林長平 教授
植物細菌學研究室
專長:
植物細菌
實驗室:
一號館 218 室 (辦公室)、一號館 306 室 (實驗室)
電話:
02-33664596 (辦公室)、02-33664103 (實驗室)
傳真:
02- 23661980
E-mail:
cplin@ntu.edu.tw
研究室簡介

本研究室以研究篩管及導管侷限性病原細菌為主。研究工作包括以單元抗體、核酸探針、及PCR技術從事病害偵測,亦進行此類病原細菌之分子遺傳學、生物學及生態學之探討,以及此類病原菌與寄主之間交互作用之研究。研究之病原細菌包括:引花生簇葉病、梨衰弱病、翠菊黃萎病等病害之植物菌質體(Candidatus Phytoplasma), Xylella fastidiosa, Leifsonia xyli subsp. xyli等。目前本研究室亦進行nitrogen-fixation rhizobacteria及各種 PGPR之開發應用。

目前研究4大主軸:
一、植物菌質體病害之診斷與病害生態調查
民國83年6月,台灣中部東勢、和平兩地陸續發生疑似梨衰弱病之病害。經長期田間發病生態之調查,確定其病原為屬於第十群以及第二群之植物菌質體,且於田間存在兩群複合感染情形。此外亦證實黔梨木蝨及中國梨木蝨為此病害之媒介昆蟲並完成帶菌梨葉與蟲體之電顯觀察及梨接穗嫁接日日春試驗。本研究室針對兩群台灣梨衰弱病菌質體,開發出一系列PCR 及 real-time PCR 檢測工具,可應用於田間單一及複合感染病株之快速檢測、分類地位鑑定及菌量定量分析。本研究室亦於民國94年首次發現桃園縣大園鄉花卉栽種區之新興植物菌質體病害,並將其命名為日日春葉片黃化病 (periwinkle leaf yellowing),經序列比對後發現其屬於第一群植物菌質體 (16SrI group)。該群植物菌質體又稱為翠菊黃萎病群 (aster yellows group)植物菌質體,為廣泛分佈於世界各地之重要病原。針對此一病害,本研究室已開發專一性引子對 AYTWf1/AYTWr1 可應用於快速檢測。經過長期生態調查,已確認7種寄主植物及4種媒介昆蟲。
Diagnosis techniques for phytoplasma diseases:

  1. Pear decline
  2. Periwinkle leaf yellowing
  3. Peanut witches' broom

二、植物菌質體基因體之研究
本研究室曾針對植物菌植體特定基因加以選殖與分析,包括 RNA 聚合酵素之 sigma factor: gyrAgyrBrecAdnaJ,dnaK polCrpoC; plasmid: matRdnaB1dnaGrpsTserShflB 以及膜蛋白相關基因等,用來研究該等基因之序列及功能,以瞭解該等基因在植物菌質體中所扮演的角色,並將成果運用於植物菌質體之檢測與分群,發展出以 PCR 或 real-time PCR 對罹病植株及進口農作物進行快速檢測之系統,以確認植株是否罹病及鑑定植物菌質體的種類。
本研究室目前亦與與中研院植微所郭志鴻博士合作,進行 peanut witches' broom (PnWB) phytoplasma之基因體解序,並進行以下子題之研究: 
Genomics of peanut witches’broom (PnWB) phytoplasma:

  1. Whole genome sequencing/ bioinformatics
  2. Host specificity, vectorship, effectors and virulence factors, metabolism and cultivability.

三、植物菌質體與寄主植物交互作用之研究
因植物菌質體尚無法人工培養,其細胞的純化仍有許多困難,因此相關之生理、生化特性研究仍屬有限,而有關植物菌質體與寄主植物交互作用之分子生物學探討更是付之闕如。本研究室利用花生簇葉病植物菌質體 (peanut witches' broom phytoplasma, PnWB phytoplasma) 及日日春葉片黃化病植物菌質體 (periwinkle leaf yellowing phytoplasma, PLY phytoplasma) 與模式植物日日春 (periwinkle, Catharanthus roseus) 進行其交互作用之探討。除以差異表現分析法 (differential display) 篩選日日春受到花生簇葉病植物菌質體感染後表現量具有差異性之基因外,亦針對花器及花色變異相關基因進行探討。此外我們亦利用次世代定序 (next generation sequencing, NGS)進行 high-throughput transcriptome 與 small RNA profile 之分析。再藉由建立基因傱q系統 (gene silencing)與基因過量表現系統 (over-expression system),期能對該基因做後續的研究,以瞭解植物菌質體造成植物各種病徵之機制。針對植物菌質體部分我們亦進行了 effectors and virulence factors之篩選與探討。
本研究室在此一主題上與本校生技所林詩舜博士及陳仁治博士合作,針對 PnWB phytoplasma 及 PLY phytoplasma 與日日春植物之交互反應,進行之以下子題之研究:
Molecular Plant-phytoplasma interactions:

1. Differential display: (陳武揚, 2011, NTU)

Periwinkle vs. PnWB phytoplasma
3 down-regulated genes’ cDNA obtained:
ML domain protein
Eukaryotic translation initiation factor SUI1
Plastidic aldolase

2. Flower organ identity and pigment synthesis-related genes: (蘇意婷, 2010, NTU)

Flower organ identity genes
------B-class (AP3 and PI), C-class (AG), E-class (SEP3)
Flower pigment synthesis genes
------Chalcone synthase (CHS), chalcone isomerase (CHI)
Periwinkle vs. PnWB phytoplasma or PLY phytoplasma
Down-regulated

3. High-throughput transcriptome and small RNA of PnWB phytoplasma analyzes with next generation sequencing (NGS) (曾欣怡, 2011, NTU; 張天昫, 2012, NTU)

4.Effectors and virulence factors/ proteomics------Aux/IAA family genes analyses

5.Gene silencing & overexpression system

四、PGPR/ nitrogen-fixation rhizobacteria之開發應用

本研究室目前與本校生技所劉啟德博士進行此一研究主題之合作。

研究室成員及動態:
研究生及研究主題:

  1. 楊喬茵:The role of microRNA396-SVP regulation PHYL1pn-mediated leafy flower transition
  2. 陳鈺凌:Using periwinkle leaf yellowing infected Nicotiana benthamiana as model to study genetic regulation of plant resistance
  3. 張偉俊:Transcriptomic analysis using RNA-seq to dissect plant defense against PLY phytoplasma in Catharanthus roseus

研究生畢業論文主題:(2009後)

  • 黃耀徵 (2009):Study on the etiology and insect vectors of periwinkle leaf yellowing disease
  • 程諭揚 (2010):Cloning and analysis of phytoplasmal immunodominant membrane protein genes imp, idpA and amp
  • 蔡孟旅 (2010):Phylogenetic analyses of periwinkle leaf yellowing phytoplasma based on combined analyses of rRNA, rplV-rpsC, secY and tuf gene sequences
  • 蘇意婷 (2010):The correlation of symptom development in phyllody and virescence with gene expressions in floral organ identity and pigment synthesis and with phytoplasma accumulation in phytoplasma-infected Catharanthus roseus
  • 陳泠伶 (2010):Cloning and analysis of plasmids of periwinkle leaf yellowing phytoplasma
  • 陳武揚 (2011):Characterization of Catharanthus roseus genes regulated differentially by peanut witches’ broom phytoplasma infection
  • 曾欣怡 (2011):High-throughput transcriptome and small RNA analysis for studying phytoplasma infection on Catharanthus roseus using Next Generation Sequencing
  • 張天昫 (2012):Identification of conserved microRNAs and their targets in PnWB phytoplasma induced leafy flower of Cantharanthus roseus
  • 邱敏慈 (2012):The stability of P1 of Turnip mosaic virus impacts the HC-Pro suppression of miRNA pathway
  • 戴肇鋒 (2012):The Effects of Auxin Treatment on Disease Development in Periwinkle Leaf Yellowing Phytoplasma-Infected Periwinkle Plants
  • 宋宜璋 (2013):Dissecting the role of NPR1 in defense against periwinkle leaf yellowing phytoplasma in Catharanthus reseus using virus-indeced gene silencing
  • 黃禹馨 (2013):MiR396-mediated SVP gene expression in floral transtion reprogramming and its involvement with phytoplasma effector SAP54
  • 黃Z甯 (2014):Screening soil bacterial strains for controlling tomato bacterial wilt in Taiwan
  • 賴建閔 (2014):Construction of PR-related defense network using VIGS in Catharanthus roseus
  • 羅孝甫 (2014):Understanding of transcriptional regulation of PR1 gene in Catharanthus roseus
  • 林晏宇 (2014):Integrated-multiple NGS approaches for transcriptome and miRNA studies in Catharanthus roseus
研究領域
◆ 植物保護
◆ 分子生物學
◆ 植物細菌性病害之分子診斷鑑定
◆ 植物病原細菌致病機制及與寄主植物交互作用之探討
個人學歷
1982-1985 美國紐澤西州立羅格斯大學 (Rutgers University) 博士
1978-1980 國立台灣大學植物病蟲害學系研究所碩士
1974-1978 國立台灣大學植物病蟲害學系 學士
個人經歷
2010迄今國立臺灣大學植物醫學學程教授(研究生學程)
2006迄今國立臺灣大學生物科技研究所教授
2002迄今國立臺灣大學植物病理與微生物學系暨研究所教授
2011至2014國立臺灣大學生物資源暨農學院副院長
2005至2008國立臺灣大學生物技術研究中心主任
1998至2001國立臺灣大學植物病理學系系主任
1990至2002國立臺灣大學植病系病理組教授
1986至1990國立臺灣大學植病系病理組副教授
1985至1986美國農業部(USDA)博士後研究員Appalachian Fruit Research Center,USDA, WV, USA
1982至1985美國紐澤西州立羅格斯大學(Rutgers) 植病研究所研究生獎學金 (fellowship)
教授課程
◆ 微生物學 (more)
◆ 植物細菌學
◆ 細菌學特論
研究成果及論文
三年內期刊論文
01.
Lin, Y. Y., Fang, M. M., Lin, P. C., Chiu, M. T., Liu, L. Y., Lin, C. P., and Lin, S. S. 2013. Improving initial infectivity of the Turnip mosaic virus (TuMV) infectious clone by a mini binary vector via agro-infiltration. Botanical Studies 54 (2): 22-31 (SCI).
02.
Chung, W. C., Chen, L. L., Lo, W. S., Lin, C. P., and Kuo, C. H. 2013. Comparative analysis of the peanut witches'-broom phytoplasma genome reveals horizontal transfer of potential mobile units and effectors. PLoS ONE 8(4): e62770. (SCI).
03.
Chiu, M. T., Lin, C. P., Lin, P. C., and Lin, S. S. 2013. Enhancement of IgG purification by FPLC for a serological study on the Turnip mosaic virus P1 protein. Plant Pathol. Bull. 22: 21-30.
04.
Tai, C. F., Lin, C. P., Sung, Y. C., and Chen, J. C. 2013. Auxin promotes symptom development in shoots infected with periwinkle leaf yellowing phytoplasmas but delays their colonization in neighboring shoots in periwinkle. Annals of Applied Biology 163: 420-429 (SCI) (Lin, C. P.: Co-first author).
05.
Sung, Y. C., Lin C. P., and Chen, J. C. 2014. Optimization of virus-induced gene silencing in Catharanthus roseus. Plant Pathology 63: 1159-1167 (SCI) (Lin, C. P.: Co-first author).
06.
Tseng, H. I., Lin, C. P., and Lin, S. S. 2014. Characterization and identification of Catharanthus roseus epigenetic-related genes that in response to peanut witches’-broom phytoplasma-mediated infection. Plant Pathology Bulletin 23: 67-77 (Lin, C. P.: Co-first author).
07.
Liu, L.Y., Tseng, H. I., Lin, C. P., Lin, Y. Y., Huang, Y. H., Huang, C. K., Chang, T.H., and Lin, S. S. 2014. High-throughput transcriptome analysis of the leafy flower transition of Catharanthus roseus induced by peanut witches'-broom phytoplasma infection. Plant and Cell Physiology 55: 942-957 (SCI) (Lin, C. P.: Co-first author) (cover image)
08.
Kung, Y. J., Lin,P. C., Yeh, S. D., Hong, S. F., Chua, N. H., Liu, L. Y., Lin, C. P., Huang, Y. H., Wu, H. W., Chen, C. C., and Lin, S. S. 2014. Genetic analyses of the FRNK motif function of Turnip mosaic virus uncover multiple and potentially interactive pathways of cross-protection. MPMI 27: 944-955. (SCI)
09.
Hu, S. F., Huang, Y. H., Lin, C. P., Hong, S. F., Yang, C. Y., Lo, H. F., Chen, W. Y., Lin, S. S. 2015. Development of a mild viral expression system for gain-of-function study of phytoplasma effector in planta. PLoS ONE: 10(6): e0130139. doi:10.1371/journal.pone.0130139 (SCI) (Lin, C. P.: Co-first author).
 
三年內研討會論文
01.
Chiu, M. T., Lin, C. P., and Lin, S. S. 2012. The function analysis of unstable viral suppressor P1/HC-Pro in post-transcriptional gene silenicng (PTGS) and virus infection.  Plant Pathol. Bull. 21: in press.
02.
Chang, T. H., Tseng, H. I., Kao, S. M., Chao, K. H., Lin, C. P., and Lin, S. S.  2012. The mechanism study of miRNA-mediated reprogramming in floral transition of periwinkle during the PnWB phytoplasma infection. Plant Pathol. Bull. 21: in press.
03. Lin, Y. Y., Fang, M. M., Lin, P. C., Chiu, M. T., Liu, L. Y., Lin, C. P., and Lin, S. S. 2012. Improving the Turnip mosaic virus (TuMV) infectious clone infectivity by redesigning the binary vector and agro-infiltration. Plant Pathology Bulletin 21: 131-192.
04.
戴肇鋒、林長平、陳仁治. 2012. 以生長素處理感染植物菌質體之日日春其生理反應之研究。植物病理學會刊 21: in press.
05.
Sung, Y. C., Lin, C. P., Chen, J. C. 2013. Dissecting the role of NPR1 in defense against periwinkle leaf yellowing phytoplasma in Catharanthus roseus using virus-induced gene silencing. Plant Pathol. Bull. 22: in press.
06.
Lin, Y. Y., Lin, S. S., and Lin, C. P. 2013.  Integrated-multiple NGS approaches fortranscriptome and miRNA studies in Catharanthus roseus. Plant Pathol. Bull. 22: in press.
07.
Huang, Y. S., Lin, C. P., and Lin, S. S. 2013. The functional analysis of phytoplasma effector SAP54 in miRNA396-mediated SVP gene expression in floral transition reprogramming. Plant Pathol. Bull. 22: in press.
08.
Sung, Y. C., Lin, C. P., Chen, J. C. 2013. Dissecting the role of NPR1 in defense against periwinkle leaf yellowing phytoplasma in Catharanthus roseus using virus-induced gene silencing. 10th International Congress of Plant Pathology: p. 204-205.
09.
Lin, Y. Y., Lin, C. P., and Lin, S. S. 2014.  Integrated-multiple high-throughput approaches for studying miRNA regulated transcriptome in phytoplasma-infected Catharanthus roseus. Plant Pathol. Bull. 23: in press.
10.
Lo, H. F., Lai, C. M., Lin, C. P., Chen, J.C. 2014.  Screening CrPR1a-related transcription factors using virus-induced-gene-silencing. Plant Pathol. Bull. 23: in press.
11. Lai, C. M., Lo, H. F., Lin, C. P., and Chen, J. C. 2014. Construction of regulatory network of CrPR1a-related defense against phytoplasma in Catharanthus roseus. Plant Pathol. Bull. 23: in press.
 
12. Yang, C. Y., Huang, Y. H., Lin, Y. Y., Lin, C. P., and Lin, S.S. 2015. The role of microRNA396-SVP regulation in PHYL1Pn-mediated leafy flower transition.Plant Pathol. Bull. 24: in press.
 
13. Chang, W. C., Tai, C. F., Sung, Y. C., Yeh, T. J., Hsu, H. J., Lin, C. P., and Chen, J. C. 2015. Transcriptomic analysis using RNA-seq to dissect plant defense against PLY phytoplasma in Catharanthus roseus. Plant Pathol. Bull. 24: in press.
 
專書及專書論文
01.
林長平(主編).  2007.  動植物疫病害蟲診斷鑑定作業流程-植物疫病.  農委會動植物防疫檢疫局  國立台灣大學出版,台北市
02.
林長平、劉瑞芬、莊榮輝、丁詩同、李宣書(編著).  2009.  生物技術概論.  448 pp.  華杏出版股份有限公司出版,台北市
03.
許輔、林長平.  2009.  生物技術的範疇.  生物技術概論. (林長平、劉瑞芬、莊榮輝、丁詩同、李宣書等編著).  華杏出版股份有限公司出版,台北市  p. 1-24.
04.
陳武揚、林長平.  2010.  台灣地區重要植物菌質體病害診斷鑑定技術之發展與應用.  動植物防疫檢疫季刊26: 66-69.
05.
洪挺軒、林長平.  2011.  台灣農作物重要植物菌質體病害研究現況.  農作物害蟲及其媒介病害整合防治技術研討會專刊 p. 63-70.
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