Zhang Yan-Min, Liu Zi-Hui, Yang Rui-Juan, Li Guo-Liang, Guo Xiu-Lin, Zhang Hua-Ning, Zhang Hong-Mei, Di Rui, Zhao Qing-Song, Zhang Meng-Chen
Institute of Genetics and Physiology, Plant Genetic Engineering Center of Hebei Province, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang, 050051, China.
Institute of Food and Oil Crops, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang, 050035, China.
Plant Cell Rep. 2016 Jun;35(6):1259-71. doi: 10.1007/s00299-016-1958-2. Epub 2016 Mar 9.
Antagonists and sonication treatment relieved the structural barriers of Agrobacterium entering into cells; hindered signal perception and transmission; alleviated defense responses and increased cell susceptibility to Agrobacterium infection. Soybean gene expression analysis was performed to elucidate the general response of soybean plant to Agrobacterium at an early stage of infection. Agrobacterium infection stimulated the PAMPs-triggered immunity (BRI1, BAK1, BZR1, FLS2 and EFR) and effector-triggered immunity (RPM1, RPS2, RPS5, RIN4, and PBS1); up-regulated the transcript factors (WRKY25, WRKY29, MEKK1P, MKK4/5P and MYC2) in MAPK pathway; strengthened the biosynthesis of flavonoid and isoflavonoid in the second metabolism; finally led to a fierce defense response of soybean to Agrobacterium infection and thereby lower transformation efficiency. To overcome it, antagonist α-aminooxyacetic acid (AOA) and sonication treatment along with Agrobacterium infection were applied. This novel method dramatically decreased the expression of genes coding for F3'H, HCT, β-glucosidase and IF7GT, etc., which are important for isoflavone biosynthesis or the interconversion of aglycones and glycon; genes coding for peroxidase, FLS2, PBS1 and transcription factor MYC2, etc., which are important components in plant-pathogen interaction; and genes coding for GPAT and α-L-fucosidase, which are important in polyesters formation in cell membrane and the degradation of fucose-containing glycoproteins and glycolipids on the external surface of cell membrane, respectively. This analysis implied that AOA and sonication treatment not only relieved the structural membrane barriers of Agrobacterium entering into cells, but also hindered the perception of 'invasion' signal on cell membrane and intercellular signal transmission, thus effectively alleviated the defense responses and increased the cell susceptibility to Agrobacterium infection. All these factors benefit the transformation process; other measures should also be further explored to improve soybean transformation.
拮抗剂和超声处理解除了农杆菌进入细胞的结构障碍；阻碍了信号感知和传递；减轻了防御反应并增加了细胞对农杆菌感染的敏感性。进行了大豆基因表达分析，以阐明大豆植株在感染早期对农杆菌的一般反应。农杆菌感染刺激了模式识别受体触发的免疫反应（BRI1、BAK1、BZR1、FLS2和EFR）和效应子触发的免疫反应（RPM1、RPS2、RPS5、RIN4和PBS1）；上调了丝裂原活化蛋白激酶（MAPK）途径中的转录因子（WRKY25、WRKY29、MEKK1P、MKK4/5P和MYC2）；增强了次生代谢中黄酮类化合物和异黄酮类化合物的生物合成；最终导致大豆对农杆菌感染产生强烈的防御反应，从而降低转化效率。为克服这一问题，将拮抗剂α-氨基氧乙酸（AOA）和超声处理与农杆菌感染一起应用。这种新方法显著降低了编码F3'H、HCT、β-葡萄糖苷酶和IF7GT等基因的表达，这些基因对异黄酮生物合成或糖苷配基与糖苷的相互转化很重要；编码过氧化物酶、FLS2、PBS1和转录因子MYC2等基因的表达，这些基因是植物-病原体相互作用中的重要组成部分；以及编码GPAT和α-L-岩藻糖苷酶的基因的表达，它们分别在细胞膜聚酯形成和细胞膜外表面含岩藻糖糖蛋白和糖脂的降解中起重要作用。该分析表明，AOA和超声处理不仅解除了农杆菌进入细胞的结构膜障碍，还阻碍了细胞膜上“入侵”信号的感知和细胞间信号传递，从而有效减轻了防御反应并增加了细胞对农杆菌感染的敏感性。所有这些因素都有利于转化过程；还应进一步探索其他措施以提高大豆转化效率。
