GFP在病原菌及其与植物互作中的应用

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Application of GFP in fungal research
Isolation of transformed cells or specific cell types from populations cells (细胞分离) Quantification of gene expression of individual cells within whole organisms (定量分析基因表达) Assessment of the dispersal and biomass of organisms in complex environments, such as in animal of plant hosts, in biofilms, in fermentors, on leaf surfaces, or in soils (分析真菌的分布与生物量)
F. sambucinum hyphae
C. magna germinated conidia with germ tube and appressoria (附着胞)
B. Cinerea Conidiophores (分生孢子梗)
A. alternata hyphae and conidia (菌丝与分生孢子)
GFP在病原菌及其与 植物互作研究中的应用
李洪杰 2009年1月7日
2008年诺贝尔化学奖－GFP
Osamu Shimomura 下村修 美国Woods Hole 海洋生物学实验室 名誉退休教授 1960年日本名古屋大学 有机化学博士 首次分离出GFP 发现GFP在紫外线下 发出明亮的绿光
Martin Chalfie 马丁·沙尔菲 美国哥伦比亚大学教授 1977年美国哈佛大学 神经生物学博士
GFP expression in different structures of fungi
P. tritici-repentis (小麦褐斑病菌) 分生孢子
V. Dahliae (棉花黄萎病菌) 微菌核
分生孢子
C. sativa (麦根腐旋胞腔菌)
GFP expression in different structures of fungi
Roger Tsien 钱永键 美国加州大学 圣地亚哥分校教授 1977年英国剑桥大学 生理学博士 明确了GFP发光机制 拓展出绿色之外 的其他颜色
证明了GFP作为多种 生物学现象的发光 遗传标记的价值
Story of GFP research
1955, D Davenport: Describe green fluorescent substance in jellyfish 1962, O Shimomura: Identify GFP as a protein 1969, JW Hasting, JG Morin: Named green fluorescent protein (GFP) 1979, O Shimomura: Characterized structure of chromophore 1992, D Prasher: Cloned GFP 1993, CW Cody: Confirmed structure of GFP chromophore 1994, M Chalfie: GFP expressed in E. coli and C. elegans 1995, R Heim: The origin of enhanced GFP (EGFP) 1996, M Ormo: First crystal structures of wild-type and enhanced GFP. Tsien designed yellow fluorescent GFP T203Y mutant 1999, MV Matz, SA Lukyanov: Red fluorescent proteins discovered in anthozoan corals 2000, A Terskikh: Fluorescent timer protein 2002, RE Campbell: Monomeric DsRed (mRFP) 2004, NC Shaner, L Wang: New GFPs generated by in vitro and in vivo 2007, D Shcherbo: mKate, bright far-red FP 2008, AS Mishin: The first mutant of the Aequorea victoria GFP that forms a red chromophore reported 2008 Nobel Prize
GFP expression in host plants infected with fungus
A mint root infected with GFP-labeled fungus
References
Nesher I, Barhoom S, Sharon A. 2008. Cell cycle and cell death are not necessary for appressorium formation and plant infection in the fungal plant pathogen Colletotrichum gloeosporioides. BMC Biology, 6: 9 Sukno SA, Garcia V, Shaw BD, Thon MR. 2008. Root infection and Systemic colonization of maize by Colletotrichum graminicola. Applied Environmental Biology, 74: 823-832.
---in combination with fluorescence-activated cell sorting, confocal microscopy or quantitative image analysis techniques.
GFP expression in fungi
1996, Spellig T: Ustilago maydis (玉蜀黍黑粉菌) 1997, Suelmann R: Aspergillus nidulans (构巢曲霉) 1997, Vanden Wymelenberg AJ: Aureobasidium pullulans (出芽短梗霉) ----12 genera 16 species of filamentous fungi: Colletotrichum (炭疽菌 ), Mycosphaerella (球腔 菌属), Magnaporthe (大角间座壳属), Cochliobolus (玉圆斑菌属), Trichoderma (木霉 菌), Podospora (柄孢壳菌属), Sclerotinia (核盘菌), Schizophyllum (木霉), Aspergillus (曲霉属), and Phytophthora (疫霉)
O Shimomura
DC her
M Chalfie
S Lukyanov
RY Tsien
NC Shaner, GH Patterson, MW Davidson (2008) FHC Wong, DS Banks, A Abu-Arish, C Fradin (2007) DD Deheyn, K Kubokawa, JK McCarthy, A Murakami, M Porrachia, GW Rouse, ND Holland (2007) D Shcherbo, EM Merzlyak, TV Chepurnykh, AF Fradkov, GV Ermakova, EA Solovieva, KA Lukyanov, EA Bogdanova, AG Zaraisky, S Lukyanov, DM Chudakov (2007) L Wang, WC Jackson, PA Steinbach, RY Tsien (2004) NC Shaner, RE Campbell, PA Steinbach, BNG Giepmans, AE Palmer, RY Tsien (2004) DM Chudakov, VV Belousov, AG Zaraisky, VV Novoselov, DB Staroverov, DB Zorov, S Lukyanov, KA Lukyanov (2003) GH Patterson, J Lippincott-Schwartz (2002) H Mizuno, TK Mal, KI Tong, R Ando, T Furuta, M Ikura, A Miyawakil (2003) RE Campbell, O Tour, AE Palmer, PA Steinbach, GS Baird, DA Zacharias, RY Tsien (2002) LA Gross, GS Baird, RC Hoffman, KK Baldridge, RY Tsien (2000) A Terskikh, A Fradkov, G Ermakova, A Zaraisky, P Tan, AV Kajava, XN Zhao, S Lukyanov, M Matz, S Kim, I Weissman, P Siebert (2000) MV Matz, AF Fradkov, YA Labas, AP Savitisky, AG Zaraisky, ML Markelov, SA Lukyanov (1999) K Brejc, TK Sixma, PA Kitts, SR Kain, RY Tsien, M Ormo, SJ Remington (1997) A Miyawaki, O Griesbeck, R Heim, RY Tsien (1999) A Miyawaki, J Llopis, R Heim, JM McCaffrey, JA Adams, M Ikura, RY Tsien (1997) F Yang, LG Moss, GN Phillips (1996) M Ormo, AB Cubitt, K Kallio, LA Gross, RY Tsien, SJ Remington (1996) R Heim, A Cubitt, RY Tsien (1995) R Heim, DC Prasher, RY Tsien (1994) S Inouye, F Tsuji (1994) M Chalfie, Y Tu, G Euskirchen, WW Ward, DC Prasher (1994) CW Cody, DC Prasher, WM Westler, FG Pendergast, WW Ward (1993), D Prasher RO DC Prasher, VK Eckenrode, WW Ward, FG Pendergast, MJ Cormie (1992) McCann, MJ Cormier (1985) O Shimomura: (1979) H Morise, O Shimomura, FH Johnson, J Winant (1974) JG Morin, JW Hastings (1971) JW Hasting, JG Morin (1969) FH Johnson, Y Saiga (1962) O Shimomura (1962) D Davenport, JAC Nicol (1955) (151 scientists)