Ezawa Tatsuhiro, Hayatsu Masahito, Saito Masanori
Graduate School of Agriculture, Hokkaido University, Sapporo, Japan.
Mol Plant Microbe Interact. 2005 Oct;18(10):1046-53. doi: 10.1094/MPMI-18-1046.
The mycorrhiza-responsive phosphatase of Tagetes patula in symbiosis with Glomus etunicatum was detected by electrophoresis, was purified by column chromatography, and was characterized as acid phosphatase that was secreted into rhizosphere. The N-terminal amino acid sequence was determined by a gas-phase sequencer, and a cDNA fragment of the phosphatase gene (TpPAP1) was amplified by degenerate primers designed based on the N-terminal amino acid sequence. The full-length cDNA was obtained by the rapid amplification of cDNA ends technique. The TpPAP1 was of host origin, and the cDNA was 1,843 bp long with a predicted open reading frame of polypeptide of 466 amino acids. Phylogenetic analysis revealed that the gene fell into the cluster of plant high-molecular-weight purple acid phosphatase. Expression analysis of the TpPAP1 in T. patula in symbiosis with Archaeospora leptoticha showed that the levels of transcripts increased eightfold by mycorrhizal colonization. Western blot analysis revealed that the 57-kDa protein corresponding to the mycorrhiza-responsive phosphatase increased by mycorrhizal colonization. The present study proposes a new strategy for acquisition of P in arbuscular mycorrhizal associations in which the fungal partner activates a part of the low-P adaptation system of the plant partner, phosphatase secretion, and improves the overall efficiency of P uptake.
通过电泳检测了与幼套球囊霉共生的孔雀草中菌根响应性磷酸酶,经柱色谱法纯化,并鉴定为分泌到根际的酸性磷酸酶。通过气相测序仪测定了其N端氨基酸序列,并基于该N端氨基酸序列设计简并引物,扩增出磷酸酶基因(TpPAP1)的cDNA片段。通过cDNA末端快速扩增技术获得了全长cDNA。TpPAP1来自宿主,cDNA长1843 bp,预测的开放阅读框编码466个氨基酸的多肽。系统发育分析表明,该基因属于植物高分子量紫色酸性磷酸酶簇。对与细凹无梗囊霉共生的孔雀草中TpPAP1的表达分析表明,菌根定殖使转录本水平提高了八倍。蛋白质免疫印迹分析表明,与菌根响应性磷酸酶对应的57 kDa蛋白因菌根定殖而增加。本研究提出了一种在丛枝菌根共生关系中获取磷的新策略,即真菌伙伴激活植物伙伴低磷适应系统的一部分——磷酸酶分泌,从而提高磷吸收的整体效率。