Balestrini Raffaella, Gómez-Ariza Jorge, Lanfranco Luisa, Bonfante Paola
Istituto Protezione Piante, CNR and Dipartimento di Biologia Vegetale, Università di Torino, Viale Mattioli, 25-10125 Torino, Italy.
Mol Plant Microbe Interact. 2007 Sep;20(9):1055-62. doi: 10.1094/MPMI-20-9-1055.
The establishment of a symbiotic interaction between plant roots and arbuscular mycorrhizal (AM) fungi requires both partners to undergo significant morphological and physiological modifications which eventually lead to reciprocal beneficial effects. Extensive changes in gene expression profiles recently have been described in transcriptomic studies that have analyzed the whole mycorrhizal root. However, because root colonization by AM fungi involves different cell types, a cell-specific gene expression pattern is likely to occur. We have applied the laser microdissection (LMD) technology to investigate expression profiles of both plant and fungal genes in Lycopersicon esculentum roots colonized by Glomus mosseae. A protocol to harvest arbuscule-containing cells from paraffin sections of mycorrhizal roots has been developed using a Leica AS LMD system. RNA of satisfactory quantity and quality has been extracted for molecular analysis. Transcripts for plant phosphate transporters (LePTs), selected as molecular markers for a functional symbiosis, have been detected by reverse-transcriptase polymerase chain reaction assays and associated to distinct cell types, leading to novel insights into the distribution of LePT mRNAs. In fact, the transcripts of the five phosphate transporters (PTs) have been detected contemporaneously in the same arbusculated cell population, unlike from the neighboring noncolonized cells. In addition, fungal H(+)ATPase (GmHA5) and phosphate transporter (GmosPT) mRNAs were found exclusively in arbusculated cells. The discovery that five plant and one fungal PT genes are consistently expressed inside the arbusculated cells provides a new scenario for plant-fungus nutrient exchanges.
植物根系与丛枝菌根(AM)真菌之间建立共生相互作用需要双方都经历显著的形态和生理变化,最终产生互惠互利的效果。最近在分析整个菌根根系的转录组学研究中描述了基因表达谱的广泛变化。然而,由于AM真菌对根系的定殖涉及不同的细胞类型,可能会出现细胞特异性的基因表达模式。我们应用激光显微切割(LMD)技术来研究被摩西球囊霉定殖的番茄根系中植物和真菌基因的表达谱。利用徕卡AS LMD系统开发了一种从菌根根系石蜡切片中收获含丛枝细胞的方案。已提取出数量和质量令人满意的RNA用于分子分析。通过逆转录聚合酶链反应检测了被选为功能共生分子标记的植物磷酸盐转运蛋白(LePTs)的转录本,并将其与不同的细胞类型相关联,从而对LePT mRNA的分布有了新的认识。事实上,与相邻的未被定殖的细胞不同,在同一批含丛枝的细胞群体中同时检测到了五种磷酸盐转运蛋白(PTs)的转录本。此外,真菌H(+)ATP酶(GmHA5)和磷酸盐转运蛋白(GmosPT)的mRNA仅在含丛枝的细胞中被发现。在含丛枝的细胞中一致表达五个植物PT基因和一个真菌PT基因这一发现为植物 - 真菌营养交换提供了一个新的情况。