Development and Environmental Stress Employ Different Mechanisms in the Expression of a Plant Gene Family.

Ribulose bisphosphate carboxylase small subunit (RbcS) genes in the common ice plant, as in all higher plants, constitute a multigene family. We have measured transcription activity and steady state mRNA levels of individual members of the family, six RbcS genes, in the ice plant with emphasis on the transition from C3 photosynthesis to Crassulacean acid metabolism (CAM), which this plant undergoes during development and under environmental stress. Four RbcS genes are differentially expressed in leaves but are regulated in a coordinate fashion. A developmentally engrained, sharp decline in the steady state mRNA levels, which is observed during the juvenile-to-adult growth phase transition, coincides with the time interval when the C3-to-CAM switch occurs. Developmental down regulation of RbcS is due to down regulation of transcription. In contrast, NaCl stress specifically affected RbcS transcript accumulation post-transcriptionally, resulting in decreased RbcS mRNA levels. Antagonistic regulatory programs are apparent in stress/stress relief experiments. The results indicate complex controls, affecting both transcriptional and post-transcriptional processes, that act differentially during plant development, stress, and recovery from stress.

The six genes of the Rubisco small subunit multigene family from Mesembryanthemum crystallinum, a facultative CAM plant.

The nucleotide sequences of the entire gene family, comprising six genes, that encodes the Rubisco small subunit (rbcS) multigene family in Mesembryanthemum crystallinum (common ice plant), were determined. Five of the genes are arranged in a tandem array spanning 20 kb, while the sixth gene is not closely linked to this array. The mature small subunit coding regions are highly conserved and encode four distinct polypeptides of equal lengths with up to five amino acid differences distinguishing individual genes. The transit peptide coding regions are more divergent in both amino acid sequence and length, encoding five distinct peptide sequences that range from 55 to 61 amino acids in length. Each of the genes has two introns located at conserved sites within the mature peptide-coding regions. The first introns are diverse in sequence and length ranging from 122 bp to 1092 bp. Five of the six second introns are highly conserved in sequence and length. Two genes, rbcS-4 and rbcS-5, are identical at the nucleotide level starting from 121 bp upstream of the ATG initiation codon to 9 bp downstream of the stop codon including the sequences of both introns, indicating recent gene duplication and/or gene conversion. Functionally important regulatory elements identified in rbcS promoters of other species are absent from the upstream regions of all but one of the ice plant rbcS genes. Relative expression levels were determined for the rbcS genes and indicate that they are differentially expressed in leaves.

Phosphoribulokinase from ice plant: Transcription, transcripts and protein expression during environmental stress.

The expression of PRK (phosphoribulokinase, E.C.2.7.1.19) in ice plant (Mesembryanthemum crystallinum) during development and under environmental stress was studied. cDNA clones were isolated and full-length cDNAs were characterized. Ice plant PRK is contained in a 1520 nucleotide transcript including a 126 nucleotide leader sequence, a 175 nucleotide 3'-end and a 20-30 nucleotide polyA(+)-stretch. The coding region, 397 codons, specifies a protein of Mr 44 064. The mature sequence is preceded by a transit peptide of approximately 46 amino acids. The mature portion of ice plant PRK is 86.4% identical to that of spinach and, e.g., 16.2% identical to PRK from Xanthomonas flavus. Under salt stress or cold adaptation conditions, the amount of mRNA declined by a factor of approximately three within days, followed by an increase to approximately pre-stress levels. The fluctuation in mRNA amount is not reflected on the level of transcription of the gene, suggesting post-transcriptional control, nor is PRK protein amount affected significantly over the short stress period. The recovery of transcript levels for photosynthesis-related proteins after stress appears to be a general response to environmental stresses that affect water status in ice plant. We suggest that the photosynthetic machinery in this facultative halophyte is effectively buffered from damage caused by such environmental stress.