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通过生物射弹轰击法转化三倍体狗牙根(Cynodon dactylon x C. transvaalensis cv. TifEagle)

Transformation of triploid bermudagrass (Cynodon dactylon x C. transvaalensis cv. TifEagle) by means of biolistic bombardment.

作者信息

Zhang G, Lu S, Chen T A, Funk C R, Meyer W A

机构信息

Department of Plant Biology and Pathology, Rutgers University, New Brunswick, NJ 08901, USA.

出版信息

Plant Cell Rep. 2003 Jun;21(9):860-4. doi: 10.1007/s00299-003-0585-x. Epub 2003 Mar 27.

Abstract

A transformation system for triploid bermudagrass ( Cynodon dactylon x C. transvaalensis cv. TifEagle) was established with a biolistic bombardment delivery system. Embryogenic callus was induced from stolons and maintained on Murashige and Skoog's medium supplemented with 30 microM dicamba, 20 microM benzylaminopurine, and 100 mg/l myo-inositol. Using the hygromycin phosphotransferase ( hpt) gene as the selectable marker gene, we obtained 75 transgenic lines from 18 petri dishes bombarded. Integration of the hpt gene into genomic DNA and transcription of hpt was confirmed by Southern and Northern blot analyses, respectively. Through suspension culture screening, we obtained homogeneously transformed plants showing stable transcription of the hpt gene.

摘要

利用生物枪轰击递送系统建立了三倍体狗牙根(Cynodon dactylon x C. transvaalensis cv. TifEagle)的转化体系。从匍匐茎诱导出胚性愈伤组织,并在添加了30微摩尔麦草畏、20微摩尔苄氨基嘌呤和100毫克/升肌醇的Murashige和Skoog培养基上进行培养。以潮霉素磷酸转移酶(hpt)基因作为选择标记基因,我们从18个经轰击的培养皿中获得了75个转基因株系。分别通过Southern杂交和Northern杂交分析证实了hpt基因整合到基因组DNA中以及hpt的转录。通过悬浮培养筛选,我们获得了显示hpt基因稳定转录的均匀转化植株。

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