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Morphological characterization and in vitro germination of heat-treated pollen in Eucommia ulmoides

Abstract Polyploid breeding has the potential to increase the economic secondary metabolites of Eucommia ulmoides. However, pollination with induced ploidy-mixed pollen has failed to produce polyploids (GAO, 2006). In this investigation, the morphological characterization and in vitro germination of heat-induced ploidy-mixed pollen of E. ulmoides were analysed to determine why there is no polyploid production. Heat-treated pollen grains were easily distinguished as large and small according to their length. The large pollen grains were significantly longer than both untreated and heat-treated small samples, suggesting that they were probably 2n pollen. Rather than the three germinal pores in small pollen, the large grains typically had four pores and, in some cases, shallow furrows, which might affect their germination. Although the maximal germination rates of the treated small and large pollen were not significantly different, the large pollen germinated tardily during the early stages of incubation. The small pollen maintained its growth during the incubation, but the tube growth of large pollen almost stopped after 24 h incubation. Both vegetative and generative nuclei in the large pollen moved into tubes later than in small pollen and the frequency of mitosis in generative nuclei of large pollen was low. Therefore, the tardy germination, poor tube growth, and weak activity of both vegetative and generative nuclei probably caused the poor competition of large pollen in certation. Finally, techniques to increase the competition of highploidy pollen and the prospect of 2n female gamete induction in the polyploid breeding program of E. ulmoides are discussed.