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Chloroplast-Expressed MSI-99 in Tobacco Improves Disease Resistance and Displays Inhibitory Effect against Rice Blast Fungus
Authors:Yun-Peng Wang  Zheng-Yi Wei  Yu-Ying Zhang  Chun-Jing Lin  Xiao-Fang Zhong  Yue-Lin Wang  Jing-Yong Ma  Jian Ma  Shao-Chen Xing
Affiliation:1.Faculty of Agronomy, Jilin Agricultural University, No. 2888 Xincheng Street, Changchun 130118, China; E-Mails: (Y.-P.W.); (J.-Y.M.);2.Agro-Biotechnology Research Institute, Jilin Academy of Agricultural Sciences, No. 1363 Shengtai Street, Changchun 130033, China; E-Mails: (Z.-Y.W.); (Y.-Y.Z.); (C.-J.L.); (X.-F.Z.); (Y.-L.W.);3.College of Biological Sciences, China Agricultural University, No. 2 West Yuanmingyuan Road, Beijing 100094, China
Abstract: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.
Keywords:antimicrobial peptide  chloroplast transformation  rice blast  tobacco
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