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ISHS Acta Horticulturae 1143: VII International Symposium on Edible Alliaceae

Recent aspects of Allium cryopreservation in the federal German genebank

Authors:   E.R.J. Keller, A. Senula
Keywords:   garlic, shallot, Allium obliquum, immature inflorescences, pollen, cryo-microscopy, molecular analysis
DOI:   10.17660/ActaHortic.2016.1143.6
Abstract:
The genus Allium has been one of the main research objects of the Gatersleben institute for many years. Due to the high proportion of accessions that had been collected from the original sites a comprehensive living collection was established. Maintenance of living material was performed with a spectrum of methods, namely field culture, seed and in vitro storage, and cryopreservation. Vegetatively propagated material is very vulnerable both in the field and also in vitro as recent data on field loss and endophytic threat show. Endophytes may influence plant development in vitro and regeneration of explants without their directly visible appearance. Resulting from this experience, permanent storage in liquid nitrogen turned out to be the safest conservation method in the long term. Suppression of dangerous outbreaks after rewarming is a possible strategy to store material with high bacterial load. Various antibiotics were applied to rescue these explants. A case study of an accession lost in the field and re-established from liquid nitrogen storage is illustrating the value of cryopreservation. The main method of cryopreservation is storage of explants from shoot tips out of the bulbs, cloves, bulbils, or in vitro plants. More recently other organ sources were introduced such as basal parts of young inflorescences. This was performed with garlic. To support the taxonomic research collection, which is difficult to maintain, this method was also adopted for cryopreservation of wild Allium species. Furthermore, the maintenance of the latter very complex collection is supported by pollen cryopreservation. Pollen is a very suitable object for investigations by cryo-microscopy, which is recently a main research topic in the cryopreservation unit. Storage of pollen of the botanical family Amaryllidaceae to which Allium belongs seems to be facilitated by its low water content, which was demonstrated by Differential Scanning Calorimetry. Resulting from the accumulated experience in cryopreservation, recommendations will be given how to apply the various techniques in the complex genebank collection.

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