Zygotic embryo cell wall responses to drying in three gymnosperm species differing in seed desiccation sensitivity
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Plant cell walls (CWs) are dynamic in that they can change conformation during ontogeny and in response to various stresses. Though seeds are the main propagatory units of higher plants, little is known of the conformational responses of zygotic embryo CWs to drying. This study employed cryo-scanning electron microscopy to compare the effects of desiccation on zygotic embryo CW morphology across three gymnosperm species that were shown here to differ in seed desiccation sensitivity: Podocarpus henkelii (highly desiccation-sensitive), Podocarpus falcatus (moderately desiccation-sensitive), and Pinus elliottii (desiccation-tolerant). Fresh/imbibed (i.e. fresh Podocarpus at shedding and imbibed Pi. elliottii) embryos showed polyhedral cells with regular walls, typical of turgid cells with an intact plasmalemma. Upon desiccation to c. 0.05 g g−1 (dry mass basis), CWs assumed an undulating conformation, the severity of which appeared to depend on the amount and type of dry matter accumulated. After desiccation, intercellular spaces between cortical cells in all species were comparably enlarged relative to those of fresh/imbibed embryos. After rehydration, meristematic and cotyledonary CWs of P. henkelii and meristematic CWs of P. falcatus remained slightly undulated, suggestive of plasmalemma and/or CW damage, while those of Pi. elliottii returned to their original conformation. Cell areas in dried-rehydrated P. henkelii root meristem and cotyledon were also significantly lower than those from fresh embryos, suggesting incomplete recovery, even though embryo water contents were comparable between the two states. Electrolyte leakage measurements suggest that the two desiccation-sensitive species incurred significant plasmalemma damage relative to the tolerant species upon desiccation, in agreement with the CW abnormalities observed in these species after rehydration. Immunocytochemistry studies revealed that of the four CW epitopes common to embryos of all three species, an increase in arabinan (LM6) upon desiccation and rehydration in desiccation-tolerant Pi. elliottii was the only difference, although this was not statistically significant. Seed desiccation sensitivity in species like P. henkelii and P. falcatus may therefore be partly based on the inability of the plasmalemma and consequently CWs of dried embryos to regain their original conformation following rehydration.
KeywordsCell wall Cryo-SEM Desiccation sensitivity Gymnosperm Seed Zygotic embryo
We thank Vishal Bharuth and Nelisha Murugan of the Microscopy and Microanalysis Unit, University of KwaZulu-Natal, for the assistance with the cryo-SEM. Thanks are also due to Sappi Seed Centre for the donation of Pi. elliottii seeds and to Minoli Appalasamy for the assistance with the statistics.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflicts of interest.
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