Megaspore Chromosome Doubling in Eucalyptus urophylla S.T. Blake Induced by Colchicine Treatment to Produce Triploids

Triploids generally provide an advantage in vegetative growth in forest trees. However, the technique of triploid breeding is still an open field in the Eucalyptus tree species. This study aims to explore the colchicine treatment technique for megaspore chromosome doubling to establish triploids in this tree species. Cytological observation on microsporogenesis and megasporogenesis was carried out to guide megaspore chromosome doubling induced by colchicine treatment. Ploidy level in progenies was detected by flow cytometry and somatic chromosome counting. A relationship between microsporogenesis and megasporogenesis was established to guide the colchicine treatment. Seven triploids were obtained in the progenies, and the highest efficiency of triploid production was 6.25% when the flower buds underwent a 0.25% colchicine solution treatment for 6 h using an aspiration method seven days after the first observation of leptotene during microsporogenesis on the floral branch. Cytological analysis showed that the megasporocyte from leptotene to diakinesis may be the optimal period for megaspore chromosome doubling by colchicine treatment. Plant height, ground diameter, leaf area, and the photosynthetic parameter of triploid eucalypt were significantly higher than those of the diploid plant at 6 months old. Hybridization with 2n megaspores induced by colchicine treatment is an effective way for Eucalyptus triploid breeding. These results should accelerate the development of advanced germplasms in this tree species.

High temperature-induced triploid production during embryo sac development in Populus

Triploid breeding plays an important role in cultivar improvement in the genus Populus L. A novel approach for triploid production with colchicine during embryo sac development was reported recently by Wang et al. (2010). In the present investigation, female catkins of Populus pseudo-simonii × P. nigra ‘Zheyin3#’ during embryo sac development were exposed to high temperature to assess the effectiveness of high temperature for induction of triploid production. In the progeny, 45 triploids were determined by both flow cytometric analysis and somatic chromosome counting. The period 66-72 h after pollination was the most suitable for high temperature-induced triploid production during embryo sac development in the ‘Zheyin3#’. Cytological analysis showed that the frequency of eight-nucleate embryo sacs rose at an increased rate during 66-78 h after pollination, which suggested that the third mitosis during embryo sac development could be the optimal stage for high temperature-induced triploid production. The highest frequency of triploid production was 40%, which was obtained in the 44°C for 2 h treatment 72 h after pollination. In view of both triploid number and production efficiency, treatments with 41°C for 4-6 h or with 44°C for 2 h during 66-72 h after pollination were both effective for triploid induction in ‘Zheyin3#’. Statistical analysis showed that the growth of triploids and diploids was not significantly different. However, highly significant differences were observed for all leaf characteristics. Finally, the significance of high temperature treatment in Populus triploid breeding programs is discussed.