UNIVERSITI PUTRA MALAYSIA STUDIES ON …psasir.upm.edu.my/10417/1/FP_1998_7_A.pdf · universiti...
Transcript of UNIVERSITI PUTRA MALAYSIA STUDIES ON …psasir.upm.edu.my/10417/1/FP_1998_7_A.pdf · universiti...
UNIVERSITI PUTRA MALAYSIA
STUDIES ON INCOMPATIBILITY POLLINATION AND FLORAL
MORPHOLOGICAL POLYMORPHISM OF CARAMBOLA (AVERRHOA CORAMBOLA L)
NOBUHIKO FUSHIMI
FP 1998 7
STUDIES ON INCOMPATIBILITY POLLINATION AND AND FLORAL MORPHOLOGICAL POLYMORPHIS M OF CARAMBOLA
(Averrhoa coramlJola L)
By
NOBUHIKO FUSHIMI
Thesis submitted in fulfilment of the Requirements for Degree of Master of Agricultural Science
in the Faculty of Agricultural Scien ce Universiti Putra Malaysia
January 1998
ACKOWLEDGEMENTS
I am grateful to Professor Dr. Wong Kai Choo, Chainnan of supervisory
committee, Dr. Sayed Mohd Zain Sayed Hasan and Dr. Awang Soh Mamat,
members of SupetVisory Committee, Universiti Putra Malaysia (UPM), for their
valuable guidance, encouragement and support throughout the research and
preparation of the thesis.
I particularly would like to thank Dr. Yasuo Ota, Professor of the
Department of International Agricultural Development Science, Tokyo University of
Agriculture, for his approva� support and encouragement on these studies.
I would like to thank Dr. Hitoshi Uematsu, Senior lecturer of the Department
of Agricultural Science, Tokyo University of Agriculture, Junior College, for his
approval, encouragement and support on these studies.
I would like to thank Mr. Hoe Oi Kuan, Ms.Aminah Jusoh and Ms Agilah
Abdul Salil in Electron Microscopy Unit, Faculty of Veterinary and Animal Science,
UPM, for their guidance, encouragement and support throughout this research.
I am grateful to Dr. Naohiro Ikeda, Associate Professor of the Department of
Agronomy, Tokyo University of Agriculture, for critical reading of manuscript and
making valuable suggestions throughout this research.
11
I would like to thank Dr. Kiyomi Kosaka, Resident Representative of Japan
International Research Centre for Agricultural Sciences (JIRCAS) Malaysia Office,
for encouragement and financial support on this research.
I would like to express my sincere gratitude to the Supporting Organisation of
Japan Overseas Cooperation Volunteers for the scholarship award and support.
Finally, I am indebted to my mother, brother and sister for their infinite love,
encouragement and support.
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TABLE OF CONTENTS
Page
ACKOWLEGE.MENTS. .......... .......... ...... ...... ............ ........ .................... ....... 11
LIST OF CF TABLES.................................................................................... vi
LIST OF PI...ATES...................................................................................... . .. IX
LIST OF FIGtJRES ................ ....... . .. ............ ........ ................................ . . . . . .. , xiv
ABSTRACT..................................................................................................... xv
ABSTRAK.......................... ........................................................................... . . XVl1
CHAPTER
I
II
INTRODUCTION .............................................................................. .. 1
REVIEW OF LITERATtJRE ........................................................... . 5
The Carantbola................................... ........ . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . 5 General....................................................................................... 5 Tree Characteristics............. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 Flower Characteristics... .. .............. ........ . . . . . . . . . . . . .. . . . . .. . . .. . . . . . . . . . . . . 6 Fruit Characteristics................................................................... 7 Oones....................................................................................... 8 Flowering and Pollination...... . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . .. .. 10 Nutritive Value and Uses... ......... . . .. .. . . . . . . . . . . . . . . . . .. . . .. . . . . .. . . . . . . . . . . . . . 1 1
Self-incompatibility Systent.......... . . . . . . . . . . . . . . . . . . . . .. .. . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12 Definition................................................................................. . . 12 Oassification of Self-incompatibility System... ................... . . . . . . 13
Heteromorphic Incompatibility.............................................................. 14 Introduction.................... . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . .. .. .. . . . . . . . . . . . 14 Style and Stanten I..ength.................. ........ . . .. . . . . . . .. . . .. .. .. . . . . . . . . . . . . . 15 Pollen Characteristics............................................................... . 16 Stigma Characteristics... .... . . . . . . . . . . . . .. . . . . . . . . .. . .. . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . 17 Patterns of Heteromorphic Incompatibility................................ 19 Pollen-pistil Interaction Associated with Heteromorphic Incompatibility........................................................................... 20
Previous Experimental Methodology for Self-incompatibility Study.... 23 Field Pollination Study.................................. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23 Fluorescence Microscopy........................................................... 24 Scanning Electron Microscopy........... . . . . .. . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
iv
III MATERIALS AND METIIODS.......................................................... 26
Plant Materials........................................................................................ 26 Study 1. Florescence and Scanning Electron Microscopic Studies
on Compatible and Incompatible Pollinations of Pin and Thnun Morphs........................................................................... 31
Study 2. Field Pollination Studies on Compatible and Incompatible Pollinations of Pin and Thnun Morphs........................ ..... . . . . . . . .. 33
Study 3. Studies on Morphological Polymorphism of Pin and Thrum Morphs.................................................................................... . .. 34
Statistical Analysis and Experimental Designs......... .................. . .. . . . . . ... 36
IV RESULTS........................................................................................... . . 38
V
VI
Study 1. Florescence and Scanning Electron Microscopic Studies on Compatible and Incompatible Pollinations of Pin and Thnun Morphs.......................................................................... 38
Study 2. Field Pollination Studies on Compatible and Incompatible Pollinations of Pin and Thnun Morphs............ ........ .... .. .. .. . ... .. 52
Study 3. Studies on Morphological Polymorphism of Pin and Thrum Morphs...................................................................................... 64
DISCUSSION ...................................................................................... . 84
Floral Dimorphism Associated with Heteromorphic Incompatibility... 84 Pollen-pistil Interaction Associated with Heteromorphic Incompatibility. 86
Pollen Adhesion and Gennination... .......... . ...... . . .. . . . ..... ...... ...... 86 Inhibition Site of Pollen-tube Growth....................................... 87 Fruit Set Associated with Self-incompatibility Response........ 89 Fruit Parameters Associated with Self-incompatibility Response 90
SUl\fMARY AND CONCLUSION .................................................. .. 92
REFEREN"CES....................... . . . ..... . .. . . . . . ... . . . .. .. . . . . ........ . . . ... .. ...... . . . . .. . . .... . .. . ..... 95
APPENDICES
A Additional Tables.............................................................................. .... 102 B Additional Figures..... ................ .... . . . . . . . . . . . . ...... ... . ........ . . . . . . . . . . . . . . .. .. . . .... . . . 121
VITA.............................................................................................................. . . . . 123
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LIST OF TABLES
Table Page
1 Characteristics of Carambola Clones in Florida, USA. .. . . . . . . . . . . . .. . . . . .... . . . . . 9
2 List of Registered Carambola Clones in DOA, Serdang, Malaysia .. . . . . . .. . . . . 9
3 Sites of Inhibition of Male Gametophyte in lntramorph Pollinations of Some Heterostylous Plants. . . . . . . . . . . . . . . .. .. . . .. .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
4 List of Carambola Clones Used in Study 1 . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
5 List of Carambola Clones Used in Study 2................. .... . ....... . . . . . . . . . . . . . . . 27
6 List of Carambola Clones Used in Study 3.. . .. .. .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28
7 Mean Value of Percentage Pollination Success for Six Pollination Treatments . . . . . . . .. . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . 52
8 Percentage of Nonnal Fruit in Four Pollination Treatments. . . . . . . . . . . . . . . . . . . . . . . 55
9 General Characteristics of Nonnal Fruit in Four Pollination Treatments . . . 56
10 Mean Values of Fruits Characteristics obtained from Intennoxph Pollination in Pin Morph .......................... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . '" 59
1 1 Mean Values of Fruits Characteristics obtained from Self Pollination in Pin Morph . ...... ... . .... . . . ..... .... . . . . . . . . . . . ... ...... . . . . . . . . . . . . . . . . . . " . . . . . . . . . . . . . . . 59
12 Mean Values of Fruits Characteristics obtained from Intramorph Pollination in Pin Morph . ... .. .. .. . . .. .. ....... . . ..... .. .. . . . . . . . . . . . . . . . . . . . . , . . . . . . . . . . . . . . . . . . . . . . . . . 60
13 Mean Values of Fruits Characteristics obtained from Abnonnal Form in Pin Morph. . . ... . .. .. . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60
14 Mean Values of Fruits Characteristics obtained from Intermorph Pollination in Thrum Morph. ...... .. ...... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62
1 5 Mean Values of Fruits Characteristics obtained from Abnonnal Form in Thrum Intermorph Pollination. . . . ... . ... . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
16 Measurements of Floral Parts in Two Floral Morphs of Averrhoa carambola L. .. .. . . . . . . . . . . . . . . . . .. . . . . . . .. .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65
17 Measurements of Pistil and Stamen Length among Clones of Averrhoa carambola L. . .. .... . ... . . ........ . ....... .... .. . . .. . . . . .... . . ....... . . . . ... ..... .. . 67
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1 8 Measurements of Pollen Diameter among Clones of Averrhoa carambola L.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
19 Measurements of Stigma among Clones of Averrhoa carambola L. . . . . . . . . 72
20 Measurements of Stigmatic Vesicle among Clones of Averrhoa carambola L. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74
21 Percentage of Pollination Success in Intennorph Pollination in Pin Morph ... ... . .. . . ... . . . . . .. . . . . . . . . . . . . . . . .. . . . .. . . . . . . . . . . ... . ... ..... . . . . . . . . . . . . . 102
22 Percentage of Pollination Success in Self-Pollination in Pin Morph . . . . . . 103
23 Percentage of Pollination Success in Intramorph Pollination in Pin Morph . . . . . .. .. . . . . . . . ....... . . . . . ..... . . . . . . . . . . . . . . . . . . . . . . . .. . ..... . . . .. . . . . . . .. . . 103
24 Percentage of Pollination Success in Intennorph Pollination in Thrum Morph . .. . . . . .. . . . . . . . .. . . . . . . . . . . . . . .. . ... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . .. 104
25 Percentage of Pollination Success in Self-Pollination in Thrum Morph .... 105
26 Percentage of Pollination Success in Intramorph Pollination in Thrum Morph . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 106
27 Results of Analysis of Variance from Mean Values of Pollination Percentage for Six TreatInents ...... . ................ . ............. ...... .......... ....... . . . . . 107
28 Results of Analysis of Variance from Mean Values of Fruit Parameters (Nonnal Formed) ................... ........................................................ . . . . . . . . . . 108
29 Results of Analysis of Variance from Mean Seed Number per Fruit. . .. . . . . 109
30 Results of Analysis of Variance from Mean Fruit Weight.. . .. .................. . 1 10
3 1 Results of Analysis of Variance from Mean Fruit Length....... .... .. . . . . .. . . . . . 1 1 1
32 Results of Analysis of Variance :from Mean Fruit Width . . . . . . . . . . . . . . . . . . . .. . . . . . 1 12
33 Results of T -test on Pistil and Stamen Length......................... . . . . . . . . . . . . . . . . . 1 13
34 Results of T -test on Pollen Size........................................................... . . . . . 1 14
35 Results of T-test on Stigma Size.......................................................... . . ... 1 15
36 Results of T -test on Stigmatic Vesicle Size............................................ . . 1 16
37 Results of Analysis of Variance on Pistil and Stamen Length.................. 1 17
38 Results of Analysis of Variance on Pollen S ize.................................... .... 118
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39 Results of Analysis of Variance on Stigma Size....................... .......... . . ... 119
40 Results of Analysis of Variance on Stigmatic Vesicle Size...................... 120
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LIST OF PLATES
Plate Page
1 Source of Plant Material in Study 1 (Field 2, UPM, Serdang). . . . . . . . . ... . . . . . . . 29
2 Source of plant material in Study 2 (Horticultural unit, UPM, Serdang).. . . . 30
3 Source of plant material in Study 3 (DOA, Serdang).. . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . 30
4
5
6
Scanning Electron Micrograph of Pollen-tube Behaviour of Averrhoa carambola L. at 0.5 h after Thrum Intennorph Pollination (BI0 xB2) . ....... ........ ........................................ .......... ........................... .
Scanning Electron lvlicrograph of Pollen-tube Behaviour of Averrhoa carambola L. at 1 h after Thrum Intennorph Pollination (BI0 x B2) . . .. . . . . .... .. ..... ... . . ...... .... .... .. .. . . . ... ... . .. ... . .......... .... . . . ... . . .. .. .. . . . . .. . .
Scanning Electron Micrograph of Pollen-tube Behaviour of Averrhoa carambola L. at 2 h after Thrum Intermorph Pollination (BI0 xB2) . . . . . . . . . . .. . . .. . . . . . . . . . . . . . . . . . . .... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . .. . .
39
39
40
7 Fluorescence Micrograph of Pollen-tube Behaviour of Ave"hoa carambola L. at 2 h after Pin Intermorph Pollination (B2 x BlO) (X 100) . . . . . . . . . . . . . . 41
8 Fluorescence Micrograph of Pollen-tube Behaviour of Averrhoa carambola L. at 4 h after Pin Intennorph Pollination (B2 x BI0) (X 100). .. . . .. . . . . . . . . . . . 41
9 Fluorescence Micrograph of Pollen-tube Behaviour of Ave"hoa carambola L. at 8 h after Pin Intermorph Pollination (B2 x BI0) (X 100) . . . . . . . . . . . . . . . . . 42
10 Fluorescence Micrograph of Pollen-tube Behaviour of Ave"hoa carambola L. at 2 h after Pin SeltPollination (B2 x B2) (X 100) . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . 43
11 Fluorescence Micrograph of Pollen-tube Behaviour of Ave"hoa carambola L. at 4 h after Pin self Pollination (B2 x B2) (X 100). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
12 Fluorescence Micrograph of Pollen-tube Behaviour ofAverrhoa carambola L. at 8 h after Pin SeltPollination (B2 x B2) (X 100).. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
13 Fluorescence Micrograph of Pollen-tube Behaviour of Averrhoa carambola L. at 24 h after Pin SeltPollination (B12 x B12) (X 100).. . . . . . . . . . . . . . . . . . . . . . . . . 45
14 Fluorescence Micrograph of Pollen-tube Behaviour of Averrhoa carambola L. at 2 h after Thrum Intennorph Pollination (BI0 x B2) (X 100). . . . . . . . . . . . 47
15 Fluorescence Micrograph of Pollen-tube Behaviour of Averrhoa carambola L. at 4 h after Thrwn Intermorph Pollination (BI0 x B2) (X 100). . . . . . . . . . . . . 47
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16 Fluorescence Micrograph of Pollen-tube Behaviour ofAverrhoa carambola L. at 8 h after Thrum Intennorph Pollination (BI0 x B2) (X 100). .. . . . . . . . . . 48
1 7 Fluorescence Micrograph of Pollen-tube Behaviour of Averrhoa carambola L. at 2 h after Thrum Self Pollination (BlO x BlO) (X 100). . . . . . . . . . . .. . . . . . . . . . 49
1 8 Fluorescence Micrograph of Pollen-tube Behaviour of Averrhoa carambola L. at 4 h after Thrum Self Pollination (BI0 x B10) (X 100). . . . . . . . . . . . . . . . . . . . . . . 49
19 Fluorescence Micrograph of Pollen-tube Behaviour ofAverrhoa carambola L. at 8 h after Thrum Self Pollination (B lOx B 10) (X 100). . .. . . . . . . . . . . . . . .. . . . 50
20 Fluorescence Micrograph of Pollen-tube Behaviour ofAverrhoa carambola L. at 4 h after Thrum Intramorph Pollination (B10 x B17) (X 100). . . . . . . . . . . 50
21 Fluorescence Micrograph of Pollen-tube Behaviour of Averrhoa carambola L. at 8 h after Thrum Intramorph Pollination (B10 x B17) (X 100) . . . . . . . . . . . . 5 1
22 Scanning Electron Micrograph of Averrhoa carambola L. Pollen (x 3000) Clone B 1 (Pin). . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . 75
23 Scanning Electron Micrograph of Averrhoa carambola L. Pollen (X 3000) Clone B 3 (Thrum).. . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75
24 Scanning Electron Micrograph of Averrhoa carambola L. Pollen (X 3000) Clone B 2 (Pin). . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . 75
25 Scanning Electron Micrograph of Averrhoa carambola L. Pollen (X 3000) Clone B 4 (Thrum). . . . . . . .. . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . 75
26 Scanning Electron Micrograph of Averrhoa carambola L. Pollen (X 3000) Clone B 5 (Pin). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75
27 Scanning Electron Micrograph of Averrhoa carambola L. Pollen (X 3000) Clone B 8 (Thrum)... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75
28 Scanning electron Micrograph of Averrhoa carambola L. Pollen (X 3000) Clone B 6 ( Pin). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76
29 Scanning Electron Micrograph of Averrhoa carambola L. Pollen (X 3000) Clone B 9 (Thnun)... . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76
30 Scanning Electron Micrograph of Averrhoa carambola L. Pollen (X 3000) Clone B 7 (Pin) . . . . . . . . . . . . . . . .. . . . . . . . . . . . .. . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . 76
3 1 Scanning Electron Micrograph of Averrhoa carambola L. Pollen (X 3000) Clone B 10 (Thrum). . . . .. . .. . . . . . . . . . . . . . . . . . . .. .. . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . 76
x
32 Scanning Electron Micrograph of Averrhoa carambola L. Pollen (X 3000) Clone B 1 1 (Pin) . . .. . .. . .. . . . . .. ... . .. . . .. .. . .... . .. . . .. .... . . . .. ... .. . . . . . . . .. . '" 76
33 Scanning Electron Micrograph of Averrhoa carambola L. Pollen (X 3000) Clone B 12 ( Pin) . . .. . .. . .. . ........ . .. ... . .. . . .. ........... . . .... ... ... . . .... . . .. ... 77
34 Scanning Electron micrograph of Averrhoa carambola L. Pollen (X 3000) Clone B 13 ('Thrum)... ... ... . ... .. . . . .. . .. ... . .. .. . . . . . . .. . ... . . .. . . . . .. . . . . . . .. .. 77
35 Scanning Electron Micrograph of Averrhoa carambola L. Pollen (X 3000) Clone B 16 (Pin) . . .. . . .... .. .. .. . . .... . . . .. . . ... ... . . . . . . . . ... .... ..... . ..... . . . . . .. 77
36 Scanning Electron Micrograph of Averrhoa carambola L. Pollen (X 3000) Clone B 17 (Thrum)......... . . . . . ... . .. . . . . . .. . . .. . . ... . .... .... . . . . .... . .. . . . .. . 77
37 Scanning Electron Micrograph of Averrhoa carambola L. Pollen (X 3000) Clone BP 1 (Pin).......... ... . . . . . .. .. ... .. . .. . . . . . . . . . ... . . .. .. . . . .. . . .. . . . . . . . . .. . 77
38 Scanning Electron Micrograph of Averrhoa carambola L. Pollen eX 3000) Clone BP 2 (pin). . .. .. . ... . . . .. . . .. . .. .. . . . . . .... ... . . .... . . .. .... .. . . . . . . .. . . . . . .. . 77
39 Scanning Electron Micrograph of Averrhoa carambola L. Stigma (X 80) Clone B 1 (Pin)...... .. . . . . .. . . . ..... . .. . . ... .. . .. . . . . .. . . . .. . . . . .. . . . . . . . . .. .... ..... . .. . 78
40 Scanning Electron Micrograph of Averrhoa carambola L. Stigma (X 80) Clone B 3 (Thrum)..... .. ... ... . . . . . . .... .. . . . . . . .. ...... ... . .. . .. . . . . . . . ... . .. .. . . . . . . 78
41 Scanning Electron Micrograph of Averrhoa carambola L. Stigma (X 80) Clone B 2 (Pin).. . .. . . . . . . . . . .. . . . ... .... . . . . . . . .. . . . . . .. .. .. . . . . . . .. .. . .. . . . . . . . . . .. . . . . 78
42 Scanning Electron Micrograph of Averrhoa carambola L. Stigma (X 80) Clone B 4 (1brum)..................... . ... .. .. . . . . . . .. ...... ..... . . . . . . . . ... . .. . .. . . .. 78
43 Scanning Electron Micrograph of Averrhoa carambola L. Stigma (X 80) Clone B 5 ( Pin). . . . . . . . . . . . . . .... .. .. . ... . . .. . . .. ..... .. . .. . . . . . ... .. . . . . . .... . . .. . . .. . . . 78
44 Scanning Electron Micrograph of Averrhoa carambola L. Stigma (X 80) Clone B 8 ('Thrum). .. ... . . ... . . . . . . . .. . . . . . . . . ... . . . . .. . ... . . .... ..... . ... . . . .. . .. .. . .. 78
45 Scanning Electron Micrograph of Averrhoa carambola L. Stigma (X 80) Clone B 6 (Pin).. . . . . . . . . .. . . . . .. ... . . . . . . . .. . ... .. .. . .. .. . .. . . . . .. . . ..... ... .. . . .. . . .. . .. 79
46 Scanning Electron Micrograph of Averrhoa caramboJa L. Stigma (X 80) Clone B 9 (lbrum)..................................................................... 79
47 Scanning Electron Micrograph of Averrhoa caramboJa L. Stigma (X 80) Clone B 7 (Pin). . ... . .. . . . . .. . . . . . ...... . . .. .. . . . . ... .. . . .. . ... ....... ... .. ..... . . . . . . . .. . 79
48 Scanning Electron Micrograph of Averrhoa carambola L. Stigma (X 80) Clone B 10 (Thrum)..... . .. ... . . .. .. . .. . .. . . . . .. . . . . . . . . . .. . .. . . . . .. . . . . .. . .. . . .. . . . . 79
xi
49 Scanning Electron Micrograph of Averrhoa carambola L. Stigma (X 80) Clone B 11 (Pin)...... . . .. . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . ... . . . 79
50 Scanning Electron Micrograph of Averrhoa carambola L. Stigma (X 80) Clone B 12 (Pin)......................................................................... 80
5 1 Scanning Electron Micrograph of Averrhoa carambola L. Stigma (X 80) Clone B 13 (Thrunt)................................................................... 80
52 Scanning Electron Micrograph of Averrhoa carambola L. Stigma (X 80) Clone B 16 (Pin).......................................................................... 80
53 Scanning Electron Micrograph of Averrhoa carambola L. Stigma (X 80) Clone B 17 ('fhrurn)..................... .......... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 80
54 Scanning Electron Micrograph of Averrhoa carambola L. Stigma (X 80) Clone BP 1 (Pin.).................... .. ............ . . . . .. . . . . . . . . . . . . . . . . .. . . . . . . . .. . . . . . . 80
55 Scanning Electron Micrograph of Averrhoa carambola L. Stigma (X 80) Clone BP 2 (Pin). ............... . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . 80
56 Scanning Electron Micrograph of Averrhoa carambola L. Stigmatic Vesicles (X 1000) Clone B 1 (Pin).. .......... .. . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . 8 1
57 Scanning Electron Micrograph of Averrhoa carambola L. Stigmatic Vesicles (X 1000) Clone B 3 eThrum)...................................................... . . . . . . . ... 81
58 Scanning Electron Micrograph of Averrhoa carambola L. Stigmatic Vesicles (X 1000) Clone B 2 (Pin)............................................................ . . . . . . . ... 8 1
59 Scanning Electron Micrograph of Aver rho a carambola L. Stigmatic Vesicles (X 1000) Clone B 4 (Thnun) ... ........ .................................. .......... . . .. . . . ... 8 1
60 Scanning Electron Micrograph of Averrhoa carambola L. Stigmatic Vesicles (X 1 000) Clone B 5 (Pin)............................................................. . .... ... 8 1
61 Scanning Electron Micrograph of Averrhoa carambola L. Stigmatic Vesicles (X 10(0) Clone B 8 (Thrum)............................................. ................... 8 1
62 Scanning Electron Micrograph of Averrhoa carambola L. Stigmatic Vesicles (X 10(0) Clone B 6 (Pin)................................................................. . . .. 82
63 Scanning Electron Micrograph of Averrhoa carambola L. Stigmatic Vesicles (X 1000) Clone B 9 (Thrurn)....................................................... . . ....... 82
64 Scanning Electron Micrograph of Averrhoa carambola L. Stigmatic Vesicles (X 1000) Clone B 7 (Pin).......................... .................... ................. ....... 82
xii
65 Scanning Electron Micrograph ofAverrhoa carambola L. Stigmatic Vesicles (X 1000) Clone B 10 (fhrwn)... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . .. . . . . . . . . 82
66 Scanning Electron Micrograph of Averrhoa carambola L. Stigmatic Vesicles (X 1000) Clone B 1 1 (Pin).... .. .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . .. . . . .. . . . . . .. . . . . . . 82
67 Scanning Electron Micrograph of Averrhoa carambola L. Stigmatic Vesicles (X 1000) Clone B 12 ( Pin). . . . .. . .. . . . . .. . .. . . . . .. . . . . .. . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . .. . . . . . 83
68 Scanning Electron Micrograph of Averrhoa carambola L. Stigmatic Vesicles (X 1000) Clone B 13 (I1ll1lm).. . . . . .. . . . . . . . . . . . . . . . . . . . ... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83
69 Scanning Electron Micrograph of Averrhoa carambola L. Stigmatic Vesicles (X 1000) Clone B 16 (Pin).......................................................... . . . . . ... 83
70 Scanning Electron Micrograph ofAverrhoa carambola L. Stigmatic Vesicles (X 1000) Clone B 17 (l1u11m)... . . .. .. . . .. .. ... . .. .. . . . . .. . . . . . . . . . . . .. . . .. . . . . . . . . .. . . . . . . . 83
71 Scanning Electron Micrograph ofAverrhoa carambola L. Stigmatic Vesicles (X 1000) Clone BP 1 (Pin).............. ........ .... . . ... . . . . .. . ... ..... . . .. . . . . . . . . . . . . .. . . . 83
72 Scanning Electron Micrograph of Averrhoa carambola L. Stigmatic Vesicles (X 1000) Clone BP 2 (Pin).......................................................... ... ....... 83
xiii
LIST OF FIGURES
Figures
1 Diagram Showing the Measurement of Pistil and Stamen of
Page
Avverhoa carambola L.. .. ....... ... . ..... ...................................................... 121
2 Diagram Showing the Measurement of Stigma of Avverhoa carambola L.......................................................................... 121
3 Diagram Showing the Measurement of Vesicle of Avverhoa carambola L..... . . . . . . . ...... . . . .. . .. ................. . ... . .. ...... ... . . . ........ 122
4 Diagram Showing the Measurement of Pollen grains of Avverhoa carambola L.. . . .. . . . . . . . . . ... . . . ... ................ . . . ....... .... ..... ......... ....... 122
xiv
Abstract of thesis submitted to the Senate of Universiti Putra Malaysia in fulfIlment of the requirements
for Degree of Master of Agricultural Science
STUDIES ON COMP ATffiLE AND INCOMP ATmLE POLLINATIONS AND FLORAL MORPHOLOGICAL POLYMORPffiS M OF PIN AND TIIRUM
MORPHS OF CARAMBOLA (Al'errhoa clD'ambolll L)
By
NOBUlllKO FUS HIMI
January 1 998
Chairman: Professor Dr. Wong Kai Choo
Faculty : Agriculture
The objectives of this study were to elucidate the physiological basis of self-
incompatibility system and the floral morphological polymorphism in carambola
(Averrhoa carambola L.).
Within 30 minutes after pollination, pollen degradation and hydration
commenced on the stigma surface. Pollen tubes subsequently emerged from colpal
furrow and crawled into apertures of vesicles. The pollen tubes reached the base of
the transmitting tract of the style on legitimate pollinations in pin and thrum morphs
xv
at 8.0 h after pollinations. Pollen tubes were Wlifonnly inhibited at the middle of the
style in pin illegitimate pollinations, however, tubes occasionally penetrated to the
base of the transmitting tract of the style. fu thrum illegitimate pollinations, tubes
were inhibited at the junction between stigmatic and stylar tissues. Result of
illegitimate pollination confinned the partial loss of incompatibility in pin as
opposed to the strong incompatibility in thrum as shown by both fluorescence
microscopic observation and the field controlled pollinations study. Ripen fruits
from pin illegitimate pollinations were significantly smaller and contained less seed
compared to legitimate pollinations.
Carambola flower consisted of 5 rose-pink sepals, a pistil, 5 stamens and 5
staminoids. Pistil length was significantly longer in pin than in thrum morphs. On
the other hand, the stamen length was significantly longer in thrum than in pin
morphs. Pollen grain shape of pin and thrum morphs was oblate spherical and
tricolporate. Pollen diameter was significantly larger in thrum than in pin mOl-phs.
Foveolate pattern of pollen exine sculpturing was more intense in pin than in thrum
morphs. Each of the five stylar tissues tenninated in a capitate stigma. The stigma
receptive surface surrounded the stylar tip, except for a strip along the abaxial part
between the middle and base of stylar tissues. Vesicle was a dewdrop-shaped cell.
Stigma and vesicles sizes were significantly greater in pin than in thrum morphs.
Result of measurements of the various floral morphological parts revealed highly
significant variations even among the various clones within the same mOl-phs.
xvi
Abstrak tesis dikemukakan kepada Senat Universiti Putra Malaysia wttuk memenuhi keperluan untuk Ijazah Master Sains
KAJIAN KEATAS KETIDAK SERASIAN PENDEBUNGAAN DAN POLIMOFISMA BUNGA BELIMBING (Averrhoa caramiJoia L.)
Oleh
NOBUIllKO FUSlllMI
Januari 1998
Pengerusi: Professor Dr. Wong Kai Choo
Faculti : Pertanian
Objektif kajian ini ialah untuk memahami asas flSiologi dari sistem
ketidakserasian sendiri dan polimorfisma morfologi bWlga dalam belimbing
(Averrhoa carambola L.).
Dalam masa tiga putoh minit selepas pendebungaan, penghidratan dan
degradasi debwtga bermula diatas permukaan stigma, dan diikuti dengan
kemunculan tiub-tiub dari kerutan colpal yang kemudianya merayap masuk kedalam
bukaan vesikel. Tiub-tiub debunga tersebut kemudianya sampai kekawasan
penghantar dipangkal stit dalam rnasa Japan jam selepas pendebungaan berbantu.
Tiub debunga secara seragam disekat dipertengahan stit bunga pada pokok pin yang
xvii
didebungakan seeara terbuka, walau bagaimanapun, maseh ada diantara tiub-tiub
tersebut yang menyerap masuk kekawasan perhantaran dipangkal stil. Dalam pokok
thrum yang didebunga seeara terbuka, tiub-tiub debunga tersekat ditisu yang
menyambungkan diantara stigma dan stil. Hasil dati pendebungaan terbuka
mengesahkan bahawa kehilangan separuh dari ketidakserasian berlaku dalam pokok
pin, manakala ketidakserasian yang penuh terjadi dalam pokok thrwn, dan ini juga
telah diperjelaskan lagi dari kedua-dua permerhatian mikroskop fluoresen dan
pendebungaan terkawal diladang. Buah matang yang terjadi pada pokok pin yang
didebungakan seeara terbuka lebih keeil dan mengandungi kurang biji berbanding
dengan buah dari pokok yang didebungakan seeara terkawal.
Bunga belimbing mempunyai 5 sepal berwarna jingga-ros, 5 stamen, 5
staminoda dan satu pistil. Stil pokok pin seeara bennakna lebih panjang dari stil
pokok thrum. Disebaliknya pula stamen pokok thrum lebih panjang dari stamen
pokok pin. Bentuk butir debunga dari kedua-dua pokok pin dan thrum adalah bulat
oblate dan trieolporate. Garispusat butir debunga adalah seeara bermakna lebih
panjang dalam pokok thrum dari pokok pin. Corak 'foveolate' dari ukiran eksin
dipermukaan butir debunga adalah lebih halus dalam pokok pin berbanding dengan
pokok thrum. Semua kelima-lima stit berakhir dengan hujung yang reseptif dalam
kesemua jenis morfs. Permukaan stigma yang sempurna adalah besar dan melingkugi
hujung stil. Vesikel adalah satu sel yang berbentuk titisan embun. Saiz stigma dan
vesikel secara bermakna lebih besar dalam pokok pin berbanding pokok thrum. Hasil
ukuran dari berbagai bahagian-bahagian bunga menunjukan terdapatnya variasi
sangat bererti diantara pelbagai klon-klon belimbing.
xviii
CHAPTER I
INTRODUCTION
Carambola (Averrhoa carambola L.) belongs to the family Oxalidaceae. It is
believed to have been originated from Indonesia (Purseglove, 1968). It is a non
seasonal crop. As it is flowering throughout the year, fruit set may be found at any
time and at different stages of development on the tree. The fleshy, five-angled fruit
has sweet to sour taste and is eaten fresh or used for drinks. It is now found growing in
the region of sub-tropics and tropics. In Malaysia, carambola is popular fruit, and is
found growing in almost every part of the country. The crop is cultivated extensively
with a total area of more than 1,900 ha in 1993 (Anon, 1994). It has become an
important fruit crop in Malaysia, where it is now grown commercially for export. In
1994, Malaysia exported carambola to the amount of 8,218 tonnes, which was
equivalent to RM$ 23.82 million. This values were equal to 11.6 % of fruit export in
Malaysia (Anon, 1997). In recent years carambola fruits are becoming more popular
in many countries especially in developed countries such as Japan, USA and Europe
(Watson et at., 1988).
Carambola is a heterostylous species, with individual trees having either the
long styled (pin) or the short styled (thrum) flowers which are compatible with each
other but are self- and intramorph incompatible (Wong et at., 1994). Carambola
2
produces as many as 4,760-1 3,781 flowers per tree per month. Despite the production
of large number of flowers, matured fruit set is considerably low, with only 5 - 198
flowers setting fruits (Saad et al., 1993). For satisfactory fruit production, it is
necessary to plant a mixture of the two morphs, one of which acts as a pollenizer. The
presence of pollenizer is sometimes achieved by bud grafting to a branch of an existing
established commercial clone grown in the field. Such a practice can be an extra
burden to the grower (Wong et al., 1994).
Heterostyly is a genetically-controlled floral polymorphism that is known in
1 55 genera under 24 families of flowering plants (Gander, 1979). The floral morphs
differ reciprocally in style and stamen lengths. The flowers also differ in pollen size
and production, and may differ in pollen exine sculpturing, pollen colour and presence
of starch in pollen, stigma size, stigmatic papillae or corolla size or morphology
(Dulberger, 1992). There are two morphs in distylous plants (distyly) and three
mOlphs in tristylous plants (tristyly). The floral polymorphism is usually genetically
linked with diallelic, sporophytic self-incompatibility system (Gander, 1979). In
distylous plants the supergene determining floral morphology also controls a diallelic,
sporophytic self-incompatibility system associated with floral polymorphism, so that
pollinations between different morphs are compatible. Tristylous plants usually
possess a two-locus diallelic, sporophytic self-incompatibility system associated with
floral polymorphism, but whether or not a supergene is involved has not been
demonstrated genetically (Gander, 1 979).
The physiological fimction of various morphological components of the
heterostylous syndrome is not well understood. Darwin (1 877) suggested that the
3
complementary placing of stigmas and anthers in the floral morphs is a mechanical
device for promoting insect-mediated pollinations between the morphs, which is
reinforced by a distinct physiological mechanism of illegitimacy. Alternatively, Mather
and De Winton (1941) suggested that the real significance of the morphological
differences in pistils and stamens shown by heterostylous flowers was that they were
causally and developmentally related to physiologically self-incompatibility system.
Furthermore, Dulberger (1975a) has argued that heteromorphic characters of stigma
and pollen grains participate in the physiological mechanism of incompatibility. She
has hypothesized that the differential growth of styles and stamens in the morphs
mediated the production of the incompatibility substances, and that metabolism related
to cell wall extension played an important role in incompatibility. However, the
significance of this association has been unclear (Dulberger, 1975a).
Richards(1986) has suggested that incompatibility in some heterostylous
species, i.e., Primula sp, Linum sp, Armeria sp is controlled by factors, perhaps
glycoproteins, which arise from the tapetum on the male side, and are located on the
stigma surface on the female side. Furthermore, Dulberger (1992) has recently
concluded that if the association between pollen size heteromorphism and differing
inhibition sites has a functional basis, then neither the pollen storage product nor the
pollen grain size proper are likely to be involved, although it must be strongly linked
with pollen morph-specific size or pollen dry weight. She has suggested that a protein
or growth factor deriving from an early premeiotic stage of microsporogenesis, during
differential growth of the sporogenous cells or the pollen mother cell, may be involved.
4
Since the earliest report of Darwin (1862), the phenomenon of heterostylous
syndrome has attracted considerable attention to many biologists (Gander, 1979).
Most of researches on heteromorphic incompatibility have been devoted to studies on
the genetics and ecology of natural populations in an effect to understand the
evolution, adaptive, significance and breakdown of heterostylous syndrome. Very little
is known about the physiological basis of incompatibility in heterostylous plants.
The present study is, therefore, carried out to provide a comprehensive
understanding of the incompatibility system of the distylous carambola. Floral
morphological polymorphisms are investigated in an attempt to relate their
participation in the physiological mechanism of the in compatibility system.
Objectives
The objectives of this present study are :
1. To study the compatibility relationship among different morphs of carambola,
2. To study floral morphological polymorphisms with objectives of relating their
participation in the physiological mechanism of the self-incompatibility system
in carambola.
Hypothesis
Self-incompatibility in heterostylous species is closely associated with floral
morphological differences between mating types. These morphological polymorphisms
play an important role in the mechanism of self-incompatibility in the carambola.
General
CHAPTER II
REVIEW OF LITERATURE
The Carambola
Averrhoa carambola, locally known as "beJimbing besi" or starfruit or
carambola has been grown in Malaysia for more than 100 years. It is believed to have
been originated from Indonesia (Purseglove, 1968). It is now found growing in the
region of sub-tropics and tropics. It has been reported to be cultivated in Taiwan
(Green, 1977), Australia (Watson et aI., 1988), USA (Campbell et aI., 1985, Crane,
1992), Brazil (Donadio, 1990), Guyana (Ramsammy, 1989), Trinidad, Canary Island
(Galan-Sauco et aI., 1989), Tabago (Andrew, 1989) and Surinam (Lewis and
Groeizam, 1989).
In Malaysia, carambola is a popular fruit. It is found growing in almost every
part of the countIy. The crop is cultivated extensively with a total area of more than
1,900 ha in 1993 (Anon, 1994). The main carambola cultivation areas include Sungai
5
6
Besi - Serdang - Seri Kembangan - Kajang (Se1angor region), Seremban - Port Dikson
(Negri Sembilan region), Muara - Batu Pahat (Johore region) and Kuching (Sarawak
region) (Bong, 1984).
In recent years carambola is becoming more popular as a tropical fruit
especially in developed countries such as Japan, USA and Europe (Watson et al.,
1988). It has become an increasingly important fruit crop in some tropical countries, in
particular Malaysia, where it is now grown commercially for export. In 1994, Malaysia
exported carambola amounting to 8,218 tonnes, which was equivalent to RM$ 23. 181
million. This value was equivalent to 1 1 .6 % of fruit export in Malaysia (Anon, 1997).
Tree Characteristics
Carambola belongs to the OxaJidaceae family. The Oxalidaceae family contains
primarily herbaceous genera. It is pyramidal in shape when young but rounded at the
tops with an open canopy when mature (Watson et al., 1988). Carambola is a low tree
or arborescent shrub, 5-12 m height, with a short or crooked, low branched trunk, 20-
35 cm in diameter (Ochese et aI. , 1961). Carambola is normally vegetatively
propagated by bud-grafting but tissue culture technique was also investigated as an
alternative propagation method (Litz and Conover, 1980).
Flower Characteristics
Oxalidaceae is known as one of the families in heterostylous species for having
both distyly and tristyly (Ganders, 1979). Carambola is a distylous species, with