A new tool for plant cell biology: in vivo antibody uptake in plant protoplasts

Abstract

We report on the in vivo uptake of antibodies into plant protoplasts. It is shown by immunofluorescence that when protoplasts of sunflower, Arabidopsis or tobacco were incubated in vivo with an antibody, this antibody was detected in the cytoplasm and/or the nucleus, depending on the location of the target protein. Furthermore, when protoplasts were cultured in the presence of antibodies specific effects were observed. Incubation with antibodies raised against p34 cdc2 led to a strong inhibition of the division rate and a decrease of the average DNA content of protoplasts. With antibodies against HaWLIM1, a LIM domain protein of the CRP type, a negative effect on actin organization was observed. It is concluded that antibodies can penetrate in vivo into plant protoplasts and thus may be used as powerful tools for the study of protein function.

Plant protoplast technology: Current status

“…"Blot ting" pro -ce dures (South ern, north ern and west ern) are routine to con firm gene in te gra tion and ex pres sion in trans gen ic plants (Har ding andMillam 2000, Bhat andSrinivasan 2002). Bričre et al (2004) have shown that an ti bod ies can pen e trate into liv ing plant proto plasts and, there fore, may be pow er ful tools in eval u at ing pro tein func tion.…”

Fluorescent Staining of Living Plant Cells

“…Although semipermeable, plant primary cell walls only allow molecules and proteins up to 60-120 kDa in size to enter [9,10], making them impermeable to antibodies exceeding this size limit. Thus, to increase the access of antibody, cell walls can be enzymatically removed and the released protoplasts studied [11]. For live-cell imaging, the expression of biological fluorochromes has become a valuable and powerful alternative to immunofluorescence.…”