Abstract
Marchantia polymorpha is a species of basal land plant that sheres the basic mechanism of morphogenesis with other land plants. There are several advantages to using M. polymorpha for forward genetic analysis: a relatively small genome size (280 Mb), low gene redundancy and a haploid-dominant life cycle. In addition, molecular genetic resources, such as an Agrobacterium-mediated transformation technique, an EST database and a genetic database, have recently been developing rapidly for M. polymorpha. To establish our procedure of forward genetic analysis, we generated T-DNA-inserted lines of M. polymorpha using the binary vector pCAMBIA1300. We screened 10,000 lines and isolated 25 lines that show various morphological phenotypes, such as loss of air-chamber and loss of gemma formation. Southern blot analysis indicated that the average of 2.4 tags were inserted, and 10 single-tag lines were obtained. Then, by analyzing T-DNA flanking sequence, in 10 lines it appeared that T-DNA tags were inserted into an ORF or upstream of a gene. Currently, we are analyzing the linkage between the morphological phenotype and the inserted T-DNA tag and performing a complementation test.
