Induction of Diploid Eggs With Colchicine During Embryo Sac Development in Populus

Diploid (2n) eggs were induced by treating developing embryo sacs of Populus with colchicine solution, in order to produce triploid plants. The optimal pollinated time of female catkins was confirmed as timing point for each treatment. When female catkins of P. pseudo-simonii x P. nigra ‘Zheyin3#’ had become 5.62 ± 0.13 cm long 84 h after they emerged from their bract scales and all stigmas were exposed, pistils all over the entire catkin had optimal stigma receptivity. Observation of paraffin sections showed that embryo sac development of ‘Zheyin3#’, which initiated 12 h before pollination and finished 132 h after pollination, was a successive and asynchronous process. Generative cell division of pollen of the male parent P. x beijingensis took place 3-16 h after pollination. Catkins of 18-96 h after pollination of ‘Zheyin3#’ were treated with colchicine solution. In the progeny, twenty three triploids were detected by chromosome counting and the highest rate of triploids was 66.7% in one treatment. The rate of triploid yield was positively correlated with the frequency of four-nucleate embryo sacs (r = 0.6721, p = 0.0981) and was not significantly correlated with the percentages of uni-, twoand eight-nucleate embryo sac (r = -0.1667, p = 0.7210, r = -0.3069, p = 0.5031 and r = 0.0189, p = 0.9679, respectively), suggesting that the third mitotic division of embryo sac may be the effective stage to induce 2n eggs. Through this approach, completely homozygous 2n eggs can be produced. Its significance for plant breeding is discussed.