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r. howard berg, ph.d. director, integrated microscopy facility, associate member

Te Integrated Microscopy Facility has a repertoire of instrumentation for live cell light microscopy and electron microscopy, striving to furnish state of the art imaging for plant scientists within the institute and regionally.

Structural studies in plant cell biology are commonly oriented to the use of fluorescence microscopy, optical sectioning, and 3D and 4D digital reconstructions from these data. Te resulting imagery is often outstanding and provides compelling evidence for plant cell function. Te Danforth Center’s Integrated Microscopy Facility’s array of optical sectioning light microscopes produce high quality data for users and collaborators. Our Zeiss 510 Meta confocal and multiphoton micro- scope provides 3D and 4D live cell imaging for many ddpsc labs.

Proper analysis of plant cell biology requires not only live cell imaging but also the ability to resolve cellular structures on the molecular level. Tis is implemented in the facility by the use of thin section transmis- sion electron microscopy (tem) of specimens prepared by ultra-rapid freezing, using our Bal-Tec high pressure freezer. Te facility has pro- cessed over 5,500 frozen samples of a large variety of plant tissues, pro- viding experience in analysis of plant ultrastructure that has a significant impact on cell biology studies at the ddpsc. Resin sections of freeze substituted samples are imaged using our Leo 912 energy filter tem. Te energy filter is an electron spectrometer that is used to improve image contrast and can be used to conduct electron energy loss spectroscopy analysis of specimen elemental composition. Current tem projects in- clude analysis of the ultrastructure of tmv viral replication complexes in leaf tissue, soybean root nodule cells, Arabidopsis anther development, oil body development in seeds of several plant species, and altered cell wall development in the moss Physcomitrella.

Tat the facility has significant impact in plant cell biology is shown by me being invited to organize a platform session on advances in plant cell biology at the annual meeting of the Microscopy Society of America and also by being invited to write a review for the journal Advances in Agronomy on the use of microscopy in the plant sciences today.

Te facility is moving forward in technology via grant proposals seeking funding to upgrade the Zeiss lsm with an even more sensitive and ver- satile instrument. Furthermore, I seek funding to equip this instrument with a fluorescence lifetime detector for sophisticated detection of the mo- lecular milieu of fluorophores. Tis will also include the ability to measure molecular kinetics and diffusion constants in vivo due to the detector also being able to perform fluorescence correlation spectroscopy.

Cover illustration, produced by the Integrated Microscopy Facility, for the February 5, 2010 issue of the Journal of Biological Chem- istry. Tis image of epicuticular wax in the petunia flower (green, red is chlorophyll from chloroplasts) highlighted the article from the Jaworski lab in the journal, on petunia flower stigma exudate molecular biology, by Han et al. (Te cytochrome P450 CYP86A22 is a fatty acyl-CoA ω-hydroxylase essential for estolide synthesis in the stigma of Petunia hybrida, JBC 285: 3986-39960).

I am also seeking funding, in collaboration with scientists at Washing- ton University, to acquire an intermediate voltage tem that will bring to scientists in the St. Louis region the ability to do electron tomography. Current projects using this technology are conducted by acquiring raw data at the University of Chicago; funding of our grant will bring instrumentation to Washington University.

recent publications: Refereed Publications Huifen Zhu, Guo-Jing Li, Lei Ding, Xiangqin Cui, Howard Berg, Sarah M. Assmann, Yiji Xia. (2009) Arabidopsis Extra Large G Protein 2 (XLG2) interacts with the G beta subunit of heterotrimeric G protein and functions in disease resistance. Molecular Plant, 2: 513-525. Manjula Govindarajulu, Sung-Yong Kim, Marc Libault, R. Howard Berg, Kiwamu Tanaka, Gary Stacey, Christopher G. Taylor. (2009) GS52 ecto- apyrase plays a critical role during soybean nodulation. Plant Physiology, 149: 994-1004.

Book Berg, R. Howard and C.G. Taylor (editors). (2009) Cell biology of plant nematode parasitism. Plant Cell Monographs, Volume 15, Springer: Berlin. 273 pages. Book Chapter Berg, R. Howard, Thomas Fester, and Christopher G. Taylor. (2009) Development of the root-knot nematode feeding cell. In: R.H. Berg and C.G. Taylor (eds.), Cell biology of plant nematode parasitism, Springer, pp 115-152.

2009 scientific report the donald danforth plant science center page 25

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