甘蓝型油菜小孢子再生植株染色体倍数检测研究

Studies on the Determination of Ploidy Level of Microspore-derived Plants in Brassica napus

采用流式细胞光度法 (FCM )对甘蓝型冬油菜 4个F1代杂种的小孢子再生植株 ,在幼苗期 (取再生株新长出的叶片嫩组织 )进行了染色体倍数检测。其矩形图上清晰地检测出单倍体、二倍体、三倍体和四倍体以及单倍体加二倍体二倍体加四倍体和三倍体加六倍体等嵌合体。小孢子再生植株中 ,单倍体和自发二倍体比率分别为 5 0 %～85 % (平均为 6 0 % )和 2 0 %～ 5 0 % (平均为 35 % )。FCM倍数检测结果与在油菜花蕾期进行的花器形态鉴别相一致。本文还讨论了采用FCM倍数检测的优势及其检测进展。

Flow cytometric (FCM) analysis of ploidy level of microspore derived plants was made in four winter oilseed rape F 1 hybrids. Ploidy could be well determined from fresh, newly born leaf blades at the seedling stage. An unequivocal identification of haploids, diploids, triploids, and tetraploids as well as chimerics such as haploid plus diploid, diploid plus tetraploid, and triploid plus hexaploid, was possible from their discrete peaks of histograms. 50%-85% (with the average of 60%) of haploid percentage and 20%-50% (with the mean of 35%) of spontaneous doubling efficiency were obtained from microspore derived plants. These results were well confirmed by close inspection of the morphological characteristics of rape flowers at flowering stage. The advantage and development of FCM analysis were also discussed in the present paper.