Zhou A, Xia G, Zhang X, Chen H, Hu H
School of Life Sciences, Shandong University, Jinan, People's Republic of China.
Mol Genet Genomics. 2001 May;265(3):387-93. doi: 10.1007/s004380000430.
Intergeneric somatic hybridization between wheat (cv. Jinan 177) protoplasts that have 24-28 chromosomes and Haynaldia villosa protoplasts containing 11-14 chromosomes was carried out by the polyethylene glycol (PEG) method. A high frequency of hybrid calli and plants were obtained from the fusion products, as revealed by cytological and biochemical techniques and by PCR analysis of 5S rDNA spacer sequences. GISH (genomic in situ hybridization) analysis confirmed the presence of chromosomes from both parents in the hybrid clones and the common occurrence of translocations between them. The RFLP analysis of the organellar DNA using mitochondrion- and chloroplast-specific probes revealed that mitochondria from both parents existed in the cells of hybrid calli and their recombination, whereas chloroplasts segregated and recombined randomly. The gross morphology of hybrid plants resembled that of wheat, but the gross morphology of their ovaries and anthers were intermediate between those of the two parents. The relationship between hybrid plant regeneration and the balance of genetic materials in hybrid clones is discussed.
采用聚乙二醇(PEG)法,对具有24 - 28条染色体的小麦(品种济南177)原生质体与含有11 - 14条染色体的簇毛麦原生质体进行了属间体细胞杂交。通过细胞学、生化技术以及5S rDNA间隔序列的PCR分析表明,融合产物中获得了高频的杂种愈伤组织和植株。基因组原位杂交(GISH)分析证实杂种克隆中存在双亲的染色体,且它们之间普遍发生易位。使用线粒体和叶绿体特异性探针进行的细胞器DNA的RFLP分析表明,双亲的线粒体存在于杂种愈伤组织细胞中并发生重组,而叶绿体则随机分离和重组。杂种植物的总体形态与小麦相似,但其子房和花药的总体形态介于双亲之间。文中讨论了杂种植物再生与杂种克隆中遗传物质平衡之间的关系。