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標題: Genetic Fingerprint Analysis of Pineapple Germplasm
作者: 林伯耀
關鍵字: 生物技術, 園藝
摘要: This project aims to set up an effective AFLP fingerprint system and to establish the phylogenetic relation of 72accessions of pineapple germplasm which have been maintained at Chayi Agriculture Experimental Station.In the past two years, we have successfully established an effective AFLP system and characterized 50accessions of germplasm.The project of this year is to complete the analysis of the remainder accessions.The AFLP fingerprint system includes three steps: after extraction from young leaves, genomic DNA were subject to digestion of restriction enzymes, ligation with adopters, selective PCR amplification with 32P end-labelling primers and finally banding analysis in a denaturing gel.The resulting banding pattern was used to analyze the phylogenic relation, using Jaccard's system ( 1908 ).The genetic similarity was calculated by formula Nab/( Na+Nb-Nab ), where Na and Nb stands for total number of bands associated with the two compared samples, respectively, and Nab common bands of the two samples.The data of phylogenic relation of 72accessions, plotted by UPGMA of NTSYS, will then be used to establish a core group of germplasm for the long term maintenance and for breeding application.This core group is of particular importance, since pineapple is an self-incompatible species; its genetic conservation relies solely on vegetative propagation, which requires continuous consumption of land and manpower.
本計畫之目標為建立鳳梨AFLP遺傳指紋分析系統, 並以此系統分析嘉義農試分所之72種鳳梨種原, 以取得其親緣關係.過去二個年度已經成功建立鳳梨AFLP指紋分析系統, 且分析50個種原, 本年度目標為完成所有72個種原的分析, 並確定其核心種原.鳳梨AFLP指紋分析系統, 包含鳳梨染色體組DNA ( genomic DNA )之抽取, AFLP分析和親緣分析.AFLP分析是應用限制酵素切割配合聚合酵素連鎖反應( PCR )的技術.其操作程序包括三個基本步驟: ( 1 )先將染色體組DNA以限制酵素切割後, ( 2 )以尖端具32P標示之引子, 進行選擇性的重複複製( Selective PCR amplification )這些片段, 最後, ( 3 )利用變性膠片之電泳, 分析訊號片段.鳳梨種原之親緣關係在本計畫中, 採用Jaccard在1908年所提出的方法, 遺傳的相似性=Nab/( Na +Nb -Nab ).其中Na是樣本a的條帶數, Nb是樣本b的條帶數, Nab是樣本a和b共有的條帶數.群叢分析則是利用NTSYS軟體所提供UPGMA方法來分析.建立嘉義農試分所所有72個鳳梨種原之親緣關係後, 可依據此資料歸納出其所含之核心種原.將來只針對核心種原做種原保存, 可以大幅降底維持鳳梨種原的成本.同時AFLP指紋也可以提供種原鑑定的依據, 有助於保護商業品種的專利權.
其他識別: 90農科-2.1.1-糧-Z1(12)
Appears in Collections:分子生物學研究所



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