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In vitro shoot regeneration from internode stem segments and internode-derived callus of blueberry (Vaccinium spp.)
Abstract
A protocol was developed to regenerate viable shoots from internode stem segments in blueberry. Explant response to the treatments varied and included: no response, callus growth only, callus growth and subsequent shoot formation originating from the callus mass, and adventitious shoot formation directly from the internode segment without an intervening callus. The most efficient shoot regeneration was obtained using zeatin at 25 $\mu$M. Up to as many as 25 microshoots could be produced from a single internode stem segment. Thidiazuron induced callus formation followed by the induction of shoots that originated from the callus mass. Optimum regeneration occurred when either of the top 2 (youngest) internodes were selected as explants, rather than older internode stem segments (3 through 8) for all cytokinins studied. Blueberry explant material tended to lower medium pH over a 6-week culture period, the extent of which depended upon the degree of tissue development. Individual internode stem segments did not appear to influence medium pH until after 3 or 4 weeks in culture, corresponding to the time when such tissue began to develop into a small shoot mass or callus mass with small bud primordia. Internode regeneration was influenced by 7 medium pH, with significantly higher regeneration scores obtained at lower medium pH. The use of a pH buffer in the medium appeared to negatively affect internode regeneration significantly over and above that which may have been accounted for by the buffer's influence on medium pH. The use of a "rooting" medium containing 12.3 $\mu$M 2iP, 5 $\mu$M IBA, and no adenine sulfate, proved unfavorable for the induction of in vitro adventitious shoots and unfavorable for the induction of in vitro roots, but favorable for the ultimate establishment of shoots in a soil mix for shoots first cultured on the "rooting" medium compared to the "standard" medium containing 73.5 $\mu$M 2iP.
Subject Area
Plant propagation|Botany
Recommended Citation
Hruskoci, James David, "In vitro shoot regeneration from internode stem segments and internode-derived callus of blueberry (Vaccinium spp.)" (1997). ETD collection for University of Nebraska-Lincoln. AAI9812355.
https://digitalcommons.unl.edu/dissertations/AAI9812355