STABLY TRANSFORMED HERBICIDE RESISTANT CALLUS OF SUGARCANE VIA MICROPROJECTILE BOMBARDMENT OF CELL-SUSPENSION CULTURES AND ELECTROPORATION OF PROTOPLASTS
STABLY TRANSFORMED HERBICIDE RESISTANT CALLUS OF SUGARCANE VIA MICROPROJECTILE BOMBARDMENT OF CELL-SUSPENSION CULTURES AND ELECTROPORATION OF PROTOPLASTS
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DOI：
10.1007/bf00236264
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发表时间：
1992-09-01
影响因子：
6.2
通讯作者：
VASIL, IK
中科院分区：
文献类型：
--
作者：
CHOWDHURY, MKU;VASIL, IK
以甘蔗杂交种CP 72 -1210为材料，采用高速基因枪轰击悬浮培养细胞，原生质体电穿孔，获得了稳定的转化愈伤组织。用包被有pBARGUS质粒DNA的金颗粒轰击三天龄的细胞悬浮培养物（SC 88），所述质粒DNA含有β-葡糖醛酸酶（GUS）报告基因和赋予除草剂basta抗性的bar选择基因。pBARGUS质粒也电穿孔到另一个细胞系（SCPP）的原生质体中。从两个来源中均回收到对basta具有抗性的菌落。通过Southern分析证实了bar基因在抗性细胞系中的稳定整合。此外，膦丝菌素乙酰转移酶（PAT）的活性也被证明在转化的细胞系。
Stably transformed callus of a hybrid sugarcane cultivar (Saccharum species hybrid, CP72-1210) was achieved following high velocity microprojectile bombardment of suspension culture cells, and electroporation of protoplasts. A three-day old cell suspension culture (SC88) was bombarded with gold particles coated with pBARGUS plasmid DNA containing the beta-glucuronidase (GUS) reporter gene and the bar selectable gene that confers resistance to the herbicide basta. The pBARGUS plasmid was also electroporated into the protoplasts of another cell line (SCPP). Colonies resistant to basta were recovered from both sources. Stable integration of the bar gene in the resistant cell lines was confirmed by Southern analysis. In addition, phosphinothricin acetyltransferase (PAT) activity was also demonstrated in the transformed cell lines.