van Aarle Ingrid M, Viennois Gaëlle, Amenc Laurie K, Tatry Marie-Violaine, Luu Doan T, Plassard Claude
Biogéochimie du Sol et de la Rhizosphère, UMR 1222 INRA/SupAgro, 2 Place Viala, 34060, Montpellier Cedex 1, France.
Biochimie et Physiologie Moléculaire des Plantes, UMR 5004 INRA/CNRS/SupAgro/UM II, 2 Place Viala, 34060, Montpellier Cedex 1, France.
Mycorrhiza. 2007 Sep;17(6):487-494. doi: 10.1007/s00572-007-0127-4. Epub 2007 May 23.
Expression of a mycorrhizal fungal-specific phosphate (P) transporter gene (HcPT1) was studied in mycelium of the ectomycorrhizal fungus Hebeloma cylindrosporum, by in situ reverse transcriptase polymerase chain reaction using amplification of complementary DNA sequences. The expression of HcPT1 was visualised under two different P treatments. Mycelium was transferred to liquid medium with or without P and incubated for 5 days. Under P starvation, mycelium growth and vitality was reduced and the expression of HcPT1 up regulated. Enzyme-labelled fluorescent substrate was used to detect gene expression in situ with epi-fluorescence microscopy and to visualise it at the level of the individual hyphae both in starved and non-starved hyphae. Up-regulation of HcPT1 was observed as a more intense fluorescent signal and from the larger proportion of hyphae that showed expression.
通过使用互补DNA序列扩增的原位逆转录聚合酶链反应，研究了外生菌根真菌圆柱齿杯菌菌丝体中菌根真菌特异性磷酸盐（P）转运蛋白基因（HcPT1）的表达。在两种不同的磷处理下观察HcPT1的表达情况。将菌丝体转移至含磷或不含磷的液体培养基中培养5天。在磷饥饿条件下，菌丝体生长和活力降低，HcPT1的表达上调。使用酶标记的荧光底物，通过落射荧光显微镜原位检测基因表达，并在饥饿和未饥饿菌丝体的单个菌丝水平上观察其表达情况。观察到HcPT1的上调表现为更强的荧光信号以及显示表达的菌丝比例更高。
