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Establishment and analysis of core sets of primers among rice varieties in Taiwan.
|摘要:||研究水稻品種間的親源關係有助於日後在育種上選擇親本時的考量，本實驗主要以逢機擴增多型性DNA（random amplified polymorphic DNA）來探討台灣栽培稻種間的關係，並由使用的引子中篩選核心引子；且針對特定的RAPD產物片段作分析，嘗試建立特定RAPD產物與核心引子間的關係。
在RAPD產物分析方面，取三段在秈稻中特有且明顯的片段，命名為IJ131800；IK16600；ID51500；將片段分別定序後與基因庫比對的結果，其中IJ13與ID5都與水稻第一條染色體有相似的片段，其中IJ13比對到一段相同度91％，ID5比對到三段相同度分別為82％、83％及96％。OPK-16的片段則是與水稻Ra基因相似，推衍氨基酸序列有約70％的相似度。Ra基因是水稻的一個轉錄因子，已知可調節花青素的生合成，目前已知位置在水稻第四條染色體上。更進一步設計gene specific primer擴增水稻基因組DNA，結果發現絕大多數的秈稻均含有此三段片段，而稉稻的基因組則缺乏此序列。
The knowledge of relationship between rice varieties is helpful in selecting parents for hybridization breeding program. The objectives of this study were to investigate the genetic relationship among rice varieties in Taiwan by RAPD ( random amplified polymorphic DNA), identify the core primers that are informative in estimating relationship among varities, and analyze the RAPD's fragment to evaluate the relationship between core primer and the RAPD's products. The genetic diversity of 45 rice varieties were evaluated by RAPD including 41 varieties provided by Taiwan Agricultural Research Intsitute, the Milyang 23 collected from Japan, and Koshihikari of three seed sources from Chia-Yi Agricultural Experiment Station. There were 13 Indica varieties and 32 Japonica varieties. Of the 120 primers screened, 19 were sufficient to produce 100 reproducible banding patterns. When all 19 primers were taken into consideration, 62% of banding patterns were polymorphic among all rice varieties, 33.7% were polymorphic among 32 Japonica varieties, and 52% were polymorphic among 13 Indica varieties. Based on the clustering dendrograph, the OPJ-12 was the most important core primer to differentiate varieties within the rice varieties tested. Based on the heterozygosity estimates, the OPK-16 was the core factor to affect the varieties clustering between Japonica and Indica subspecies. To analyze the RAPD's band patterns revealing potency in Indica varieties, three fragments IJ13, 1800 bp;IK16, 600 bp;and ID5, 1500 bp were cloned and sequenced. The search of the Genebank by BLAST revealed that IJ13 and ID5 had four fragments to align with on the rice chromosome 1 and the identity was 91%, 82%, 83%, and 96%. The fragment of IK16 showed 70% similarity at deduced amino acid sequence to rice transcriptional activator Ra. Ra gene encodes a transcriptional activator that regulates the anthocyanin biosynthetic pathway and has been mapped on rice chromosome 4. The primer OPK-16 not only is the core factor affecting heterozygosity, but also the OPK-16 fragment is located in the coding region of a regulatory gene. We speculated that the different morphology between Japonica and Indica may be the result of differential gene expression regulated by this transcription factor. Future studies will be focused on the novel relationship between rice phenotypes and the expression of Ra.
|Appears in Collections:||生命科學系所|
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