| Project/Area Number |
14560001
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Breeding science
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| Research Institution | Ibaraki University |
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Principal Investigator |
KUBOYAMA Tsutomu Ibaraki University, the college of Agriculture, Associate Professor, 農学部, 助教授 (10260506)
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| Co-Investigator(Kenkyū-buntansha) |
TSUCHIYA Tohru Mie University, Life Science Research Center, Associate Professo, 生命科学研究支援センター, 助教授 (30293806)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2003: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2002: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | pollen / anther / sterile / petunia / transposon mediated gene disruption / β-1,3 glucanase / cDNA microarray / rice / 花粉管 / 雄生不稔 |
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| Research Abstract |
Two β-1,3 glucanase genes (PhPOGLU, PhPTGLU) found in EST of petunia pollen were targeted for transposon mediated gene disruption. 1024 plants of W138 line were screened by 3 dimensional PCR (Koes et al. 1995). One dTph1 insertion mutant of PhPOGLU was found. This mutant should lose the glucanase activity because dTph1 was inserted in 56th codon of PhPOGLU and this position was relatively near the aminoterminal of PhPOGLU (491 a.a.). However, this mutant did not show any phenotypic change in pollen tube elongation. Pollen fertility was decreased in the, mutant (68.4%), but heterozygous plants showed severer pollen sterility (ca. 30%) than homozygous mutants. Therefore, it is not clear whether the cause of this decreased pollen fertility is due to the mutation of PhPOGLU or not. When we made plant population, some pink flowered female sterile plants (pfs line) were observed. The plants of pfs line had an1 as hetero zygous. One was an1 which had dTphl insertion in the intron exon junction (an1-W138) and the other was an1 which had a 5bp footprint remained after excision of dTphl from an1-W138 allele (an1-pfs). This hererozygous state should be the cause of the female sterile because plants that had homozygous an1-pfs or homozygous an1-w138 did not show female sterility.
In addition, we performed the analyses of the cDNA microarray to monitor gene expression patterns during anther development in rice. Microarray analysis of 4,304 cDNA clones revealed that 259 cDNA clones (156 non redundant groups) were specifically or predominantly expressed in anther tissues and were regulated by developmental stage-specific manners in the anther tissues. These co-regulated genes would be important for development of functional anther tissues. Furthermore, we selected several clones for RNA in situ hybridization analysis and transformation mediated gene silencing. Four lines derived from two chimeric gene constructs show male sterility.
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