Direct Tissue and Cell Analysis of Tulip Plants by Combining Pressure Probe and UV-MALDI-MS

HIROSHI NONAMI; YOUSEF GHOLIPOUR; ROSA ERRA-BALSELLS

Lisboa, Portugal, August 22-27.

Congreso; 28th International Horticulture Congress, Science and Horticulture for People.; 2010

International Horticulture Society

Direct Tissue and Cell Analysis of Tulip Plants by Combining Pressure Probe and UV-MALDI-MS   Nonami, Hiroshi (1,2); Gholipour, Yousef (2); Erra-Balsells, Rosa (3)   1: Plant Biophysics/Biochemistry Research Laboratory, Faculty of Agriculture, Ehime University, Matsuyama, Japan; 2: The United Graduate School of Agricultural Sciences, Ehime University, Matsuyama, Japan; 3: CIHIDECAR-CONICET, Departamento de Quimica Organica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina   Keywords: tulip bulb, cell pressure probe, UV-MALDI-MS, carbon nanotube   Abstract In the case of single-cell analyses, because of very low (usually subpicoliter) volume of extracts, precise extraction techniques are required. A cell pressure probe technique has been used to measure single plant cell turgor and extract cell solution to determine osmotic pressure. Such a pressure probe technique can be combined together with UV-MALDI-MS for analyses of single cell solution. Single-cell cytoplasm (1-10 pL) of tulip leaf and bulb cells was extracted by a home-made cell pressure probe equipped with a glass microcapillary tube and micro-manipulator, mixed with 1ìL water in a pipette and deposited on air-dried layer of matrices including 2,5-hydroxybenzoic acid (DHB), 2,4,6-trihydroxyacetophenone (THAP), nor-harmane (9H-pyrido[3,4-b]indole; nHo), and carbon nanotubes (CNTs) and analyzed with Voyager-DE STR TOF MS (Applied Biosystems) equipped with nitrogen laser (337 nm, 3 ns pulse width) and with 20 kV of accelerating potential. Tissue slices from tulip leaves and bulbs were made by a razor blade and located on the probe, immediately air-dried, and covered by matrices mentioned above and directly analyzed with the MALDI MS after re-drying.     The positive ion mode showed superior reproducibility of neutral carbohydrate signals when both single-cell cytoplasm and/or tissues were analyzed. Sucrose and fructans (up to 15Hex when THAP was used, 11Hex with DHB and 7Hex with CNTs) were detected in tulip bulb cell cytoplasm samples. In the case of tissues, only CNTs could efficiently desorb/ionize neutral carbohydrate-related signals from the surface. Hexose, sucrose, and fructan (up to 9Hex) were detected in bulb tissues. Fructans signals were considerably similar in both single-cell and tissue analyses. In leaf samples, hexoses, sucrose and a triose were detected in both single-cell cytoplasm and tissues.