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电融合法产生骆驼刺与鹰嘴紫云英属间体细胞杂种
引用本文:张改娜,贾敬芬,孔祥生,胥华伟. 电融合法产生骆驼刺与鹰嘴紫云英属间体细胞杂种[J]. 生物工程学报, 2010, 26(5): 635-642
作者姓名:张改娜  贾敬芬  孔祥生  胥华伟
作者单位:1. 河南科技大学农学院,洛阳,471003;西北大学生命科学学院,陕西省生物技术重点实验室,西安,710069
2. 西北大学生命科学学院,陕西省生物技术重点实验室,西安,710069
3. 河南科技大学农学院,洛阳,471003
摘    要:采用电融合法获得了骆驼刺和鹰嘴紫云英属间体细胞杂种。骆驼刺原生质体来自发根农杆菌A4转化系细胞,并经碘乙酰胺处理;鹰嘴紫云英原生质体从甲硫氨酸抗性系细胞分离。双亲及同源融合产物均不能在无激素的筛选培养基上持续分裂,融合后的杂种细胞由于双亲生理互补效应可恢复持续分裂能力而被筛选出来。本实验优化了电融合参数,如直流脉冲、交流脉冲和脉冲次数。融合产物经培养获得的杂种细胞系经形态学、染色体数目检查、生化及随机扩增多态性DNA分析,鉴定出10个杂种克隆,并从3个杂种克隆再生了小植株。

关 键 词:骆驼刺,鹰嘴紫云英,体细胞杂交
收稿时间:2009-10-27

Intergenus somatic hybridization between Alhagi pseudalhagi and Astragalus cicer by electroporation
Gaina Zhang,Jingfen Ji,Xiangsheng Kong and Huawei Xu. Intergenus somatic hybridization between Alhagi pseudalhagi and Astragalus cicer by electroporation[J]. Chinese journal of biotechnology, 2010, 26(5): 635-642
Authors:Gaina Zhang  Jingfen Ji  Xiangsheng Kong  Huawei Xu
Abstract:We obtained intergenus somatic hybrid between Alhagi pseudalhagi and Astragalus cicer by using electroporation. Agrobacterium rhizogenes A4-transformed A. pseudalhagi protoplasts were treated with iodoacetamide so that they were unable to sustain divisions. A. cicer protoplasts were isolated from a methionine-resistant mutant and did not survive in the medium without plant growth regulator. The intergenus somatic hybrid cells were selected based on physiological complementation between the two parents. We optimized some parameters of electroporation, such as direct current impulse, alternating current impulse and the impulse number. We identified ten hybrid clones by morphological observation, checking the chromosome numbers, isoenzyme analysis and random amplified polymorphic DNA analysis, and obtained regenerated plantlets from three hybrid clones.
Keywords:Alhagi pseudal hagi   Astragalus cicer L.   somatic hybridiation
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