1. 蝴蝶蘭的分子細胞遺傳學研究
2. 菸草異源染色體添加系的分子細胞遺傳學研究

1. 本研究室以菸草Nicotiana plumbaginifolia-sylvestris異源染色體添加系(monosomic alien addition lines)為材料，將64個RFLP標誌定位在N. sylvestris正常之第2、4、5、6、7、8、9、10、12條染色體上。植株中有4株在N. sylvestris染色體間發生轉座(translocation)，3株染色體發生缺失(deletion)，以及6株含中節片段(centric fragments)染色體的添加系，依核型分析、64個RFLP標誌定位、和25S rDNA、5S rDNA螢光原位雜交(fluorescence in situ hybridization；簡稱FISH)，繪製出N. sylvestris之第5、6、7、8、10、12條染色體DNA標誌分佈圖。以基因組原位雜交(genomic in situ hybridization; 簡稱GISH)技術檢測變異型染色體添加系，發現N. plumbaginifolia與N. sylvestris二基因組間未發生重組(recombination)，為單純的異源染色體添加系，另以菸草的端粒(telomere)重複性序列(TTTAGGG)n為探針，與帶缺失染色體的添加系做螢光原位雜交，發現所有染色體斷裂處皆有新生的端粒序列。

2. 我們曾觀察九種蝴蝶蘭(Phalaenopsis spp.)和一種朵麗蘭(Doritis pulcherrima)有絲分裂早期及中期的染色體，發現蝴蝶蘭染色體分大、中、小三型，且染色體的大小與異染色質(heterochromatin)的多寡成正比，即染色體較大的蝴蝶蘭含較多的異染色質。由於異染色質是重複性(repetitive)DNA所組成，而微衛星體(microsatellites)是以1-6 bp為組成單位的簡單重複性序列，本研究室李宜學同學以六種人工合成的微衛星體序列為探針，與七種蝴蝶蘭和一種朵麗蘭進行螢光原位雜交，發現(GA)11在六種蝴蝶蘭集中於染色體中節處，其餘五種微衛星體序列則散佈於染色體上。我們亦以數種蝴蝶蘭雜交種為材料，進行南方氏基因組雜交(Southern genomic hybridization)，結果顯示具大型染色體的蝴蝶蘭較小型染色體的蝴蝶蘭有較多的重複性DNA，但小型染色體的蝴蝶蘭也含本身特殊的重複性DNA。研究室李文靜同學利用FISH技術將45S和5S rDNA實質定位於七種蝴蝶蘭染色體上，解決過去無法用傳統染色法觀察到的困擾。林志忠同學利用GISH技術觀察八種蝴蝶蘭雜交種的基因組組成，藉以探討不同蝴蝶蘭間親緣關係。另外高玉馨同學以5S rDNA之IGS序列而趙芷瑩同學以45S rDNA之ITS序列分別探討28種蝴蝶蘭的親緣關係。

Recent publications

1. Suen, D. F., C. K. Wang, R. F. Lin, Y. Y. Kao , F. M. Lee and C. C. Chen. 1997. Assignment of DNA markers to Nicotiana sylvestris chromosomes using monosomic alien addition lines. Theoretical and Applied Genetics 94: 331-337. (SCI)
2. Chen, C. M., C. T. Wang, C. J. Wang, C. H. Ho, Y. Y. Kao and C. C. Chen. 1997. Two tandemly repeated telomere-associated sequences in Nicotiana plumbaginifolia . Chromosome Research 5: 561-568. (SCI)
3. Chen, C. C., C. M. Chen, J. T. Yang and Y. Y. Kao . 1998. Localization of a repetitive DNA sequence to the primary constrictions of maize pachytene chromosomes. Chromosome Research 6: 236-238. (SCI)
4. Chen, C. C., C. M. Chen, F. C. Hsu, C. J. Wang, J. T. Yang, Y. Y. Kao . 2000. The pachytene chromosomes of maize as revealed by fluorescence in situ hybridization with repetitive DNA sequences. Theoretical and Applied Genetics 101: 30-36. (SCI)
5. Kao, Y. Y ., S. B. Chang, T. Y. Lin, C. H. Hsieh, Y. H. Chen, W. H. Chen and C. C. Chen. 2001. Differential accumulation of heterochromatin as a cause for karyotype variation in Phalaenopsis orchids. Annals of Botany 87: 387-395. (SCI)
6. Lin, S., H. C. Lee, W. H. Chen, C. C. Chen, Y. Y. Kao , Y. M. Fu, Y. H. Chen and T. Y. Lin. 2001. Nuclear DNA contents of Phalaenopsis sp. and Doritis pulcherrima . Journal of the American Society for Horticultural Science 126: 195-199. (SCI)
7. Lin, T. Y., Y. Y. Kao , S. Lin, R. F. Lin, C. M. Chen, C. H. Huang, C. K. Wang, Y. Z. Lin and C. C. Chen. 2001. A genetic linkage map of Nicotiana plumbaginifolia / N. longiflora based on RFLP and RAPD markers. Theoretical and Applied Genetics 103: 905-911. (The first two authors contribute equally.) (SCI)
8. Chen, C. C., Y. Y. Kao , F. M. Lee and R. F. Lin. 2001. Somatic hybridization between Nicotiana sylvestris Speg. & Comes and N. plumbaginifolia Viv. In : T. Nagata and Y. P. S. Bajaj (eds.) Biotechnology in Agriculture and Forestry, Vol. 49. Somatic hybridization in crop improvement. II. Springer-Verlag , Berlin , Heidelberg , New York . pp. 292-303.
9. Chen, C. C., S. K. Chen, M. C. Liu and Y. Y. Kao* . 2002. Mapping of DNA markers to arms and sub-arm regions of Nicotiana sylvestris chromosomes using aberrant alien addition lines. Theoretical and Applied Genetics 105: 8-15. (*corresponding author) (SCI)
10. Chen, C. C. and Y. Y. Kao . 2002. Chromosomal fluorescence in situ hybridization: techniques and applications in plant genetics. Scientific Agriculture 50: 69-77. (in Chinese)
11. Kao, Y. Y . and C. C. Chen. 2002. A model system for molecular genetics: alien chromosome additions. Scientific Agriculture 50: 351-357. (in Chinese)
12. Lin, C. C., Y. H. Chen, W. H. Chen, C. C. Chen and Y. Y. Kao* . 2005. Genome organization and relationships of Phalaenopsis orchids inferred from genomic in situ hybridization. Bot. Bull. Acad. Sin. 46: 339-345. (*corresponding author) (SCI)
13. Kao, Y. Y., Lin, C. C., Huang, C. H., and Li, Y. H. 2007. The cytogenetics of Phalaenopsis orchids. In: Chen, W. H., and Chen, H. H. (eds) Orchid biotechnology. World Scientific, Singapore, pp 115-128.