Method for culturing of unculturable phytoplasma by a hybrid-genome strategy

• Project/Area Number: 23681043
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Single-year Grants
• Research Field: Applied genomics
• Research Institution: National Institute of Advanced Industrial Science and Technology
• Principal Investigator: KAKIZAWA shigeyuki 独立行政法人産業技術総合研究所, 生物プロセス研究部門, 主任研究員 (10588669)
• Project Period (FY): 2011-11-18 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥15,080,000 (Direct Cost: ¥11,600,000、Indirect Cost: ¥3,480,000); Fiscal Year 2013: ¥7,540,000 (Direct Cost: ¥5,800,000、Indirect Cost: ¥1,740,000); Fiscal Year 2012: ¥7,540,000 (Direct Cost: ¥5,800,000、Indirect Cost: ¥1,740,000)
• Keywords: ゲノム / 細菌

Research Abstract

A vector system for the whole genome cloning was established. A previously reported circular YAC (yeast artificial chromosome) vector was modified and used for genome cloning. Using this new system, high efficiency of positive clones was observed. A method to extract phytoplasma genome from infected plant tissues was also established. In addition, a multiplex-PCR system was established to confirm cloning of large genomic fragment. Strain-identification of phytoplasmas in infected host was also performed using the multiplex-PCR system.

Research Products

• [Journal Article] A Multiplex-PCR Method for Strain Identification and Detailed Phylogenetic Analysis of AY-Group Phytoplasmas (2014)
  • Author(s): Shigeyuki Kakizawa and Yoichi Kamagata
  • Journal Title: Plant Disease Volume: Volume 98, Number 3 Issue: 3 Pages: 299-305
  • DOI: 10.1094/pdis-03-13-0216-re
  • Peer Reviewed
• [Journal Article] Unique morphological changes in plant pathogenic phytoplasma-infected petunia flowers are related to transcriptional regulation of floral homeotic genes in an organ-specific manner (2011)
  • Author(s): Himeno et al.
  • Journal Title: PLANT JOURNAL Volume: 67 Pages: 971-979
  • Peer Reviewed
• [Journal Article] Dramatic Transcriptional Changes in an Intracellular Parasite Enable Host Switching between Plant and Insect (2011)
  • Author(s): Oshima et al.
  • Journal Title: PLoS One Volume: 6-8
  • Peer Reviewed
• [Journal Article] Cloning, expression analysis, and sequence diversity of genes encoding two different immunodominant membrane proteins in poinsettia branch-inducing phytoplasma (2011)
  • Author(s): Neriya et al.
  • Journal Title: FEMS MICROBIOLOGY LETTERS Volume: 324 Pages: 38-47
  • Peer Reviewed
• [Presentation] マイコプラズマの全ゲノム操作技術を用いた難培養性細菌の研究 (2014)
  • Author(s): 柿澤茂行、鎌形洋一
  • Organizer: 第8回日本ゲノム微生物学会年会
  • Place of Presentation: 東京都世田谷区
  • Year and Date: 2014-03-08
• [Presentation] マイコプラズマの全ゲノム操作技術を用いた難培養性細菌の研究 (2014)
  • Author(s): 柿澤茂行、鎌形洋一
  • Organizer: 第8回 日本ゲノム微生物学会年会
  • Place of Presentation: 東京都世田谷区
• [Presentation] Recent progress on phytoplasma-host interactions and future of phytoplasma genomics (2013)
  • Author(s): 柿澤茂行、鎌形洋一
  • Organizer: 10th International Congress of Plant Pathology
  • Place of Presentation: Beijing, China
• [Presentation] マイコプラズマの全ゲノム操作技術を用いた難培養性細菌の研究 (2012)
  • Author(s): 柿澤茂行、鎌形洋一
  • Organizer: 第28回日本微生物生態学会大会(JSME 2012)
  • Place of Presentation: 愛知県豊橋市
  • Year and Date: 2012-09-20
• [Presentation] マイコプラズマの全ゲノム操作技術を用いた難培養性細菌の研究 (2012)
  • Author(s): 柿澤茂行、鎌形洋一
  • Organizer: 第39回日本マイコプラズマ学会総会
  • Place of Presentation: 岩手県盛岡市
  • Year and Date: 2012-05-24
• [Presentation] Multiplex-PCR for epidemiology of phytoplasma (2012)
  • Author(s): 柿澤茂行、鎌形洋一
  • Organizer: 19th Congress of the International Organization for Mycoplasmology (IOM)
  • Place of Presentation: Toulouse, France
• [Presentation] マイコプラズマの全ゲノム操作技術を用いた難培養性細菌の研究 (2012)
  • Author(s): 柿澤茂行、鎌形洋一
  • Organizer: 第28回日本微生物生態学会大会 (JSME 2012)
  • Place of Presentation: 愛知県豊橋市