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2001 Fiscal Year Final Research Report Summary

An evaluation system of resistance ability of Japanese black pine clones against pine wood nematode based on parentage diagnoses using DNA molecular markers

Research Project

Project/Area Number 12556024
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field 林学
Research InstitutionKYUSHU UNIVERSITY

Principal Investigator

SHIRAISHI Susumu  Kyushu University, Faculty of Agriculture, Professor, 大学院・農学研究院, 教授 (70226314)

Co-Investigator(Kenkyū-buntansha) MIYAHARA Fumihiko  Fukuoka Prefecture Forest Research and Extension Center, Senior Researcher, 専門研究員
Project Period (FY) 2000 – 2001
Keywordsforest tree breeding / DNA molecular marker / parentage diagnosis / Japanese black pine / pollen management / sequence polymorphism / microsatellite DNA / paternal inheritance marker
Research Abstract

To establish the evaluation system in commercial forests of the strongness of resistance Japanese black pine (Pinus thunbergii) clones against pine wood nematode, a parentage diagnosis was developed as a basic technique for the system. Since chloroplast DNA (cpDNA) is paternally inherited in coniferous species, useful cpDNA markers were developed for the determination of the paternal parent of the progenies. The results were as follows ;
1. For screening sequence polymorphisms of cpDNA, 18,569 bp-sequence constructed with 52 non-coding regions (13,360 bp) and coding regions (5,209 bp) were sequenced in 16 clones. The length is one seventh of the whole cpDNA. As the results, seven variations in five non-coding regions were recognized. Two mutations of them were single nucleotide polymorphism (SNP), three were simple sequence repeat polymorphism (SSR), one was insertion, and one was insertion / deletion (indel).
2. To evaluate the length polymorphism in cpDNA SSR, twelve SSR regions were investigated. Eleven variations were recognized in the five regions.
3. Chloroplast DNA markers inherited from its paternal parent were actually applied into the parentage diagnosis of the seeds produced in seed orchards. The DNA types of the embryo and endosperm isolated from a seed were analyzed separately and the maternal parent and paternal parent were detected from their cpDNA types. The contribution of each clone was estimated from the parentage diagnosis data. The result indicated that a few clones product the next generation (seeds) in seed orchards. This suggests a serious problem that must be overcome in seed orchard management.
4. An evaluation system of resistance ability of 16 clones is available by using these cpDNA markers and nuclear SSRs together.

  • Research Products

    (13 results)

All Other

All Publications (13 results)

  • [Publications] Shiraishi, S. et al.: "Incomplete paternal inheritance of chloroplast DNA recognized in Chamaecyparis obtusa using an intraspecific polymorphism of the trnD-trnY intergenic spacer region"Theor. Appl. Genet.. 102. 935-941 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 井田和彦: "Non-Isotopic RNASE Cleavage Assay(NIRCA)によるクロマツ、アカマツ、リュウキュウマツ間のrbcL塩基配列多型の検出"日本林学会九州支部論文集. 54. 43-44 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 村上春樹: "AFLP法に使用するDNAポリメラーゼの検討"日本林学会九州支部論文集. 54. 45-46 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Seido, K. et al.: "Pollen dispersal in a Hinoki (Chamaecyparis obtusa) seed orchard detected using a chloroplast DNA marker"J. For. Res.. 6. 7-12 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Seido, K. et al.: "Determination of the selfing rate in a Hinoki (Chamaecyparis obtusa) seed orchard by using a chloroplast PCR-SSCP marker"Silvae Genet.. 49. 165-168 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 渡辺敦史: "ITS領域を利用したマツ属・ヒノキ属植物における雑種識別"日本林学会九州支部論文集. 53. 57-58 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shiraishi, S., Maeda, H., Toda, T., Seido, K., Sasaki, Y.: "Incomplete paternal inheritance of chloroplast DNA recognized in Chamaecypan's obtusa using an intraspecific polymorphism of the trnD-trnY intergenic spacer region"Theor. Appl. Genet.. 102. 935-941 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ida, K., Shiraishi, S.: "Detection of mutation in the rbcL gene among Pinus thunbergii, P. densiflora and P. luchuensis using Non-Isotopic RNASE Cleavage Assay (NIRCA)"Kyushu J. For. Res.. 54. 43-44 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Murakami, H., Watanabe, A., Shiraishi, S.: "Screening of DNA polymerase for AFLP analysis"Kyushu J. For. Res.. 54. 45-46 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Seido, K., Maeda, H., Shiraishi, S.: "Pollen dispersal in a Hinoki (Chamaecyparis obtusa) seed orchard detected using a chloroplast DNA marker"J. For. Res.. 6. 7-12 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Seido, K., Maeda, H., Shiraishi, S.: "Determination of the selfing rate in a Hinoki (Chamaecyparis obtusa) seed orchard by using a chloroplast PCR-SSCP marker"Silvae Genet.. 49. 165-168 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Seido, K., Yamagishi, K., Ida, K., Murakami, H., Shiraishi, S.: "Investigation of selective fertilization in hinoki (Chamaecyparis obtusa) using a chloroplast DNA marker"J. Jpn. For. Soc.. 82. 380-383 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Watanabe, A., Shiraishi, S.: "The ITS region as a marker to detect hybrids of related species in the genera Pinus and Chamaecyparis"Kyushu J. For. Res.. 53. 57-58 (2000)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2003-09-17  

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