Turrini A, Sbrana C, Pitto L, Ruffini Castiglione M, Giorgetti L, Briganti R, Bracci T, Evangelista M, Nuti M P, Giovannetti M
Dipartimento di Chimica e Biotecnologie Agrarie, Università di Pisa, via del Borghetto 80, I-56124 Pisa, Italy.
Istituto di Biologia e Biotecnologia Agraria, CNR, Sezione di Pisa, via del Borghetto 80, I-56124 Pisa, Italy.
New Phytol. 2004 Aug;163(2):393-403. doi: 10.1111/j.1469-8137.2004.01107.x.
• Transformed aubergine plants constitutively expressing the Dm-AMP1 antimicrobial defensin (from Dahlia merckii) were generated and characterized. • Transgenic plants were selected on kanamycin and screened by polymerase chain reaction analysis. The expression of Dm-AMP1 in plant tissues and its release in root exudates were detected by Western blot analyses. Dm-AMP1 localization was performed by immunohistochemical experiments. • Dm-AMP1 expression ranged from 0.2% to 0.48% of total soluble proteins in primary transformants and from 0.16% to 0.66% in F plants. Transformed clones showed resistance to the pathogenic fungus Botrytis cinerea, whose development on leaves was reduced by 36-100%, with respect to controls. The protein was released in root exudates of the transformed plants and was active in reducing the growth of the co-cultured pathogenic fungus Verticillium albo-atrum, whereas it did not interfere with recognition responses and symbiosis establishment by the arbuscular mycorrhizal fungus Glomus mosseae. • Dm-AMP1 transformants may represent a useful model to study the interactions between genetically modified plants and pathogenic fungi or beneficial nontarget microorganisms.
• 构建并鉴定了组成型表达Dm-AMP1抗菌防御素(来自大丽花)的转基因茄子植株。
• 在卡那霉素上筛选转基因植株,并通过聚合酶链反应分析进行检测。通过蛋白质印迹分析检测植物组织中Dm-AMP1的表达及其在根系分泌物中的释放。通过免疫组织化学实验进行Dm-AMP1定位。
• 在初级转化体中,Dm-AMP1的表达占总可溶性蛋白的0.2%至0.48%,在F植株中为0.16%至0.66%。转化克隆对致病真菌灰葡萄孢表现出抗性,与对照相比,其在叶片上的生长减少了36%至100%。该蛋白在转基因植株的根系分泌物中释放,并能有效抑制共培养的致病真菌黑白轮枝菌的生长,而不干扰丛枝菌根真菌摩西球囊霉的识别反应和共生建立。
• Dm-AMP1转化体可能是研究转基因植物与致病真菌或有益非靶标微生物之间相互作用的有用模型。