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Fourier Transform Infrared Microspectroscopy Detects Changes in Protein Secondary Structure Associated with Desiccation Tolerance in Developing Maize Embryos1

Willem F. Wolkers*, Adriana Bochicchio, Giuseppe Selvaggi, and Folkert A. Hoekstra

Department of Plant Physiology, Wageningen Agricultural University, Arboretumlaan 4, NL-6703 BD Wageningen, The Netherlands (W.F.W., F.A.H.); and Dipartimento di Agronomia e Produzioni Erbacee, Piazzale delle Cascine 18, 50144 Firenze, Italy (A.B., G.S.)

Isolated immature maize (Zea mays L.) embryos have been shown to acquire tolerance to rapid drying between 22 and 25 d after pollination (DAP) and to slow drying from 18 DAP onward. To investigate adaptations in protein profile in association with the acquisition of desiccation tolerance in isolated, immature maize embryos, we applied in situ Fourier transform infrared microspectroscopy. In fresh, viable, 20- and 25-DAP embryo axes, the shapes of the different amide-I bands were identical, and this was maintained after flash drying. On rapid drying, the 20-DAP axes had a reduced relative proportion of alpha -helical protein structure and lost viability. Rapidly dried 25-DAP embryos germinated (74%) and had a protein profile similar to the fresh control axes. On slow drying, the alpha -helical contribution in both the 20- and 25-DAP embryo axes increased compared with that in the fresh control axes, and survival of desiccation was high. The protein profile in dry, mature axes resembled that after slow drying of the immature axes. Rapid drying resulted in an almost complete loss of membrane integrity in the 20-DAP embryo axes and much less so in the 25-DAP axes. After slow drying, low plasma membrane permeability ensued in both the 20- and 25-DAP axes. We conclude that slow drying of excised, immature embryos leads to an increased proportion of alpha -helical protein structures in their axes, which coincides with additional tolerance of desiccation stress.

1   This research was supported by the Life Sciences Foundation, which is subsidized by the Netherlands Organization for Scientific Research (W.F.W.), and by European Community (EC) grant PL 920248 from the EC AIR program to A.B. and G.S.
*   Corresponding author; e-mail wim.wolkers{at}algem.pf.wau.nl; fax 31-317-484740.

Plant Physiol. (1998) 116: 1169-1177
Copyright Clearance Center:   0032-0889/98/116/1169/09
© 1998 American Society of Plant Physiologists




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