Guether Mike, Neuhäuser Benjamin, Balestrini Raffaella, Dynowski Marek, Ludewig Uwe, Bonfante Paola
Department of Plant Biology, University of Torino and Istituto per la Protezione delle Piante/Consiglio Nazionale delle Ricerche, 10125 Torino, Italy.
Plant Physiol. 2009 May;150(1):73-83. doi: 10.1104/pp.109.136390. Epub 2009 Mar 27.
In mycorrhizal associations, the fungal partner assists its plant host by providing nitrogen (N) in addition to phosphate. Arbuscular mycorrhizal (AM) fungi have access to inorganic or organic forms of N and translocate them via arginine from the extra- to the intraradical mycelium, where the N is transferred to the plant without any carbon skeleton. However, the molecular form in which N is transferred, as well as the involved mechanisms, is still under debate. NH(4)(+) seems to be the preferential transferred molecule, but no plant ammonium transporter (AMT) has been identified so far. Here, we offer evidence of a plant AMT that is involved in N uptake during mycorrhiza symbiosis. The gene LjAMT2;2, which has been shown to be the highest up-regulated gene in a transcriptomic analysis of Lotus japonicus roots upon colonization with Gigaspora margarita, has been characterized as a high-affinity AMT belonging to the AMT2 subfamily. It is exclusively expressed in the mycorrhizal roots, but not in the nodules, and transcripts have preferentially been located in the arbusculated cells. Yeast (Saccharomyces cerevisiae) mutant complementation has confirmed its functionality and revealed its dependency on acidic pH. The transport experiments using Xenopus laevis oocytes indicated that, unlike other plant AMTs, LjAMT2;2 transports NH(3) instead of NH(4)(+). Our results suggest that the transporter binds charged ammonium in the apoplastic interfacial compartment and releases the uncharged NH(3) into the plant cytoplasm. The implications of such a finding are discussed in the context of AM functioning and plant phosphorus uptake.
在菌根共生关系中,真菌伙伴除了提供磷酸盐外,还通过提供氮(N)来协助其植物宿主。丛枝菌根(AM)真菌能够获取无机或有机形式的氮,并通过精氨酸将其从根外菌丝转运到根内菌丝,在根内菌丝中,氮在没有任何碳骨架的情况下转移到植物中。然而,氮转移的分子形式以及相关机制仍存在争议。NH₄⁺似乎是优先转移的分子,但到目前为止尚未鉴定出植物铵转运蛋白(AMT)。在这里,我们提供了一种植物AMT参与菌根共生期间氮吸收的证据。基因LjAMT2;2在转录组分析中显示,在用珠状巨孢囊霉侵染后,它是百脉根根中上调程度最高的基因,已被鉴定为属于AMT2亚家族的高亲和力AMT。它仅在菌根根中表达,而不在根瘤中表达,并且转录本优先位于丛枝细胞中。酵母(酿酒酵母)突变体互补实验证实了其功能,并揭示了其对酸性pH的依赖性。使用非洲爪蟾卵母细胞进行的转运实验表明,与其他植物AMT不同,LjAMT2;2转运NH₃而不是NH₄⁺。我们的结果表明,该转运蛋白在质外体界面区结合带电荷的铵,并将不带电荷的NH₃释放到植物细胞质中。在AM功能和植物磷吸收的背景下讨论了这一发现的意义。