TRANSGENIC EGYPTIAN WHEAT (Triticum aestivum L.) WITH Arabidopsis NPR1 GENE via BIOLISTIC BOMBARDMENT

A. Fahmy, K. E. Mangoury, Walaa Abou El-Wafa, Hoda M. S. Barakat, S. E. El-Khodary, J. Li, S. Muthukrishnan
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Abstract

AtNPR1 gene plays a crucial role in biotic resistance in plants. Immature embryo derived calli of wheat cv. Giza 164 were co-transformed with the plasmid pAHC25 containing bar and gus genes and the plasmid pJS406 containing AtNPR1 gene via Biolistic bombardment. The produced calli were subcultured on bialaphos containing medium (CIMB). Surviving calli were then regenerated on selection medium (MSRB) to produce green regenerated shoots, which were then transferred onto rooting/selection medium (FMSB). Regenerated rooted plantlets were transferred to soil pots in the control growth chamber for acclimatization. Only 48 plants succeeded in acclimatization and reached seed setting in biocontainment greenhouse. Transgene integration was confirmed by PCR and dot-blot analyses. The overall transformation frequency was 1.9%.
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转基因埃及小麦(Triticum aestium L.)拟南芥NPR1基因的生物轰击
AtNPR1基因在植物生物抗性中起着至关重要的作用。小麦愈伤组织的未成熟胚。采用生物轰击法将Giza 164与含有bar和gus基因的质粒pAHC25和含有AtNPR1基因的质粒pJS406共转化。将产生的愈伤组织在含双磷培养基(CIMB)上传代培养。将存活愈伤组织在选择培养基(MSRB)上再生成绿色再生芽,再将其转移到生根/选择培养基(FMSB)上。将再生的生根苗移栽到对照生长室内的土盆中进行驯化。在生物防护温室中,只有48株驯化成功并达到结实率。通过PCR和点印迹分析证实了基因整合。总体转换频率为1.9%。
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