Molecular cytogenetic analysis of the genome of the potato late blight fungus Phytophthora infestans
Project/Area Number |
18580043
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Research Category |
Grant-in-Aid for Scientific Research (C)
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Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Plant pathology
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Research Institution | Okayama University |
Principal Investigator |
TAGA Masatoki Okayama University, Graduate School of Natural Science and Technology, Associate Professor (80236372)
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Project Period (FY) |
2006 – 2007
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Project Status |
Completed (Fiscal Year 2007)
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Budget Amount *help |
¥3,700,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥300,000)
Fiscal Year 2007: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2006: ¥2,400,000 (Direct Cost: ¥2,400,000)
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Keywords | potato late blight / Phytophthora / chromosome / genome / Oomycete / molecular cytogenetics / ploidy / ジャガイモ疫病菌 / ジャガイモ / 疫病菌 / 細胞遺伝子 |
Research Abstract |
Phytophthora infestans is the causal pathogen of the most devastating potato disease called late blight. Although genome sequence project of this fungus is under way, basic genome information from cytogenetics is very scarce. In this study, we conducted molecular cytogenetic analyses to determine the karyotype and ploidy of P. infestans. 1. Karyotype analysis We developed a method to prepare mitotic chromosome specimens by combining the germ tube burst method and chemical treatment with colchicine, colcemid, and cyclohexamide. Chromosomes prepared with this method were stained with DAPI, Giemsa, and silver nitrate for karyotyping. Chromosome number of a standard diploid strain was 2n=10-14 in most specimens including one or two NOR chromosomes. Sister chromatids in a coiled shape and heterochromatines could be observed in not-fully condensed chromosomes. 2. Fluorescence in situ hybridization (FISH) Probes were cloned and FISH was performed to identify specific chromosomes and analyze ploidy cytologically. With rDNA as a probe, NOR was located near the end of chromosome. Using single copy genes, NiaA and Piexol, as probes, the number of signals per nucleus was shown to indicate ploidy level. 3. Flow cytometry (FCM) The method to measure nuclear DNA contents of P. infestans by FCM was established. Using Arabidopsis thaliana as the internal reference, genome size of P. infestans was estimated to be 250-300Mb. The survey of world-wide collection of ca. 60 strains showed that strains can be grouped into haploids, dipoids, triploids, aneuploids, and heterkaryons. Formerly reported tetraploids could not be detected in our collection.
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Report
(3 results)
Research Products
(7 results)
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[Presentation] 日本産ジャガイモ疫病菌の倍数性解析2008
Author(s)
有本英里香, 金子真美, 多賀正節
Organizer
平成20年度日本植物病理学会大会
Place of Presentation
島根県松江市
Year and Date
2008-04-26
Description
「研究成果報告書概要(和文)」より
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