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題名 | 單雜交青割玉米品種之RAPD標識在種子純度鑑定的應用=The Application of RAPD Markers of Single cross Silage Corn Varieties on Seed Purity Testing |
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作者 | 莊淑貞; 王小華; 胡澤寬; Chuang, Su-jean; Wang, Hsiao-hua; Hu, Tzer-kaun; |
期刊 | 中華農學會報 |
出版日期 | 19990900 |
卷期 | 187 1999.09[民88.09] |
頁次 | 頁54-68 |
分類號 | 434.11 |
語文 | chi |
關鍵詞 | 青割玉米; 種子純度檢定; 逢機擴增核酸序列多型性; Silage corn; Purity testing; Random amplified polymorphic DNAs; |
中文摘要 | 本試驗目的在應用RAPD標識對青割玉米臺農2號及臺南19號進行單株樣品及混合樣品的純度鑑定的研究;在單株樣品方面篩選出逢機引子OPF9(CCAAGCTTCC)對臺農2號之RAPD識別條帶中,可從大小為1170 bp及525 bp的出現與否,在兩親本及雜交一代間作完全識別。逢機引子OPJ6(TCGTTCCGCA)及OPJ7(CCTCTCGACA)之RAPD識別條帶中分別可從2000bp 和380 bp及650 bp和420 bp之出現與否而對臺南19號之兩親本與雜交一代間作完全識別。在混合樣品方面篩選出逢機引子OPB1(GTTTCGCTCC)及OPE17(CTACTGCCGT)之RAPD識別條帶分別在1000 bp及1420 bp和970 bp處,各為臺農2號之母本及父本所特有;而引子OPF 15(CCAGTACTCC)之RAPD識別條帶在1570 bp處則為臺南19號雜交一代所特有且父本從缺,經檢測這些條帶表現的強度,其靈敏度皆達0.1;即以10為基數,親本與雜交一代依不同比例的葉圓片混合樣品之識別條帶經ethidium bromide的染色表現上具有目視強度的變化。利用此研究結果進行此兩品種之種子純度檢定時,不但可作為未知樣品檢出親本混雜的識別標準,而且可同時檢定出父本及母本混雜之比率,當應用在大量樣品種子純度鑑定時,混合樣品檢定方法更可在檢定費用的支出上節省達九倍之多。 |
英文摘要 | The objective of this study was to apply the RAPD markers to seed purity test in two silage corn varieties Tainon No.2 and Tainan No.19 by using single-plant sample and bulked sample. Two DNA fragments, 1170bp/525bp, from RAPD with primer OPF9 for their presence/absence could be used to distinguish female, male parent and F1 hybrid of variety Tainon No.2 in single-plant sample. Same as the former result, two pairs of DNA fragments, 2000bp/380bp and 650bp/420bp, were RAPD products of OPJ6 and OPJ7, respectively, and for their presence/absence also could be used to distinguish female, male parent and F1 hybrid of variety Tainan No.19 in single-plant sample. Bulked sample was collected by 10 leaf-discs mixture of female parent and F1 hybrid of male parent and F1 hybrid. In case of Tainon No.2, the ethidium bromide stains of fragment 1000bp with primer OPB1, unique to female parent from bulked samples showed visible varying persistence as its prevalence increase in proportion of female parent in bulked samples and could be used as standard. On the other hand, the ethidium bromide stains of the DNA fragments 1420bp and 970bp with primer OPE 17 unique to male parent from bulked samples also showed visible varying persistence as their prevalence increase in proportion of male parent in bulked samples and also could be used as a standard. Unknown sample from seed lot could be divided into several bulked samples and could be tested simultaneously by using primer OPB1 and OPE17. So, the contaminate rate by male parent and female parent in a seed lot could be calculated. In case of Tainan No.19, the female parent's seed coat was white color different from male parent's and F1 hybrid's and could be directly used as a marker to pick out contamination by female parent. As to the contamination by male parent, there were four DNA fragments with primer OPF15 belonged to male parent and F1 hybrid. Their ethidium bromide stains also showed visible varying persistence as their increase in proportion of male parent or F1 hybrid in bulked samples could be used as a standard. Unknown sample from seed lot could be divided into several bulked samples and could be tested simultaneously by white seed coat color and using primer OPF15. So, the contaminate rate by male parent and female parent in seed lot could be calculated. |
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