| Project/Area Number |
16380025
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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 |
Horticulture/Landscape architecture
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| Research Institution | Kyoto University |
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Principal Investigator |
YONEMORI Keizo Kyoto University, Graduate School of Agriculture, Professor, 農学研究科, 教授 (10111949)
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| Co-Investigator(Kenkyū-buntansha) |
YAMADA Masahiko NARO, NIFTS, Grape and Persimmon Breeding and Physiology Research Team, Chief Researcher, ブドウ・カキ研究チーム, 上席研究官 (00355439)
TAO Ryutaro Kyoto University, Graduate School of Agriculture, Associate Professor, 農学研究科, 助教授 (10211997)
KITAJIMA Akira Kyoto University, Graduate School of Agriculture, Associate Professor, 農学研究科, 助教授 (70135549)
KANZAKI Shinya Kinki University, Faculty of Agriculture, Lecturer, 農学部, 講師 (20330243)
HABU Tsuyoshi Kyoto University, Graduate School of Agriculture, Assistant Professor, 農学研究科, 助手 (60335304)
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| Project Period (FY) |
2004 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥15,700,000 (Direct Cost: ¥15,700,000)
Fiscal Year 2006: ¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2005: ¥3,800,000 (Direct Cost: ¥3,800,000)
Fiscal Year 2004: ¥8,600,000 (Direct Cost: ¥8,600,000)
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| Keywords | PCNA / tannin biosynthesis / gene exploration / genomic library / chromosome walking / Luo Tian Tian Shi / タンニン蓄積遺伝子 / SSH分析 / AFLPマーカー / プロアントシアニジン / カキ / タンニン / 遺伝子発現 / フラボノイド / SSH |
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| Research Abstract |
The analysis was performed to identify key genes for regulating tannin accumulation into fruits of pollination constant non-astringent (PCNA)-type of persimmon originated in Japan and China. For this purpose, we conducted the following three experiments:
1)We constructed genomic library from closely related diploid species, D. lotus, and a seed clone was screened from the library with a probe tightly linked to the trait of natural astringency-loss of Japanese-type PCNA fruits. Then, we constructed ca. 300 kbp contig of D. lotus. Due to the sequence analysis of this contig, we found that the sequences are very homologous to those of D. kaki and it can use for exploring the gene of persimmon (D. kaki). In addition, primer sets designed from the sequences of this contig were able to be applicable to distinguish Japanese-type PCNA from others. We could make a universal primer set to select PCNA persimmon.
2)We conducted the expression analysis of mRNA to identify the genes involved in tannin accumulation of persimmon. For this purpose, suppression subtractive hybridization (SSH) analysis was performed between astringency-removed fruits with ethanol treatment on the tree and non-treated fruits. According to this analysis, we were able to obtain several candidates for the genes involved in tannin accumulation into the fruits.
3)As for investigation of a dominant gene conferring the trait of Chinese-type PCNA, we looked for molecular markers by AFLP method combined with bulked segregant analysis (BSA) using progenies derived from a Chinese PCNA 'Luo Tian Tian Shi'. By this analysis, we found three markers closely linked to the Chinese-PCNA trait. Using a marker among them, we screened genomic library of `Luo Tian Tian Shi' and obtained a genomic clone as the seed for constructing contig for the survey of the gene.
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