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INFORMATION TO USERS This reproduction was made from a copy of a document sent to us for microfilming. While the most advanced technology has been used to photograph and reproduce this document, the quality of the reproduction is heavily dependent upon the quality of the material submitted. The following explanation of techniques is provided to help clarify markings or notations which may appear on this reproduction. 1. The sign or "target" for pages apparently lacking from the document photographed is "Missing Page(s)". If it was possible to obtain the missing page(s) or section, they are spliced into the film along with adjacent pages. This may have necessitated cutting through an image and duplicating adjacent pages to assure complete continuity. 2. When an image on the film is obliterated with a round black mark, it is an indication of either blurred copy because of movement during exposure, duplicate copy, or copyrighted materials that should not have been filmed. For blurred pages, a good image of the page can be found in the adjacentframe. If copyrighted materials were deleted, a target note will appear listing the pages in the adjacent frame. 3. When a map, drawing or chart, etc., is part of the material being photographed, a definite method of "sectioning" the material has been followed. It is customary to begin filming at the upper left hand corner of a large sheet and to continue from left to right in equal sections with small overlaps. If necessary, sectioning is continued again-beginning below the first row and continuing on until complete. 4. For illustrations that cannot be satisfactorily reproduced by xerographic means, photographic prints can be purchased at additional cost and inserted into your xerographic copy. These prints are available upon request from the Dissertations Customer Services Department. 5. Some pages in any document may have indistinct print. In all cases the best available copy has been filmed. University MicrOfilms International 300 N. Zeeb Road AnnArbor,MI48106 8508799 Thammasiri, Kanchit FLOWER PIGMENTS IN YELLOW DENDROBIUM SPECIES AND HYBRIDS University of Hawaii PH.D. 1984 University Microfilms International 300 N. Zeeb Road, Ann Arbor, MI48106 PLEASE NOTE: In all cases this material has been filmed in the best possible way from the available copy. Problems encountered with this document havebeen identified here with a check mark __..j_. 1. Glossy photographs or pages --..L..... 2. Colored illustration'), paper or print__ 3. Photographs with dark background V 4. Illustrations are poor copy __ 5. Pages with black marks, not original copy__ 6. Print shows through as there is text on both sides of page__ 7. Indistinct, broken or small print on several pages ~ 8. Print exceeds margin requirements __ 9. Tightly bound copy with print lost in spine__ 10. Computer printout pages with indistinct print __ 11. Page(s) lacking when material received, and not available from school or author. 12. Page(s) seem to be missing in numbering only as text follows. 13. Two pages numbered . Textfollows. 14. Curling and wrinkled pages __ 15. Other _ University Microfilms International FLOWER PIGMENTS IN YELLOW DENDROBIUM SPECIES AND HYBRIDS A DISSERTATION SUBMITTED TO THE GRADUATE DIVISION OF THE UNIVERSITY OF HAWAII IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PllILOSOPHY IN HORTICULTURE DECEMBER 1984 By Kanchit Thammasiri Dissertation Committe: Haruyuki Kamemoto, Chairman Richard W. Hartmann Kent D. Kobayashi Charles L. Murdoch Chung-Shih Tang iii ABSTRACT Carotenoids and chlorophylls in yellow petals of Dendrobium species and hybrids were examined by high-performance liquid chromatography (HPLC). Six carotenoids, neoxanthin, violaxanthin, antheraxanthin, lutein, zeaxanthin, and s-carotene, and chlorophyll a and b were identified. Lutein, zeaxanthin, and 8-carotene were the major pigments responsible for yellow flower color in several dendrobiums. Color was affected by the numbers and concentrations of pigments. The degradation of flavonoids, carotenoids, and chlorophylls in the flowers at different stages of maturity within a raceme was examined for three progeuies, K528 (pale yellow), K637 (white-purple), and K650 (bright yellow). The amounts of carotenoids and chlorophylls in crosses K528 and K637 decreased rapidly after blooming, while cross K650 maintained a high carotenoid content in all stages of maturity. Flavonol content in all three crosses did not show any change over time. Carotenoid and chlorophyll changes in growth and development from the bud stage to flower maturity were determined qualitatively and quantitatively in crosses K528 and K637. All 6 carotenoids and 2 chlorophylls in both crosses declined continually from the bud stage to 2-4 weeks after blooming. iv The inheritance of yellow flower color in Dendrobium appears to be complex, since at least 6 carotenoids and chlorophyll a and b have been shown to co-exist. Degradation of yellow pi~ments is not uncommon, and polyploidy further complicates the analysis of color inheritance. Since both carotenoid and chlorophyll concentrations in hybrids often fall between those of the parents, it appears that yellow color is mostly quantitatively inherited although interactions with modifying genes can be significant. v TABLE OF CONTENTS PAGE ABSTRACT •••• .. .. .. ·. .... .. iii LIST OF TABLES • .. ... ·.... ... vii LIST OF FIGURES • . •• • .. .. .. ... I• INTRODUCTION . ••• .. 1. 11. REVIEW OF LITERATURE • . ... • I. •• • •• 3 2.1 Yellow flower pigments. ••••••••• •• 3 2.2 Biosynthesis of flower'pigments ••••• •• 6 2.2.1 Carotenoids ••• '. • • • • • • • • •• 6 2.2.2 Chlorophylls ••••••••••• •• 8 2.2.3 Flavonoids... ••••••••• •• q ~.3 Function of flower pigments. •••••• •• 12 2.4 Degradation of flower pigments. ••••••• 11 ~.5 Color inheritance. ••••••••••• •• 18 2.5.1 Flavonoids... ••••••••• •• 18 2.5.2 Carotenoids •••••••••••••• 20 2.6 Color inheritance in orchids. • • • • • • • • 22 III. MATERIALS AND METHODS •• · . .. ... 27 3.1 Carotenoid and chlorophyll identification in flowers of Dendrobium species and hybrids 27 3.1.1 Extraction of pigments. ••••• •• ~7 3.1.2 RPLC system •••••••••••••• 11 1.1.3 Identification of pigments. ••• •• 32 1.1.4 Quantitative determination. ••• •• 1~ 1.2 n~gradation of clower pigments in Dendrobium hybrids. •••••••••••••••• •• 34 3.2.1 Pigment changes at different stages of flowers in a single raceme •••• •• 35 3.2.2 Carotenoid and chlorophyll changes in growth and development of flowers • •• 35 3.3 Cytology and yellow flower color inheritance. 36 3.3.1 Cytology. • ••••••••••••• '36 1.3.2 Yellow flower color inheritance. • • • 36 IV. RESULTS AND DISCUSSION ••••••••..•• •• 4.1 'Carotenoid and chlorophyll identification in Elowers of Dendrobium species and hybrids •• 38 4.1.1 Discussion.. •••••••••• •• 63 4.2 Degradation of flower pigments in Dendrobium hybrids ••••••••••••••••••• 68 vi 4.2.1 Pigment changes at different stages of f.lowers in a single r a c e me , fiB 4.~.2 Carotenoid and chlorophyll ch~nges in growth and development of flowers •• 76 4.2.3 Discussion. ••••••••••• 7B 4.3 Cytology and yellow flower color inheritance. 82 4.3.1 Cytology. •••••••••••• 82 4.3.2 Yellow flower color inheritance. 82 4.3.1 Discussion.. •••••••••• qO APPENDIX • 95 LITERATURE CITED • 114 vii LIST OF TABLES TABLE PAGE 1 Petal color of Dendrobium species and hybrids used for carotenoid and chlorophyll determination in their petals. 28 2 Petal color of Dendrobium cultivars and selections used for carotenoid and chlorophyll determination in their petals. Specific extinction coeffIcients at A max of carotenoids and chlorophylls used for quantitative determination. 33 4 Petal color of Dendrobium plants used for yellow flower color inheritance. 5 Identification of carotenoids and chlorophylls from petals of the genus Dendrobium. 19 6 Quantitative distribution of carotenoids and chlorophylls in petals of Q. macrophyllum at two different time. 49 7 Quantitative distribution of carotenoids and chlorophylls in petals of some Dendrobium species • Ouantitative distrihution of carotenoIds and ch l 0 r 0 ph Y11 sIn pet a 1 S 0 f some 1) end rob ill'': hybrids • Ii 1 9 Percentage and total amounts of carotenoids and chlorophylls in relation to petal color of some yellowish-flowered Dendrobium species. 64 10 Absorbance of flower pigments in three Dendrobium crosses at different stages of Elo~ers in a single raceme • 11 Quantitative distrihution of carotenoids and chlorophylls at different stages of growth and development of flowers in two Dendrobium crosses • 77 12 Chromosome numbers in some Dendrobium accessIons. viii 13 Segregation of offspring of crosses which have ~. Jaquelyn Thomas 'Y166-1' as an ancestor • ~6 14 Quantitative distribution of carotenolds and chlorophylls in petal of some nendroblum crosses with their parents • 89 ix LIST OF FIGURES FIGURE PACiE 1 Absorption spectra of neoxanthin in me t h a noLye c e t on Lt r L'le (25:75 v/V) and with addItion of RCI. • •••• ... 40 2 Absorption spectra of violaxanthin in methanol/acetonitrile (25:75 V/V) and with addition of RCI ••••••••••• 41 3 Absorption spectra of a n t h e r a xa n t h Ln in methanol/acetonitrile (25:75 V/v) and with addition of RCI. •••• • •••• 4~ 4 Absorption spectra of lutein In methanol/acetonitr.ile (25:75 v/V) and with a d d l t Lo n of ReI. • •••• 41 'i Absorption spectra of zeaxanthin in methanol/acetonitrile (25:75 V/V) and with addItion of RCI. • ••••••• 44 Absorption spectrum of chlorophyll b in methanol/acetonitrile (25:75 V/V). 45 7 Absorption spctrum of chlorophyll a in methanol/acetonitrile (25:75 V/V) 46 8 Absorption spectra of B-carotene in methanol/acetonitrile (25:75 V/V) and with a d d Lt Lon of Hel. •••• • ••• 47 Q 1<' 1. f) '01~ r s 0 f ~. a g g r ~ g '-t t 11:n • QPLC chromato~ram of a total pigment extract of ~. aggregatum ••• '52 11 Flowers of n. helix ••••••••• 53 12 HPLC chromatogram of a total pigment extract of D. helix. ••• . .•.••.•• '54 13 Flowers of D. strehloceras • '55 14 HPLC chromato~ram of a total pigment extract of n.
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    Appendix: Orchid potting mixtures - An abridged historical review 1 T. J. SHEEHAN Introduction There is little doubt that potting media development over time has been the salvation of orchid growers (Bomba, 1975). When epiphytic orchids were first introduced into England and other European countries in the 18th century growers could not envision plants growing in anything but soil. '"Peat and loam' were good for everything and frequently became the mass murderers of the first generation of epiphytic orchids," Hooker is believed to have said around the end of the 19th century; England had become the graveyard of tropical orchids. Undoubtedly this was in reference to the concern individuals were having over the potting media problems. This problem also drew the attention of such noted individuals as John Lindley and Sir Joseph Paxton, as well as the Gardener's Chronicle, who noted that "The Rule of Thumb" had nothing to say about orchid growing; it was only effective in orchid killing (Bomba 1975). Fortunately, the ingenuity of growers solved the problem as innovative potting mixes evolved over the years. After visiting a number of orchid growing establishments it immediately becomes obvious to any orchid grower, professional or hobbyist, that orchids, both epiphytic and terrestrial, will grow in a wide variety of media. It has often been stated that epiphytic orchids can be grown in any medium except soil as long as watering and fertilization are adjusted to fit the mix being used. Ter­ restrial orchids seem to thrive in any medium that contains 40% or more organic matter. Reading cultural recommendations from the early days of orchid growing is most interesting and highly recommended.
  • The Use of the Hypervariable P8 Region of Trnl (UAA) Intron for Identification of Orchid Species: Evidence from Restriction Site Polymorphism Analysis

    The Use of the Hypervariable P8 Region of Trnl (UAA) Intron for Identification of Orchid Species: Evidence from Restriction Site Polymorphism Analysis

    RESEARCH ARTICLE The use of the hypervariable P8 region of trnL (UAA) intron for identification of orchid species: Evidence from restriction site polymorphism analysis Rajkumar Kishor¤*, G. J. Sharma Department of Life Sciences, Manipur University, Imphal, Manipur, India ¤ Current address: Kwaklei and Khonggunmelei Orchids Pvt. Ltd., Sagolband Vijaygovind, Imphal, Manipur, a1111111111 India a1111111111 * [email protected] a1111111111 a1111111111 a1111111111 Abstract The P8 stem-loop region of the trnL intron, which is known to be hypervariable in size with multiple repeat motifs and created difficulties in alignment, is always excluded in phyloge- netic as well as barcode analyses. This region was investigated for species discrimination in OPEN ACCESS 98 taxa of orchids belonging to the tribe Vandeae using in silico mapping of restriction site Citation: Kishor R, Sharma GJ (2018) The use of polymorphism. The length of the P8 regions varied from 200 nucleotides in Aerides rosea to the hypervariable P8 region of trnL(UAA) intron for identification of orchid species: Evidence from 669 nucleotides in Dendrophylax sallei. Forty two taxa had unique lengths, while as many restriction site polymorphism analysis. PLoS ONE as eight shared a common length of 521 nucleotides. Of the 35 restriction endonucleases 13(5): e0196680. https://doi.org/10.1371/journal. producing digestions in the P8 regions, three, viz., AgsI, ApoI and TspDTI turned out to pone.0196680 have recognition sites across all the 98 taxa being studied. When their restriction data were Editor: Serena Aceto, University of Naples Federico combined, 92 taxa could be discriminated leaving three taxon pairs. However, Acampe II, ITALY papillosa and Aeranthes arachnites despite having similar restriction sites differed in their Received: January 17, 2018 P8 lengths.
  • Orchid Inventory in Bantimurung-Bulusaraung National Park, South Sulawesi, Indonesia

    Orchid Inventory in Bantimurung-Bulusaraung National Park, South Sulawesi, Indonesia

    BIODIVERSITAS ISSN: 1412-033X Volume 18, Number 1, January 2017 E-ISSN: 2085-4722 Pages: 341-350 DOI: 10.13057/biodiv/d180145 Orchid inventory in Bantimurung-Bulusaraung National Park, South Sulawesi, Indonesia DWI MURTI PUSPITANINGTYAS Center for Plant Conservation-Bogor Botanical Gardens, Indonesian Institute of Sciences. Jl. Ir. H. Djuanda No. 13, Paledang, Bogor 16122, West Java, Indonesia. Tel. +62-251-8322187, Fax. +62-251-8322187, ♥email: [email protected] Manuscript received: 3 November 2016. Revision accepted: 18 January 2017. Abstract. Puspitaningtyas DM. 2017. Orchid inventory in Bantimurung-Bulusaraung National Park, South Sulawesi, Indonesia. Biodiversitas 18: 341-350. Bantimurung-Bulusaraung National Park, commonly abbreviated as Babul National Park, is in South Sulawesi. It occupies an area of 43,750 hectares between 119o34'17"-119o55'13" East and 4o42'49"-5o06'42" South. Babul National Park is an area in the transition zone between Asia and Australia and therefore has a unique flora and fauna. The study reported here aimed to inventory the orchid species in the Babul National Park area and to determine the orchid diversity in the area. The results of the study recorded approximately 60 orchid species found in Babul National Park. These were representative of 32 genera and consisted of 42 species of epiphytic orchids and 18 species of terrestrial orchids. The terrestrial orchid Habenaria beccarii and the epiphytic orchid Aerides inflexa were the most common orchids found, and were spread evenly throughout the Babul National Park area. Coelogyne celebensis and Aerides inflexa are endemic orchids of Sulawesi found within the Park. Three species of the genus Nervilia, i.e.