Preparation Of Monoclonal Atibody Against T-ZR And Analysis Of Plant Hormones Involved In The Early-stage Interactions Between Wheat And Powdery Mildew

The outcome of successful infections of plants by pathogens often leads to severe outbreaks of disease symptoms such as lesion spots, etiolation, senescence and abscission. The involvement of plant hormones in the interactions between plants and microorganisms, which has been confirmed by the studies from Molecular Pathology and Physiology, is one of the key mechanisms adopted by plants for perception and signal ing of pathogens to cause the final incompatible or compatible responses.For the purpose of performing the assay of CTK in this study, firstly the monoclonal antibody against t-ZR was prepared. The antibody showed high specificity to t-ZR but relalively low cross-readivity with some compounds structurally similar to t-ZR. Then, based on this antibody, we established a direct enzyme-linked immunosorbent assay (ELISA) with high specificity, sensitivity and precision for quantification of t-ZR in plant tissues. The detect limit and linear detect ranges are 6. 3fmol and 0.051-12.5 pmol/50 ul. The correlation coefficient of the standard curve is more than 0.99. The coefficient of variation within plates is less than 2. 47% and at between plates is less than 3.4 %.After being stored in liquid nitrogen for 5 months, the re-cultured hybrid cell t-ZR4-D1 showed the capability of secreting t-ZR McAb steadily.The inoculation of whea! (Su 3) with the spores of powdery mildew caused the formation green islands in the leaves of wheat 8d in dark. This phenomenon has been a typical model for the study of the interactions between wheat and powdery mildew in the pathological and physiological fields.In this study, the roles of pant hormones were investigated by analysisof the fluctuations of hormones in the leaves at the early stage of the infection. The contents of t-ZR and iPAs in the infected leaves were increased obviously. After 2-d of inoeuhit ion, t~ZR reached a peak of 1154.4pmol/g. FW, about 5 times higher than that in healthy leaves. The sum of t-ZR and iPAs was maintained higher level in 72h. So CTKs should play an important role in the early stage of infection. The increasing CTK level in the host might aid the growth and proliferation of the pathogen or help the defense of the host against the pathogen.The content of IAA showed little affected in the leaves by the attack ofthe pathogen, which suggested that auxin may not be involved in the early interactions between wheat loaves and the powdery mildew.