1998 Fiscal Year Final Research Report Summary
Linkage mapping of peach genome by AFLP analysis.
| Project/Area Number |
09460019
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
園芸・造園学
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| Research Institution | Kobe University |
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Principal Investigator |
NOMURA Keiichi Kobe University, Faculty of Agriculture Associate Professor, 農学部, 助教授 (60252798)
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| Co-Investigator(Kenkyū-buntansha) |
YAMAGUCHI Masami National institute of Fruit Tree Science Department of Breeding Chief, 果樹試験場・育種部, 室長
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| Project Period (FY) |
1997 – 1998
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| Keywords | peach / AFLP / polymorphism |
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| Research Abstract |
The aim of this research was construction of the linkage map of peach genome by AFLP (Amplified Fragment Length Polymorphism) analysis.
Prior to mapping reproducibility and efficiency of AFLP analysis using non-RI labeling (FITC (fluorescein isothiocyanate) or biotin) were confirmed. In addition, AFLP analysis produced 2.5 polymorphic hand per primer between 'Akame' and 'Juseitou'. This corresponded to 20 times more than polymorphism obtained by RAPD (Random Amplified Polymorphic DNA) analysis. Thus, it was concluded that AFLP analysis could provide useful genetic markers to construct the linkage map.
Next, the possibility of AFLP analysis using a combination of AFLP analysis kit obtained from Life Technologies^<TM>, Inc. and Eco primer obtained from ALOKA Co., Ltd. The results suggested that the sequence of Eco RI adapter-primers supplied by Life Technologies^<TM>, Inc. were modified form the original report because no amplified fragments was observed at the step of selective PCR.Therefore, we decided to purchase the Eco primer from Perkin-Elmar Japan Co. Ltd. and to use the model 310 DNA sequencer.
Finally, AFLP analysis was carried out on 'Akatsuki', 'Ogonto' and 45 their F.hybrids. Fifteen out of 51 primers sets exhibited polymorphisms between the parents cultivars. Using the 15 primers sets, the specific fragments, which are characteristic for flesh color and cling- or free-stone, were screened by bulk-analysis. It was found that only one primers set E/ACA and M/CAC, showed polymorphism corresponded to flesh color, yellow or white.
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