Department of Agronomy, University of Kentucky, 40546-0091, Lexington, KY, USA.
Theor Appl Genet. 1984 Dec;69(2):187-92. doi: 10.1007/BF00272893.
Hybridization of Petunia axillaris and P. parodii with Nicotiana tabacum was attempted using the method of in vitro pollination and fertilization. Seedlings were produced when the Petunia species and N. tabacum were used as the maternal parents; however, most of these had the identical somatic chromosome complement of the maternal parent. With crosses involving P. axillaris as the maternal parent, a low frequency of haploids was also produced. Due to the potential of haploids in basic and applied genetic research, additional experiments were carried out to determine whether in vitro pollination was necessary to stimulate haploid production and to more closely define the optimal time for ovule excision and culture. Four treatments were applied to accomplish these objectives. They were: placentas cultured prior to the time of anthesis, with and without pollination, and placentas cultured after the time of anthesis, with and without pollination. In vitro pollination had no effect on the frequency of haploids produced. Placenta attached ovules cultured prior to the time of anthesis produced significantly more haploids than those cultured after anthesis. The preanthesis treatment produced a frequency of 6.5 haploids per 100 ovaries cultured. The culture of placenta attached ovules provides an alternative to anther culture as a means for haploid production.
采用离体授粉和受精的方法，尝试将矮牵牛（Petunia axillaris）和矮牵牛（P. parodii）与烟草（Nicotiana tabacum）进行杂交。当矮牵牛和烟草作为母本时，会产生幼苗，但这些幼苗的体细胞染色体组型与母本完全相同。当涉及到以矮牵牛为母本的杂交时，也产生了较低频率的单倍体。由于单倍体在基础和应用遗传研究中的潜力，还进行了额外的实验，以确定离体授粉是否有必要刺激单倍体的产生，并更准确地定义胚珠切除和培养的最佳时间。为了实现这些目标，应用了四种处理方法：在开花前进行胎盘培养，有和没有授粉，以及在开花后进行胎盘培养，有和没有授粉。离体授粉对单倍体的产生频率没有影响。在开花前进行胎盘附着胚珠培养产生的单倍体数量明显多于开花后进行培养的胚珠。在开花前处理中，每 100 个培养的卵巢中有 6.5 个单倍体。胎盘附着胚珠的培养为花药培养作为产生单倍体的方法提供了一种替代方法。
