Upload
others
View
2
Download
0
Embed Size (px)
Citation preview
A TAXONOMIC STUDY ON RAFFLESIA SPECIES FROM TANJUNG DATU NATIONAL PARK, SARAWAK
Yong Choi Yen
(39423)
QK Bachelor of Science with Honours 495 (Plant Resource Science and Management)RI5 2015 Y55 2015
l'usat Khidmar Maldumat Aka« . . UNIVERSITl MALAYSIA SARA~r; : ,
A Taxonomic Study on Rafflesia species from Tanjung Datu National Park, Sarawak
Y ong Choi Yen
39423
A report submitted in partial fulfilment of the requirement for the degree of
Bachelor of Science with Honour
(Plant Resource Science and Management)
Supervisor: Dr. Aida Shafreena Ahmad Puad
Plant Resource Science and Management
Department of Plant Science and Environmental Ecology
Faculty of Resource Science and Technology
Universiti Malaysia Sarawak
.' 2015
I
APPROVAL SHEET
Name of candidate Y ong Choi Yen (39423)
Title A Taxonomic Study on Rafflesia specIes from Tanjung Datu
National Park, Sarawak
~
(Dr. Aida Shafreena Ahmad Puad)
Supervisor
-------------------.------------------------
(Dr. Rebicca Edward)
Coordinator Plant Resource Science and Management
Department of Plant Science and Environmental Ecology
Faculty of Resource Science and Technology
DECLARATION
This Final Year Project 2014 is based on my original work except for quotation and
citation which have been acknowledged. I also declared that it has not been submitted
previously or currently to any other degree at Universiti Malaysia Sarawak or any other
institution.
(Yong Choi Yen)
Plant Resource Science and Management
Department of Plant Science and Environmental Ecology
Faculty of Resource Science and Technology
Universiti Malaysia Sarawak (UNIMAS)
III
Acknowledgement
First and most of all, I am grateful to the God for giving me strength and determination to
complete my [mal year project. Second, I also would like to express my sense of gratitude
to my supervisor, Dr. Aida Shafreena binti Ahmad Puad for valuable guidance and advices
extended to me. Without her guidance and persistent help, this thesis would not have been
possible.
In addition, I would like to express my sincere thanks to Wan Nuur Fatiha binti Wan
Zakaria and Siti Noor Aishah binti Mohd Noor, postgraduates who always helping me in
this study especially in collecting and interpreting raw data as well as valuable guidance on .
doing }ab work. I take this opportunity to record my thanks to all the faculty members of
Department of Plant Science and Environmental Ecology for their help and encouragement.
I sincerely thanks to my lab mates, Muhammad Raimi bin Mohd Fauzi, Ibrahim Izzat bin
Zabidin and Muhammad Aiman bin Mohammad Rahimi for their support and help. I also
would like to express my appreciation to all my course mates for their constant
encouragement and support . ..
Finally, I would like to express the deepest gratitude to my lovely family for their
continuous moral support and encouragement. Not to forget to all my friends for their
constant encouragement, SUppOlt and understanding.
IV
Table of Contents
Title page I
Approval II
Declaration III
Acknowledgement IV
Table of Contents V
List of Abbreviations VII
List of Appendices VIII
List of Figures IX
List of Tables X
Abstract
Chapter 1 Introduction & Objective 2
Chapter 2 Literature Review 4
2.1 Rafflesiaceae 4
2.2 Morphology of RajJlesia R. Brown 4
2.3 Distribution and Ecology of RajJlesia 7
2.4 RajJlesia species in Sarawak. 8
2.4.1 RajJlesia kethii Meijer, Blumea 30 (1984): 211 8
2.4.2 RajJlesiapricei Meijer, Blumea 30 (1984): 214 9
2.4.3 RajJlesia tuan-mudae Beccari, Atti. Soc. Ita!' Sc. Nat. J J 10
(1868): 197-198
2.5 Biology of RajJlesia 11
2.6 The Host Plant-Tetrastigma (Miq.) Planch. 14
2.7 Phylogeny of RajJlesia 14
Chapter 3 Materials and Methods 16
3.1 Taxonomic Description 16
3.2 DNA Extraction and Amplification 17
3.3 Amplification of Targeted Sequence Using Polymerase Chain 18
Reaction (PCR)
3.4 Pairwise Sequence Alignment 20
Chapter 4 Results 21
4.1 Unknown RajJlesia species in Sarawak . 21
4.2 Key to the RajJlesia in Sarawak 21
V
4.3 Description of Unknown RajJlesia species from Tanjung Datu 22
4.4 Morphological Comparison between Rafflesia hasseltii and 25
Unknown Rafflesia species from Tanjung Datu
4.5 Molecular Anaiysis of Nuclear Region (ITS) and Plastid Gene 27
(16S rDNA)
4.5.1 DNA isolation of total plant genome and amplification of 27
nuclear sequence (ITS region) and plastid sequence (16S
rDNA)
4.5.2 Pairwise sequence alignment 28
Chapter 5 Discussion 31
Chapter 6 Conclusion and Recommendations 33
References 34
Appendices 36
•.
VI
bp
CIA
CTAB
DNA
dNTP
H20
ITS
MgCh
PCR
PL
rDNA
Taq polymerase
.'
List of Abbreviations
Base pair
Chloroform-isoamylalcohol
Cetyltrimethyl ammonium bromide
Deoxyribonucleic acid
Dideoxyl nucleotide
Water
Internal Transcribed Spacer
Magnesium chloride
Polymerase Chain Reaction
Perigonelobe
Nuclear ribosomal DNA cistron
Themus aquatiqus polymerase
VII
List of Appendices
Page
Appendix 1 Measurements were taken in the laboratory. A and B. 36 Perigone lobe diameter. C. Diaphragm diameter. D. Length of upper type ramenta. E. Length of middle type ramenta. F. Length of lower type of ramenta. G. Bract diameter. H. Length of process
Appendix 2 Table showing the data for speCllTIens examined In the 37 laboratory
Appendix 3 Morphological comparison among Rafflesia keithii, Rafflesia 38 pricei, Rafflesia tuan-mudae, Rafflesia hasseltii and unknown Ra.fJlesia species from Tanjung Datu
Appendix 4 Morphological comparison between Rafflesia hasseltii and 46 unknown Rafflesia species from Tanjung Datu
•.
VIII
Figure I
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8
Figure 9
I,::
Figure 10
Figure 11
I
List of Figures
.' Page
The floral morphology of RafJlesia female flower (Adapted 4 from Nais, 2001)
Diagrammatic sketch showing three concentric zones of 5 RafJlesia: (a) outer zone, (b) middle zone, (c) innermost zone (Adapted from Nais, 2004)
The floral morphology ofRafJlesia male flower (Adapted from 6 Nais, 2001)
RafJlesia keithii (Adapted from Nais, 2001) 9
RafJlesia pricei (Adapted from Nais, 2001) 10
RafJlesia tuan-mudae (Adapted from Nor-Izah, unpublished) 11
The life cycle of RafJlesia (Adapted from Nais, 2001) 13
ITS region that used for phylogenetic analysis (Adapted from 18 Soltis & Soltis, 1998)
RafJlesia species is showing A. Young and mature bud. B. 24 Open flower with 5 peri gone lobes. C. Diaphragm (shown with black arrow) and window (shown with white arrow) . D. Disk (shown with arrow). E. (a) Lower type (b) Middle type (c) Upper type (shown with arrow). F. Middle type ramenta. G. Annulus with presence of anther shown with arrow
Sequence alignment for ITS region between R. hasseltti and 29 RafJlesia sp. from Tanjung Datu National Park
Sequence alignment for Raffl6S between R. hasseltti and 30 RafJlesia sp. from Tanjung Datu National Park
.'
IX
List of Tables
Page
Table 1 Distribution of Rafflesia species in Southeast Asia 7
Table 2 Differences in floral characteristics between the unknown Rafflesia hasseltii and Rafflesia species from Tanjung Datu
26
Table 3 GenBank accession number for unknown Rafflesia species from Tanjung Datu National Park
27
.'
x
A Taxonomic Study on Rafflesia species from Tanjung Datu National Park, Sarawak
Y ong Choi Yen Plant Resource Science and Management Programme
Faculty of Resource Science and Technology Universiti Malaysia Sarawak
ABSTRACT
Rafflesia (Rafflesiaceae) is the largest flower in the world and it is an iconic species in Sarawak. Three Rafflesia species can be found in Sarawak, namely R. keithii, R. pricei, and R. tuan-mudae. An unknown Rafflesia species was discovered from Tanjung Datu National Park, Sarawak. This species morphologically resembled to R. hasseltii from Sumatra, Indonesia. In order to identify the unknown Rafflesia species, a morphological and molecular analysis were conducted. In this study, a detailed description of the unknown Rafflesia species and a taxonomic key to the Rafflesia species were made. Molecular analyses were carried out by sequencing two DNA regions (ITS and l6S rDNA) of the unknown Rafflesia species and compare with sequences of R. hasseltii. There are some morphological differences between the unknown Rafflesia species and R. hasseltii. In addition, molecular analyses showed that genetically these two species are quite distinct. Therefore, the unknown Rafflesia species may probably be a novel species. However, further molecular analysis is needed to support these [mdings.
Key words: Rafflesia, Raffiesiaceae, ITS, l6S rDNA, Tanjung Datu
ABSTRAK
Rafflesia (Rafflesiaceae) merupakan bunga yang terbesar di dunia dan ia adalah spesies .flora yang ikonik di Sarawak. riga spesies Rafflesia boleh didapati di Sarawak, iaitu R. keithii. R. pricei. dan R. tuan-mudae. Spesies Rafflesia yang tidak dikenali telah ditemui dari Taman Negara Tanjung Datu. Sarawak. Spesies ini menyerupai kepada R. hasseltii dari Sumatera dalam segi m01fologi. Untuk men genal pasti spesies Rafflesia yang tidak diketahui. analisis morfologi dan molekul telah dijalankan. Dalam kajian ini, penerangan terperinci meng~nai spesies Rafflesia yang tidak diketahui dan kunci taksonomi kepada spesies Rafflesia telah dibuat. Analisis molekul telah dijalankan dengan membuat urutan pada dua bahagian DNA (ITS dan 16S rDNA) spesies Rafflesia yang tidak diketahui dan bandingkan dengan urutan R. hasseltii. Terdapat beberapa perbezaan antara spesies Rafflesia yang tidak diketahui dan R. hasseltii dalam sefi morfologi. Di samping itu, analisis molekul menunjukkan bahawa genetik kedua-dua spesies adalah agak berbeza. Oleh itu, spesies Rafflesia yang tidak diketahui berkemungkinan adalah spesies baru. Walau bagaimanapun. analisis molekul yang lebih lanjut diperlukan untuk menyokong penemuan ini.
Kata kunci: Rafflesia. Rafflesiaceae, ITS, 16S rDNA, Tanjung Datu
I
Chapter 1
Introduction
Tropical rainforest is well known as the most complex terrestrial ecosystem on Earth and
rich in biodiversity (Nais, 2004). Therefore, tropical rainforest becomes a typical habitat
for many flora and fauna. Rafflesia is one of the unique and amazing flora species that is
endemic to tropical rainforest in Southeast Asia (Nais, 2004). Rafflesia is the largest flower
in the world (Nais, 2001). Rafflesia is rare and it is threatened with extinction mainly by
habitat destruction (Nais, 2004). Thus, it becomes an icon of plant ecology conservation
and protection especially tropical rainforests (Nais, 2004).
Rafflesia plant consists of flower without stem, leaves and roots (Nais, 2004). However,
Rafflesia has haustoria to penetrate the stem or root tissues of its host plant (Nais, 2004).
Rafflesia is an endophytic holoparasitic plant that grows completely embedded within its
host, Tetrastigma (Vitaceae), which is also widely distributed in tropical rainforests
(Barkman et ai., 2008). Rafflesia obtains nutrients and water from its host plant since this
parasitic plant is without photosynthetic pigments to carry out photosynthesis (Nais, 2001).
The pollination of Rafflesia depends on pollinators such as carrion flies through the
emission of rotten, smell from its flower (Beaman et ai., 1988). Rafflesia also relies upon
dispersal agents such as squirrel and tree-shrew to disperse its seeds for establishing the
next generation (Nais, 2001).
2
I
I
The rarity and uniqueness of Rajjlesia arises the interests of botanists and ecologists to
conduct research on Rajjlesia for scientific and education purposes (Nais, 2001). There are
many Rajjlesia species that have been described and studied since 1821. Out of the 18
species of Rajjlesia that have been described and studied before 2002, eight species were
discovered in Borneo and five in Sumatra, two from Peninsular Malaysia, three from Java
and two from the Philippines, and one species can be found in Thailand (Nais, 2001).
However, since 2002, eleven more new species were described and studied, making the
total number ofrecognised Rajjlesia species becomes 32 (Barcelona et al., 2009; Balete et
al. , 2010; Haji-Adam et al., 2013; Wong et al. , 2009).
Problem Statement and Objective
In Sarawak, there are three Rajjlesia species recorded, namely R. tuan-mudae, R. keith ii,
and R. pricei (Nais, 2004). Recently, an ullknown Rajjlesia species was discovered at
Tanjung Datu National Park, Sarawak. The floral morphology of unknown Rajjlesia
species is almost similar to R. hasseltii. Rajjlesia hasse/iii is found in Sumatra and is
believed to also be found in Samunsam, Sarawak. However, there is lack of well
documentation of this species found in Sarawak been published. Therefore, detailed
morphological characteristics and molecular analysis are necessary to detennine this
.' unknown Rajjlesia species. Therefore, the objective of this study is to identify the
unknown Rajjlesia species from Tanjung Datu National Park based on morphological and
molecular data.
3
Chapter 2
Literature Review
2.1 Rafflesiaceae
Rafflesiaceae is a holoparasitic plant family that contains eight genera which are
Apodanthes, Bdallophyton, Cythinus, Mitrastemol1 , Pilostyles, Rizanthes, Sapria, and
Rafflesia (Nais, 2001; Nais, 2004). All representatives of Rafflesiaceae are holoparasitic
plants, which mean they are without chlorophyll (Barkman et aI., 2004; Bendisksby et aI.,
2010). They totally depend on their host plant for water and nutrients (Barkman et aI.,
2004; Bendisksby et aI., 2010). Members of Rafflesiaceae also exist as endophytes (Nais,
2001). They grow completely embedded within the stem or root tissues of their host plant,
Tetrasiigma (Vitaceae) (Barkman et aI., 2004; Bendisksby et aI., 2010).
2.2 Morphology of Rafflesia R. Brown.
Rafflesia is exclusively characterised with its floral morphology as shown in Figure 1 (Nais,
2001; Nais, 2004). Rafflesia flower basically has three concentric zones: outer zone,
middle zone and innermost zone (Figure 2) (Nais, 2001; Nais, 2004).
Diaphragl'T1
Stigmatic are a ~(---~
Cupula ~---....".-
Perigonelobe
~-~ Ramenta
--::-lO""'--.;:-+--~)o Central col umn
Figure 1. The floral morphology of Rafflesia female flower (Adapted from Nais, 2001)
4
Figure 2. Diagrammatic sketch showing three concentric zones of Rafflesia: (8) outer zone, (b) middle zone, (c) innermost zone (Adapted from Nais, 2004)
The outer zone is made up of perigone lobes resembling petal, usually in five and
sometimes six, or even seven. The colour and pattern of perigone lobes indicates the
differences among species of Rafflesia (Nais, 2001; Nais, 2004).
The middle zone is called diaphragm that is ornamented by various pattern of white warts,
which differ among species. The lower face of diaphragm is covered by ramenta. Ramenta
differs among species based on their position at perigone tube of Rafflesia flower. There
are three types of ramenta: upper type, middle type and lower type. The upper type is
positioned near or on the lower surface of the diaphragm, whereas the middle type is on the
vertical centre of the perigone tube. The lower type is located on or near the base of the
perigone tube (Nais, 2001; Nais, 2004).
The innermost zone is known as diaphragm aperture. The lower face feature of aperture,
around the opening is called window. Window also ornamented by white warts, whereby
5
the arrangement pattern is important to characterise different Rafflesia species (Nais, 2001;
Nais, 2004).
The centre of the flower has a central column supporting a circular disc. The top of the
circular disc has projecting processes (Mat-SaUeh, 1991). According to Mat-Salleh (1991),
the central colurrm bears the sexual reproductive organs of Rafflesia, holding either ovaries
(Figure 1) or anthers (Figure 3).
Anther ~-----+'I"""""--'r--:iII .....~"'--dil'-->'i'---~ Annulus interior
Bristles
Figure 3. The floral morphology of Rafflesia male flower (Adapted from Nais, 2001 )
6
2.3 Distribution and Ecology of Rafflesia
Rajjlesia is restricted to tropical rainforests of Southeast Asia. The 32 species of Rajjlesia
are distributed from Thailand, throughout Peninsular Malaysia, Sumatra, Java, Borneo to
the Philippines (Bendiksby et aI., 2010). The distribution of Rajjlesia species in Southeast
Asia is presented in the following table (Table l).
Table 1. Distribution ofRajJIesia species in Southeast Asia (Adapted from Nais, 2001)
Localities Species
Borneo
Java
Peninsular Malaysia
Philippines
Sumatra
Thailand
R. arnoldii R. Brown, R. borneensis Koorders, R. ciliata Koorders, R. keithii Meijer, R. pricei Meijer, R. tengkuadlinii Mat Salleh & Latiff, R. tuan-mlldae Beccari, R. witkampii Koorders
R. patma Blume, R. rochussenii Teijsmann & Binnendjik, R. zollingeriana Koorders
R. cantleyi Solms, R. kerrii Meijer, R. azlanii Latiff & M. Wong, R. su-meiae Wong, N ais, & Gan, R. sharifahhapsahiae Adam, Mohamed, Aizat-Juhari & Wan
R. balatei Barcelona & Cajano, R. banahaw Barcelona, Pelser & Cajano, R. leonardi Barcelona & PeIser, R. lobata R. Galang & Madulid, R. manillana Tescehemacher, R. mira Fernando & Ong, R. philippensis Blanco, R. schadenbergiana Geoppert, R. speciosa Bercelona & Fernando, R. verrucosa Balete, PeIser, Nickrent & Barcelona
R. arnoldii R. Brown, R. gadutensis Meijer, R. hasseltii Suringar, R. micropylora Meijer, R. patma Blume, R. rochussenii TeijsmaIpl & Binnendjik
R. ken'ii Meijer
7
RafJlesia is often found in both primary and secondary forests (Nais, 2001). Rafflesia
distribution is reliant on that of the typical habitat of Tetrastigma which is also found in
primary and secondary forests (Nais, 2001). Ralflesia populations are usually found near to
the stream banks and high elevation areas (Nais, 2001). Nais (2001) stated that the
altitudinal distribution of Rafflesia differs according to species, from as low as 5 m above
sea level (R. patma in Java), to as high as 1400 m or above (R. pricei in Sabah).
2.4 RaJjlesia species in Sarawak
In Sarawak, there are three species of Rafflesia present, namely R. keithii, R. pricei and R.
luan-mudae (Nais, 2004). Rafflesia keithii was found at Gunung Hose, whereas R. pricei
was discovered at Bario Highlands in Sarawak (Nais, 2001). Ralflesia tuan-mudae which is
an endemic species of Sarawak was first discovered at Mount Pueh in 1868 by Odoardo
. Beccari (Nais, 2001). At present, this species can be found in Gunung Gading National
Park, Lundu (Sarawak) (Nais, 2004). Rajjlesia keithii and R. pricei were also found in
Sabah.
2.4.1 Rafflesia keithii Meijer, Blumea 30 (1984): 211
Type: Madani s.n. (SAN); Sabah, Sungai Melaut, along the Ranau-Tambunan road.
Rafflesia keithii is one of the larger-flowered species. Its open flower can reach up to 80-94
cm in diameter. Ralflesia species commonly having five perigone lobes but R. keithii has
been discovered with six perigone lobes at Kampung Kokob, Ranau (Sabah). Perigone
warts of R. keitlzii are interspaced to each other. White warts on diaphragm are contrasting,
isolated and in 5 concentric rings. The number of anthers is always 40. Rafflesia keithii is
8
-------
widely distributed in Borneo (Sabah; probably also Kalimantan and Sarawak). It grows in
primary and secondary hill dipterocarp forest, normally near streams or rivers (Nais, 200 1).
Figure 4. RafJlesia keithii (Adapted from Nais, 200 1)
. 2.4.2 Rafflesia pricei Meijer, Blumea 30 (1984): 214
Type: Price S.n. (K), Sabah, Mount Kinabalu, Mamut Copper Mine, 1300 m a.s.l., 4 May
1967.
Rafflesia pricei is the smallest among RajJlesia species in Sarawak. The diameter of open
flower is about 25-30 cm, with aperture about 5-6 cm wide. The perigone warts of R. price;
are white, matted-like and not interspaced to each other. The diaphragm has 4-5 irregular
rings of white warts smaller than those of the peri gone lobes. The number of anthers is 20.
This species usually found in Sabah and probably also in north-eastern Kalimantan and
Sarawak (Borneo). The typical habitats of R. pricei are in primary and secondary hill
dipterocarp forest, normally near streams or rivers (Nais, 200 I).
9
--------
Figure 5. Ra.fflesia pricei (Adapted from Nais, 200 I)
2.4.3 Rafflesia tuan-mudae Beccari, Atti. Soc. Ital. Sc. Nat. 11 (1868): 197-198
Type: Beeeari S.n. (FI-B), Gunung Poe (Pueh), Sarawak.
RajJlesia tuan-mudae blooms up to 56 cm in diameter, with aperture about 15-18 cm wide.
However, the diameter can reach up to 72 cm as recorded by Nor-Izah (unpublished).
RajJlesia tuan-mudae has distinct brick-red colouration of its perigone lobes, and the apex
tapering into an obtuse tip, giving it an oblong shape, makes it easy to identify. The white
warts on perigone lobes are less numerous and interspaced to each other. The warts on
diaphragm arranged in 2 concentric rings. The number of anthers is 25. This species is
endemic to Sarawak but it also can be found in Kalimantan (eagar Alam Gunung Rara
Pasi near Singkawang). RajJlesia tuan-mudae grows in lowland and hill dipterocarp forest
(Nais, 2001).
10
Figure 6. RajJlesia tuan-mudae (Adapted from Nor-Izah, unpublished)
2.5 Biology ofRafj1esia
The biological system of Rafflesia is unique and incredible (Nais, 2004). Rajjlesia is not
only endophytic and holoparasitic, but also depends upon animals for pollination and
dispersal to establish next generation (Nais,.2004).
Blooming Rafflesia flower emits rotten smell to attract the pollinators (Nais, 2004). The
main pollination agent of Rafflesia is carrion flies from the genera of Lucilia and
Chrysoma (Mat-Salleh, 1991). Carrion flies are lured by the smell and crawl into the anther
grooves in male flowers to find food sources (Mat-Salleh, 1991). They are precisely guided
by ridges armed with bristles (Beaman et aI., 198~). In the process of crawling upward
searching for food, the back of the flies come into contact with the pollen, and the sticky
mass attached to its back (Nais, 2001). However, there is no such food provided by
Rafflesia for the pollinators but deceives them by an apparent offering of food and possibly
brood place (Beaman et aI., 1988). Hence, pollen-loaded flies leave the male flower, and
searching for another flower. If they visit the female flower, the flies enter the sulcus of
female flower where the stigmatic area is positioned (Beaman et al., 1988). On entering the
11
,.
sulcus, the sticky mass pollen attached at flies' back comes into contact with the stigmatic
area, and rubbed offthe pollen, affecting pollination (Nais, 2001).
The dispersal of Rafflesia is dependent upon that of the small animals such as squirrel and
tree-shrew (Mat-Salleh, 1991). Species of squirrel and tree-shrew that has been reported to
disperse seeds of Rafflesia is Callsociurus notatus and Tupaia tana (Nais, 2004).
According to Nais (2004), the remaining fruit and seeds that attached at the mouth and feet
of squirrel and tree-shrew will be anchored on stem or root of Tetrastigma when they are
climbing the plant. The seed then wiU be germinated, to start over the life cycle of
RajJlesia (Nais, 2004).
RajJlesia takes 46 months to complete its life cycle (Nais, 2004). The life cycle of Rafflesia
follows a sequence of stages: seed germination, development of the Rafflesia plant inside
the host plant, emergence of flower buds, flower buds development, flowering, pollination,
fruit and seed set, seeds dispersal, and seed germination for the establishment of next
generation (Nais, 2001) (Figure 7).
12
~
I'
I'
I
Seed ge
Stage VIII
nnmation & inoculation 1'----7of host
(46 months)
Stage VII
Seed dispersal
(1-2 days)
Stage VI
Mature fruits
(6-8 months)
Stage V
Flo wer fully open for
4~8 days to open
Figure 7. The life eye Ie of RajJlesia (Adapted from Nais, 200 I)
13
Stage I
Host organ swollen
(6 days)
Stage II
Emergence from host
(160 days)
\
Stage III
Bract abscission
(8 days)
Stage IV
Flower beginning
(24-48 hours)