State Key Laboratory of Eco-environment and Bio-resource of Three Gorges Reservoir Region, Key Laboratory of Eco-environments of Three Gorges Reservoir Region, Ministry of Education, Institute of Resources Botany, School of Life Sciences, Southwest University, No. 1 Tiansheng Road, Beibei, Chongqing 400715, China.
Tree Physiol. 2010 Dec;30(12):1599-605. doi: 10.1093/treephys/tpq093. Epub 2010 Nov 17.
The antimicrobial protein gene LJAMP2 is a plant non-specific lipid transfer protein from motherwort (Leonurus japonicus). In this study, it was introduced into Chinese white poplar (Populus tomentosa Carr.) via Agrobacterium-mediated transformation with neomycin phosphotransferase II gene conferring kanamycin resistance as selectable marker. A total of 16 poplar lines were obtained, and polymerase chain reaction (PCR) analysis established the stable integration of transgenes in the plant genome. Reverse transcription-PCR detected LJAMP2 expression in transgenic plants. Resistance to fungal pathogens Alternaria alternata (Fr.) Keissler and Colletotrichum gloeosporioides (Penz.) of transgenic poplar lines was tested. In vitro inhibitory activity against the fungal pathogens was evident from the crude leaf extracts from the transformants. In vivo assays showed that, after infection with both A. alternata (Fr.) Keissler and C. gloeosporioides (Penz.), there was a significant reduction in disease symptoms in transgenic poplar plants compared with the control. These results suggest that constitutive expression of the LJAMP2 gene from motherwort can be exploited to improve resistance to fungal pathogens in poplar.
抗菌蛋白基因 LJAMP2 来自益母草(Leonurus japonicus),是一种植物非特异性脂质转移蛋白。本研究采用根癌农杆菌介导法将其转入毛白杨(Populus tomentosa Carr.),以赋予卡那霉素抗性的新霉素磷酸转移酶 II 基因为选择标记。共获得 16 株杨树株系,通过聚合酶链反应(PCR)分析证实外源基因已稳定整合到植物基因组中。反转录 PCR 检测到转基因植物中 LJAMP2 的表达。对转基因杨树株系进行了真菌病原菌Alternaria alternata(Fr.)Keissler 和 Colletotrichum gloeosporioides(Penz.)的抗性测试。从转化体的粗叶提取物中明显看出其对真菌病原菌的体外抑制活性。体内试验表明,与对照相比,在感染 A. alternata(Fr.)Keissler 和 C. gloeosporioides(Penz.)后,转基因杨树植株的病症明显减轻。这些结果表明,益母草 LJAMP2 基因的组成型表达可用于提高杨树对真菌病原菌的抗性。