Marasek Agnieszka, Hasterok Robert, Wiejacha Katarzyna, Orlikowska Teresa
Department of Biotechnology of Ornamental Plants, Research Institute of Pomology and Floriculture, Skierniewice, Poland.
Hereditas. 2004;140(1):1-7. doi: 10.1111/j.1601-5223.2004.01721.x.
In the genus Lilium, plants obtained from crosses, especially between distant relatives, are not always hybrids because embryos can develop as a result of apomixis. These plants constitute genetic material of the maternal parent only. In this study, verification of hybrid status of plants which have been obtained from the crosses 'Marco Polo'xLilium henryi and 'Expression'xL. henryi was performed through the use of cytological and molecular cytogenetic methods. According to cytological analyses, all genotypes tested had 2n = 2x = 24 chromosomes. Genomic in situ hybridisation (GISH) was used for hybrid verification. In hybrid plants, this method distinguished all paternal and maternal chromosomes at the stage of somatic metaphase and prophase. For GISH, paternal genomic DNA was used as a probe and maternal DNAs were used as blocks. Fluorescence in situ hybridisation (FISH) with 5S rDNA and 25S rDNA probes was used as the second method of hybrid verification. Selected chromosome markers based on genome-specific localisation of rDNA loci were used for analysis of the F1 hybrids obtained from the crosses 'Marco Polo'xL. henryi and 'Expression'xL. henryi. The presence of marker chromosomes characteristic for each of the paternal genotypes was a confirmation that the plants obtained were hybrids.
在百合属中，通过杂交获得的植株，尤其是远缘杂交的后代，并不总是杂种，因为胚胎可能通过无融合生殖发育而成。这些植株仅包含母本的遗传物质。在本研究中，利用细胞学和分子细胞遗传学方法对‘马可波罗’×湖北百合以及‘风采’×湖北百合杂交所得植株的杂种状态进行了验证。根据细胞学分析，所有测试基因型均具有2n = 2x = 24条染色体。基因组原位杂交（GISH）用于杂种验证。在杂种植株中，该方法在体细胞中期和前期阶段能够区分所有父本和母本染色体。对于GISH，父本基因组DNA用作探针，母本DNA用作封阻。使用5S rDNA和25S rDNA探针进行荧光原位杂交（FISH）作为杂种验证的第二种方法。基于rDNA位点的基因组特异性定位选择的染色体标记用于分析‘马可波罗’×湖北百合以及‘风采’×湖北百合杂交所得的F1杂种。每个父本基因型特有的标记染色体的存在证实所获得的植株为杂种。
