The Map-based Cloning Of Arabidopsis Hydra3 Mutant

Map-based cloning techniques are widely used to identify the mutation gene. Sequencing of the Arabidopsis (Columbia-0 ) genome and Ler's DNA fragments give this approach a great boost. The accession of a great number of sequence polymorphisms and molecular markers makes polymerase chain reaction-based methods more effective. As a result if has become possible to complete positional cloning projects in a short time and with relatively little effort.We gain a mutant, induced by EMS, which is characterized by having short root, little development of the hypocotyls and cabbage-like leaves. It is similar to sterol-deficient mutant, hydral (hydl, Topping et al., 1997 ), and named hydra3 after hydral and hydra2 (fackel, jk). Promoter-reporter gene analysis showed misexpression ofIAA14 promoter. But other promoter, such as DR5 .IAA2, A UX1 . CHL1 and SCR, express normally.Besides, We do some research for map- based cloning ofhyd3. By 16 simple sequence length polymphisms (SSLP) markers, we located hyd3 at the end of the chromosome 5. hi the fine-mapping experiment, we make use of 6 SSLP markers, 4 cleaved amplified polymorphic sequences (CAPS) markers, which are got from TAIR database, and 2 new designed SSLP markers and position hydS in the area of Arabidopsis chromosome, from 24860kb to 25494kb.