1993 Fiscal Year Final Research Report Summary
Studies on breeding of self-fruitful varieties transformed with apomixis genes in apples.
| Project/Area Number |
03454034
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Breeding science
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| Research Institution | HIROSAKI University |
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Principal Investigator |
SAITO Ken-ichi HIROSAKI Univ., Fac.of Agric., Professor, 農学部, 教授 (70003423)
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| Co-Investigator(Kenkyū-buntansha) |
ISHIKAWA Ryuji HIROSAKI Univ., Fac.of Agric., Associate Professor, 農学部, 助教授 (90202978)
HARADA Takeo HIROSAKI Univ., Fac.of Agric., Associate Professor, 農学部, 助教授 (10228645)
OKUNO Toshikatu HIROSAKI Univ., Fac.of Agric., Professor, 農学部, 教授 (60003571)
NIIZEKI Minoru HIROSAKI Univ., Fac.of Agric., Professor, 農学部, 教授 (40001490)
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| Project Period (FY) |
1991 – 1993
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| Keywords | apomixis / self-compatibility / fingerprinting / transgenic plant / self-fruitfulness / M.hupehensis |
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| Research Abstract |
M.hupehensis has 51 chromosomes (3n) and shows complete male sterility, but female fertility was proved by the elongation of pollen tubes in the style after cross pollination.
No variations in peroxidase zymogram and traits such as pollen fertility, the morphology of flower, fruit and leaf, and the degree of apomixis, were observed in the offspring of M.hupehensis crossed with 'Megumi' and in the offspring derived from the apomictic seeds of M.hupehensis.
The embryo formation of M.hupehensis was observed at the beginning of August, about two months later than that of 'Indo'. The embryo culture was successful after the end of August. Low temperature treatment at 4ﾟC in the dark for 80 days produced normal plantlets in embryo culture.
Repeatability values of self-fruitfulness (SF) were 0.59-0.89 depending upon experimental materials, and the values of stimulative parthenocarpy (SP) were smaller than that of SF.
Because any of single correlation coefficients among self-compatability (SC), aut
… More
omatic parthenocarpy (AP) and SP was not statistically significant, it was estimated that genetic factors controlling SC, AP and SP differ with one another.
The variations within clonal lines and those between years for fruit-set percentage of parthenocarpic fruit, were remarkably large in comparison with that of cross-pollinated fruit.
DNA amplification fingerprinting using arbitrary primer(s)was applied to the identification of Malus species. Highly varaible DNA fragment patterns were clearly detected by polyacrylamide gel electrophoresis of the amplified extension products, although three sports of 'Delicious' exhibited the same fingerprint as the original cultivar. We applied this fingerprinting to paternity analysis of an apple cultivar of which the pollen parent was unknown. By using 5 arbitrary primers and RFLP analysis of the amplified products, one cultivar was singled out for paternity among six putative candidates.
The leaf segments of an apple root stock (Malus prunifolia Borkh. var. ringo Asami) were inoculated with a strain Agrobacterium tumefaciens into which a herbicide resistant gene (bar) was introduced as a binary vector. The segments inoculated with the bacteria produced callus on the selection medium containing the herbicide (bialaphos), and regenerated shoots were obtained from the callus. The presence of the gene, inserted into the host genome by the vector, was confirmed by PCR analysis. These transgenic plantlets showed complete resistance to the herbicide. Less
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