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Simple strategy for the in vitro conservation on Alnus glutinosa (L.) Gaertn. Germplasm.

AuthorsSan José, M.C.; Janeiro, L. V.; Corredoira, E.
Alnus glutinosa
cold storage
medium-term storage
Issue Date2015
CitationTrees - Structure and Function 29: 539-549 (2015).
AbstractThe aim of this study was to develop a simple method for the medium-term storage of Alnus glutinosa (L.) Gaertn. explants obtained from trees aged 20-30 years. Several parameters were evaluated, including type of explant (shoot apex or nodal segments), pre-storage treatment (0 or 10 days after the last subculture) and duration of cold storage (3-24 months) at 2-4ºC. Explants were maintained at this temperature under dim lighting on Woody Plant Medium supplemented with 0.1 mg l-1 6-benzyladenine and 0.5 mg l-1 indole-3-acetic acid. Under these conditions, a high percentage (75-87%) of cultures remained viable after 18 months in cold cabinets. The stored material was successfully recovered and multiplied normally in the same medium, showing good growth and developing into normal shoots that were morphologically similar to those of non-stored controls. At the histological level, the main change observed was the accumulation of starch granules in cells of the shoot apex, as well as in cells located close to the vascular bundles, after 3 months of cold storage. As the duration of cold storage increased, the number and size of the starch granules decreased but cell plasmolysis and the content of lipid droplets increased. Cold damage was generalized after 24 months at 4ºC. This study provides new insights into the changes occurring in A. glutinosa during cold storage. Key words alder, Alnus glutinosa, cold storage, histology, micropropagation, medium-term storage Author Contribution Statement MC San José and E Corredoira designed the research, analyzed the data and wrote the paper. LV Janeiro conducted the research. All authors have read and approved the final manuscript. Key Message This study provides a simple protocol for storage of alder clones. The technique described is useful for in vitro germplasm collections, decreasing the risk of genetic changes and associated costs.
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