小麦赤霉病穗组织中亚洲镰刀菌定量分析及其在品种(系)抗性评价中的应用

Real-time PCR Quantification of Fusarium asiaticumin Inoculated Head Tissues of Wheat and Its Application Value on Evaluation of Cultivar(Line)Resistance

为探明小麦穗中亚洲镰刀菌Fusarium asiaticum积累菌量与品种赤霉病抗性的关系,并探讨该菌定量分析方法在品种抗性评价中的应用价值,用TaqMan探针实时荧光定量PCR法对接种5d和10d后的9个小麦品种(系)穗组织中的F.asiaticum DNA进行测定,并与相应品种(系)田间抗性调查结果进行比较。结果表明,在F.asiaticum DNA定量检测中,Taq Man探针实时PCR技术具有高度的稳定性和可重复性。小麦品种间F.asiaticumDNA含量差异显著,接种10d后比接种5d后的DNA检测结果更能真实地反映不同品种(系)之间抗性水平的差异。在田间表现中抗水平的品种(系)之间接种10d后F.asiaticumDNA含量也有显著差异。与田间鉴定结果比较,定量分析具有较好的区分作用。

To elucidate the relationship between the accumulation of Fusarium asiaticurn in inoculated head tissues and the resistance of wheat cultivars, also to investigate the application value of quantifi cation analysis in evaluating the resistance of wheat cultivar, real-time quantitative polymerase chain reaction （RT-Q-PCR） based on Taq Man technology was used for the quantification of F. asiaticum DNA in the head tissues of 9 wheat varieties at 5 d and 10 d after fungal inoculation. TaqMan-based RT-Q-PCR assay was found to be highly reliable and reproducible in quantifying the F. asiaticurn DNA. The amount of F. asiaticum DNA varied in different wheat varieties, quantification data at 10 d after inoculation （dai） was better than that at 5 dai in reflecting the cultivarrs resistance. The amount of F. asiaticurn DNA varied in wheat varieties which were proved to be the same middle resistance lev- el in the field. Compared with the resistance assessment in the field, quantification analysis was betterin the discrimination of resistance level.