CSB353H1 Lecture Notes - Lecture 6: Bacterial Effector Protein, Flagellin, Haustorium
28 Apr 2016
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Culture host plant cells in liquid media, add pathogen. When cell recognize pathogen, start to activate pti. Produce ros, and release k+ into media, which becomes alkaline, making ph increase. Can use ph change as indicator of recognition of pathogen. When boil bacteria, see stronger alkalization epitope was more exposed by denaturing by heat. The activity of heat-treated crude preparations was sensitive to treatment with trypsin or proteinease, indicating that elicitor was protein. There"s a conserved 22 aa sequence important for flagellin function; this motif triggered pti. Genetic approach find mutant host that can"t recognize this epitope, see what gene was mutated (forward genetics) When added to plant, plant growth was arrested after pathogen induction shows defense system is activated. Experiment: if treat flg22 (which induces pti) pathogen has much less growth compared to wildtype. Experiment: used fsl2 ko plant, add flg22, plant is insensitive one day before infection (pretreat),
