Roy-Barman Subhankar, Sautter Christof, Chattoo Bharat B
Department of Microbiology and Biotechnology Centre, Faculty of Science, The M. S. University of Baroda, Vadodara 390 002, India.
Transgenic Res. 2006 Aug;15(4):435-46. doi: 10.1007/s11248-006-0016-1.
To enhance fungal disease resistance, wheat plants (cv. Bobwhite) were engineered to constitutively express the potent antimicrobial protein Ace-AMP1 from Allium cepa, driven by a maize ubiquitin promoter along with its first intron. The bar gene was used for selection of putative transformants on medium containing phosphinothricin (PPT). Transgene inheritance, integration and stability of expression were confirmed over two generations by PCR, Southern, northern and western blot analyses, respectively. The levels of Ace-AMP1 in different transgenic lines correlated with the transcript levels of the transgene. Up to 50% increase in resistance to Blumeria graminis f. sp. tritici was detected in detached leaf assays. In ears of transgenic wheat inoculated with Neovossia indica, Ace-AMP1 intensified expression of defense-related genes. Elevated levels of salicylic acid and of transcripts of phenylalanine ammonia lyase (PAL), glucanase (PR2) and chitinase (PR3) in the transgenic plants indicated manifestation of systemic acquired resistance (SAR).
为增强小麦对真菌病害的抗性，对小麦植株（品种：Bobwhite）进行基因工程改造，使其由玉米泛素启动子及其第一个内含子驱动，组成型表达来自洋葱的强效抗菌蛋白Ace-AMP1。bar基因用于在含有草丁膦（PPT）的培养基上筛选推定的转化体。分别通过PCR、Southern、Northern和Western印迹分析在两代中证实了转基因的遗传、整合和表达稳定性。不同转基因株系中Ace-AMP1的水平与转基因的转录水平相关。在离体叶片试验中，检测到对小麦白粉病菌的抗性提高了50%。在接种印度腥黑粉菌的转基因小麦穗中，Ace-AMP1增强了防御相关基因的表达。转基因植物中水杨酸以及苯丙氨酸解氨酶（PAL）、葡聚糖酶（PR2）和几丁质酶（PR3）转录本水平的升高表明系统获得性抗性（SAR）的表现。
