臺灣大學: 農藝學研究所謝兆樞陳玄Chen, HsuanHsuanChen2013-03-272018-07-112013-03-272018-07-112010http://ntur.lib.ntu.edu.tw//handle/246246/253678大豆 (Glycine max (L.) Merr.) 是世界上最重要的豆科 (Leguminosae) 作物，大豆屬野生物種為育種上相當重要的遺傳資源。台灣是一年生野生大豆分佈之南界，同時也是多年生野生種大豆分佈之北界，其中包括四種基因組之野生物種，G. soja (GG 基因組)、G. tomentella (DDD1D1 基因組)、G. dolichocarpa (A6A6DD 基因組) 及G. tabacina (Tab074；A6A6B3B3 基因組) 。 大豆屬物種的染色體數量多 (2n=40)，大小相似又缺乏明顯的形態特徵，對於其基因組內重複性序列的分佈位置及扮演角色的認識又相當缺乏，因此，大豆屬物種之基因組分類及染色體核型分析的研究都受到相當大的限制。利用螢光原位雜交 (fluorescent in situ hybridization, FISH) 技術可用於標定特定基因組或特定染色體區域，藉以了解物種基因組差異及並輔助染色體核型分析，解決分類爭議及染色體研究困難的問題。 本研究首先利用生物資訊的方式，於大豆全基因組定序 (whole-genome shotgun；WGS) 計畫資料庫內找到三個新的重複性序列，分別命名為SBRS1、SBRS2及SBRS3。SBRS1可做為大豆屬物種GG基因組之標幟。SBRS2、SBRS3可當成標記對應於第9、13、14及20條虛擬染色體之GG基因組染色體的標幟。比較這三個重複性序列在資料庫中的分佈與螢光原位雜交之結果，證實該資料庫可用於預測重複性序列在染色體上可能分佈的位置，但不適合用於評估基因組內重複性序列之重複數。 利用點轉漬雜合篩選出兩個有用的ISSR (inter simple sequence repeat) 擴增片段，其中SBRS4可用於GG基因組之染色體核型分析。而SBRS5可做為G. tomentella (DDD1D1) 基因組及其中兩對染色體之標幟。本實驗同時指出，物種間之非專一重複性片段因其在不同基因組內分佈狀況的差異，仍能用於標定特定基因組及染色體，解決部份大豆屬物種染色體研究困難的問題。Soybean (Glycine max (L.) Merr.) is the most important crop in the Leguminosae families. And the wild Glycine species are very momentous breeding resources for soybean improvement. Taiwan is the northern boundary of the annual wild soybeans, and the southern boundary of the perennial wild soybeans. They are the annual one G. soja (GG genome) , also the three perennial ones G. tomentella (DDD1D1 genome) , G. dolichocarpa (A6A6DD genome) and G. tabacina (A6A6B3B3 genome) . The basic chromosome number of the Glycine species is 2n=40. Their chromosomes are similar in size and lacks of obvious morphological characters. Little is known about the repeat sequences composition in the genomes of the Glycine species, hence the research about genome taxonomy and chromosome karyotyping are limited. Using fluorescent in situ hybridization (FISH) method, target sequences can be labeled with repeat sequences, and thus labeling specific genomes or the particular regions of chromosomes. FISH can also be used to reveal the genome diversity between species, and perform karyotyping. Thus, it is helpful to solve the problems among taxonomy controversies or chromosome or studies. Three G. max repeat sequences were isolated by bioinformatics analysis using soybean whole-genome shotgun (WGS) sequences, and designated SBRS1、SBRS2 and SBRS3. SBRS1 could be used on the special specific probe of the GG genome. SBRS2 and SBRS3 could be used to label the pseudomolecules 9, 13, 14 and 20 in the GG genome. The searches were carried out, and then the compared with the result of FISH. Thus, it indicated that the WGS data could be used for predicting the distribution of repeats, while not suitable for estimating the amount of these repeat sequences. Two useful ISSR amplified fragments are selected by Dot blotting hybridization. The SBRS4 could be used for GG genomic chromosomes karyotyping. And the SBRS5 could be a specific probe of the G. tomentella genome and would label two pairs of chromosomes. It is thus suggested that, the unspecific sequences still may offer the potential to be applied as the genomic or chromosome markers.1411857 bytesapplication/pdfen-US大豆大豆屬染色體螢光原位雜交染色體標幟基因組sybeanGlycine maxGlycinechromosomeFISHfluorescent in situ hybridizationchromosome markergenomethesishttp://ntur.lib.ntu.edu.tw/bitstream/246246/253678/1/ntu-99-R96621122-1.pdf