Biotechnology Centre, Indian Agricultural Research Institute, 110012, New Delhi, India.
Plant Cell Rep. 1992 Jun;11(5-6):318-21. doi: 10.1007/BF00235090.
Intergeneric somatic hybrids have been produced between Brassica juncea (2n=36, AABB) cv. RLM-198 and Moricandia arvensis (2n=28, MM) by protoplast fusion. Hypocotyl protoplasts of B. juncea were fused with mesophyll protoplasts of M. arvensis using polyethylene glycol. Fusion frequency, estimated on the basis of differential morphological characterstics of parental protoplasts was about 5%. Of the 156 calli obtained, four calli produced shoots intermediate in morphology between the parents. Hybrid nature of the plants was confirmed using wheat nuclear rDNA probe. Hybridization of total DNA with a mitochondrial DNA probe carrying 5s-18s rRNA genes of maize showed that the mitochondria of the somatic hybrids were derived from the wild species M. arvensis. Meiosis in the only hybrid that produced normal flowers revealed the occurrence of 64 chromosomes, the sum of chromosomes of parental species. Inspite of complete pollen sterility, siliquas were produced in this hybrid by back-crossing with B. juncea. These siliquas on in vitro culture produced 12 seeds.
通过原生质体融合,已经在芸薹属(2n=36,AABB)cv.RLM-198 和芝麻菜(2n=28,MM)之间产生了种间体细胞杂种。用聚乙二醇融合了芸薹属的下胚轴原生质体和芝麻菜的叶肉原生质体。根据亲本原生质体的差异形态特征估计融合频率约为 5%。在获得的 156 个愈伤组织中,有 4 个愈伤组织产生的形态介于亲本之间的芽。利用小麦核 rDNA 探针证实了这些植株的杂种性质。用携带玉米线粒体 5s-18s rRNA 基因的线粒体 DNA 探针进行总 DNA 杂交表明,体细胞杂种的线粒体来自野生种芝麻菜。唯一产生正常花朵的杂种减数分裂显示出 64 条染色体的存在,这是双亲种染色体的总和。尽管花粉完全不育,但通过与芸薹属回交,这个杂种仍产生了荚果。这些荚果在体外培养中产生了 12 颗种子。