Transgenic rice plants expressing a Bacillus subtilis protoporphyrinogen oxidase gene are resistant to diphenyl ether herbicide oxyfluorfen.

Protoporphyrinogen oxidase (Protox), the penultimate step enzyme of the branch point for the biosynthetic pathway of Chl and hemes, is the target site of action of diphenyl ether (DPE) herbicides. However, Bacillus subtilis Protox is known to be resistant to the herbicides. In order to develop the herbicide-resistant plants, the transgenic rice plants were generated via expression of B. subtilis Protox gene under ubiquitin promoter targeted to the cytoplasm or to the plastid using Agrobacterium-mediated gene transformation. The integration and expression of the transgene were investigated at T0 generation by DNA and RNA blots. Most transgenic rice plants revealed one copy transgene insertion into the rice genome, but some with 3 copies. The expression levels of B. subtilis Protox mRNA appeared to correlate with the copy number. Furthermore, the plastidal transgenic lines exhibited much higher expression of the Protox mRNA than the cytoplasmic transgenic lines. The transgenic plants expressing the B. subtilis Protox gene at T0 generation were found to be resistant to oxyfluorfen when judged by cellular damage with respect to cellular leakage, Chl loss, and lipid peroxidation. The transgenic rice plants targeted to the plastid exhibited higher resistance to the herbicide than the transgenic plants targeted to the cytoplasm. In addition, possible resistance mechanisms in the transgenic plants to DPE herbicides are discussed.

Characterization of a glutathione S-transferase gene ATGST 1 in Arabidopsis thaliana.

A glutathione S-transferase (GST) gene was cloned in Arabidopsis thaliana. The gene, designated ATGST 1, contained the entire transcription unit in three exons interrupted by two introns. The combined sequence of three exons had an open reading frame which predicted a GST protein of 208 amino acids. Gene transcription has been reported to be induced by pathogen attack and dehydration. In the present study northern blot analysis using a gene-specific DNA probe in the 3' untranslated region revealed that expression of the gene was also rapidly induced by other environmental stresses such as wounding, low temperature, high salt and DPE herbicide treatment. The promoter region of the gene contained the sequence motif ATTTCAAA that is known to be present in ethylene-responsive elements and other motifs that are highly conserved amongst stress-inducible gene promoters.

Generation of Resistance to the Diphenyl Ether Herbicide, Oxyfluorfen, via Expression of the Bacillus subtilis Protoporphyrinogen Oxidase Gene in Transgenic Tobacco Plants.

In an effort to develop transgenic plants resistant to diphenyl ether herbicides, we introduced the protoporphyrinogen oxidase (EC 1.3.3.4) gene of Bacillus subtilis into tobacco plants. The results from a Northern analysis and leaf disc assay indicate that the expression of the B. subtilis protoporphyrinogen oxidase gene under the cauliflower mosaic virus 35S promoter generated resistance to the diphenyl ether herbicide, oxyfluorfen, in transgenic tobacco plants.

Structure and expression of the AWI 31 gene specifically induced by wounding in Arabidopsis thaliana.

Differential screening of an Arabidopsis cDNA library constructed from the plant tissues harvested 1 h after wounding resulted in the isolation of wound-inducible cDNA clones (Kim et al., 1994). The cDNA clones could be broadly classified into two groups according to the expression time of their transcripts. Nine clones from the 10 different wound-inducible cDNAs were rapidly induced, reaching a maximum level in approximately 1-1.5 h and then were progressively reduced after wounding. The cDNA clone AWI 31 showed steady accumulation of the transcripts and reached the maximum value at a later time point of 2.5 h and then started to decline. The corresponding gene of the AWI 31 in which the coding region was interrupted by an intron, had an open reading frame that predicted a protein of 386 amino acids. However, the gene product did not show any significant homology to other known proteins in the database. Northern hybridization study using the cDNA probe revealed that the gene was not regulated by other environmental stresses such as drought, high salt, low temperature, or a DPE herbicide treatment, indicating that the cDNA clone AWI 31 was specifically induced by wounding.