Lanfranco Luisa, Novero Mara, Bonfante Paola
Dipartimento di Biologia Vegetale, Università di Torino, 10125 Turin, Italy.
Plant Physiol. 2005 Apr;137(4):1319-30. doi: 10.1104/pp.104.050435. Epub 2005 Mar 4.
A full-length cDNA showing high similarity to previously described CuZn superoxide dismutases (SODs) was identified in an expressed sequence tag collection from germinated spores of the arbuscular mycorrhizal fungus Gigaspora margarita (BEG 34). The corresponding gene sequence, named GmarCuZnSOD, is composed of four exons. As revealed by heterologous complementation assays in a yeast mutant, GmarCuZnSOD encodes a functional polypeptide able to confer increased tolerance to oxidative stress. The GmarCuZnSOD RNA was differentially expressed during the fungal life cycle; highest transcript levels were found in fungal structures inside the roots as observed on two host plants, Lotus japonicus and Medicago truncatula. These structures also reacted positively to 3,3'-diaminobenzidine, used to localize H2O2 accumulation. This H2O2 is likely to be produced by CuZnSOD activity since treatment with a chelator of copper ions, generally used to inhibit CuZnSODs, strongly reduced the 3,3'-diaminobenzidine deposits. A slight induction of GmarCuZnSOD gene expression was also observed in germinated spores exposed to L. japonicus root exudates, although the response showed variation in independent samples. These results provide evidence of the occurrence, in an arbuscular mycorrhizal fungus, of a functional SOD gene that is modulated during the life cycle and may offer protection as a reactive oxygen species-inactivating system against localized host defense responses raised in arbuscule-containing cells.
在丛枝菌根真菌珠状巨孢囊霉(BEG 34)萌发孢子的表达序列标签文库中,鉴定出一个与先前描述的铜锌超氧化物歧化酶(SOD)高度相似的全长cDNA。相应的基因序列命名为GmarCuZnSOD,由四个外显子组成。通过酵母突变体中的异源互补试验表明,GmarCuZnSOD编码一种功能性多肽,能够赋予对氧化应激的耐受性增强。GmarCuZnSOD RNA在真菌生命周期中差异表达;在两种宿主植物百脉根和蒺藜苜蓿的根内真菌结构中发现转录水平最高。这些结构对用于定位H2O2积累的3,3'-二氨基联苯胺也呈阳性反应。这种H2O2可能是由铜锌SOD活性产生的,因为用通常用于抑制铜锌SOD的铜离子螯合剂处理后,3,3'-二氨基联苯胺沉积物显著减少。在暴露于百脉根根分泌物的萌发孢子中也观察到GmarCuZnSOD基因表达的轻微诱导,尽管在独立样本中反应存在差异。这些结果提供了证据,证明在丛枝菌根真菌中存在一个功能性SOD基因,该基因在生命周期中受到调控,并可能作为一种活性氧灭活系统,对含丛枝细胞中引发的局部宿主防御反应提供保护。