In Vitro Binding of Agrobacterium tumefaciens to Plant Cells from Suspension Culture.
In Vitro Binding of Agrobacterium tumefaciens to Plant Cells from Suspension Culture.
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根癌农杆菌与悬浮培养植物细胞的体外结合。
DOI：
10.1104/pp.63.2.382
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发表时间：
1979
期刊：
影响因子：
7.4
通讯作者：
L. Fowke
中科院分区：
文献类型：
--
作者：
Kanji Ohyama;L. Pelcher;Angelika Schaefer;L. Fowke
采用毒力强的农杆菌B6菌株（32）p标记细胞与悬浮培养的曼陀罗细胞进行体外结合实验。结合动力学表明，细菌对曼陀罗细胞的粘附在前60分钟逐渐增加，并在120分钟内达到最大水平。pH为6.0时结合最大。Ca（2+）和Mg（2+）的存在略微降低了结合，EDTA在0.1 ~ 10毫摩尔浓度下几乎没有影响。细菌与曼陀罗细胞的结合是温度依赖性的。大肠杆菌、鼠伤寒沙门氏菌、日本根瘤菌和溶嗜微球菌在与曼陀罗细胞结合方面与强毒的瘤胃杆菌B6没有竞争关系。无毒株IIBNV6的混合物增强了有毒株B6对曼陀罗细胞的粘附性。章鱼碱对致病菌B6与曼陀罗细胞的结合没有影响，而10毫摩尔的菜豆碱则有抑制作用。当精氨酸浓度高于0.1毫摩尔时，细菌的粘附能力增强。与DNase、RNase和脂肪酶孵育不影响结合，但蛋白酶刺激细菌粘附到曼陀罗细胞。豆毒苷A和大豆凝集素对其结合作用不大，卵磷脂和溶卵磷脂对其结合作用略有增强。聚赖氨酸显著促进了细菌与植物细胞的粘附。豌豆、豌豆和大豆悬浮培养细胞的结合能力比曼陀罗细胞高2- 3倍，而小麦、玉米、水稻和高粱细胞与毒力强的瘤胃分枝杆菌B6的结合能力要低得多。通过放射自显影和电子显微镜证实了细菌对植物细胞的粘附。放射自显像分析显示细菌与细胞壁相关，并且细菌的结合通常是局部的。电子显微照片清楚地表明，毒力强的结核菌菌株B6细胞与曼陀罗细胞壁紧密相关。
In vitro binding experiments were carried out using (32)P-labeled cells of the virulent Agrobacterium tumefaciens strain B6 and Datura innoxia cells from suspension culture. Binding kinetics showed that adherence of bacteria to Datura cells increased gradually during the first 60 minutes and attained a maximum level within 120 minutes of incubation. Maximum binding occurred at pH 6.0. The presence of Ca(2+) and Mg(2+) reduced binding slightly and EDTA had little effect at concentrations of 0.1 to 10 millimolar. The binding of bacteria to Datura cells was temperature-dependent. Escherichia coli, Salmonella typhimurium, Rhizobium japonicum, and Micrococcus lysodeikticus did not compete with virulent A. tumefaciens strain B6 for binding to Datura cells. The admixture of avirulent A. tumefaciens strain IIBNV6 enhanced adherence of virulent A. tumefaciens strain B6 to Datura cells. Octopine had no effect on the binding of virulent A. tumefaciens strain B6 to Datura cells, but 10 millimolar canavanine was inhibitory. Arginine enhanced the adherence of the bacteria at concentrations higher than 0.1 millimolar. Incubation with DNase, RNase, and lipase did not affect the binding, but protease stimulated the adherence of bacteria to Datura cells. Concanavaline A and soybean lectin had little effect whereas lecithin and lysolecithin enhanced binding slightly. Poly-l-lysine markedly stimulated the bacteria-plant cell adherence. Cells from suspension cultures of pea, vetch, and soybean had a 2- to 3-fold higher binding capacity than Datura cells, whereas cells from wheat, corn, rice, and sorghum had a considerably lower affinity for binding with virulent A. tumefaciens strain B6. Bacterial adherence to plant cells was confirmed by autoradiography and electron microscopy. Autoradiographic analysis showed that bacteria were associated with the cell wall, and that often binding of bacteria was localized. Electron micrographs clearly illustrated a tight association of virulent A. tumefaciens strain B6 cells to the Datura cell wall.