Wang Yun-Peng, Wei Zheng-Yi, Zhang Yu-Ying, Lin Chun-Jing, Zhong Xiao-Fang, Wang Yue-Lin, Ma Jing-Yong, Ma Jian, Xing Shao-Chen
Faculty of Agronomy, Jilin Agricultural University, No. 2888 Xincheng Street, Changchun 130118, China.
Agro-Biotechnology Research Institute, Jilin Academy of Agricultural Sciences, No. 1363 Shengtai Street, Changchun 130033, China.
Int J Mol Sci. 2015 Mar 2;16(3):4628-41. doi: 10.3390/ijms16034628.
Rice blast is a major destructive fungal disease that poses a serious threat to rice production and the improvement of blast resistance is critical to rice breeding. The antimicrobial peptide MSI-99 has been suggested as an antimicrobial peptide conferring resistance to bacterial and fungal diseases. Here, a vector harboring the MSI-99 gene was constructed and introduced into the tobacco chloroplast genome via particle bombardment. Transformed plants were obtained and verified to be homoplastomic by PCR and Southern hybridization. In planta assays demonstrated that the transgenic tobacco plants displayed an enhanced resistance to the fungal disease. The evaluation of the antimicrobial activity revealed that the crude protein extracts from the transgenic plants manifested an antimicrobial activity against E. coli, even after incubation at 120 °C for 20 min, indicating significant heat stability of MSI-99. More importantly, the MSI-99-containing protein extracts were firstly proved in vitro and in vivo to display significant suppressive effects on two rice blast isolates. These findings provide a strong basis for the development of new biopesticides to combat rice blast.
稻瘟病是一种主要的毁灭性真菌病害，对水稻生产构成严重威胁，提高稻瘟病抗性对水稻育种至关重要。抗菌肽MSI-99被认为是一种能赋予对细菌和真菌病害抗性的抗菌肽。在此，构建了一个携带MSI-99基因的载体，并通过粒子轰击将其导入烟草叶绿体基因组。获得了转化植株，并通过PCR和Southern杂交验证为同质性叶绿体转化体。在植物体内试验表明，转基因烟草植株对真菌病害表现出增强的抗性。抗菌活性评估显示，转基因植株的粗蛋白提取物即使在120℃孵育20分钟后仍对大肠杆菌表现出抗菌活性，表明MSI-99具有显著的热稳定性。更重要的是，首次在体外和体内证明含MSI-99的蛋白提取物对两种稻瘟病菌株具有显著的抑制作用。这些发现为开发防治稻瘟病的新型生物农药提供了有力依据。
