Control of expression of a gene encoding an extensin by phytochrome and a blue light receptor in spores ofAdiantum capillus‐venerisL.

Abstract

SummaryIn the present study, using a newly developed fluorescent differential display technique, we have carried out largescale screening for genes whose expression was regulated by phytochrome and antagonistically by a blue light receptor in the spores of the fern Adiantum capillus-veneris L. Spores after imbibition were briefly irradiated with red, red/blue or blue light and collected 8 h after the irradiation. Total RNA was isolated from each sample and used to make cDNA with an oligo-dT primer. The cDNA was then used as a template for PCR with the oligo-dT primer and 80 arbitrary primers. The resulting PCR products were analyzed by an automated fluorescent DNA sequencer. Among 8000 displayed bands, we identified 15 upregulated and four down-regulated bands by red light, and this red light effect was irreversibly reversed by blue light. We cloned one of the up-regulated cDNA fragments and used it to screen a cDNA library prepared from the spores. The isolated insert is predicted to encode Ser-(Pro) n repeats and showed homology with cell wall-associated extensins. The expression of this cDNA was induced 8 h after a red light treatment and the red light induction was photoreversibly prevented by far-red light and photoirreversibly by blue light. The mRNA of this gene was detectable 4 h after red light irradiation and gradually increased in germinating spores.

Global Analysis of Proline-Rich Tandem Repeat Proteins Reveals Broad Phylogenetic Diversity in Plant Secretomes

“…(A) Dendrogram showing the evolutionary relationships among major plant divisions as well as the distribution of EXTAs identified in this study and in previous literature [14], [25], [26]. (To reflect results of a recent phylogenetic analysis [52], the large divisions, super-rosids and super-asterids are shown rather than rosids, asterids, and other phylogenetic groups).…”

“…Our global analysis identified YxY-containing extensins, referred to as Extensin type alpha (EXTA) (Text S1), throughout most land plant divisions (Figures 2A, 3A, and Figure S4). In addition to the core eudicots and ferns [14], [25], [26], EXTAs are found in at least some non-grass monocots, including onion, orchids, and asparagus, and in a diversity of non-flowering vascular plants, ranging from gymnosperms to Selaginella , a member of the oldest extant vascular plant group [27]. Furthermore, although missing in mosses, transcripts encoding EXTAs were found in the non-vascular liverwort species Marchantia polymorpha demonstrating that EXTAs predate the evolution of plant vasculature (Text S3, also see PlantPro20Fam).…”

Fluorescent Differential Display Identifies Circadian Clock-Regulated Genes in Arabidopsis thaliana

“…The total RNA samples were used to make single-stranded cDNA using an anchored dGpolydTdG primer that had a fluorescent tag according to the methods described (Uchida et al, 1998). Total RNA samples for use in FDD were DNAse treated, extracted with phenol/chloroform and chloroform, ethanol precipitated, and resuspended in H 2 O.…”

“…PCR products were analyzed using PAGE and a fluorescence detector system (Hitachi SQ 5500 DNA Sequencer) according to the methods described (Uchida et al, 1998). PCR products that consistently showed a cycling phenotype were separated on polyacrylamide gels, and the cycling bands were visualized using a fluorimager (Takara FMBIO) and excised.…”

“…Ectopic expression of these two putative transcription factors affected the expression of known ccgs, suggesting that CCA1 and LHY are trans factors involved in clock-regulated gene expression. Recent modifications to this technique, such as the use of fluorescent tags instead of radiolabels together with reduced primer size, have enhanced the utility of this method for screening for ccgs (Ito et al, 1994;Liang et al, 1994;Uchida et al, 1998). Liu et al (1995) used a luciferase reporter system to screen for clock-controlled transcription in cyanobacteria.…”