School of Life Sciences, Shandong University, Jinan, China.
Protoplasma. 2012 Jan;249(1):197-205. doi: 10.1007/s00709-011-0278-1. Epub 2011 Apr 12.
In our early experiments, a variety of Bupleurum scorzonerifolium-like somatic hybrid plants were obtained from protoplast fusion between Arabidopsis thaliana and UV-treated/untreated B. scorzonerifolium. To compare the effects of UV and γ-ray irradiation on the B. scorzonerifolium partner and obtain Arabidopsis-like hybrids, we designed a novel combination of somatic hybridization between A. thaliana and B. scorzonerifolium. Before protoplast isolation and fusion, the suspension cells of B. scorzonerifolium were irradiated by gamma ray ((60)Co, 50 Gy with 1.3 Gy min(-1)). Both parental protoplasts lost regeneration capacity, but over 100 somatic hybrids restored the capacity and developed to Arabidopsis-like inflorescences and flowers with some characteristics of B. scorzonerifolium. Some hybrid flowers showed yellow sepal, petal, or carpel, whose color was similar to the petal of B. scorzonerifolium; the others had silique of Arabidopsis with angularity of B. scorzonerifolium, and their parts possessed five stamens, the same as B. scorzonerifolium. Cytological analysis showed that three hybrids had Arabidopsis-like karyotypes. Random Amplified Polymorphic DNA (RAPD) and Simple Sequence Repeats (SSR) profiles revealed that both parental fragments were amplified from these hybrids. These results indicated chromatin introgression from B. scorzonerifolium to A. thaliana, which may be related to the complementation of hybrid inflorescence and flower generation.
在我们的早期实验中,通过拟南芥和经紫外线处理/未经处理的柴胡原生质体融合,获得了多种柴胡样体细胞杂种植物。为了比较紫外线和γ射线辐照对柴胡供体的影响,并获得拟南芥样杂种,我们设计了拟南芥和柴胡体细胞杂交的新组合。在原生质体分离和融合之前,柴胡悬浮细胞被γ射线((60)Co,50 Gy,1.3 Gy min(-1))辐照。双亲原生质体均失去再生能力,但超过 100 个体细胞杂种恢复了能力,并发育成具有柴胡某些特征的拟南芥样花序和花朵。一些杂种花表现出黄色的萼片、花瓣或心皮,其颜色与柴胡的花瓣相似;其他的则具有拟南芥的角果和柴胡的棱角,其部分具有五个雄蕊,与柴胡相同。细胞学分析表明,三种杂种具有拟南芥样的核型。随机扩增多态性 DNA(RAPD)和简单序列重复(SSR)图谱显示,这些杂种都扩增了双亲片段。这些结果表明,来自柴胡的染色质渗入到拟南芥中,这可能与杂种花序和花的生成有关。