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|標題:||Establishment and application of CL-repeat display markers in maize B chromosome
|關鍵字:||maize;B-chromosome;display technique;CL-repeat;玉米;B染色體;顯示技術;CL-repeat||引用:||林倩宇。2013。利用玉米B-10L易位染色體進行B染色體斷裂點及CL-repeat之相互定位。碩士論文。台中：國立中興大學農藝學系。 高國維。2013。實體定位與結構分析四個玉米B染色體專一性RAPD標誌。碩士論文。台中：國立中興大學農藝學系。 Alfenito, M.R. and J.A. Birchler. 1993. Molecular characterization of a maize B chromosome centric sequence. Genetics 135: 589-597. Beckett, J.B. 1978. B-A translocations in maize I. Use in locating genes by chromosome arms. J. Hered. 69: 27-36. Beckett, J.B. 1991. Cytogenetic, genetic and plant breeding applications of B-A translocations in maize. pp. 493-529 in Chromosome engineering in plants: Genetics, Breeding, Evolution. Part A, edited by P.K. Gupta and T. Tsuchiya. Elsevier, Amsterdam. Bouck, A.C., R. Peeler, M.L. Arnold and S.R. Wessler. 2005. Genetic mapping of species boundaries in Louisiana irises using IRRE retrotransposon display markers. Genetics 171: 1289-1303. Carlson, W.R. 1969. Factors affecting preferential fertilization in maize. Genetics 62: 543-554. 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Randolph, L. 1941. Genetic characteristics of the B chromosomes in maize. Genetics 26:608-631. Roman, H. 1947. Mitotic nondisjunction in the case of interchanges involving the B-type chromosome in maize. Genetics 32: 391-409. Roman, H. 1948. Directed fertilization in maize. Proc. Natl. Acad. Sci. USA 34: 36-42. Saghai-Maroof, M.A., K.M, Soliman, R.A. Jorgensen and R.W. Allard. 1984. Ribosomal DNA spacer-length polymorphisms in barley: Mendelian inheritance, chromosomal location, and population dynamics. Proc. Natl. Acad. Sci. USA 81: 8014-8018. Stark, E.A., I. Connerton, S.T. Bennett, S.R. Barnes, J.S. Parker and J.W. Forster. 1996. Molecular analysis of the structure of the maize B-chromosome. Chromosome Res. 4: 15-23. Vos, P., R. Hogers, M. Bleeker, M. Reijans, T. Lee, M. Hornes, A. Friters, J. Pot, J. Paleman and M. Kuiper. 1995. AFLP: A new technique for DNA fingerprinting. Nucleic Acids Res. 23: 4407-4414. Ward, E.J. 1973. Nondisjunction: localization of the controlling site in the maize B chromosome. Genetics 73: 387-391. Williams, J.G., A.R. Kubelik, K.J. Livak, J.A. Rafalski and S.V. Tingey. 1990. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 18: 6531-6535.||摘要:||
為了獲得大量玉米B染色體專一性的分子標誌，利用B染色體專一性序列CL-repeat廣泛分佈在B染色體的特性，並參考鳶尾逆轉位子顯示技術，建立玉米B染色體CL-repeat顯示技術。總共獲得26個B染色體專一性標誌，並利用5個B-10L易位染色體進行標誌定位，再利用13個位於異染色質區的B-10L易位染色體進行相互定位，藉以確定易位染色體斷裂點的位置並精細定位各個CL-repeat顯示標誌。將26個標誌進行選殖與序列分析，依序列特性可分成CL-repeat序列標誌、B染色體中節序列標誌、同時帶有CL-repeat與B染色體中節序列標誌及未知序列標誌此4大類。接著將1個B染色體中節序列標誌、2個同時帶有CL-repeat與B染色體中節序列標誌及2個未知序列標誌轉換成SCAR (sequence characterized amplified region)標誌，並利用B-10L易位染色體進行定位，其中帶B染色體中節序列之3i-08-SCAR標誌出現在非預期的區域，將不同區域的標誌進行選殖與定序，並製成無根樹演化圖，以探討玉米B染色體中節序列在中節與遠端異染色質區的差異。另外，以限制酵素HindIII進行CL-repeat顯示技術，期望獲得更多不同的B染色體專一性的CL-repeat顯示標誌。
For isolating a large number of maize B-chromosome specific molecular markers, we used the B-chromosome specific sequence, CL-repeat which are widely distributed on the B-chromosome, and referred to the IRRE (iris retroelement) retrotransposon display technique to establish the CL-repeat display technique of maize B-chromosome. A total of 26 B-chromosome specific markers were obtained and mapped by five B-10L translocations. Additonal 13 B-10L translocations with breakpoints on the distal heterochromatic region were used to verify breakpoints of these translocations and to finely map each CL-repeat display marker. All of the 26 markers were cloned and sequenced. On the basis of sequence characteristics, those sequence could be divide into four categories: markers with CL-repeat sequence, markers with B-centromere sequence, markers with CL-repeat and B-centromere sequences, and markers with unknown sequence. Subsequently, one marker with B-centromere sequence, two markers with CL-repeat and B-centromere sequences, and two markers with unknown sequence were selected to convert into SCAR (sequence characterized amplified region) markers, and mapped by B-10L translocations. Among these five markers, the 3i-08-SCAR marker with B-centromere sequence were mapped to unexpected regions. Those unexpected SCAR markers were cloned and sequenced from different regions, and used to constructed an unrooted evolution tree for exploring the differentiation of maize B-centromere sequences between the B-centromere and the distal heterochromatin region. Furthermore, we used restriction enzyme HindIII to perform the CL-repeat display technique and hoped to get more different B-chromosome specific CL-repeat display markers.
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