Cryogenic strategy for the establishment of a germplasm bank of pineapple (Ananas comosus var. Comosus)
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Abstract
protocols based on a vitrification procedure. However,
the influence of some technical factors is still unknown
to achieve a strategy cryogenic routine application to a
wide range of genotypes. Furthermore, there has been no
histological analysis to display any structural changes during
a process of cryopreservation. In the present investigation
different technical aspects of a cryogenic strategy are
determined to increase survival levels from the apexes to
immersion in liquid nitrogen. These were: type of apex,
consisting of 3-4 leaf primordia with an approximate size
of 2,5-3mm; preculture 0,3 mol L-1 sucrose; application of
the loading solution 0,4 mol L-1+2 0,4 mol L-1 mol glycerol;
application temperature of the vitrification solution PVS2 at
0 °C for 420 min. The analysis of the display of structural
changes in the cryopreserved apices revealed that the
meristematic cells in the apical dome and leaf primordia
in formation, suffered few cellular alterations and kept
almost intact its morpho-physiological characteristics in
the best conditions of survival. The method of vitrification
was applied successfully for 9 accessions of the in vitro
genebank of Bioplant center. This is an important step for
the establishment of a gene bank for long-term cultivation
of pineapple.
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