Drought Insensitive Mutant Screening And Guard Cell Protoplast Isolation In Arabidopsis

Plants often encount many kinds of environmental stresses. Knowledge about molecular mechanism of drought-resistant and its genetics, especially to expressions of main gene of drought-resistant and controls of upstream signal transduction. Mutant, a powerful tool of analyzing genetic process, has already been extensively used for research in plant growth,metabolism and signal transduction.Firstly we maked sure method of isolating Arabidopsisis mutant by thermal imagings in many ways on trial, optimize conditions for isolation, and confirm the good period of seedlings and fine density for isolation.Secondly we obtain an M2 population derived from 0.4%ethyl methanesulfonate (EMS)-mutagenized wild type Arabidopsis thaliana. With the tiny difference of temperature leaf between mutant and wild type for index sign,we isolate M2 seedlings by 8-10 days drought-stress treatment by infrared thermography in a large scale.A great deal of doi and dos Arabidopsis mutant were isolated (the difference of leaf temperature between candidate mutant and wild type is very obvious,about 0.6-1.2℃).In our experiments,we use drought insensitive mutant doi21 which was selected by far-red imaging methods and their corresponding wild type C24 as materials.Based on the traits of patch clamp,we fumbled a successful method for planting mutant and wildtype materials and a successful method for guard cell protoplast preparation.