Development of new ornamental Allium though interspecific hybridization and survey of RAPD markers by PCR
| Project/Area Number |
05454057
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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 |
園芸・造園学
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| Research Institution | Kagoshima University |
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Principal Investigator |
ARISUMI Kenichi Kagoshima University, Faculty of Agriculture, Professor, 農学部, 教授 (40035100)
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| Co-Investigator(Kenkyū-buntansha) |
ETOH Takeomi Kagoshima University, Faculty of Agriculture, Professor, 農学部, 教授 (10041659)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥4,900,000 (Direct Cost: ¥4,900,000)
Fiscal Year 1994: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1993: ¥3,000,000 (Direct Cost: ¥3,000,000)
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| Keywords | Apomixis / Embryo rescue / Incompatibility / Interspecific hybridization / Ornamental Allium / PCR / Pull-style pollination / RAPD polymorphism / 和合性 / 胚培養 / 培養下での倍数体化 |
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| Research Abstract |
In the genus Allium it has been difficult to obtain interspcific hybrids through standard pollination because of widely prevailing incompatibility. However, when pull-style pollination was used for cutflowers and then embryo rescue was conducted before the commencement of embryo abortion, the efficiency in obtaining interspecific hybrids became much higher than that of common pollination technique. A number of hybrids thus obtained are growing in our laboratory.
On the other hand, based on the analyzes of RAPD polymorphisms after the amplification of DNA by PCR,the following facts were clarified ;
(1). After establishing the extraction of DNA from young leaves and the amplification of DNA by PCR,RAPD profiles of 10 species of ornamental Allium were compared and it was revealed that the deduction of mutual phylogenetic relationships among these species was feasible.
(2). Based on the newly modified DNA microextraction procedure which was applicable to minute (2-4 mg) leaf samples, the conf
… More
irmation of interspecific hybridity became feasible in much younger seedling stage through the analysis of RAPD profiles.
(3). Taq DNA polymerase is the enzyme of choice for PCR,but it often leads to the formation of misincorporated nucleotides and thus complicates the analysis of RAPD polymorphisms. However, it was revealed that the addition of small amount of Pfu polymerase into Taq polymerase substantially improved the RAPD pattern obtained.
(4). Based on the improvement of RAPD profiles through mixing different polymerases, the combination of Taq or Tth DNA polymerase with Pfu DNA polymerase was used to estimate the extent of apomixis in A.tuberosum. The results obtained indicated 90-95% apomixis.
(5). For the first time in higher plants, the amplification of DNA from single pollen grains by PCR has been accomplished, although there are previous reports on DNA amplification using single sperm cells or fungal spores. This method is expected to have wide application in various research fields. Less
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Report
(3 results)
Research Products
(12 results)
