波缘烟草和枸杞属间原生质体融合再生杂种小植株

Intergeneric Protoplast Fusion between Nicotiana undulata L. and Lycium barbarum L. and Regeneration of Hybrid Plantlets

分别利用碘乙酰胺(IOA)和^(60)Co-γ射线失活波缘烟草和枸杞的原生质体,然后用改良PEG方法诱导融合。结果表明,异种融合频率为3％左右。利用超氧歧化酶对50个随机选取的细胞系进行同工酶分析发现,9个细胞系为可能的杂种细胞系。 对其中4个分化出小植株的可能杂种细胞系进行叶片酯酶同工酶分析,证实它们均为体细胞杂种,杂种小植株在形态上兼有双亲特点且难以生根。细胞学观察指出,这4个杂种细胞系中的3个为可能的二细胞融合物,其平均染色体数目为62.3,59.1和68.2;1个为可能的多细胞融合物,其平均染色体数目为123.6。对一个杂种细胞系进行rDNA Southern杂交分析,结果表明,杂种中含有双亲特有的DNA片段且出现了双亲没有的新片段。

An improved PEG method was used to induce protoplast fusion between N. undulata and L. barbarum , which had been respectively inactivated by IOA and 60Co-γ irradiation. Results showed that the frequency for hetero-fusion was ca. 3%. Superoxide dimutase isozyme analysis on randomely-chosen 50 cell lines suggested that 9 of them were probably hybrids. Esterase analysis on leaf material from regenerants of parental and 4 putative hybrid cell lines confirmed that the 4 cell lines were all somatic nuclear hybrids. The hybrid plantlets were morphologically intermediate of the parents and recalcitrant to regenerate root, cytological observation indicated that 3 of the 4 hybrid cell lines were probably binary-fusion products with average chromosome numbers 62. 3, 59.1, and 68. 2 respectively; one was possibly multi-cell fusion product with average chromosome number 123. 6, Ribosomal DNA Southern analysis showed that the hybrid cell line contained DNA fragments specific to both parents. In addition, it had u-nique DNA fragments suggesting that inter-genomic recombination (or translocation) might had occured.