Ethylene influences a number of developmental processes and responses to stress in higher plants. The molecular basis for the action of ethylene was investigated in mutants of Arabidopsis thaliana that have altered responses to ethylene. One mutant line, which has a dominant mutation at a locus designated etr, lacks a number of responses to ethylene that are present in the wild-type plant. These include inhibition of cell elongation, promotion of seed germination, enhancement of peroxidase activity, acceleration of leaf senescence, and feedback suppression of ethylene synthesis by ethylene. These diverse responses, which occur in different tissues of Arabidopsis, appear to share some common element in their transduction pathways--for example, a single receptor for ethylene. Results of ethylene binding experiments in vivo indicate that this receptor may be affected by the etr mutation.

We have identified a number of genes of the flowering plant Arabidopsis thaliana that are abundantly expressed during, embryogenesis. In this paper we discuss four of these genes, which comprise a gene family: complete genomic nucleotide sequence of two of the genes and partial sequence of the other two shows that they are all homologous to the 12S globulin seed storage protein genes of other angiosperms. The four genes fall into three subfamilies, as defined by cross-hybridization. One subfamily contains two genes in the Landsberg erecta strain, but only a single gene in the Columbia strain of Arabidopsis. The other two of these 12S gene subfamilies contain only single genes in both strains. Thus, the seed storage protein gene family in Arabidopsis appears much simpler than that in other higher plants. These genes are expressed during the latter half of embryogenesis, a period in which abscisic acid (ABA) is thought to play a role in gene regulation, and known to play a role in seed physiology. We observed no significant difference in the expression profiles of these four genes in ABA-deficient and ABA-insensitive mutants of Arabidopsis, except that the onset of detectable expression of all of the transcripts is slightly delayed in both types of mutants.

Nucleotide sequences for the nuclear genes encoding chloroplast (GapA and GapB) and cytosolic (GapC) glyceraldehyde-3-phosphate dehydrogenases (GAPDHs) from Arabidopsis thaliana were determined. Comparison of nucleotide sequences indicates that the divergence of chloroplast and cytosolic GAPDH genes preceded the divergence of prokaryotes and eukaryotes. In addition, some intron-exon junctions are conserved among GapB, GapC, and chicken GAPDH genes. These results provide evidence at the molecular level to support the idea that introns existed before the divergence of prokaryotes and eukaryotes.

The primary structure of the alpha 1 tubulin gene of Arabidopsis thaliana was determined and the 5' and 3' ends of its transcript were identified by S1 nuclease mapping experiments. The information obtained was used to (i) predict the amino acid sequence of the alpha 1-tubulin, (ii) deduce the positions of introns within the alpha 1-tubulin gene, and (iii) construct 3' noncoding gene-specific hybridization probes with which to study the pattern of alpha 1-tubulin transcript accumulation in different tissues and at different stages of development. The predicted amino acid sequence of the alpha 1-tubulin has 92% identity with the predicted product of the previously characterized A. thaliana alpha 3-tubulin gene. The coding sequence of the alpha 1-tubulin gene is interrupted by four introns located at positions identical to those of the four introns in the alpha 3 gene. RNA blot hybridization studies carried out with an alpha 1-tubulin gene-specific probe showed that the alpha 1 gene transcript accumulates primarily in flowers, with little transcript present in RNA isolated from roots or leaves. In order to investigate the pattern of alpha-tubulin gene expression in developing flowers, RNA was isolated from flowers at five different stages of development: flower buds, unopened flowers with pollen, open flowers, flowers with elongating carpels, and green seed pods. RNA blot hybridizations performed with 3' noncoding gene-specific probes showed that the alpha 3 tubulin gene transcript is present in flowers at all stages of development, whereas the alpha 1-tubulin gene transcript could only be detected in RNA from unopened flowers with pollen, open flowers, and flowers with elongating carpels.

The multigene family encoding the small subunit polypeptides of ribulose-1,5-bisphosphate carboxylase/oxygenase in the crucifer Arabidopsis thaliana has been isolated and the organization and structure of the individual members determined. The family consists of four genes which have been divided into two subfamilies on the basis of linkage and DNA and amino acid sequence similarities. Three of the genes, designated ats1B, ats2B, and ats3B, reside in tandem on an 8 kb stretch of the chromosome. These genes share greater than 95% similarity in DNA sequence and encode polypeptides identical in length and 96.7% similar in amino acid sequence. The fourth gene, ats1A, is at least 10 kb removed from, or completely unlinked to the B subfamily. The B subfamily genes are more similar to each other than to ats1A in nucleotide and amino acid sequence. All four genes are interrupted by two introns whose placement within the coding region of the genes is conserved. The introns of the B subfamily genes are similar in length and nucleotide sequence, but show no similarity to the introns of ats1A. Comparison of the DNA sequences within the immediate 5' and 3' flanking sequences among the genes revealed only limited regions of homology. S1 analysis shows that all four genes are expressed.