ovigera (slender).
Plowers
The flowers of most species are nocturnal, they open at night and close
before sunrise. Some species e.g. i? cucumerina are diurnal, with flowers also
open a large part of the day.
The flowers in Trichosanthes are unisexual, showy and large, with corolladiameter (excluding petal threads) about 2.5-7 cm diam., among which those of
T. auriculata, T. beccariana, T. cucumerina, T. ovigera, T. rotundifolia, T.
wawrae (from Java), are species with comparatively small flowers. In general, the
male and female flower (corollas) are about the same size.
In the present study, the characteristics of the flowers were often ignored
because of the incomplete material available, and also because the flowers are
very fragile, and not easy to be analysed on dry material.
Pedicel - The pedicel is slender, mostly pubescent. The pedicel of a solitary
male or female flower is much longer than that of flower in a raceme.
Receptacle tube - The receptacle is tubifonn, several centimeters long, narrow,
mostly widened towards the apex, usually with long white hairs inside. In some
species, the receptacle in male flowers is swollen at the base (pseudo-ovary), and
contains 3 cylindrical disks-parts which are possibly rudiments of style and
stigma.
Sepals -The sepals are 5 and free,usually densely (finely and inconspicuously)
pubescent, rarely glabrous, subulate, or narrowly tciangular, lanceolate, or
narrowly ovate, with long acute apex, with the margin entire, dentate or lacineate.
In some species the sepals of the female flowers are entire, but dentate or incised
in the male flower.
Corolla (Fig. 2) - All species have 5 free petals with conspicuous thread-like
(rarely short) long-filiform appendages on the margin (fimbriate), the entire part
(which the size is given in the description) is very thin and delicate, ovate-oblong
to obovate-rhomboid, usually pubescent rarely glabrous, white as greenish white,
rarely pinkish (T. rubriflos, not in Malesia), or with pinkish thread-tips (T.
pubera). The precise shape of the petals are not known in all species, because of
the scanty material available in the herbarium.
Male flowers (Stamen) - The three stamens of all species are inserted in the
hypanthium tube towards the throat. The filaments are free, (very) short, glabrous
or pubescent. The anthers have S-shaped anther cells, two anthers are 2-thewus,
one 1-thecous, and all are united into an elongated f truncate synandrium,
included or hardly exterted. The yellow pollen, together with the often yellow
bases of the petals forming a yellow "eye" in the centre of open corolla, which
possibly has a function with (nocturnal) pollination by insects.
Fig. 2. Type of petal of male and female flowers: a. T. cucumerina var. anguina
(elliptic-oblong), b. T. pubera (pinkish thread with obovat-rhomboidshape of petal), c. T. trimpidata (obovate-rhomboid, male flower), d. T.
tricaspidata (obovate-rhomboid, female flower).
Female flowers (Pistil)
- The
styles are long, mostly slender, and either
glabrous or pubescent. The stigma is 3. The ovary is wholly hairy or glabrescent,
globose or ellipsoid, one celled with three placentas, and numerous ovules.
Fruit
The fruits are indehiscent, various in size and shape, from narrowly
ellipsoid, mostly ovoid or ellipsoid, to subglobose or globose. Most species from
New Guinea have longish cucumber or lagenaria-like fruit (Fig. 3). In some
species the died off base of the receptacle tube remains as a short beak on the apex
of the fruit. The pericarp is thin or thick, with the exocarp smooth, leathery,
mostly red or orange-red, rarely yellow (only in T. villosa), with yellow or pale
longitudinal markings (flames) in some species; the mesocarp is yellow or whitish
and soft.
Fruit stalk
-This is the fruiting pedicel. It is striated or smooth, slender, or thick
(T. globosa, T. valida, T. sepilobnsis and T. montana, T. longspicata). Some
species (like in T. borneensis, T. ernarginata) it is of "two-toned" appearance
because of the presence of alternating smooth and striated parts (Fig. 3). The
smooth part in fact is the narrow basal portion of the fruit.
Seed
The seeds are numerous, imbedded in a mostly f a p , greenish-black, very
bitter pulp; in some species the pulp is orange-red, sweet testing (T. cucumerina
var. anguina and T. beccariana); the pulp of T. pendula and T. villosa is creamywhite and especially in T. villosa of a sweet taste, whereas in some species fiom
New Guinea (T. edulis, T. h t a t a , T. shlechteri) the pulp is whitish-red. The
seeds are mostly smooth, sometimes with a longitudinal groove at the middle part.
Their apex is rounded, obtuse or acute, or rarely emarginate; the edge of the seed
is entire, rarely undulate, or it may form a broad or narrow and sometimes
thickened band, called 'margin'; their base is broadly rounded, truncate or
cuneate, rarely f acuminate. The colour of the seed is black or brown, rarely pale
brown in drying (T. villosa).
The seed characters are important taxonomic evidences at sectional and
species levels. The seeds can be classified into several groups: 1) turgid with two
inflated lateral sides (i? ovigera, ?i pendula, T. beccariana, T. mucronata), 2) flat
with undulate margin (T. cucumerim), 3) flat (broadly) elliptic or subcircular,
with broad margin (the margin radiate in T. auriculata, not radiation in T.
rotundifolia, T. coriacea, 2: villosa), 4 ) flat with crenulate edge and narrow
margin, notched at one ends or both ends (most species from New Guinea), 5)
flat, with chisel-shaped pointed base, without margin (T. kinabaluensis, T.
planiglans, T. quinquangulata), 6 ) flat, narrowly oblong or lanceolate, without
margin (T. globosa and its related species), 7) flat, broadly ovate-oblong, without
margin (T. elmeri and its related species), 8) elliptic, without or with faint margin,
rounded