University of Fribourg, Department of Biology, Pérolles, 1700 Fribourg, Switzerland.
Mol Plant Pathol. 2003 Jul 1;4(4):237-48. doi: 10.1046/j.1364-3703.2003.00168.x.
SUMMARY Induced resistance was studied in the model pathosystem Arabidopsis-Phytophthora brassicae (formerly P. porri) in comparison with the agronomically important late blight disease of potato caused by Phytophthora infestans. For the quantification of disease progress, both Phytophthora species were transformed with the vector p34GFN carrying the selectable marker gene neomycine phosphotransferase (nptII) and the reporter gene green fluorescent protein (gfp). Eighty five per cent of the transformants of P. brassicae and P. infestans constitutively expressed GFP at high levels at all developmental stages both in vitro and in planta. Transformants with high GFP expression and normal in vitro growth and virulence were selected to quantify pathogen growth by measuring the in planta emitted GFP fluorescence. This non-destructive monitoring of the infection process was applied to analyse the efficacy of two chemical inducers of disease resistance, a functional SA-analogue, benzothiadiazole (BTH), and beta-aminobutyric acid (BABA) which is involved in priming mechanisms of unknown nature. BABA pre-treatment (300 microm) via soil drench applied 24 h before inoculation completely protected the susceptible Arabidopsis accession Landsberg erecta (Ler) from infection with P. brassicae. A similar treatment with BTH (330 microm) did not induce resistance. Spraying the susceptible potato cultivar Bintje with BABA (1 mm) 2 days before inoculation resulted in a phenocopy of the incompatible interaction shown by the resistant potato cultivar Matilda while BTH (1.5 mm) did not protect Bintje from severe infection. Thus, in both pathosystems, the mechanisms of induced resistance appeared to be similar, suggesting that the Arabidopsis-P. brassicae pathosystem is a promising model for the molecular analysis of induced resistance mechanisms of potato against the late blight disease.
摘要 本研究以拟南芥-芸薹根肿菌(原为芸薹根肿菌)模式体系为对象,与由致病疫霉引起的重要农业病害——马铃薯晚疫病进行了比较,研究了诱导抗性。为了量化疾病的进展,我们将携带选择标记基因新霉素磷酸转移酶(nptII)和报告基因绿色荧光蛋白(gfp)的载体 p34GFN 转化为两种芸薹根肿菌和致病疫霉。85%的根肿菌和致病疫霉转化体在体外和体内的所有发育阶段都能高水平、持续表达 GFP。选择 GFP 表达水平高、体外生长和毒性正常的转化体,通过测量体内发出的 GFP 荧光来定量病原体的生长。这种对感染过程的非破坏性监测被应用于分析两种化学诱导抗病剂的效果,一种是功能 SA 类似物——噻二唑(BTH),另一种是β-氨基丁酸(BABA),它涉及到性质不明的启动机制。BABA(300µM)预处理(接种前 24 小时通过土壤浇灌)可完全保护易感拟南芥品系 Landsberg erecta(Ler)免受芸薹根肿菌的感染。类似地,用 BTH(330µM)处理不能诱导抗性。在接种前 2 天用 BABA(1mm)喷洒易感马铃薯品种 Bintje 可导致与抗性马铃薯品种 Matilda 不相容的互作表现出表型模拟,而 BTH(1.5mm)不能保护 Bintje 免受严重感染。因此,在这两个体系中,诱导抗性的机制似乎相似,这表明拟南芥-芸薹根肿菌体系是一个很有前途的模式系统,可用于对马铃薯晚疫病的抗性诱导抗性机制进行分子分析。