ISSR-PCR technique: a useful method for characterizing new allotetraploid somatic hybrids of mandarin

Somatic hybrids between Redblush and Duncan grapefruit (Citrus paradisi Macfadyen) with Avana, Tardivo di Ciaculli mandarin (Citrus deliciosa Tenore) and Fortune mandarin (Citrus reticulata Blanco) were obtained by symmetric protoplast fusion. Isozyme banding pattern analysis and flow cytometry were used for early screening. DNA of the hybrids was then extracted and subjected to inter-simple sequence repeat polymerase chain reaction (ISSR-PCR) using different primers anchored at the 5’ and 3’ ends. PCR products were resolved on 1.5% agarose gel and stained with ethidium bromide. All the bands from the parental cultivars were present in the somatic hybrids. The usefulness and efficiency of the ISSR-PCR method as a screening technique for Citrus somatic hybrids are discussed.