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題 名 | RAPD標識在番茄雜交一代雜種種子純度鑑定的應用=Purity Identification of Fl-hybrid Tomato (Lycopersicon Esculentum L.) Cultivars by RAPD Markers |
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作 者 | 莊淑貞; 王小華; 胡澤寬; | 書刊名 | 農林學報 |
卷 期 | 48:2 1999.06[民88.06] |
頁 次 | 頁103-112 |
分類號 | 435.272 |
關鍵詞 | 雜交番茄; 一代雜種種子純度檢定; 逢機擴增核酸序列多型性; Tomato; F1-hybrid genetic purity testing; Random amplified polymorphic DNAs; |
語 文 | 中文(Chinese) |
中文摘要 | 對雜交種番茄台中亞蔬4號、花蓮亞蔬5號及種苗5112號篩選出Operon公司的 核甘酸引子OPI11(ACATGCCGTG)可對三個品種之母本進行識別,即條帶大小為bp 530者 均出現在各品種組合內之一代雜種及父本,而母本則缺條帶bp 530。另核甘酸引子 OPK14(CCCGCTACA)可對三個品種之父本進行識別,亦即條帶大小為bp 656者均出現在 各品種組合內之一代雜種及母本,而父本則缺條帶bp656。因此在各品種組合內檢定一代雜 種種子純度須聯合核甘酸引子OPI11檢定母本混雜率及核甘酸引子OPK14檢定父本混雜率 後,非父本及非母本的交集方為一代雜種種子的比率。即1個樣品須經2次單獨的PCR增 埴反應才可檢出一代雜種的純度。利用混合樣品經條帶強度之測試結果,檢定條帶bp 530 的靈敏度為0.11,即混合樣品可以9為基數,母本與一代雜種依不同比例的葉圓片混合樣品 之識別條帶bp530經ethidium bromide的染色表現具有可目視的強度變化。同樣地,混合樣 品檢定條帶bp656的靈敏度為0.1,即混合樣品可以10為基數,父本與一代雜種依不同比例 的葉圖片混合樣品,同樣有可目視的強度變化。利用此研究結果對上述三個一代雜種種子進 行純度檢定時,除可作為未知樣品檢出親本混雜的識別標準,而且可同時檢定父、母本混雜 之比率。當應用在大量樣品種子純度檢定時,混合樣品檢查方法更可在檢定成本的支出節省 達8倍、9倍之多。 |
英文摘要 | DNA fragments 530 bp and 656 bp from PAPD with primers OPI11 and OPK14, respectively, were used as markers for genetic purity testing of three tomato varieties (Taichung Yasu no. 4, Hualien Yaasu no.5 and TSS-5112). By their presence/absence it could be used to investigate the contamination of male parent and female parent from F1-hybrid. Bulked sample was collected by 10 and 9 leaf-discs mixture of male parent/F1-hybrid and female parent/F1-hybrid, respectively. The ethidium bromide stains of fragment 530 bp with primer OPI11 showed visible varying persistence as its prevalence increase in proportion of female parent in bulked samles and could be used as a standard. On the other hand, the ethidium bromide stains of the DNA fragment 656 bp with primer OPK14 from bulked samples also showed visible varying persistence as its prevalence increase in proportion of male parent in bulked samples and also could be used as a standard. By means of bulked sample testing, the samples (100) from seed lot could be divided inot 100/9 and 100/10 bulked samples and tested simultaneously by suing primer OPI11 and OPK14, respectively. So, the contamination rate by female parent of male parent in seed lot could be determined, mean while, the cost for seed lots testing could be decreased more than 8 and 9 times. |
本系統中英文摘要資訊取自各篇刊載內容。