The Effects of Plant Hormones Abscisic Acid, 植物激素脱落酸的影响.pptVIP

The Interactions of Plant Hormones and rop Genes By Malem Gutema Mentor Dr. John Fowler Summer 2004 Rop Proteins Plants possess a special subfamily of GTPases denoted by Rop (Rho of Plants) proteins Rop proteins have been first identified in pea (Yang and Watson), where they contribute to control tip growth in pollen tubes The plant hormone Abscisic Acid (ABA) ABA’s main roles - Regulating transpiration - Stress response - Slow down growth - Germination of seeds. The Plant hormone Auxin Auxin’s promotes initiation of - lateral roots - new leaves Auxin regulates elongation of - stem - root cells Part I - Purpose Rop proteins are thought to interact with plant hormone signals, particularly during root growth. Hypothesis and Prediction We know ABA, Auxin We Hypothesize ABA, Auxin Our prediction ABA, Auxin Part I - Experiments Maize rop mutants There are nine rop genes in maize I used samples with mutation of the rop 2, rop 6 and rop 7 genes How do you get the mutation? rop mutation Maps  Root growth assay Question? Would additional ABA and Auxin affect root growth? How are the wild-type and mutated samples affected? Experiment set up Germinate seedlings in a water soaked paper towel Measure root length before treatment Add auxin or ABA Measure after 24 and 48hrs Average the length Auxin – rop 6/7 Auxin rop 2 ABA rop 2 ABA- rop 6/7 Summary of Results Working Model Part II Do same root assay on a segregating population of rop2-m1 mutants Take samples from an ear that has both homozygote and heterozygote genotypes Do genotyping What does segregating mean? rop2-m1/+ Χ rop2-m1 / rop2-m1 rop2-m1 / rop2-m1 a homozygote rop2-m1 / + a heterozygote Population is 50:50 homozygotes:heterozygotes Procedure Collect leaf samples Extract DNA Do a PCR reaction with rop 2 primers Analyze the PCR product on an agarose gel to c