In vitro regeneration of wild pear (<Emphasis Type="Italic">Pyrus pyraster</Emphasis> Burgsd) clones tolerant to Fe-chlorosis and somaclonal variation analysis by RAPD markers |
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Authors: | Maria Antonietta Palombi Beatrice Lombardo Emilia Caboni |
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Institution: | (1) CRA–Istituto Sperimentale per la Frutticoltura, Via di Fioranello 52, 00134 Rome, Italy |
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Abstract: | An in vitro adventitious regeneration system under selective pressure was established in Pyrus pyraster Burgsd to obtain somaclones with higher adaptability to calcareous soils. P. pyraster is important species, both for its relative closeness to cultivated pear and for reforestation of marginal farmland and for
the production of timber. Shoot regeneration was induced from leaves and vegetative apices of in vitro-grown shoots on a modified
LP medium supplemented with naphtaleneacetic acid (1.07 μM) and benziladenine (BA, 8.9 μM). After 30 days, explants were transferred
to an expression medium consisting of the same basal medium with only BA present. Selective treatments utilized MS medium
with Fe-EDTA replaced by equimolar amount of FeSO4 with either KHCO3 or NaHCO3. Through the selection process 11 putatively tolerant lines were obtained from vegetative shoot apices. RAPD analysis was
performed on these lines to allow comparison to the mother clone. A total of seven 10-mer primers were used to amplify all
the genotypes and 74 scorable fragments were produced. These were analysed using the Dice similarity index, showing genetic
variability among the 11 regenerated clones and between them and the mother clone. |
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Keywords: | Adventitious shoot regeneration Crops wild relatives Fe-limiting conditions Genetic variability In vitro cultures |
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