Isolation of RNA and protein from guard cells of Nicotiana glauca

被引:4
作者
Smart, LB
Nall, NM
Bennett, AB
机构
[1] SUNY Coll Environm Sci & Forestry, Fac Environm & Forest Biol, Syracuse, NY 13210 USA
[2] Univ Calif Davis, Mann Lab, Dept Vegetable Crops, Davis, CA 95616 USA
关键词
epidermal peel; extraction; gene expression; stomata; tree tobacco;
D O I
10.1023/A:1007602017492
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Stomatal guard cells are critical for maintenance of plant homeostasis and represent an interesting cell type for studies of leaf cell differentiation and patterning. Here we describe techniques for the isolation of guard cell RNA and protein from blended epidermal peels of Nicotiana glauca. The RNA isolation procedure is a modification of the hot berate method, which is particularly well-suited for recalcitrant tissues. Protein was extracted by disrupting guard cell-enriched epidermis with a French(R) press. This system offers the following advantages: relatively high yield, low or no contamination by other cell types, fresh tissue as a source of RNA and protein rather than protoplasts, and a plant species that is readily transformable. These techniques will allow for cloning and analysis of genes expressed in guard cells, application of traditional biochemical techniques to guard cell proteins, as well as characterization of genetic manipulation of guard cell function in transgenic plants.
引用
收藏
页码:371 / 383
页数:13
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