草木樨状黄芪和木本霸王的科间体细胞杂交

Interfamilial Somatic Hybridization Between Astragalus melilotoides and Zygophyllum xanthoxylum

在成功培养原生质体的基础上,用改进的PEG-高pH高钙法诱导草木樨状黄芪(Astragalus melilotoides)和木本霸王(Zygophyllum xanthoxylum)原生质体融合,得到了科间体细胞杂种融合细胞。采用罗丹明-6G预处理草木樨状黄芪原生质体以及UV-B辐照霸王原生质体,使双亲原生质体及其同源融合产物均不能持续分裂而死亡,融合后的杂种细胞由于生理互补可恢复持续分裂能力而被筛选出来。融合产物经培养分裂获得了2个杂种细胞系,其中1个分化出芽。染色体计数和分子鉴定证明了杂种的真实性。初步比较了杂种细胞系及亲本对盐分和水分胁迫的耐受性,结果表明杂种细胞系对盐分和水分胁迫的耐受性介于两个亲本之间。

On the basis of protoplast culture, we performed protoplast fusion between Astragalus melilotoides Pall. and Zygophyllum xanthoxylum Maxim using a modified PEG high-pH, high-Ca^2＋ method. The protoplasts of A. melilotoides were pretreated with Rhodamine-6G and those of Z. xanthoxylum with UV radiation. Both parent protoplasts were unable to undergo sustained division, but the interfamilial somatic hybrid cells could be selected because fusion cells renewed their sustained division capability owing to physiological complementation between the two parents. We obtained two hybrid cell lines through this selected procedure. Few buds were induced from only one hybrid cell line. Chromosome checking and molecular biological characterization confirmed the hybrid characters of the two hybrid cell lines. Physiological analyses revealed the salt and drought tolerance of the hybrid cell lines.