The Screening Of Male Sterile Mutants St368 And St534 And Their Functional Analysis In Arabidopsis Thaliana

Arabidopsis thaliana is a small plant of Cruciferae family with short growth cycle and relatively small genome is a typical self-cross plant. It is mode plant to study plant gene function. With the complete genome sequence of Arabidopsis available in 2000, gene functional investigation has become an frontier research area in life science. In this work, anther and pollen development were investigated by forward genetics through male-sterile mutant isolation from T-DNA tagged lines. Genetic analysis indicated that the mutant st368 was controlled by a single recessive gene. To identify the corresponding ST368 gene, a genomic DNA fragment that flanked the left border of T-DNA was recovered by thermal asymmetric interlaced PCR (TAIL-PCR). Sequencing of the TAIL-PCR products suggested that the T-DNA is inserted in the last intron of a predicted open reading frame (At1g07080). ST368 encodes a gamma interferon responsive lysosomal thiol reductase (GILT) family protein that comprises 261 amino acids with an estimated molecular weight of 35.8 kDa. Cytological observations of st368 mutant and the wild type plant showed that both male and female organs are abnormal. Arabidopsis thaliana is a small plant of Cruciferae family with short growth cycle and relatively small genome is a typical self-cross plant. It is mode plant to study plant gene function. With the complete genome sequence of Arabidopsis available in 2000, gene functional investigation has become an frontier research area in life science. In this work, anther and pollen development were investigated by forward genetics through male-sterile mutant isolation from T-DNA tagged lines. Genetic analysis indicated that the mutant st534 was controlled by a single recessive gene. To identify the corresponding ST534 gene, a genomic DNA fragment that flanked the left border of T-DNA was recovered by thermal asymmetric interlaced PCR (TAIL-PCR). Sequencing of the TAIL-PCR products suggested that the T-DNA is inserted in the last intron of a predicted open reading frame (At2g44770). Sequence analysis indicated that it encodes a phagocytosis and cell motility protein. Cytological observations of st534 mutant and the wild type plant showed that its male organ is abnormal.