埃及小麦的遗传转化几丁质酶基因通过微弹轰击

A. Fahmy, K. E. Mangoury, Walaa Abou El-Wafa, Hoda M. S. Barakat, S. E. El-Khodary, S. Muthukrishnan
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引用次数: 2

摘要

植物病害是由包括真菌在内的多种植物病原体引起的,其管理需要使用转基因技术和遗传学等技术。几丁质酶基因,已知在真菌疾病防御中起重要作用,被引入埃及小麦cv。吉萨164通过生物轰炸。未成熟胚愈伤组织在含双磷的CIMB培养基上继代培养。6周后,将存活的胚性愈伤组织置于MSRB再生选择培养基上。最后将获得的绿色再生芽转移到FMSB选根培养基上。两周后,在生根选择培养基下建立健康的生根苗,然后转移到土盆中,然后在对照生长室内培养适应,然后转移到温室直到结籽。通过PCR和点印迹分析证实,该基因的整合率为1.8%。
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GENETIC TRANSFORMATION OF EGYPTIAN WHEAT (Triticum aestivum L.) WITH CHITINASE GENE VIA MICROPROJECTILE BOMBARDMENT
Plant diseases are caused by a variety of plant pathogens including fungi, and their management requires the use of techniques like transgenic technology and genetics. The chitinase gene, known to have a vital role in fungal disease defense, was introduced into the Egyptian wheat cv. Giza 164 via biolistic bombardment. Immature embryo derived calli were sub-cultured on CIMB medium containing bialaphos. After six weeks, the survived embryogenic calli were placed on MSRB regeneration selection medium. Finally, the green regenerated shoots produced were then transferred onto FMSB rooting selection medium. After the two weeks, healthy rooted plantlets were established under the rooting selection medium, and were then transferred into soil pots, then incubated in control growth chamber for acclimatization and subsequently trans-ferred to greenhouse until seed setting. In-tegration of the transgene with a transfor-mation frequency of 1.8% was confirmed by PCR and dot-blot analyses.
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