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Deciduous azaleas have been gaining popularity because of their showy floral displays and adaptability to adverse environmental conditions. However, an absence of distinguishing morphological characteristics, combined with the wide variability present in most species, has created difficulties in efforts to unambiguously identify the different species. Various DNA isolation protocols were tested in order to determine the most effective methods for isolation of DNA from 22 taxa of  Rhododendron  for subsequent PCR amplification. DNA yields from the various isolation methods varied widely. A minimum of 50 ng/μL of template DNA was necessary for PCR amplification under standard amplification conditions. Results indicated that the effect of tissue age on the efficiency of DNA isolation was taxa-dependent. For most species, extraction of DNA from freshly harvested young leaf tissue resulted in the highest DNA yields. However, DNA yields from  R. serrulatum, R. atlanticum,  and  R. viscosum  `Lemon Drop' were highest when mature leaf tissue was used. Primers designed to amplify the internal transcribed spacer (ITS) region of the nuclear ribosomal genes and the  psbD, trnK , and  16 S chloroplast genes were tested in various PCR reaction mixes in order to optimize reaction conditions for amplification. Primers to both the ITS and the  psbD  gene resulted in satisfactory amplification in the presence of 1.5 mM MgCl 2  and 50 ng template DNA.

Development of DNA Isolation and Amplification Procedures for Sequence Comparisons Among Deciduous Azaleas