成功大學博碩士論文系統

簡易檢索 / 詳目顯示

回結果列表

研究生：	曾欣晨 Tseng, Hsin-Chen
論文名稱：	第一個發現的蘭花中節染色體標誌 The first centromeric marker found in orchids
指導教授：	張松彬 Chang, Song-Bin
學位類別：	碩士 Master
系所名稱：	生物科學與科技學院 - 生命科學系 Department of Life Sciences
論文出版年：	2018
畢業學年度：	106
語文別：	英文
論文頁數：	39
中文關鍵詞：	台灣白花蝴蝶蘭、蘭嶼姬蝴蝶蘭、螢光原位雜合、中節、粗絲期染色體
外文關鍵詞：	P. aphrodite, P. equestris, Fluorescent in situ hybridization (FISH), centromere, Pachytene chromosome
相關次數：	點閱：103 下載：0
分享至:	分享至facebook 分享至twitter

查詢本校圖書館目錄查詢臺灣博碩士論文知識加值系統勘誤回報

中節(Centromere)是染色體中重要的結構區域，位在染色體長短臂間的高濃縮DNA序列的特殊區域主縊痕(primary constriction)。負責在細胞分裂時，著絲點(kinetochore)與其結合，著絲點的外層再與紡錘絲微管連結，進而將複製的染色體分離到個別的子細胞中；植物的中節特色有：常有重複性序列、由於演化快速，不同物種之間的差異性大、同物種的保守性高，因此可確保物種在繁衍或生長時，異常細胞分裂不易發生，防止功能喪失或死亡。目前已有許多植物如：玉米(Zea mays)、阿拉伯芥(Arabidopsis thaliana)、番茄(Solanum lycopersicum)、禾本科植物(Poaceae)、棉花屬(Gossypium)…等發現染色體中節區域特定DNA序列可做為標誌，來幫助了解其物種的中節結構與細胞分裂時的運作，以及物種演化上相關的鑑定依據。本篇研究是第一個發現蘭花(Orchid)的中節特定區域的DNA標誌，我們在分析台灣白花蝴蝶蘭的遺傳連鎖片段DL_09_401-3，發現此擴增產物經螢光原位雜合技術定位在台灣白花蝴蝶蘭和蘭嶼姬蝴蝶蘭的中節位置。這第一個新中節DNA標誌開啟蘭科植物的中節結構與演化研究。

Centromeres are important construction on the chromosomes, which locate at the specific region. They are between long and short arms which are called primary constriction. And the spindle fibers assemble up to the outer layer of kinetochores that is separating the pair of sister chromatids of daughter cells. There are repetitive sequences in centromere of plants. Due to the fast evolution, the difference of distinct species and high conservative of same species. Therefore, it can guarantee preventing from the abnormal cell division and preventing from malfunction and mortality. Currently, there are many reports which are about DNA marker of plant centromeric DNA markers such as maize (Zea mays), Arabidopsis thaliana, tomato (Solanum lycopersicum), Poaceae, Gossypium etc., because they make us know about the function, structure, and division of centromeres. This is the study which first discover Orchid's DNA markers of centromere region. By coincidence, we isolated the amplification product of a primer pair of Scaffold_DL_09_401-3 of Phalaenopsis aphrodite, then defined the site by fluorescent in situ hybridization (FISH). The results evidence that the gene fragment exits in centromeres in both P. aphrodite and P. equestris. The first and new DNA marker of centromere open the centromere structure and the evolution study of Orchid.

摘要	I
Abstract	II
致謝	III
Index	IV
List of Figures	VI
Chapter 1 Introduction	1
1.1 Phalaenopsis orchids	1
1.2 centromere	2
1.2.1 The significance of centromere	2
1.2.2 Marker of centromeric region in plant	2
1.3 Cytogenetics in plant genomics	3
1.3.1 Application of Cytogenetics in plant	3
1.3.2 Fluorescence in situ hybridization (FISH)	4
1.3.3 The resolution of FISH signal	5
1.4 Aim of the study	6
Chapter 2 Materials and Methods	7
2.1 Plant materials	7
2.2 Preparation of pachytene or metaphase chromosome	7
2.2.1 Chromosome stage of pollen mother cells (PMCs) examination	7
2.2.2 Preparation of acid-clean slides	8
2.2.3 Prepare chromosome spread by drop method (Phalaenopsis aphrodite)	8
2.2.4 Prepare chromosome spread by drop method (Phalaenopsis equestris)	9
2.3 Preparation of DNA probe	10
2.3.1 Extraction of P. aphrodite and P. equestris genomic DNA	10
2.3.2 Polymerase chain reaction (PCR) and electrophoresis examination	11
2.3.3 Probe labeling by nick translation	12
2.3.4 Probe quantification (dot blotting)	12
2.4 Fluorescence in situ hybridization (FISH)	13
2.5 Cytological measurements and analysis	14
2.6 Sub-cloning of the sequences from PCR products	15
2.6.1 Ligation	15
2.6.2 Transformation by heat-shock method	15
2.6.3 Quick screening	16
2.6.4 Plasmid DNA extraction	16
Chapter 3 Results	18
3.1 The centromeric DNA maker in P. aphrodite	18
3.1.1 FISH mapping of centromeric DNA at pachytene stage	18
3.1.2 FISH mapping of centromeric DNA at meiotic metaphase stage	18
3.2 The centromeric DNA maker in P. equestris at pachytene stage	18
3.3 Identification and sub-cloning the sequence from PCR amplification product	19
Chapter 4 Discussion	20
4.1 The same primer used to amplify sequence in P. aphrodite and P. equestris genomic	20
4.2 The FISH mapping of centromeric DNA marker in P. aphrodite and P. equestris	20
4.3 Sequences analysis of the sequences from PCR products by sub-cloning	21
4.4 Conclusion	21
Chapter 5 Reference	23
Chapter 6 Figures	28
Chapter 7 Appendix	34
Appendix 1. The information of Scaffold_401-3 in DL_09	34
Appendix 2. The primer pair used in amplification of a DNA marker	35
Appendix 3. The FISH mapping of four scaffolds of linkage group DL_09 on pachytene chromosomes in P. aphrodite	36
Appendix 4. Vector information	37
Chapter 8 Supplement	38



                                    

陳文輝 (2002)。蝴蝶蘭的品種改良。科學發展。351:32-39

薛豪彥 (2012)。蘭嶼姬蝴蝶蘭粗絲期染色體螢光原位雜交與核型分析。國立台灣大學農藝學研究所論文

葉志新 (2014)。台灣阿嬤-台灣白花蝴蝶蘭的故事。桃園區農業專訊。88: 3-5

蔡雅閔 (2016) Colinearity of genetic linkage groups 1 and 17 of Phalaenopsis aphrodite subsp. formosana on chromosomes of Phalaenopsis equestris. 國立成功大學生命科學研究所碩士論文

中華民國對外貿易發展協會 (Taiwan External Trade Development Council, TAITRA) http://www.taitra.org.tw/News_Detail.aspx?id=9849

Ananiev E, Phillips R, and Rines H. 1998. Chromosome-specific molecular organization of maize (Zea mays L.) centromeric regions. Proceedings of the National Academy of Sciences. 95 (22):13073-13078

Burnham CR, Stout JT, and Weinheimer WH et al. 1972. Chromosome pairing in maize. Genetics. 71 (1):111-126

Chandler SF, Sanchez C. 2012. Genetic modification; the development of transgenic ornamental plant varieties. Plant biotechnology journal. 10 (8):891-903

Chang S-B, Anderson LK, and Sherman JD et al. 2007. Predicting and testing physical locations of genetically mapped loci on tomato pachytene chromosome 1. Genetics. 176 (4):2131-2138

Chang S-B and Stephen M. 2005. A novel tomato centromere-specific retrotransposon. RESEARCH REPORTS TGC REPORT 55

Chao YT, Chen WC, and Chen CY et al. 2018. Chromosome‐level assembly, genetic and physical mapping of Phalaenopsis aphrodite genome provides new insights into species adaptation and resources for orchid breeding. Plant biotechnology journal. 10:1-15

Chase MW, Cameron KM, and Freudenstein JV et al. 2015. An updated classification of Orchidaceae. Botanical journal of the Linnean Society. 177 (2):151-174

Cheng Z, Dong F, and Langdon T et al. 2002. Functional rice centromeres are marked by a satellite repeat and a centromere-specific retrotransposon. The Plant Cell. 14 (8):1691-1704

Cheng Z, Presting GG, and Buell CR et al. 2001. High-resolution pachytene chromosome mapping of bacterial artificial chromosomes anchored by genetic markers reveals the centromere location and the distribution of genetic recombination along chromosome 10 of rice. Genetics. 157 (4):1749-1757

Christenhusz M, Byng J. 2016. The number of known plants species in the world and its annual increase. Phytotaxa. 261 (3): 201–217.

Consortium TG. 2012. The tomato genome sequence provides insights into fleshy fruit evolution. Nature. 485 (7400):635

Copenhaver GP, Nickel K, and Kuromori T et al. 1999. Genetic definition and sequence analysis of Arabidopsis centromeres. Science. 286 (5449):2468-2474

de Castro Nunes R, Orozco-Arias S, and Crouzillat D et al. 2018. Structure and distribution of centromeric retrotransposons at diploid and allotetraploid Coffea centromeric and pericentromeric regions. Frontiers in Plant Science. 9:175

Dong F, Miller JT, and Jackson SA et al. 1998. Rice (Oryza sativa) centromeric regions consist of complex DNA. Proceedings of the National Academy of Sciences. 95 (14):8135-8140

Ferreira J, Mendes S, and Dall’Agnol M et al. 2012. Comparative analyses in Lotus: the cytogenetic map of Lotus uliginosus Schkuhr. Cytogenetic and genome research. 137 (1):42-49

Fonsêca A, Ferreira J, and dos Santos TRB et al. 2010. Cytogenetic map of common bean (Phaseolus vulgaris L.). Chromosome Research. 18 (4):487-502

Gong Z, Wu Y, and Koblizkova A et al. 2012. Repeatless and repeat-based centromeres in potato: implications for centromere evolution. Plant Cell. 24 (9):3559-3574. doi:10.1105/tpc.112.100511
Hall SE, Kettler G, and Preuss D. 2003. Centromere satellites from Arabidopsis populations: maintenance of conserved and variable domains. Genome research. 13 (2):195-205

Han Y, Zhang Z, and Liu C et al. 2009. Centromere repositioning in cucurbit species: implication of the genomic impact from centromere activation and inactivation. Proceedings of the National Academy of Sciences. 106 (35):14937-14941

Hoshi Y, Pląder W, and Malepszy S. 1998. New C‐banding pattern for chromosome identification in cucumber (Cucumis sativus L.). Plant Breeding. 117 (1):77-82

Hsiao Y-Y, Pan Z-J, and Hsu C-C et al. 2011. Research on orchid biology and biotechnology. Plant and Cell Physiology. 52 (9):1467-1486

Hudakova S, Michalek W, and Presting GG et al. 2001. Sequence organization of barley centromeres. Nucleic Acids Research. 29 (24):5029-5035

Iwata-Otsubo A, Radke B, and Findley S et al. 2016. Fluorescence in situ hybridization (FISH)-based karyotyping reveals rapid evolution of centromeric and subtelomeric repeats in common bean (Phaseolus vulgaris) and relatives. G3: Genes, Genomes, Genetics:g3. 115.024984

J.C. Lamb, W. Yu, F. Han and J.A. Birchler. 2008. Plant Centromeres. Plant Genomes. 4,
95-107.

Jiang J, Birchler JA, and Parrott WA et al. 2003. A molecular view of plant centromeres. Trends in plant science. 8 (12):570-575

Jin W, Melo JR, and Nagaki K et al. 2004. Maize centromeres: organization and functional adaptation in the genetic background of oat. The Plant Cell. 16 (3):571-581

Kishii M, Nagaki K, and Tsujimoto H. 2001. A tandem repetitive sequence located in the centromeric region of common wheat (Triticum aestivum) chromosomes. Chromosome Research. 9 (5):417-428

Kumekawa N, Hosouchi T, and Tsuruoka H et al. 2000. The size and sequence organization of the centromeric region of Arabidopsis thaliana chromosome 5. DNA research. 7 (6):315-321

Kurata N, Omura T, and Iwata N. 1981. Studies on centromere, chromomere and nucleolus in pachytene nuclei of rice, Oryza sativa, microsporocytes. Cytologia. 46 (4):791-800

Kursel LE, Malik HS. 2016. Centromeres. Current biology : CB 26 (12):R487-r490. doi:10.1016/j.cub.2016.05.031
Lan T, Albert VA. 2011. Dynamic distribution patterns of ribosomal DNA and chromosomal evolution in Paphiopedilum, a lady's slipper orchid. BMC plant biology. 11:126. doi:10.1186/1471-2229-11-126

Lin H-S, Chiang C-Y, and Chang S-B et al. 2011. Development of simple sequence repeats (SSR) markers in Setaria italica (Poaceae) and cross-amplification in related species. International journal of molecular sciences. 12 (11):7835-7845

Liu Qing. 2015. Research Progress on Structure and Evolution of Plant Centromeres.
Journal of Tropical and Subtropical Botany. 23(5):576 ~ 586.

Luce AC, Sharma A, and Mollere OS et al. 2006. Precise centromere mapping using a combination of repeat junction markers and chromatin immunoprecipitation–polymerase chain reaction. Genetics. 174 (2):1057-1061

Melters DP, Bradnam KR, and Young HA et al. 2013. Comparative analysis of tandem repeats from hundreds of species reveals unique insights into centromere evolution. Genome biology. 14 (1):R10

Murata M, Ogura Y, and Motoyoshi F. 1994. Centromeric repetitive sequences in Arabidopsis thaliana. The Japanese Journal of Genetics. 69 (4):361-370

Riley R.1974. Cytogenetics of chromosome pairing in wheat. Genetics. 78 (1):193-203

Round EK, Flowers SK, and Richards EJ. 1997. Arabidopsis thaliana centromere regions: genetic map positions and repetitive DNA structure. Genome Research. 7 (11):1045-1053

Singh K, Ishii T, and Parco A et al. 1996. Centromere mapping and orientation of the molecular linkage map of rice (Oryza sativa L.). Proceedings of the National Academy of Sciences. 93 (12):6163-6168

Sumner AT. 2003. Chromosomes: organization and function.

Wang C-JR, Harper L, and Cande WZ. 2006. High-resolution single-copy gene fluorescence in situ hybridization and its use in the construction of a cytogenetic map of maize chromosome 9. The Plant Cell. 18 (3):529-544

Yang X, Zhao H, and Zhang T et al. 2018. Amplification and adaptation of centromeric repeats in polyploid switchgrass species. New Phytologist. 218 (4):1645-1657

Zafar F, Okita AK, and Onaka AT et al. 2017. Regulation of mitotic recombination between DNA repeats in centromeres. Nucleic Acids Research. 45 (19):11222-11235. doi:10.1093/nar/gkx763

Zhang L, Yang X, and Tian L et al. 2016. Identification of peanut (Arachis hypogaea) chromosomes using a fluorescence in situ hybridization system reveals multiple hybridization events during tetraploid peanut formation. New Phytologist. 211 (4):1424-1439

Zhang W, Cao Y, and Wang K et al. 2014. Identification of centromeric regions on the linkage map of cotton using centromere-related repeats. Genomics. 104 (6):587-593

Zhang W, Zuo S, and Li Z et al. 2017. Isolation and characterization of centromeric repetitive DNA sequences in Saccharum spontaneum. Scientific reports. 7:41659

Zou J, Gong H, Yang T-J, and Meng J.2009. Retrotransposons - a Major Driving Force in Plant Genome Evolution and a Useful Tool for Genome Analysis. Journal of Crop Science and Biotechnology. 12 (1): 1-8

2024-02-01公開

簡易檢索 / 詳目顯示

相關論文