Localization of lily symptomless virus and cucumber mosaic virus in anther- and filament-derived calluses and effect of callus culture duration on virus-free bulblet production in Lilium × ‘Enchantment’

Abstract

Lily symptomless virus (LSV) and cucumber mosaic virus (CMV) were investigated in cultured explants of anther and filament, induced calluses, and regenerated bulblets by the enzyme-immunohistochemical method or indirect enzyme-linked immunosorbent assay (ELISA). The viruses were detected frequently in the vascular tissues of explants, anther-derived friable calluses (33% LSV; 20% CMV), filament-derived calluses (80% LSV; 60% CMV) and their regenerants but rarely in anther wall tissues, nodular calluses (10% LSV; 3% CMV) induced from anther wall tissues, and bulblets regenerated from the nodular calluses. The anther-derived nodular calluses yielded a great virus-free bulblet by 74%, as compared to 47% for anther-derived friable calluses and 35% for filament-derived calluses. Generally, the percentages of virus-free bulblets increased as the culture duration of these calluses was extended from 90–150 days. The best production of virus-free bulblets (57–100%) was achieved in 150 day-old anther-derived nodular calluses. Efficiency of virus-free bulblet production by anther culture is related to the origin and culture duration of callus in Lilium × ‘Enchantment’.