Molecular Characterization of the Soybean Alcohol Dehydrogenase Gene Family Amplified in Vitro by the Polymerase Chain Reaction

Abstract

Soybean (Glycine max) alcohol dehydrogenase (ADH) cDNAs were amplified in vitro from total RNA by the polymerase chain reaction (PCR). The amplification strategy involved first strand cDNA synthesis from anaerobic cotyledon total RNA using an 18-thymidine primer. The second strand cDNA primer was a conserved sequence near the 5' end of known plant ADH transcripts. The PCR products were ligated into a plasmid vector and unique clones were isolated on the basis of size and restriction pattern. Sequence analysis revealed three distinct classes of soybean ADH cDNAs, all of which showed high homology to Adh genes from maize and peas. RNA blot hybridization analyses showed differential expression patterns for these genes. One gene, expressed constitutively in all seedling organs, was inducible by anaerobiosis, one gene was expressed only in anaerobic organs, and the third gene was expressed predominantly in anaerobic roots.

Characteristics of soybean 1-Cys peroxiredoxin and its behavior in seedlings under flooding stress

“…To examine whether GmPer1 genes are induced by flooding stress, 48-h-old soybean seedlings germinated on sand were submerged in water for 6-24 h at 25°C in a growth chamber (flooding treatment), and then total RNA was extracted from the seedlings. To confirm that the soybean seedlings really suffered flooding stress, expression of the Adh gene of soybean (Glyma04g41990), which is one of the marker genes for hypoxia (Newman and Vantoai 1992;Russell et al 1990), was examined using Adhspecific primers (primer 9, 5Ј-ATGTCGAGCACAGC TGGCCAAG-3Ј and primer 10, 5Ј-CCTGTGCAAATTC CACAACTGAGAAC-3Ј). Expression of the Adh gene was detected by flooding treatment, whereas expression of neither the GmPer1a nor GmPer1b was detected ( Figure 3B) indicating that neither gene is induced by flooding stress.…”

Characterization of a novel flooding stress-responsive alcohol dehydrogenase expressed in soybean roots

“…Phylogenetic analysis using amino acid sequences predicted from these homologs (Fig. 3b) showed that, among 6 subgroups of soybean Adh GroES-like peptides, Adh2 were classified into the same group as reported Adh peptides (Adh-1, SOYADH57 and SOYADH73; Newman and VanToai 1992). The Adh group contained six different peptides (Fig.…”

“…Newman and VanToai (1992) described three soybean Adh genes. One of these Adh genes is constitutively expressed in all seedling organs and is induced by anaerobiosis.…”

A novel zinc‐binding alcohol dehydrogenase 2 from Arachis diogoi, expressed in resistance responses against late leaf spot pathogen, induces cell death when transexpressed in tobacco

“…Multiple sequence alignment and phylogenetic analyses displayed its close relation in plants to soybean predicted zinc‐binding ADH2‐like protein (different from ADH2 studied 27, 28, 29) with 85% sequence similarity (Fig. S2a).…”