Characterization of Two Gene Transcripts Modulated by Cytokinins in Micropropagated Apple (Malus domestica [L.] Borkh) Plantlets

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Abstract
The micropropagation of apple (Malus domestica [L.] Borkh) cultivars is usually achieved by axillary bud stimulation and requires an exogenous cytokinin supply. Two cDNA libraries were constructed from mRNA isolated from plantlets grown in vitro on medium with or without benzyladenine. One cDNA clone (pSD3), corresponding to transcripts more abundant in plantlets grown on medium containing cytokinin than on medium lacking the hormone, was isolated. It corresponds to a mRNA of about 1,800 nucleotides which codes for a proline-rich protein with a predicted mass of 31,000 daltons. Its accumulation is restricted to roots and stems of in vivo grown apple plantlets and to stems of microcuttings cultivated on medium without cytokinin. Furthermore, it accumulates to comparable levels in leaves and stems when plantlets are grown on medium containing benzyladenine. A second cDNA clone (pSD4), corresponding to transcripts down-regulated in the presence of cytokinin in the culture medium, was also characterized. Its corresponding mRNA is about 700 nucleotides in length and encodes a protein highly homologous to the precursor of the 10-kilodalton polypeptide of the photosystem II from spinach. This mRNA accumulates specifically in leaves of apple plantlets and is more abundant in leaves of plantlets grown in the absence of cytokinin compared with plantlets grown in the presence of benzyladenine.