systematics studies of the genus amomum roxb
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Systematics Studies of the Genus Amomum Roxb. (Zingiberaceae): Fruit
Morphological Aspects
Nurul Izzati Binti Azman (45958)
Bachelor of Science with Honours
(Plant Resources Science and Management)
2017
Faculty of Resource Science and Technology
UNIVERSITI MALAYSIA SARAWAK
Grade:
Please tiek (-..J)
Final Year Project Report G2J Mast.ers D PhD D
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Systematics Studies of the Genus Amomum Roxb. (Zingiberaceae): Fruit
Morphological Aspect
Nurul Izzati Binti Azman (45958)
This project is submitted in partial fulfillment of the requirement for degree of
Bachelor of Science with Honours
Plant Resource Science and Management
Plant Resources Science and Management Programme
Faculty of Resource Science and Technology
Universiti Malaysia Sarawak
2017
iii
APPROVAL SHEET
Name of Candidate: Nurul Izzati Binti Azman
Student No: 45958
Title of Project: Systematics Studies of the Genus Amomum Roxb. (Zingiberaceae): Fruit
Morphological Aspects.
------------------------------------------------
(Dr. Qammil Muzzammil Abdullah @ Meekiong Kalu)
Supervisor
------------------------------------------------
(Dr. Freddy Yeo Kuok San)
Coordinator
Plant Resource Science and Management Programme
Faculty of Resource Science and Technology
Universiti Malaysia Sarawak.
iv
DECLARATION
This project is done for the subject of STF 3015 Final Year Project 2 2016/2017. It is based
on my original work except for quotation and citation which have been acknowledged. I
declare that no portion of this research work had been submitted to support the application
of other degree or qualification at any other universities or institutions of higher learning.
__________________________
Nurul Izzati Binti Azman
Plant Resource Science and Management Programme
Faculty of Resource Science and Technology
Universiti Malaysia Sarawak
Jun, 2017
v
ACKNOWLEDGEMENTS
Alhamdulillah, I am able to finish my final year project thesis. First of all, I would like to
express my appreciation and gratitude to my supervisor, Dr. Qammil Muzzammil Abdullah
@ Meekiong Kalu for useful guidance, encouragement, suggestion, helped me to
coordinate my project and thesis and also for giving me knowledge on identifying
Amomum species while collecting samples. A special thanks to post grade student, Aimi
Syazana Binti Sidek whose willing to guides me during my project by assistance through
my learning process. Furthermore, I also like to thanks all lecturers for helping me in
finishing my thesis. I would like to acknowledge UNIMAS for providing laboratory
facilities and give me chance to apply my knowledge here. Besides, I would like to give
many thanks to my beloved parents Azman Bin Mohd Taib and Saloni Binti Suid, and also
Muhammad Amzar Wajdi Bin Mahamad Isa for staying by my side and endless support
given along the project. Not to forget my lab mate, Nurul Amanina Binti Md. Raub who
always give many help and also suggestion in finishing my project and also my course
mate for continuous help and support in making this project successful.
vi
TABLE OF CONTENT
Page
Declaration iv
Acknowledgements v
Tables of Contents vi
List of Abbreviations vii
List of Tables viii
List of Figures ix
Abstract xi
CHAPTER 1: INTRODUCTION 1
CHAPTER 2: LITERATURE REVIEW
2.1 Order Zingiberales
2.2 Family Zingiberaceae
2.3 Genus Amomum Roxb.
2.4 Fruit Morphology
2.5 Fruit Anatomy
2.6 Scanning Electron Microscope (SEM)
3
3
6
7
8
9
10
CHAPTER 3: MATERIALS AND METHODS
3.1 Materials & Sites
3.2 Methods
3.2.1 Morphology
3.2.2 Anatomy
3.2.3 Scanning Electron Microscope (SEM)
11
11
14
14
14
15
CHAPTER 4: RESULTS AND DISCUSSION 17
4.1 Amomum
4.2 Morphology
4.2.1 Type of fruit
4.2.2 Shape
4.2.3 Measurement
4.2.4 No of fruit per peduncle
4.2.5 Colour
4.2.6 Special features
4.2.7 Bract
4.2.8 Peduncle
4.2.9 Pedicel
4.3 Micro-morphological
4.3.1 Fruit ornamentation
4.4 Anatomy
4.4.1 Locule structure
4.4.2 Pericarp
4.5 Key to selected Amomum species
4.6 Description of species
17
17
17
18
18
20
20
20
21
22
22
29
29
33
33
34
37
38
CHAPTER 6: CONCLUSION 51
CHAPTER 7: REFERENCES
52
CHAPTER 8: APPENDICES
54
vii
ABBREVIATION
aff. Affinis (around)
c. Circa (approximately)
cm Centimetre
mm Milimetre
sp. Species
SAR Acronym for Herbarium Department of Forest
Sarawak
SEM Scanning Electron Microscope
HUMS Acronym for Herbarium Universiti Malaysia Sarawak
UNIMAS Universiti Malaysia Sarawak
MP Megapixel
LED Light-emitting diode
USB Universal Serial Bus
viii
LIST OF TABLE
Page
Table 1 : Morphology of Amomum fruits. 27
Table 2 : Fruit ornamentation of Amomum fruits. 32
Table 3 : Anatomy of Amomum fruits. 36
ix
LIST OF FIGURES
Figure Page
Figure 1: Phylogenetic tree of Zingiberales based on a partitioned ML of
concatenated plastid and nuclear sequence
5
Figure 2: The Map of Malaysia (Taken from Google Map)
11
Figure 3: Location of Gunung Payang, Tebedu, Serian, Sarawak
(Taken from Google Maps)
12
Figure 4: Location of Gua Musang, Kelantan (Taken from Google Maps)
12
Figure 5: Location of Dered Krian, Bau, Sarawak (Taken from Google Maps)
12
Figure 6: Location of Long Banga, Miri, Sarawak (Taken from Google Maps)
12
Figure 7: Location of Cameron Highlands, Pahang (Taken from Google Maps)
13
Figure 8: Location of Universiti Malaysia Sarawak, Sarawak
(Taken from Google Maps)
13
Figure 9: Measurement of length of fruit with calyx. (A) Amomum
sp. 9 (B) Amomum sp. 10 (C) Amomum sp. 13 (D) Amomum sp. 8
(aff. uliginosum)
19
Figure 10: Amomum spiceum on peduncle taken from the sampling site
23
Figure 11: The bract of Amomum sp. 17 fruit connected with the pedicel. 23
Figure 12: Bract of Amomum xanthoflabium (A) Three fused bract
(B) One free bract.
23
Figure 13: Surface ornamentation of bracts. (A) Sericea surface of
Amomum xanthoflebium. (B) Glabrous surface of bract Amomum
sp. 16. (C) Tomentose surface of bract Amomum laxisquamosum.
23
Figure 14: Tomentose surface of peduncle of Amomum sp. 16.
24
Figure 15: Drawing of Amomum fruits. (A) Amomum sp. 8 (aff.
uliginosum) (B) Amomum sp. 9 (C) Amomum sp. 16 (D) Amomum
sp. 17 (E) Amomum lappaceum (F) Amomum laxisquamosum.
25
Figure 16: Drawing of Amomum fruits. (G) Amomum spiceum (H)
Amomum sp. 13 (I) Amomum oliganthum (J) Amomum
xanthophlebium (K) Amomum sp. 10
26
Figure 17: Spiny ornamentation on the surface of the fruit (A)
Amomum lappaceum (B) Amomum oliganthum (C) Amomum sp. 8
(aff. uliginosum).
29
x
Figure 18: Glabrous surface ornamentation. (A) Amomum spiceum
(B) Amomum laxisquamosum.
30
Figure 19: Glabrous ornamentation of Amomum laxisquamosum
observed under Analyrtical Scanning Electron Microscope (SEM)
with magnification of 100X.
30
Figure 20: Tomentose ornamentation. (A) Amomum
xanthophlebium (B) Amomum sp. 13.
30
Figure 21: Tomentose ornamention of Amomum
xanthophlebium viewed under Analytical Scanning Electron
Microscope (SEM) using magnification of 30X and 250X.
31
Figure 22: Amomum sp. 7 hairy ornamentation
31
Figure 23: Ornamention of Amomum sp. 16 which is tomentose
but no hairy structure detected using digital microscope.
31
Figure 24: Hair structure of Amomum sp. 16 can be seen after
viewed under Scanning Electron Microscope using magnification
250X.
31
Figure 25: Locules. (A) Amomum xanthophlebium. (B) Amomum
lappaceum (C) Amomum spiceum.
33
.Figure 26: Segmented line of Amomum laxisquamosum observed under
Analytical Scanning Electron Microscope (SEM) with magnification of 30X.
35
Figure 27: Example of measurement made to the pericarp to
obtained average measurement using Cooling Tech application.
(A) Amomum oliganthum (B) Amomum xanthophlebium.
35
Figure 28: Pericarp of (A) Amomum sp. 16 and (B)
Amomum sp. 17 observed under USB Digital Microscope.
35
Figure 29: (A) Fruit and leaves of Amomum spiceum. (B) Fruit
of Amomum lappaceum. (C) Amomum oliganthum. (D) Fruit
stalk of Amomum oliganthum. (E) Inflorescences of Amomum
laxisquamosum. (F) Fruit of Amomum laxisquamosum.
48
Figure 30: (A) Fruit of Amomum xanthophlebium. (B) Fruit of
Amomum sp. 16. (C) Leaves and fruits of Amomum sp. 17. (D)
Fruits of Amomum sp. 8 (aff. uliginosum). (E) Fruit of Amomum
sp. 9. (F) Inflorescence of Amomum flavoalbum.
49
Figure 31: Fruit of Amomum sp. 10
50
Figure 32: Fruit of Amomum sp. 13 50
xi
Systematics studies of the Genus Amomum Roxb. (Zingiberaceae) in Malaysia: Fruit
Morphological Aspects
Nurul Izzati Binti Azman
Plant Resource and Management
Programme
Faculty of Science and Technology
Universiti Malaysia Sarawak
ABSTRACT
Zingiberaceae is one of the family under the order Zingiberales. Amomum is one of the largest genus under
family Zingiberaceae. Although Amomum is one of the largest genus, this genus is poorly known among
ginger. The identification of this species is still confusing although the plant already been long utilized since
there are a lot of species under genus Amomum and herbarium specimen are also incomplete. Thus, this
project studied the morphological and anatomical characteristics of fruit from the selected Amomum species
in Malaysia and use the data for species determination. The sampling areas were taken from various localities
in Malaysia and 12 species of Amomum were examined. The method is the whole observation of the sample
and all the morphology and anatomy of the fruit were observed. To ease this process, the Scanning Electron
Microscope (SEM) and USB Digital Microscope were used. Most of Amomum fruit has the common
morphology characteristics such as fruit shape in oval, capsule types of fruits with persistent calyx, presence
of bracts and the arrangement in agglutinate.
Keywords: Amomum, Zingiberaceae, fruit, morphology, anatomy, Malaysia, systematic study, Scanning
Electron Microscope (SEM).
ABSTRAK
Zingiberaceae adalah salah satu famili di bawah order Zingiberales. Amomum adalah salah satu genus
terbesar di bawah keluarga Zingiberaceae. Walaupun Amomum adalah salah satu genus terbesar, genus ini
kurang dikenali di kalangan halia. Pengenalpastian spesies ini masih mengelirukan walaupun tumbuhan ini
telah lama digunakan kerana terdapat banyak spesies di bawah genus Amomum dan spesimen herbarium
juga tidak lengkap. Oleh itu, projek ini telah mengkaji ciri-ciri morfologi dan anatomi buah daripada spesies
Amomum terpilih di Malaysia dan menggunakan data tersebut untuk menentukan spesies. Kawasan
persampelan telah diambil dari pelbagai tempat di Malaysia dan 12 spesis Amomum telah diperiksa.
Kaedahnya adalah pemerhatian keseluruhan sampel dan semua morfologi dan anatomi buah telah diperhati.
Untuk memudahkan proses ini, Mikroskop Imbasan Elektron (SEM) dan USB mikroskop digital telah
digunapakai. Kebanyakan buah Amomum mempunyai karekter morfologi yang sama seperti bentuk buah
yang bujur, jenis buah kapsul dengan kaliks yang kekal, kehadiran pelepah dan susunan buah yang melekat.
Kata kunci: Amomum, Zingiberaceae, buah, morfologi, anatomi, Malaysia, kajian sistematik, Mikroskop
Imbasan Elektron (SEM)
1
CHAPTER 1
INTRODUCTION
Amomum Roxb. is one of the largest genus in the family Zingiberaceae which also
known as ginger family. According to Kiew (1982) and Smith (1985), genus Amomum are
widely distributed from Himalayas to Northern Australia which is in Southest Asia and
until the central Pacific. Generally, species from Amomum genus are evergreen herbs occur
in wet forests (Sakai & Nagamasu, 1998). Most of species from the genus Amomum
commonly use as spices such as Amomum capulaga, vegetables, and condiment.
The identification of this species is still confusing although the plant already been
long utilized since there are a lot of species under genus Amomum and the herbarium
specimen are incomplete (Kaewsri & Paisooksantivatana, 2007). There are a lot of species
available under Amomum, in order to compare all the species under genus Amomum
properly, further study is needed to study to clarify the different between them and fruit are
chosen as parameter in this study.
In this study, the parameters included were morphology, anatomy and micro-
morphology that were observed using Scanning Electron Microscope (SEM) and also USB
digital microscope in which made this study broader. The herbarium specimen is made and
kept for the future referencing. After all data been recorded and analyse, it is expected that
the species from genus Amomum are more easy to differentiate using the fruit since fruit is
the most distinct features available in all plants.
2
The genus Amomum in Malaysia is poorly known among ginger and need more
research and as well as the species which are difficult to describe because most of the
herbarium specimen are incomplete. There are a lot of species available under Amomum
genus, in order to compare all the species under genus Amomum genus properly, further
study are needed to study to clarify the different between them and fruit are chosen as
parameter in this study. Therefore, the objectives of this study are:
a) To study the morphological and anatomical characteristics of fruit of selected Amomum
species from Malaysia.
b) To compare the morphological and anatomical characteristics of the fruit of selected
Amomum species from Malaysia.
3
CHAPTER 2
LITERATURE REVIEW
2.1 Order Zingiberales
Order Zingiberales is morphologically diverse and a species-rich order that suited
for particular studying of evolution of floral diversity. Banana, tumeric, ginger are some of
the familiar plant that included in the order of Zingiberales. Under order Zingiberales,
there are eight families which are Cannaceae, Costaceae, Heliconiaceae, Lowiaceae,
Marantaceae, Musaceae, Strelitziaceae and Zingiberaceae. In the area before 1800s, the
classification for the family under order Zingiberales are placed differently which is
different from nowadays classification. The old version was that, Costaceae are placed
under the same family as Zingiberaceae while Strelitziaceae, Lowiaceae, and
Heliconiaceae is part of family Musaceae. Cannaceae and Marantaceae are already one of
the families under order Zingiberales. Some phylogenist are managed to reconstructed the
phylogenic history where they recognized the eight families under order Zingiberales,
Cannariae and Zingiberariae as two superfamilies, and five suborders within the order. The
attempt of reconstruction of the phylogenic history is done by several phylogenist which
are Lane, Tomlimson, Dahlgren and Rasmussen (Kress, 1990). Since the day they have
been found, this order showed a great evolutionary.
According to Kirchoff et. al. (2009), most members of the order has showy and
large flower which demonstrated specialized relationship with pollinator. The stems in
Zingiberales are commonly underground and short. The flower having three sepals, petals
4
carpel and its stamen up to six in two whorl of three each, and have three carpel. The most
common stem in Zingiberales is short and below the ground. The branches arise from
underground stem which are known as rhizome when it is elongated but a sucker when it is
short. Most of the flowers has three sepals, three petals, up to six stamen in two whorl of
three each and has three carpel.
The ovary is inferior which is enclosed by the united basal portion of other flower
parts which later arise at the upper part of the ovary. Except for Zingiberaceae, the calyx is
different from the corolla from the aspect of size and shape meanwhile the sepals are free
from each other. Zingiberaceae has a tubular calyx with small free lobes. The flowers are
zygomorphic or bilaterally symmetric but in certain cases they can be asymmetric too.
Exhibit five stamen or one functional stamen in most flowers but not for all genus
(Kirchoff et al., 2009).
In the study of phylogeny of family under order Zingiberales, Sass et al., (2016)
conducted a study to capture sequences that relatively high in polymorphisms to detect the
diversification that leading to the modern families. They used used transcriptomes that
were generated as part of the Monocot Tree of Life Project (MonAToL) or One Thousand
Plant Transcriptomes together with the whole genome of Musa acuminata recorded to
design a set of probes that were printed on an Agilent microarray chip in parallel. This
parallel printing approach enables divergent taxa to be captured on a single array and
alleviates binding competition between closely related and divergent individuals. (Sass et.
al., 2016). Thus, finding on this study showed that, as shown in Figure 1, with 100%
parsimony bootstrap, Musaceae is a sister to all other families and thus proves that the
5
gingers families (Cannaceae, Costaceae, Zingiberaceae and Marantaceae) are
monophyletic (Sass et al., 2016).
Figure 1: Phylogenetic tree of Zingiberales based on a partitioned ML of concatenated plastid and nuclear
sequence.
6
2.2 Family Zingiberaceae
The members of family Zingiberaceae are rhizomatous, perennial herbs which are
aromatic in any or all parts of the plant and it is comprises of 53 genera and more than
1500 species worldwide and its centre of diversity is at the Southeast Asia which has 80%
of all known species (Lamb et. al., 2013). It is the biggest family in the order Zingiberales
which include some important spices such as Zingiber officinale Roscoe (ginger), Curcuma
longa LINN. (turmeric), and Elettaria cardamomum L. (cardamom).
According to Smith (1985), species from the Zingiberaceae family usually are
plants of the forest floor. It also diverse in its habits and habitat thriving in shaded forests,
mixed dipterocarp kerangas, riverine habitat, secondary forest or alluvial forest with few
species of montane and limestone areas. The majority of its species is also a terrestrial with
some species occurs as epiphytes or litophytes (Ibrahim et. al, 2010). The characteristics of
this family is that fusion of lateral staminodes of the inner staminal whorl into labellum,
the occurrence of a cell that containing essential or ethereal oils that are automorphies to
the family and also the presence of the two epigynous nectar glands at the base of the style
of its flower (Kress, 1990).
Stated that there are 21 genera with over 200 species have been reported from
Borneo (Poulsen, 2006) including Haniffia as a new generic record for Sarawak (Wong et.
al., 2014). Recent study according to Gobilik and Mashitah (2005), stated that Sarawak has
141 species in 18 genera, Brunei 110 species in 13 genera and Sabah 152 species in 13
genera which all include in Borneo. (Lamb et. al., 2013)
7
2.3 Genus Amomum Roxb.
Amomum is one of the genus under family Zingiberaceae. They occur from the
Himalaya to Northern Australia and extend into the central Pacific and widely distributed
in Southest Asia (Kiew, 1982; Smith, 1985). According to Wu and Larsen (2000), most of
spesis in the genus Amomum are widely creeping rhizomes, their leaf blade mostly oblong-
lanceolate in shape, the inflorescene rising from the rhizomes, and lastly the fruit is often
in capsule. According to Kaewsri and Paisooksantivatana (2007) in their study of
Amomum, with 150-180 species, Amomum become one of the largest genera in the family
Zingiberaceae.
Previous study by Schuman (1904), he published his monograph of the
Zingiberaceae in “Das Pflanzenreich”. He accepted Roxburgh‟s generic concept to remove
Linnaean species from the genus and retained the name Amomum. He also subdivided
Amomum genus into two sections and four series. Section Geanthus which distiguished by
the absence of an anther appendage and composed of series Oliganthae and Polyanthae.
Section Euamomum with an anther appendage, it comprised of series Lobulatae and
Integrae. The four members of series Oliganthae have been subsequently transferred to the
genus Etlingera Giseke and also most members of series Polyanthae, while including
several true Amomum, may be more appropriately transferred to that genus as well.
Majority of much larger section Euamomum are retained in genus Amomum. Members of
family Lobulatae are distinguished by a bilobed or triloped anther connective and series
Integrae by an entire anther connective. Besides, he purposed that 40 African Amomum
species should be excluded and exstablished in genus Aframomum. This new genus was
distinguished by the shape of fruit and the trilobed anther connective, whereas the
8
reformed genus Amomum was wholly Asiatic. Several species transferred to four other
genera such as Alpinia, Curcuma, Elettaria and Zingiber.
According to Boyce (2006), currently there is no accurate number of Amomum in
Sarawak and some suggest that the number is more than 50 species. Many species from the
genus Amomum has been utilized as medicine, spice, condiment, and vegetable. The plant
from this genus has been long utilized, however, the identification is still confusing since
there are no good specimens in many herbaria and may lead to confusion and need more
intensive study (Kaewsri & Paisooksantivatana, 2007). For instant, many species of
Amomum were transferred to the other genera such as Aframomum, Elettariopsis, Alpinia,
Etlingera, and Hornstedtia.
2.4 Fruit Morphology
Fruits are adaptation that results from the protection and the distribution of the seed.
The fruits and seeds are mostly dispersed by animals but sometimes there are also another
disperser such as wind, gravity and water. Fruits that are though and full of fibre or
sclereids, such as walnuts, brazilnuts and coconuts have a maximum protection but they
are heavy and somehow expensive metabolically. Furthermore, a fragile fruit is better for
the germination to occur as if the fruits are hard to be broken, the seed cannot be easily
germinate. If animals are going to disperse the seeds, part of the fruits must be edible or at
least attractive to attract the animals while the seed must be protected from consumption.
9
Fusion of carpels also affects the nature of the fruit, if the fruit developed from a
single ovary or the fused ovaries of one flower, it is a simple fruit. Meanwhile, if a separate
carpel of one gymnoecium fuse during the development, aggregate fruits will form and if
during the development all the individuals fruit of an inflorescences fuse into one fruit, it is
a multiple fruit (Mauseth, 2012). In fruits, there are also a several layers such as epicarp,
mesocarp, and endocarp which known as pericarp. Morphological study is the study of
external part of the plant structure. According to Sattler and Rutishauser (1997), plant
morphology are remains relevant to practically all disciplines of plant biology such as
molecular genetics, physiology, ecology, evolutionary biology and also systematics. Study
conducted by Kaewsri and Paisooksantivatana (2007) revealed the morphology of
Amomum fruit from Thailand but the study are not in detail.
2.5 Fruit Anatomy
Anatomical study is the study of internal part of the fruit and can also be called as
phytotomy. Anatomy study deals with the structural organization of the fruit. Since the
middle 20th century, the investigations of plants anatomy considered as separate distinct
field and plant anatomy refer to just internal plant structures. In study by Kaewsri and
Paisooksantivatana (2007), the fruit anatomy is obtained, but there are no detail description
on the anatomy of the fruit such as the placentation types and the colour of the pericarp
being made. Schuman (1904) that purposed 40 African Amomum species should be
excluded and exstablished in genus Aframomum, used shape of fruit to distinguished the
new genus and the trilobed anther connective.
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2.6 Scanning Electron Microscope (SEM)
Scanning Electron Microscope (SEM) can be used to view structure of the object
that cannot barely seen by the naked eyes and also examine microscopic objects at a very
fine scale. Instead of light using in the other usual microscope, SEM shot electron particles
along with its high spatial resolution which will resulted in specific and informative image
up to the scale of nanometre. SEM use electromagnets instead of lenses which result in
much control in the degree of magnification up 15,000X. Previous study conducted by
Kaewsri and Paisooksantivatana (2007) using the SEM as tool to view the pollen grains of
Amomum in details. Clear image of the pollen are obtained. There is no study of fruit from
genus Amomum using SEM found.
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CHAPTER 3
3.0 MATERIALS AND METHODS
3.1 Materials and Sites
The samples were collected from various localities in Malaysia that include
Kelantan (Gua Musang), Pahang (Cameron Highland), Sarawak (Kuching, Serian-Gunung
Payang, Bau-Dered Krian, Miri-Long Banga) and also Universiti Malaysia Sarawak
(UNIMAS). Twelve species of Amomum with fruits have been examined.
Figure 2: The Map of Malaysia (Source: Google Map)