臺灣博碩士論文加值系統

秋海棠屬(Begonia L.) 是維管束植物中第六大屬，有超過1500種原生種遍布於熱帶與亞熱帶地區。台灣共有18種已被記錄的原生秋海棠，其中的14種分別屬於3個組(section)：秋海棠組(sect. Diplocinium)、扁果組(sect. Platycentrum) 以及無翅組(sect. Sphenanthera)，另外4種為天然雜交種。台灣為世界已知秋海棠天然雜交種和物種特有率最多的地區，也是秋海棠染色體型態最複雜的亞洲地區，此外，台灣原生秋海棠具有觀賞和藥用之價值，因此物種的鑑別和多樣性研究對原生秋海棠的應用具有重要性。本試驗利用120個來自單子葉作物的SSR分子標誌進行台灣原生秋海棠的遺傳多樣性分析，以測試單子葉SSR分子標誌應用於遠緣植物的可行性，並且藉此了解台灣原生秋海棠的親緣關係。結果顯示平均60.6%的單子葉SSR分子標誌可以增幅秋海棠各原生種的基因組，而單子葉SSR分子標誌的轉移率會受到SSR重複序列的重複類型、長度、以及分布於基因區與否所影響，其中以SSR分子標誌在基因組的分布對轉移率影響最為明顯，位於基因區的SSR分子標誌增幅秋海棠基因組的能力和多型性程度顯著的高於位於非基因區的SSR分子標誌。進一步分別以基因區分子標誌、非基因區分子標誌、以及綜合以上兩者的增幅結果進行親緣性分析，可將台灣原生秋海棠成功地分成三群，而蘭嶼秋海棠(B. fenicis)、岩生秋海棠(B. ravenii)、以及裂葉秋海棠(B. palmata)與此三群原生秋海棠的親緣關係較遠。利用分子標誌分類結果與型態分類不一致，但與染色體數目相關，推測可能是由於台灣原生秋海棠發生種間雜交和平行演化的結果。本篇論文證明單子葉SSR分子標誌可跨物種轉移的可行性，未來將可提供序列資訊缺乏之植物作為種源多樣性與親緣關係研究的工具。

Begonia is the sixth-largest genus of vascular plants in the world. There are fourteen native species and four endemic natural hybrids in Taiwan in which the highest density of natural hybrids and species containing the most complex chromosome numbers in Asia have been reported. Besides, the native Begonia species in Taiwan show great potential as ornamental and medical plants. However, very few molecular tools have been conducted in Begonia species identification and diversity investigation. This study had employed 120 SSR markers designed from monocot genomes to test transferability on Begonia species in Taiwan. An average of 60.6% transferability and multi-unique loci with 100% polymorphism were obtained. In addition, the degree of transferability was influenced by SSR motif types, length and distribution in coding or noncoding regions especially. Simple and imperfect type showed higher transferability. Meanwhile, transferability values of tetra- and dinucleotide repeats were higher than those of tri-nucleotide. Markers derived from coding region revealed higher polymorphism and transferability than those of noncoding region markers. The dendrograms penetrated by data from coding region markers was more similar to total region markers than those of noncoding region markers in diversity analysis. Three groups of dendrograms based on coding, noncoding and total markers were in accordance with previous studies of phylogeny of Begonia species in Taiwan. It demonstrated that SSR markers derived from monocot genomes can be applied in genetic diversity and phylogenetic analysis of distantly related dicotyledons.

Contents

Page
摘要............................................. i
Abstract........................................ ii
Contents........................................ iii
Contents of table............................... iv
Contents of figure.............................. v
1. Introduction............................ 1
2. Materials and methods..................... 6
Plant materials.................................. 6
Selection and classification of SSR markers...... 6
Genomic DNA extraction and PCR amplification..... 7
Gel electrophoresis analysis.................... 8
Data analysis................................... 8
Genetic similarity, diversity analysis and clustering.. 10
3. Results................................... 11
Transferability and polymorphism of monocot SSR markers across native Begonia species in Taiwan ................11
Correlation between transferability of monocot SSR markers and motif categories............................. 12
Comparison of PIC values of SSR markers derived from coding and noncoding regions for Begonia genetic diversity analysis........................................ 14
Genetic relationships of Taiwan native Begonia species 15
4. Discussion............................... 18
Patterns of transferring genomic SSR markers into distantly-related taxa.................................... 18
Effects of targets species characters on transferability................................. 21
Effects of marker types and distributions on transferability................................. 24
Polymorphism and diversity of SSR markers from coding and noncoding regions................................ 27
Genetic relationship among Taiwanese Begonia species by monocot SSR markers.............................. 29
5. Conclusion............................... 33
6. References.............................. 34
7. Appendix................................ 42

Anderson, J.A., G.A. Churchill, J.E. Autrique, S.D. Tanksley, and M.E. Sorrells. 1993. Optimizing parental selection for genetic linkage maps. Genome 36: 181-186.
Asp, T., U. Frei, T. Didion, K. Nielsen, and T. Lübberstedt. 2007. Frequency, type, and distribution of EST-SSRs from three genotypes of Lolium perenne, and their conservation across orthologous sequences of Festuca arundinacea, Brachypodium distachyon, and Oryza sativa. BMC Plant Biol. 7: 36.
Bao, J.S., H. Corke and M. Sun. 2002. Microsatellites in starch-synthesizing genes in relation to starch physicochemical properties in waxy rice (Oryza sativa L.). Theor. and Appl. Genet. 105: 898-905.
Barbará, T., C. Palma‐Silva, G.M. Paggi, F. Bered, M.F. Fay, and C. Lexer. 2007. Cross‐species transfer of nuclear microsatellite markers: potential and limitations. Mol. Ecol. 16: 3759-3767.
Barkley, N.A., M.L. Roose, R.R. Krueger, and C.T. Federici. 2006. Assessing genetic diversity and population structure in a citrus germplasm collection utilizing simple sequence repeat markers (SSRs). Theor. Appl. Genet. 112: 1519-1531.
Barreneche, T., M. Casasoli, K. Russell, A. Akkak, H. Meddour, C. Plomion, F. Villani, and A. Kremer. 2004. Comparative mapping between Quercus and Castanea using simple-sequence repeats (SSRs). Theor. Appl. Genet. 108: 558-566.
Bassam, B.J., G. Caetano-Anolles, and P.M. Gresshoff. 1991. Fast and sensitive silver staining of DNA in polyacrylamide gels. Anal. Biochem. 196: 80-83.
Brennan, A.C., S. Bridgett, M.S. Ali, N. Harrison, A. Matthews, J. Pellicer, A.D. Twyford, and C.A. Kidner. 2012. Genomic resources for evolutionary studies in the large, diverse, tropical genus, Begonia. Tropical Plant Biol. 5: 261-276.
Cavagnaro, P., S. M. Chung, S. Manin, M. Yildiz, A. Ali, M. S Alessandro, M. Iorizzo, D. A Senalik and P. W Simon. 2011. Microsatellite isolation and marker development in carrot-genomic distribution, linkage mapping, genetic diversity analysis and marker transferability across Apiaceae. BMC Genomics 12: 386.
Chagné, D., P. Chaumeil, A. Ramboer, C. Collada, A. Guevara, M.T. Cervera, G.G. Vendramin, V. Garcia, J.M. Frigerio, C. Echt, T. Richardson, and C. Plomion. 2004. Cross-species transferability and mapping of genomic and cDNA SSRs in pines. Theor. Appl. Genet. 109: 1204-1214.
Chen, C. 1993. Begoniaceae. In Editorial Committee of the Flora of Taiwan (eds.), Flora of Taiwan, 2nd ed., vol. 3. Editorial Committee of the Flora of Taiwan, Second Edition, Taipei. pp. 845-854.
Chen, S.Y., Y.T. Lin, C.W. Lin, W.Y. Chen, C.H. Yang, and H.M. Ku. 2010. Transferability of rice SSR markers to bamboo. Euphytica 175: 23-33.
Chen, S.Y. 2012. Conservation of microsatellites in vascular plant and their application in genetic diversity analysis. Doctoral Dissertation, National Chung Hsing University, Taiwan.
Chen, X., Y.G. Cho, and S.R. McCouch. 2002. Sequence divergence of rice microsatellites in Oryza and other plant species. Mol. Genet. Genomics 268: 331-343.
Chiang, T.Y., K.H. Hong, and C.I Peng. 2001. Experimental hybridization reveals biased inheritance of the internal transcribed spacer of the nuclear ribosomal DNA in Begonia× taipeiensis. J. Plant Res. 114: 343-351.
Cho, Y.G., T. Ishii, S. Temnykh, X. Chen, L. Lipovich, S.R. McCouch, W.D. Park, N. Ayres, and S. Cartinhour. 2000. Diversity of microsatellites derived from genomic libraries and GenBank sequences in rice (Oryza sativa L.). Theor. Appl. Genet. 100: 713-722.
Choudhary, O.P., and S. Trived. 2010. Microsatellite or simple sequence repeat (SSR) instability depends on repeat characteristics during replication and repair. J. Cell and Mol. Biol. 8: 21-34.
Clement, W.L., M.C. Tebbitt, L.L. Forrest, J.E. Blair, L. Brouillet, T. Eriksson, and S.M. Swensen. 2004. Phylogenetic position and biogeography of Hillebrandia sandwicensis (Begoniaceae): a rare Hawaiian relict. Am. J. Bot. 91: 905-917.
Cordeiro, G.M., R. Casu, C.L. McIntyre, J.M. Manners, and R.J. Henry. 2001. Microsatellite markers from sugarcane (Saccharum spp.) ESTs cross transferable to erianthus and sorghum. Plant sci. 160: 1115-1123.
Dewitte, A., L. Leus, T. Eeckhaut, I. Vanstechelman, J.V. Huylenbroeck, and E.V. Bockstaele. 2009. Genome size variation in Begonia. Genome 52: 829-838.
Dewitte, A., A.D. Twyford, D.C. Thomas, C.A. Kidner, and J. Van Huylenbroeck. 2011. The origin of diversity in Begonia: genome dynamism, population processes and phylogenetic patterns. In O. Grillo and G. Venora (eds), The dynamical processes of biodiversity–case studies of evolution and spatial distribution. InTech. doi: 10.5772/23789.
Dirlewanger, E., P. Cosson, M. Tavaud, M.J. Aranzana, C. Poizat, A. Zanetto, P. Arús, and F. Laigret. 2002. Development of microsatellite markers in peach [Prunus persica (L.) Batsch] and their use in genetic diversity analysis in peach and sweet cherry (Prunus avium L.). Theor. Appl. Genet. 105: 127-138.
Diwan, N., and P. Cregan. 1997. Automated sizing of fluorescent-labeled simple sequence repeat (SSR) markers to assay genetic variation in soybean. Theor. Appl. Genet. 95: 723-733.
Doyle, J., and J. Doyle. 1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem. Bull. 19: 11-15.
Ellegren, H. 2004. Microsatellites: simple sequences with complex evolution. Nat. Rev. Genet. 5: 435-445.
Fay, M.F., R.S. Cowan, and I.J. Leitch. 2005. The effects of nuclear DNA content (C-value) on the quality and utility of AFLP fingerprints. Ann. Bot. 95: 237-246.
Forrest, L.L., and P.M. Hollingsworth. 2003. A recircumscription of Begonia based on nuclear ribosomal sequences. Plant Syst. Evol. 241: 193-211.
Forrest, L.L., M. Hughes, and P.M. Hollingsworth. 2005. A phylogeny of Begonia using nuclear ribosomal sequence data and morphological characters. Syst. Bot. 30: 671-682.
Garner, T.W.J. 2002. Genome size and microsatellites: the effect of nuclear size on amplification potential. Genome 45: 212-215.
Goodall-Copestake, W.P., S. Pérez‐Espona, D.J. Harris, and P.M. Hollingsworth. 2010. The early evolution of the mega‐diverse genus Begonia (Begoniaceae) inferred from organelle DNA phylogenies. Biol. J. Linnean Soc. 101: 243-250.
Guan, K.Y., H. Yamaguchi, J.X. Li, H.Z. Li, and H. Ma. 2007. Traditional uses of begonias (Begoniaceae) in China. Acta Bot. Yunnanica 29: 58-66.
Guo, W., W. Wang, B. Zhou, and T. Zhang. 2006. Cross-species transferability of G. arboreum-derived EST-SSRs in the diploid species of Gossypium. Theor. Appl. Genet. 112: 1573-1581.
Gupta, P.K., S. Rustgi, S. Sharma, R. Singh, N. Kumar, and H.S. Balyan. 2003. Transferable EST-SSR markers for the study of polymorphism and genetic diversity in bread wheat. Mol. Genet. Genomics 270: 315-323.
He, G., F. Woullard, I. Marong, and B. Guo. 2006. Transferability of soybean SSR markers in peanut (Arachis hypogaea L.). Peanut Sci. 33: 22-28.
Hendrix, R., J. S. Hauswaldt, M. Veith, and S. Steinfartz. 2010. Strong correlation between cross‐amplification success and genetic distance across all members of ‘True Salamanders’(Amphibia: Salamandridae) revealed by Salamandra salamandra‐specific microsatellite loci. Mol. Ecol. Resour. 10: 1038-1047.
Hu, J., L. Wang, and J. Li. 2011. Comparison of genomic SSR and EST-SSR markers for estimating genetic diversity in cucumber. Biol. Plant. 55: 577-580.
Huang, X.Q., A. Börner, M.S. Röder, and M.W. Ganal. 2002. Assessing genetic diversity of wheat (Triticum aestivum L.) germplasm using microsatellite markers. Theor. Appl. Genet. 105: 699-707.
Hughes, M., and P.M. Hollingsworth. 2008. Population genetic divergence corresponds with species‐level biodiversity patterns in the large genus Begonia. Mol. Ecol. 17: 2643-2651.
Hughes, M., P.M. Hollingsworth, and J. Squirrell. 2002a. Isolation of polymorphic microsatellite markers for Begonia sutherlandii Hook. f. Mol. Ecol. Notes 2: 185-186.
Hughes, M., J. Russell, and P. Hollingsworth. 2002b. Polymorphic microsatellite markers for the Socotran endemic herb Begonia socotrana. Mol. Ecol. Notes 2: 159-160.
Jaccard, P. 1908. Nouvelles recherchés sur la distribution florale. Bul. Soc. Vaudoise Sci. Nat. 44: 223-270.
Jatoi, S.A., A. Kikuchi, S.S. Yi, K.W. Naing, S. Yamanaka, J.A. Watanabe, and K.N. Watanabe. 2006. Use of rice SSR markers as RAPD markers for genetic diversity analysis in Zingiberaceae. Breed. Sci. 56: 107-111.
Jeeva, S., and J.M. Antonisamy. 2012. Anti–bacterial and phytochemical studies on methanolic extracts of Begonia floccifera Bedd. flower. Asian Pac. J. Trop. Med. 2: S151-S154.
Ji, Y., Y. Luo, B. Hou, W. Wang, J. Zhao, L. Yang, Q. Xue, and X. Ding. 2012. Development of polymorphic microsatellite loci in Momordica charantia (Cucurbitaceae) and their transferability to other cucurbit species. Sci. Hortic. 140: 115-118.
Jiang, C.B., M.J. Chang, C.L. Wen, Y.P. Lin, F.L. Hsu, and M.H. Lee. 2006. Natural products of cosmetics: Analysis of extracts of plants endemic to Taiwan for the presence of tyrosinase-inhibitory, melanin-reducing, and free radical scavenging activities. J. Food Drug Anal. 14: 346.
Jones, E.S., M.P. Dupal, R. Kölliker, M.C. Drayton and J.W. Forster. 2001. Development and characterisation of simple sequence repeat (SSR) markers for perennial ryegrass (Lolium perenne L.). Theor. Appl. Genet. 102: 405-415.
Joukhadar, R., and A. Jighly. 2012. Microsatellites grant more stable flanking genes. BMC Res. Notes 5: 556.
Kalia, R.K., M.K. Rai, S. Kalia, R. Singh, and A.K. Dhawan. 2011. Microsatellite markers: an overview of the recent progress in plants. Euphytica 177: 309-334.
Kishimoto, S., R. Aida, and M. Shibata. 2002. Agrobacterium tumefaciens-mediated transformation of Elatior Begonia (Begonia×hiemalis Fotsch). Plant Sci. 162: 697-703.
Kofler, R., C. Schlötterer, E. Luschützky, and T. Lelley. 2008. Survey of microsatellite clustering in eight fully sequenced species sheds light on the origin of compound microsatellites. BMC Genomics 9: 612.
Kono, Y., M.C. Chung, and C.I Peng. 2012. Identification of genome constitutions in Begonia×chungii and its putative parents, B. longifolia and B. palmata, by genomic in situ hybridization (GISH). Plant Sci. 185: 156-160.
Kuleung, C., P.S. Baenziger, and I. Dweikat. 2004. Transferability of SSR markers among wheat, rye, and triticale. Theor. Appl. Genet. 108: 1147-1150.
Kumar, J., and P.K. Gupta. 2008. Molecular approaches for improvement of medicinal and aromatic plants. Plant Biotechnol. Rep. 2: 93-112.
Laferrière, J.E. 1992. Begonias as food and medicine. Econ. Bot. 46: 114-116.
Li, Y.C., A.B. Korol, T. Fahima and E. Nevo. 2004. Microsatellites within genes: structure, function, and evolution. Mol. Biol. Evol. 21: 991-1007.
Mantel, N.A. 1967. The detection of disease clustering and generalized regression approach. Cancer Res. 27: 209-220.
Matus, I.A., and P.M. Hayes. 2002. Genetic diversity in three groups of barley germplasm assessed by simple sequence repeats. Genome 45: 1095-1106.
McCouch, S.R., L. Teytelman, Y. Xu, K.B. Lobos, K. Clare, M. Walton, B. Fu, R. Maghirang, Z. Li, Y. Xing, Q. Zhang, I. Kono, M. Yano, R. Fjellstrom, G. Declerck, D. Schneider, S. Cartinhour, D. Ware, and L. Stein. 2002. Development and mapping of 2240 new SSR markers for rice (Oryza sativa L.). DNA Res. 9: 199-207.
Mondini, L., A. Noorani, and M.A. Pagnotta. 2009. Assessing plant genetic diversity by molecular tools. Diversity 1: 19-35.
Moon, H.S., J.S. Nicholson, and R.S. Lewis. 2008. Use of transferable Nicotiana tabacum L. microsatellite markers for investigating genetic diversity in the genus Nicotiana. Genome 51: 547-559.
Morgante, M., M. Hanafey, and W. Powell. 2002. Microsatellites are preferentially associated with nonrepetitive DNA in plant genomes. Nature Genet. 30: 194-200.
Mun, J.H., D.J. Kim, H.K. Choi, J. Gish, F. Debellé, J. Mudge, R. Denny, G. Endré, O. Saurat, A.M. Dudez, G.B. Kiss, B. Roe, N.D. Young, and D.R. Cook. 2006. Distribution of microsatellites in the genome of Medicago truncatula: a resource of genetic markers that integrate genetic and physical maps. Genetics 172: 2541-2555.
Nakamura, K., C.J. Huang, R.R. Rubite, W.C. Leong, Y. Kono, H.A. Yang, and C.I Peng. 2012. Isolation of compound microsatellite markers in Begonia fenicis (Begoniaceae) endemic to east and southeast Asian islands. Am. J. Bot. 99: e20-e23.
Neale, S., W. Goodall-Copestake, and C.A. Kidner. 2006. The evolution of diversity in Begonia. In Teixeira da Silva J.A. (ed), Floriculture, ornamental and plant biotechnology: advances and topical issues. Global Science books. pp. 606-611.
Oginuma, K., and C.I Peng. 2002. Karyomorphology of Taiwanese Begonia (Begoniaceae): taxonomic implications. J. Plant Res. 115: 225-235.
Pandey, M.K., B. Gautami, T. Jayakumar, M. Sriswathi, H.D. Upadhyaya, M.V.C. Gowda, T. Radhakrishnan, D.J. Bertioli, S.J. Knapp, D.R. Cook, and R.K. Varshney. 2012. Highly informative genic and genomic SSR markers to facilitate molecular breeding in cultivated groundnut (Arachis hypogaea). Plant Breed. 131: 139-147.
Park, Y.J., J.K. Lee, and N.S. Kim. 2009. Simple sequence repeat polymorphisms (SSRPs) for evaluation of molecular diversity and germplasm classification of minor crops. Molecules 14: 4546-4569.
Peakall, R., S. Gilmore, W. Keys, M. Morgante, and A. Rafalski. 1998. Cross-species amplification of soybean (Glycine max) simple sequence repeats (SSRs) within the genus and other legume genera: implications for the transferability of SSRs in plants. Mol. Biol. Evol. 15: 1275-1287.
Peng, C.I, and Y.K. Chen. 1991. Hybridity and parentage of Begonia buimontana Yamamoto (Begoniaceae) from Taiwan. Ann. Missouri Bot. Gard. 78: 995-1001.
Peng, C.I, and T.Y. Chiang. 2000. Molecular confirmation of unidirectional hybridization in Begonia x taipeiensis Peng (Begoniaceae) from Taiwan. Ann. Missouri. Bot. Gard. 87: 273-285.
Peng, C.I, and S.M. Ku. 2009. Begonia× chungii (Begoniaceae), a new natural hybrid in Taiwan. Bot. Stud. 50: 241-250.
Peng, C.I, and C.Y. Sue. 2000. Begonia× taipeiensis (Begoniaceae), a new natural hybrid in Taiwan. Bot. Bull. Acad. Sin. 41: 151-158.
Peng, C.I, Y.K. Chen, and W.C. Leong. 2005. Five new species of Begonia (Begoniaceae) from Taiwan. Bot. Bull. Acad. Sin. 46: 255-272.
Plana, V. 2003. Phylogenetic relationships of the Afro-Malagasy members of the large genus Begonia inferred from trnL intron sequences. Syst. Botany 28: 693-704.
Plana, V., A. Gascoigne, L.L. Forrest, D. Harris, and R.T. Pennington. 2004. Pleistocene and pre-Pleistocene Begonia speciation in Africa. Mol. Phylogenet. Evol. 31: 449-461.
Prasad, M., R. Varshney, J. Roy, H. Balyan, and P. Gupta. 2000. The use of microsatellites for detecting DNA polymorphism, genotype identification and genetic diversity in wheat. Theor. Appl. Genet. 100: 584-592.
Prevost, A., and M.J. Wilkinson. 1999. A new system of comparing PCR primers applied to ISSR fingerprinting of potato cultivars. Theor. Appl. Genet. 98: 107-112.
Rajbhandary, S., M. Hughes, T. Phutthai, D.C. Thomas, and K.K. Shrestha. 2011. Asian Begonia: out of Africa via the Himalayas? Gard. Bull. Singapore 63: 277-286.
Rohlf, F.J. 2000. NTSYS-PC, Numerical taxonomy and multivariate analysis system, Version 2.10e. Exeter Software, Setauket, New York, USA.
Roldàn-Ruiz, I., J. Dendauw, E.V. Bockstaele, A. Depicker, and M. De Loose. 2000. AFLP markers reveal high polymorphic rates in ryegrasses (Lolium spp.). Mol. Breed. 6: 125-134.
Roy, C.B., M. Nazeer, and T. Saha. 2004. Identification of simple sequence repeats in rubber (Hevea brasiliensis). Curr. Sci. 87: 807-810.
Savadi, S.B., B. Fakrudin, H.L. Nadaf, and M.V.C. Gowda. 2012. Transferability of sorghum genic microsatellite markers to peanut. Am. J. Plant Sci. 3: 1169-1180.
Sharma, R.K., P. Gupta, V. Sharma, A. Sood, T. Mohapatra, and P.S. Ahuja. 2008. Evaluation of rice and sugarcane SSR markers for phylogenetic and genetic diversity analyses in bamboo. Genome 51: 91-103.
Stults, D.Z., and B.J. Axsmith. 2011. First macrofossil record of Begonia (Begoniaceae). Am. J. Bot. 98: 150-153.
Sundaram, R.M., B. Naveenkumar, S.K. Biradar, S.M. Balachandran, B. Mishra, M. IlyasAhmed, B.C. Viraktamath, M.S. Ramesha, and N.P. Sarma. 2008. Identification of informative SSR markers capable of distinguishing hybrid rice parental lines and their utilization in seed purity assessment. Euphytica 163: 215-224.
Tang, S., J.K. Yu, M.B. Slabaugh, D.K. Shintani, and S.J. Knapp. 2002. Simple sequence repeat map of the sunflower genome. Theor. Appl. Genet.105: 1124-1136.
Taylor, J.S., J.M.H. Durkin, and F. Breden. 1999. The death of a microsatellite: a phylogenetic perspective on microsatellite interruptions. Mol. Biol. Evol. 16: 567-572.
Tebbitt, M.C., L. Lowe-Forrest, A. Santoriello, W.L. Clement, and S.M. Swensen. 2006. Phylogenetic relationships of Asian Begonia, with an emphasis on the evolution of rain-ballist and animal dispersal mechanisms in sections Platycentrum, Sphenanthera and Leprosae. Syst. Botany 31: 327-336.
Thomas, D.C., M. Hughes, T. Phutthai, S. Rajbhandary, R. Rubite, W.H. Ardi, and J.E. Richardson. 2011. A non-coding plastid DNA phylogeny of Asian Begonia (Begoniaceae): Evidence for morphological homoplasy and sectional polyphyly. Mol. Phylogenet. Evol. 60: 428-444.
Varshney, R.K., A. Graner, and M.E. Sorrells. 2005. Genic microsatellite markers in plants: features and applications. Trends Biotechnol. 23: 48-55.
Wang, M.L., N.A. Barkley, J.K. Yu, R.E. Dean, M.L. Newman, M.E. Sorrells, and G.A. Pederson. 2005. Transfer of simple sequence repeat (SSR) markers from major cereal crops to minor grass species for germplasm characterization and evaluation. Plant Genet. Resour. 3: 45-58.
Wang, M.L., N.A. Barkley and T.M. Jenkins. 2009. Microsatellite markers in plants and insects. Part I: Applications of biotechnology. Genes, genomes and genomics 3: 54-67.
Wu, P.L., F.W. Lin, T.S. Wu, C.S. Kuoh, K.H. Lee, and S.J. Lee. 2004. Cytotoxic and anti-HIV principles from the rhizomes of Begonia nantoensis. Chem. Pharm. Bull. 52: 345-349.
Yang, C.H. 2012. The comparative analysis of genetic diversity in two different plants species based on different molecular marker systems. Master Thesis, National Chung Hsing University, Taiwan.
Yang, H.A. 2011. Hybridization and Polyploidization of Taiwanese Begonias. Master Thesis, National Taiwan University, Taiwan.
Zeid, M., J.K. Yu, I. Goldowitz, M.E. Denton, D.E. Costich, C.T. Jayasuriya, M. Saha, R. Elshire, D. Benscher, F. Breseghello, J. Munkvold, R.K. Varshney, G. Belay, and M.E. Sorrells. 2010. Cross-amplification of EST-derived markers among 16 grass species. Field Crop. Res. 118: 28-35.