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Osmotin induces cold protection in olive trees by affecting programmed cell death and cytoskeleton organization

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Abstract

Osmotin is a pathogenesis-related protein exhibiting cryoprotective functions. Our aim was to understand whether it is involved in the cold acclimation of the olive tree (Olea europaea L.), a frost-sensitive species lacking dormancy. We exposed olive trees expressing tobacco osmotin gene under the 35S promoter (35S:osm) [in the same manner as wild type (wt) plants] to cold shocks in the presence/absence of cold acclimation, and monitored changes in programmed cell death (PCD), cytoskeleton, and calcium ([Ca2+]c) signalling. In the wt, osmotin was immunolocalized only in cold-acclimated plants, and in the tissues showing PCD. In the 35S:osm clones, the protein was detected also in the non-acclimated plants, and always in the tissues exhibiting PCD. In the non-acclimated wt protoplasts exposed to cold shock, a transient decrease in phallotoxin signal suggests a temporary disassembly of F-actin, a transient increase occurred instead in 35S:osm protoplasts exposed to the same shock. Transient increases in [Ca2+]c were observed only in the wt protoplasts. However, when F-actin was depolymerized by cytochalasin or latrunculin, and microtubules by colchicine, increase in [Ca2+]c also occurred in the 35S:osm protoplasts. Successive cold shocks caused transient rises in [Ca2+]c and transient decreases in the phallotoxin signal in wt protoplasts. No change occurred in [Ca2+]c occurred in the 35S:osm protoplasts. The phallotoxin signal transiently increased at the first shock, but did not change after the subsequent shocks, and an overall signal reduction occurred with shock repetition. Following acclimation, no cold shock-induced change in [Ca2+]c levels and F-actin signal occurred either in wt or 35S:osm protoplasts. The results show that osmotin is positively involved in the acclimation-related PCD, in blocking the cold-induced calcium signalling, and in affecting cytoskeleton in response to cold stimuli.

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Abbreviations

AFs:

Actin filaments

CD:

Cytochalasin D

Gd:

Gadolinium-chloride

LA:

Latrunculin A

MTs:

Microtubules

PCD:

Programmed cell death

ROS:

Reactive oxygen species

TMB-8:

8-(N,N-di-methylamino)octyl 3,4,5-trimethoxy-benzoate

35S:osm1, osm2, osm3 :

Transgenic clones overexpressing osmotin gene under CaMV 35S promoter, collectively referred as 35S:osm1-3

wt:

Wild type

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Acknowledgments

We thank Dr. C. Scotta’ of the Università “La Sapienza” (Rome, Italy) for technical support, and Prof. E. Piccolella for helpful suggestions in flow cytometric analysis. We also thank Dr R. Bressan of Purdue University (USA) for the generous gift of the 35S:osmotin construct, Prof. E. Rugini of the Dipartimento di Produzione Vegetale, Università della Tuscia (Viterbo, Italy) for helpful suggestions in plant transformation and culture, and for providing materials for preliminary experiments, and Dr R. Malhò of Departamento de Biologia Vegetal, Lisboa (Portugal) for helpful suggestions in the evaluations of fluorescence signals. This work was supported by funds from the Università “La Sapienza” of Rome (Progetti Ateneo).

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  1. Dipartimento Biologia Vegetale, Università “La Sapienza”, P.le Aldo Moro 5, 00185, Rome, Italy

    S. D’Angeli & M. M. Altamura

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  1. S. D’Angeli
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Correspondence to M. M. Altamura.

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D’Angeli, S., Altamura, M.M. Osmotin induces cold protection in olive trees by affecting programmed cell death and cytoskeleton organization. Planta 225, 1147–1163 (2007). https://doi.org/10.1007/s00425-006-0426-6

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