Further attempts to induce "egg transformation" using irradiated pollen

Numerous pollination treatments involving heavily irradiated (40-100 krad) pollen of diverse plant species failed to produce any clear cut "egg transformants" of the type reported by Pandey in Nicotiana. Genetic stocks of pea, rapeseed, and apple, bearing multiple Mendelian markers, were employed to detect any possible transformation events. For each plant species, an optimal level of irradiation was determined which would allow normal pollen tube growth leading to fertilization, but which would prevent the formation of normal hybrids due to the "pulverized" condition of the chromosomes contributed by the irradiated pollen. Pollination treatments included selfing, pollination with donor pollen mixed with self pollen, pollination with irradiated donor pollen mixed with self pollen, pollination with irradiated pollen followed by a delayed self pollination, and pollination with irradiated pollen by itself. None of these treatments produced clearly transformed seedlings. The total number of potential transformation events screened was in excess of 6,046 including 2,268 for pea, 3,309 for rapeseed, and 469 for apple. It is concluded that if egg transformation occurs outside of Nicotiana it is a rare event, and its frequent occurrence in Nicotiana must be, at best, an isolated phenomenon.