LEAF MICROMORPHOLOGICAL CHARACTERISTICS OF SELECTED

VATICA SPP. AND COMPARISON WITH OTHER GENERA IN TRIBE

DIPTEROCARPEAE (DIPTEROCARPACEAE)

Miraadila Binti Mohd Isa

(26867)

Bachelor of Science with Honours

(Plant Resource Science and Management)

2013

Faculty of Resource Science and Technology

Leaf Micromorphological Characteristics of Selected Vatica spp. and Comparison with Other

Genera in Tribe Dipterocarpeae (Dipterocarpaceae)

MIRAADILA BINTI MOHD ISA

This project is submitted in partial fulfillment of the requirement for degree of

Bachelor of Science with Honours in

Plant Science Resource and Management

FACULTY OF RESOURCE SCIENCE AND TECHNOLOGY

UNIVERSITI MALAYSIA SARAWAK

2013

i

ACKNOWLEDGEMENT

In the Name of Allah, the Compassionate, the Merciful, Praise be to Allah,

Lord of the Universe.

I would like to express my deepest appreciation to all those who provided me the possibility

to complete this final year project. A special gratitude I give to my supervisor, Prof Dr.

Cheksum Tawan and to my co-supervisor En. Qammil Muzzammil Abdullah for the entire

guidance, advices and suggestions in preparing this project.

My gratitude and sincere thanks also goes to all my coursemate, friends as well as Unimas

staffs especially En.Safri and En. Hidir Marzuki who offers their help in making this project a

reality. Finally, special dedicated and thanks a lot to all my beloved family especially my

father En. Mohd Isa Hassan, my mother Pn. Sapiah Baba who have contributed in giving me

their moral support, encouragement and understanding.

ii

APPROVAL SHEET

Name of candidate: Miraadila Binti Mohd Isa

Title of dissertation: Leaf Micromorphological Characteristics of Selected Vatica spp. and

Comparison with Other Genera in Tribe Dipterocarpeae

(Dipterocarpaceae)

________________________________

(Prof. Dr. Cheksum@ Supiah Tawan)

Supervisor

_________________________________

(Mr. Qammil Muzzammil Abdullah)

Co-Supervisor

_________________________________

(Dr. Rebicca Edward)

Coordinator

Plant Resource Science and Management Programme

Department of Plant Science and Environmental Ecology

Faculty of Resource Science and Technology

Universiti Malaysia Sarawak.

iii

DECLARATION

I hereby declare that the project is based on my original work except for quotation and

citations which have been duly acknowledgement. No portion of the work in this dissertation

has been submitted in support of an application for another degree of qualification of this or

any other university or institution of higher learning.

...........................................

(Miraadila Binti Mohd Isa)

Plant Resource Science and Management Programme

Department of Plant Science and Environmental Ecology

Faculty of Resource Science and Technology

Universiti Malaysia Sarawak.

iv

TABLE OF CONTENT

PAGE

Acknowledgements i

Approval Sheet ii

Declaration iii

Table of Content iv

List of Abbreviation vii

List of Tables viii

List of Figures ix

List of Plate x

Abstract xi

Abstrak xii

Chapter 1 1.0 Introduction 1

Chapter 2 2.0 Literature review 3

2.1 Family Dipterocarpaceae 3

2.2 Subfamily Dipterocarpoideae 7

2.3 Genus Vatica L. 7

2.4 Scanning Electron Microscopy (SEM) 8

2.5 Morphology studies 9

2.6 Micromorphology studies 9

2.7 Epidermal studies 9

2.7.1 Epidermal cell 10

2.7.2 Stomata 10

2.8 Petiole studies 11

Chapter 3 3.0 Materials and Methods 12

3.1 Materials 12

3.2 Methodology 14

3.2.1 Morphology studies 14

3.2.1.1 Leaves measurement 14

3.2.2 Micro-morphology studies 15

v

3.2.3 Leaf epidermis studies 15

3.2.4 Petiole anatomical studies 17

Chapter 4 4.0 Results and Discussion 18

4.1 Genus Vatica L. 18

4.2 Species Description 20

4.2.1 V. badiifolia P.S. Ashton 20

4.2.2 V. brunigii P.S. Ashton 22

4.2.3 V. coriacea P.S. Ashton 24

4.2.4 V. compressa P.S. Ashton 26

4.2.5 V. glabrata P.S. Ashton 28

4.2.6 V. globosa P.S. Ashton 30

4.2.7 V. nitens King 32

4.2.8 V. pedicellata Brandis 34

4.2.9 V. rynchocarpa P.S. Ashton 36

4.2.10 V. sarawakensis F. Heim 39

4.2.11 U. borneensis Symington 41

4.2.12 A. grossivenia Slooten 44

4.2.13 A. marginata Korth. 47

4.3 Leaf morphological 49

4.3.1 Leaf shape 50

4.3.2 Leaf margin 50

4.3.3 Leaf apex 53

4.3.4 Leaf base 53

4.3.5 Secondary veins 53

4.3.6 Glandular mark 54

4.3.7 Petiole 55

vi

4.4 Leaf micro-morphological 56

4.4.1 Trichomes 58

4.4.2 Stomata 65

4.5 Leaf Anatomical 69

4.5.1 Petiole 70

4.5.2 Epidermal 74

Chapter 5 5.1 Conclusion 80

5.2 Recommendation 81

References 82

Glossary 86

Appendices 1 98

vii

LIST OF ABBREVIATION

BO : Herbarium of Bogorense

CST : Herbarium prefix number for Cheksum@ Supiah Tawan

FR : Forest Reserve

HUMS : Herbarium Universiti Malaysia Sarawak

K : Kew Herbarium

KEP : Herbarium of Forest Research Institute Malaysia

L : Herbarium of Leiden University

MK : Herbarium prefix number for Meekiong Kalu

MI : Herbarium prefix number for Miraadila Mohd Isa

MWC : Matang Wildlife Centre

NaOCl : Sodium hypochlorite

NP : National Park

Pd : Silver

S. : Herbarium prefix number for SAR

SAR : Herbarium of Sarawak Forestry Department

SEM : Scanning Electron Microscopic

Spp. : Species

UNIMAS : Universiti Malaysia Sarawak

viii

LIST OF TABLES PAGE

Table 1

:

Distribution of genera and species of Dipterocarps in the

world

6

Table 2 : List of species studied. 13

Table 3 : Leaves morphological characteristics of Vatica spp., U.

borneensis and Anisoptera spp.

52

Table 4 : Trichomes analysis of 10 selected Vatica spp., U.

borneensis and two Anisoptera spp.

59

Table 5 : Stomata analysis on the leaf surfaces of 10 selected Vatica

spp., U. borneensis and 2 Anisoptera spp.

66

Table 6 : Petiole analysis of ten selected Vatica spp., U. borneensis

and two Anisoptera spp.

71

Table 7 : Epidermal analysis of leaves of ten selected Vatica spp., U.

borneensis and two Anisoptera spp.

75

ix

LIST OF FIGURES PAGE

Figure 1 : Phylogeny of Dipterocarpaceae 4

Figure 2 : World Distribution of Dipterocarpaceae. 5

Figure 3 : Leaf measurement of Vatica species. 14

Figure 4 : Vatica badiifolia P.S. Ashton 21

Figure 5 : Vatica brunigii P.S. Ashton 23

Figure 6 : Vatica coriacea P.S. Ashton 25

Figure 7 : Vatica compressa P.S. Ashton 27

Figure 8 : Vatica glabrata P.S. Ashton 29

Figure 9 : Vatica globosa P.S. Ashton 31

Figure 10 : Vatica nitens King 33

Figure 11 : Vatica pedicellata Brandis 35

Figure 12 : Vatica rynchocarpa P.S. Ashton 37

Figure 13 : Vatica sarawakensis F. Heim 40

Figure 14 : Upuna borneensis Symington 42

Figure 15 : Anisoptera grossivenia Slooten. 45

Figure 16 : Anisoptera marginata Korth 48

Figure 17 : Comparative illustration of leaf for 10 species of Vatica, 2 species

of Anisoptera and Upuna borneensis.

51

x

LIST OF PLATES PAGE

Plate 1 : Vatica rynchocarpa P.S. Ashton. 38

Plate 2 : Upuna borneensis [specimen S.26237, herbarium specimen

deposited at SAR]

43

Plate 3 : Anisoptera grossivenia (variations of leaf characteristics). 46

Plate 4 : Trichomes present on the glandular marks. 60

Plate 5 : Surface ornamentation on the glandular marks of Vatica spp. 61

Plate 6 : Peltate trichomes on the abaxial surface 62

Plate 7 : Stellate trichomes on abaxial surface. 63

Plate 8 : Various types of stellate trichomes found on petiole surface 64

Plate 9 : Stomata found on the abaxial surface (i). 67

Plate 10 : Stomata found on the abaxial surface (ii). 68

Plate 11 : Anisoptera grossivenia. 70

Plate 12 : Transverse sections of leaf petiole of selected Vatica spp. 72

Plate 13 : Tranverse sections of leaf petiole of selected Vatica spp,

Upuna borneensis & two Anisoptera spp.

73

Plate 14 : Epidermal cells of selected Vatica species. 77

Plate 15 : Stomata [cleared leaves] (i) 78

Plate 16 : Stomata [cleared leaves] (ii) 79

xi

LEAF MICROMORPHOLOGICAL CHARACTERISTICS OF SELECTED VATICA

SPP. AND COMPARISON WITH OTHER GENERA IN TRIBE DIPTEROCARPEAE

(DIPTEROCARPACEAE)

MIRAADILA BINTI MOHD ISA

Plant Resource Science and Management Programme

Faculty of Science and Technology

Universiti Malaysia Sarawak

Abstract

The Asian Dipterocarpaceae family has been divided into two major groups; Tribe Shoreae

and Dipterocarpeae based on the arrangement of sepals and number of chromosome. Species

identification for certain genera within the tribe Dipterocarpeae are still doubtful due to

lacked of information. Therefore, detail studies on morphology, micro morphology and

anatomy of leaves for 10 selected Vatica species, Upuna borneensis and two Anisoptera

species have been conducted. Examinations on the leaf morphological have been observed

via naked eyes, lens, microscopes and Scanning Electron Microscopic. The results from leaf

morphological observation show significant difference for both generic and species levels.

Meanwhile, the results from micromorphology studies showed significant difference only at

generic level. The noticeable difference that spotted on the leaf surfaces were the presence of

trichomes. The peltate and stellate trichomes were encountered in Vatica species while

comparably, simple trichomes were observed only on U. borneensis and the two Anisoptera

species. Moreover, anatomy studies which included the petiole tranverse section and

epidermal characteristics showed no significant difference for both generic and species levels

except for V. ryncocarpa. The results from morphology, micromorphology and anatomy

studies were very useful for taxonomical aspects in Dipterocarps family. However,

comprehensive studies should be conducted to achieve precise results.

Keywords: Dipterocarpaceae, Dipterocarpeae, Vatica, Upuna borneensis, Anisoptera, leaf

morphology, micromorphology, anatomy.

xii

Abstrak

Famili Dipterocarpaceae di Asia dibahagikan kepada 2 kumpulan yang besar; iaitu tribal

Shoreae dan Dipterocarpeae berdasarkan susunan sepal dan bilangan kromosom.

Identifikasi spesies untuk beberapa genera dalam tribal Dipterocarpoideae masih banyak

dipersoalkan kerana maklumat morfologi yang kurang. Oleh itu, kajian terperinci

berdasarkan morfologi, mikromorfologi dan anatomi daun untuk 10 spesies terpilih dari

genus Vatica, Upuna borneensis dan dua spesies dari genus Anisoptera telah dijalankan.

Penelitian morfologi daun dilakukan dengan menggunakan mata kasar, kanta pembesar,

mikroskop dan secara Imbasan Mikroskopik Elektron. Keputusan dari pengamatan morfologi

daun menunjukkan signifikasi pengecaman pada peringkat antara generik dan antara spesies

di dalam genus Vatica. Manakala, keputusan daripada mikromorfologi menunjukkan

signifikasi pada peringkat generik. Perbezaan yang ketara dapat dilihat melalui jenis trikom

yang terdapat pada daun. Jenis trikom yang terdapat pada spesies Vatica adalah peltate dan

stellate manakala, trikom jenis mudah terdapat pada U. borneensis dan dua spesies

Anisoptera. Kajian anatomi yang melibatkan penelitian keratan rentas tangkai daun dan

karakter pada lapisan epidermis tidak menunjukkan perbezaan yang signifikan di antara

genus dan spesies kecuali V. ryncocarpa. Kajian morfologi, mikromorfologi dan anatomi

daun untuk genera dan spesies yang terpilih dalam ini adalah sangat berguna dalam

taksonomi famili Dipterokarp. Namun demikian,kajian menyeluruh perlu dilaksanakan untuk

mendapatkan keputusan yang lebih jitu.

Kata kunci: Dipterocarpaceae, Dipterocarpeae, Vatica, Upuna borneensis, Anisoptera,

morfologi daun, mikromorfologi, anatomi

1

CHAPTER 1

INTRODUCTION

The tropical rainforests in Southeast Asia are characterized by a high species diversity of trees

(Whitmore, 1984). The extreme floristic richness is largely due to co-occurrence of a great

number of species within the same community (Whitmore, 1998). It has been stated that Borneo

is the largest area that has long been known to have high species diversity of trees that compare

among the world’s tropical rainforests. It has also been listed as one of the key area for

endemism due to remarkably large number of species. In lowland Southeast Asian tropical

forests, dipterocarp species dominate the forest canopy (Ashton, 1988). Approximately one

quarter of all trees in most lowland forests of Borneo are dipterocarps (Ashton 1982; Sist 1996;

Slik et al., 2003). As a result of its isolation from the continent, Borneo is considered to be a

center of biodiversity in tropical Asia based to the wide variety of plant species that have been

developed earlier (Hazebroek & Morshidi, 2001).

The genus Vatica L. is one of the largest natural groups in the family Dipterocarpaceae with 71

species have been botanically described. This genus distributed from India, Sri Lanka to

Myanmar, Indochina, Sumatra, Borneo and the Philippines, many of their species are still

doubtfully identified. In general, following characteristics are used for describing leaf surface:

epidermis, venation systems, trichomes, structure of epicuticular waxes, and stomata. These

micro morphological characters of leaf are not only significant in physiological functions of

plant but they are also valuable in taxonomic studies and also useful for investigation of

phylogenetic relationships (Stace 1984; Joshi 2001). Metcalfe & Chalk (1957) reported the 2

2

importance of the stomatal types, the distribution and types of trichomes, as informative

anatomical characteristics.

This study need to be done to add the significant features on the leaf structures in the genus

Vatica. However, this is the first attempt to use micromorphological and anatomy characters

particularly for Vatica species in Sarawak and hopefully all the features in micromorphological

as well as anatomy are able to be used for supporting the identification based on morphological

data. Therefore, the aims of this study are:

1. To compare leaves morphological aspects of selected Vatica spp.with selected genera in

tribe Dipterocarpeae by comparing with the existing information.

2. To obtain the characteristics of Vatica spp. especially on micromorphological and

compare with selected genera in tribe Dipterocarpeae such as Upuna, and Anisoptera..

3. To compare leaf epidermal and petiole anatomy of selected Vatica spp. with selected

genera in tribe Dipterocarpeae.

4. To produce key of the Vatica spp. based on the morphology, micromorphological and

anatomy of selected features for identification.

3

CHAPTER 2

LITERATURE REVIEW

2.1 Family Dipterocarpaceae

Family Dipterocarpaceae was established for the first time in 1925 by Blume. At that time, this

family is considered to be related to Tiliaceae and Clusiaceae. The Dipterocarpaceae family

comprises three subfamilies (Dipterocarpoideae, Monotoideae and Paraimoideae) with about 510

species in 17 genera according to recent classification [Ashton (1982); Londono et. al., (1995);

Morton (1995); Symington (2004)], Dipterocarpaceae from tropical Asia with approximately 470

species in 13 genera, distributed in the Seychelles, Sri Lanka and India northeastwards to

southern China and the Batan Islands, and southeastwards to New Guinea and the

D’Entrecasteaux Island; Monotoideae from Africa with 39 species in two genera and the new

monotypic genus Pseudomonotes from Colombia in South Amerika, and the monotypic

subfamily Pakaraimoideae from Guyana in South Amerika (Table 1).

The Asian Dipterocarps (subfamily Dipterocarpoideae) are further divided into two tribes,

Dipterocarpeae and Shoreae (Brandis 1895). The tribe Shoreae is richer in species than

Dipterocarpaeae due to species richness in the genera Shorea and Hopea. Shoreae is the largest

and economically most important genus of Dipterocarpaceae, encompasses about 200 species in

11 sections, of which 163 species are distributed in Malesia, mostly in Indonesia, in particular on

Sumatra and Borneo (Kalimantan), while genus Hopea comprises more than 100 species.

4

Figure 1: Phylogeny of Dipterocarpaceae (Adopted from Ashton, 1982).

5

Figure 2: World Distribution of Dipterocarpaceae (Based on Symington, 2004)

6

Table 1: Distribution of genera and species of Dipterocarps in the world (Based on Symington, 2004)

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South America 1 1 2 2

Africa 30 3 2 33

Seychelles Islands 1 1 1

India (West Peninsular) 2 1 8 2 1 2 6 16

Sri Lanka 4 2 10 5 5 3 2 25 1 9 57

Bangladesh 1 1 4 1 1 5 8

India (Northern East) 1 2 4 1 4 8

Andaman Island 2 6 2 8

China 1 1 3 1 3 5 9

Myanmar (Burma) 4 5 1 2 6 1 11 2 4 9 36

Indo-China 4 4 1 4 10 2 11 1 9 9 46

Thailand 4 6 1 2 8 3 13 2 6 1 10 46

Peninsular Malaysia 17 10 10 21 1 3 11 21 1 31 3 6 23 2 14 160

Sumatra and adjacent Islands 10 9 8 23 3 6 10 25 2 4 11 2 12 113

Borneo 27 12 29 60 6 16 27 41 7 5 35 3 1 13 269

Java 1 1 4 1 3 5 10

Filipina 5 4 1 5 1 1 9 5 9 3 9 11 52

Sulawesi 2 2 1 2 4 7

Maluku 1 1 2 1 1 5 6

New Guinea 12 1 1 1 4 15

GEOGRAPHICAL

REGION

7

2.2 Subfamily Dipterocarpoideae

The Asian subfamily Dipterocarpoideae includes 13 genera and 475 species, confined to the

Asian tropics between the Seychelles Island and South Asia to New Guinea but absent in the

Lesser Sunda Island east of Sunbawa (Ashton, 1982). Most genera and species occur in the

West Malesia: Sumatra, Peninsular Malaysia, Singapore, Java, Borneo, the Philippines and

islands in between Sabah and Sarawak, which have 267 species in 9 genera, is the richest area

in the world for dipterocarp species (Ashton, 2004).

Morphologically, the Dipterocarpoideae can be divided into two groups which are the tribe

named Shoreae and Dipterocarpeae (Ashton, 1982; Maury-Lechon & Curtet, 1998). Tribe

Dipterocarpeae (Valvate–Dipterocarpi group) includes Anisoptera, Cotylelobium,

Dipterocarpus, Stemonoporus, Upuna, Vateria, Vateriopsis, and Vatica. The genera of this

group have valvate sepals in fruit, solitary vessels, scattered resin canals, and basic

chromosome number x = 11. The tribe Shoreae (Imbricate-Shoreae group) comprises four

genera which are Balanocarpus, Hopea, Parashoraea, and Shorea. All of them have

imbricate sepals, grouped vessels, resin canals in tangential bands, and basic chromosome

number x=7.

2.3 Genus Vatica L.

The genus Vatica was described by Carl Linnaeus in 1771 based on the type specimen Vatica

chinensis from China (Kostermans, 1992) . This genus Vatica is rich in species diversity with

65-71 species distributed in SriLanka, Thailand, Malesia, Burma, Indo-china, South and East

India, South China and Bangladesh (Dayanandan et al., 1999; Joshi, 2001). Distribution of

Vatica occurring particularly in mixed and upper dipterocarp forests and also non-saline

habitats at elevation from sea level up to about 1,800 m altitude. The Vatica species are

distinguished from other genera of the Dipterocarpaceae by the absence of looped intra-

8

marginal nerves on the leaves, the winged fruits enclose less than half of the nut, anthers are

glabrous and the style is stout (Ashton, 1982; Pooma & Newman, 2001; Srinual &

Thammthaworn, 2008). Sasaki (1980) said that the seed of this genus contain starch grain,

and able to tolerance to cold temperature because it can be endured at 4oC for 2 months.

2.4 Scanning Electron Microscopy (SEM)

Scanning Electron Microscopy (SEM) that was introduced about 20 years ago, has given

very fine details of surface three dimensional, and it can be appreciated this remarkable

development will be used in all kinds of scientific fields (Troughton & Donaldson ,1972).

Nowadays SEM is one of an important tool that being used in the Plant Systematic Study. It

is scientific instruments that use a beam of energetic electrons to examine objects on a very

fine scale. To perform well it required 200-1000 × or more magnification which are not

possible using current optical microscopes.

Anonymous (2010) stated that SEM provide high magnification, high resolution images of

samples at magnification up to 50,000x. This enable details structures that cannot be seen

using bare eyes were conducted. For example, Srinual & Thammthaworn, (2008) also

included the SEM images in their study on leaf anatomy of Vatica in Thailand. Melcalfe &

Chalk (1979) compiled a historical review of the plant surface and present several data

concerning observations by scanning electron microscope. Troughton & Donaldson (1972)

said that this SEM tool was made with reference to some anatomical features in plants and

discussed the relationship of these structures to physiological processes. With the

combination of higher magnification, larger depth of field, greater resolution and

crystallographic information, there is no doubt why this SEM is one of the most heavily used

instruments in academic, research areas and industry.

9

2.5 Morphology studies

Originated by Goethe in 1790, plant morphology has established for 211 years as a scientific

discipline (Donald, 2001). Plant morphology or phytomorphology is the study of the physical

form and external structure of plants (Raven et al., 2005). In other word, leaf morphology is

the study of external feature of leaves. Leaves vary greatly from plant to plant and are useful

in classification and identification. Usually, the parameter involves are the leaf parts itself for

example the blade, petiole and stipules. Leaf structure, attachment, shapes, margins, and their

venation is always being the main measure used to describe the plant. The comparative study

of leaf morphology of Asteraceae conducted by Hayakawa et al., (2012) mentioned that the

science of plant morphology still have their role in modern biology.

2.6 Micromorphology studies

Micro-morphological is the study of plant structures at a microscopic level. The significant of

micromorphological features in taxonomic considerations of Angiosperms is now well

recognized (Ramayya, 1972; Tomlison 1979; Go & Latiff, 1998; Parveen et al., 2000).

According to Pole (2010), micro-morphological studies of epidermal structures such as

epicuticular waxes, cuticle morphology or various types of hairs may provide important

insights into ecological properties of plant and often also used in taxonomic studies.

2.7 Epidermal studies

Leaf epidermal characteristics have potential on taxonomical importance (Jones, 1986;

Baronova, 1992; Ding et al., 2005). For example, study has done by Noraini & Cutler (2009)

able to reveals leaf anatomical and micromorphological characters of some Parashorea

10

(Dipterocarpaceae) species. The study includes transverse section of the lamina, transverse

sections of leaf margin, clearing of epidermal layer which emphasizes on variations of

epidermal cells, stomata complexes, trichomes and resin canals.

2.7.1 Epidermal cell

The epidermis is the outer layer of cells covering the leaf. It forms the boundary separating

the plant’s inner cells from the external world. The epidermis tissue includes several

differentiated cell types for example epidermal cells itself, the trichomes and stomata

complex which comprises guard cells and subsidiary cells. The leaf epidermal cells are of

significant taxonomic importance for example their length and width which useful for

classification (Wilkin & Sabanci, 1990; Albert & Sharma, 2013).

2.7.2 Stomata

Stomata are very minute openings formed in the epidermal layer of green parts of the plants,

especially in the leaves (Ananta & Harisha, 2012). Every stoma is enclosed by the semi lunar

cells known as the guard cells. The function of stomata is important on the physiology,

adaption and productivity of plants. According to Metcalfe & Chalk (1950), generally

stomata occur only on the lower surface; however they have recorded them on both the

surface in Empogona buxifolia, E. corollata and E. radians. Stomata are divided into 64

different types depending upon the type of guard cells and arrangement of subsidiary cells

(Pandey, 2006). But six to seven types are very common in nature such as paracytic, diacytic,

anisocytic, anomocytic, actinocytic, gramineous and coniferous stomata.