PubMed Health⌕ Search

Biomedical subjects

P E Staswick

Publications and source records attributed to P E Staswick.

18 recordsLinked to original sources
PubMed2001

Efficient down-regulation of the major vegetative storage protein genes in transgenic soybean does not compromise plant productivity.

Soybean (Glycine max L. Merr.) contains two related and abundant proteins, VSP alpha and VSP beta, that have been called vegetative storage proteins (VSP) based on their pattern of accumulation, degradation, tissue localization, and other characteristics. To determine whether these proteins play a critical role in sequestering N and other nutrients during early plant development, a VspA antisense gene construct was used to create transgenic plants in which VSP expression was suppressed in leaves, flowers, and seed pods. Total VSP was reduced at least 50-fold due to a 100-fold reduction in VSP alpha and a 10-fold reduction in VSP beta. Transgenic lines were grown in replicated yield trials in the field in Nebraska during the summer of 1999 and seed harvested from the lines was analyzed for yield, protein, oil, and amino acid composition. No significant difference (alpha = 0.05) was found between down-regulated lines and controls for any of the traits tested. Young leaves of antisense plants grown in the greenhouse contained around 3% less soluble leaf protein than controls at the time of flowering. However, total leaf N did not vary. Withdrawing N from plants during seed fill did not alter final seed protein content of antisense lines compared with controls. These results indicate that the VSPs play little if any direct role in overall plant productivity under typical growth conditions. The lack of VSPs in antisense plants might be partially compensated for by increases in other proteins and/or non-protein N. The results also suggest that the VSPs could be genetically engineered or replaced without deleterious effects.

Down-Regulation↗
PubMed1998

Jasmonate signaling mutants of Arabidopsis are susceptible to the soil fungus Pythium irregulare.

Jasmonic acid has properties of a plant hormone, including the induction of specific genes associated with plant defense. We previously described jar1-1, an Arabidopsis jasmonate response mutant that exhibits reduced sensitivity to methyl jasmonate. We have further characterized this mutant and two new alleles; jar1-2 from a gamma irradiated population, and jar1-4 from a T-DNA mutant population. Seedling root growth in jar1-1 was equally insensitive to methyl jasmonate and jasmonic acid, indicating that the defect was not in the conversion of methyl jasmonate to the acid. None of the jar1 mutants showed an altered sensitivity to auxin, cytokinin, or the ethylene precursor 1-aminocyclopropane-1-carboxylic acid, indicating that the lesion does not affect the general uptake or transport of hormones. A soil fungus, Pythium irregulare, was found to blight jar1-1. Cultures of this organism caused the symptoms in all three jar1 mutants but not in wild type, indicating that increased susceptibility was due to the lesion in the JAR1 locus. A fatty acid desaturase triple mutant that is defective in the biosynthesis of jasmonic acid (J. Browse, Washington State University) was also susceptible, confirming that jasmonate is involved in resistance. The jar1-1 locus was mapped to the lower end of chromosome 2, about 11.4 cM from as1 and 1.6 cM from cer8. These results establish that jasmonate signaling plays an important role in resistance to soil micro-organisms in plants.

Alleles↗
PubMed1994

Arabidopsis p40 homologue. A novel acidic protein associated with the 40 S subunit of ribosomes.

We have isolated a full-length cDNA clone from Arabidopsis thaliana that has extensive homology to p40 proteins from other organisms. The cDNA predicts a protein (A-p40) of 298 amino acids with a calculated molecular mass of 32.5 kDa and a pI of 4.8. Antibodies raised against an A-p40 fusion protein detected a polypeptide of about 40 kDa in Arabidopsis and about 42 kDa in corn and soybean. In two-dimensional gel electrophoresis the antibodies detected several acidic isoforms. A-p40 appears to be located in the cytoplasm in two forms: soluble, or tightly associated with 40 S subunits and polysomes. Mixing experiments with soluble A-p40 and purified corn polysomes showed that the ribosome-associated form does not result from nonspecific binding during extraction. High concentrations of KCl (1.5 M) are needed to release A-p40 from ribosomes, which suggests that it may be a true ribosomal protein. Several pieces of evidence suggest that A-p40 belongs to the acidic class of ribosomal proteins; first, its acidic pI; second, the presence of amino acid repeats conserved in other acidic ribosomal proteins; and third, the fact that it exists in the cell in two pools (free and ribosome-associated).

Amino Acid Sequence↗
PubMed1992

Methyl jasmonate inhibition of root growth and induction of a leaf protein are decreased in an Arabidopsis thaliana mutant.

Jasmonic acid and its methyl ester, methyl jasmonate (MeJA), are plant signaling molecules that affect plant growth and gene expression. Primary root growth of wild-type Arabidopsis thaliana seedlings was inhibited 50% when seedlings were grown on agar medium containing 0.1 M MeJA. An ethyl methanesulfonate mutant (jar1) with decreased sensitivity to MeJA inhibition of root elongation was isolated and characterized. Genetic data indicated the trait was recessive and controlled by a single Mendelian factor. MeJA-induced polypeptides were detected in Arabidopsis leaves by antiserum to a MeJA-inducible vegetative storage protein from soybean. The induction of these proteins by MeJA in the mutant was at least 4-fold less in jar1 compared to wild type. In contrast, seeds of jar1 plants were more sensitive than wild type to inhibition of germination by abscisic acid. These results suggest that the defect in jar1 affects a general jasmonate response pathway, which may regulate multiple genes in different plant organs.

Journal Article↗
PubMed1991

Nitrogen and methyl jasmonate induction of soybean vegetative storage protein genes.

Vegetative storage protein (VSP) and VSP mRNA levels in soybean (Glycine max) leaves correlated with the amount of NH(4)NO(3) provided to nonnodulated plants. The mRNA level declined as leaves matured, but high levels of N delayed the decline. This is consistent with the proposed role for VSP in the temporary storage of N. Wounding, petiole girdling, and treatment with methyljasmonate (MeJA) increased VSP mRNA in leaves 24 hours after treatment. The magnitude of the response depended on leaf age and N availability. N deficiency essentially eliminated the response to wounding and petiole girdling. MeJA was almost as effective in N-deficient plants as in those receiving abundant N. Inhibitors of lipoxygenase, the first enzyme in the jasmonic acid biosynthetic pathway, blocked induction by wounding and petiole girdling but not by MeJA. This supports a role for endogenous leaf jasmonic acid (or MeJA) in the regulation of VSP gene expression.

Journal Article↗
PubMed1991

Methyl jasmonate treatment eliminates cell-specific expression of vegetative storage protein genes in soybean leaves.

Soybean (Glycine max) plants accumulate a vacuolar glycoprotein in the parenchymal cells of leaves, petioles, stems, seed pods, and germinating cotyledons that acts in temporary nitrogen storage during vegetative growth. In situ immunolocalization of this vegetative storage protein (VSP) revealed that it accumulates in those parenchymal cells in close proximity to existing and developing vasculature, as well as in epidermal and cortical cells. The protein was more prevalent in younger, nitrogen-importing tissues before pod and seed development. Removal of actively growing seed pods greatly enhanced VSP accumulation, primarily in bundle sheath and paraveinal mesophyll cells. In situ hybridization of a VSP RNA probe to mRNA in leaf sections demonstrated that cell-specific mRNA accumulation corresponded with the pattern of protein localization. Treatment of leaf explants with 50 micromolar methyl jasmonate resulted in accumulation of VSP mRNA and protein in all cell types.

Journal Article↗
PubMed1989

Developmental regulation and the influence of plant sinks on vegetative storage protein gene expression in soybean leaves.

Soybeans (Glycine max L.) accumulate a storage glycoprotein which is abundant in vegetative tissues, but is only a minor component of seeds. Changes in vegetative storage protein gene expression in leaves of control and depodded plants were monitored throughout plant development. Western and Northern blot hybridization analysis of protein and mRNA levels, respectively, showed that expression of these genes was highly regulated during development. Expression correlated with periods when expected demand for mobilized leaf reserves by other plant sinks was low. Vegetative storage protein mRNA comprised about 0.5% of the total mRNA in immature leaves and declined at least 20-fold by flowering. Depodding or blockage of leaf petiole phloem transport increased these mRNAs to about 16% of the total mRNA. Transcript levels also increased dramatically after seed maturation, just before leaf senescence. Protein levels followed a similar pattern and were inversely related to the number of seed pods allowed to develop on the plants. The results support the role for these proteins as temporary storage molecules which can be rapidly synthesized or degraded according to the need for nutrients by other plant tissues.

Journal Article↗
PubMed1989

Preferential Loss of an Abundant Storage Protein from Soybean Pods during Seed Development.

A temporary vegetative storage protein, composed of similar 25 kilodalton and 27 kilodalton subunits, was found to be abundant in soybean (Glycine max (L.) Herr. var Hobbit) leaves, stems, pods, flower petals, germinated cotyledons, and less abundant in roots, nodules and seeds. Total pod protein was highest at 3 weeks after flowering and declined by 37% within 3 weeks during seed development. During this time the vegetative storage protein declined from 18% to 1.5% of the total pod protein and accounted for 45% of the protein lost from pods. This indicates that the vegetative storage protein makes a significant contribution to the pool of nutrients mobilized from pods for transport to developing seeds.

Journal Article↗
PubMed1988

Soybean vegetative storage protein structure and gene expression.

Depodded soybean (Glycine max [L] Merr. cv Williams) plants accumulate high levels of a glycoprotein in their leaves that has many features of a storage protein. The protein is found in all vegetative tissues which have been examined but not in the seeds. Translation in vitro indicated that elevated mRNA levels were at least partially responsible for the specific increase in vegetative storage protein. cDNA clones were isolated and sequenced, and an amino acid sequence was predicted. Although the amino acid composition is similar to that of seed storage proteins, no sequence similarity could be detected. Northern blot hybridization confirmed a large increase in vegetative storage protein mRNA in leaves of depodded plants. The vegetative storage proteins are represented by about four gene copies in the haploid genome.

Journal Article↗
PubMed1984

The amino acid sequence of the A2B1a subunit of glycinin.

The amino acid sequences of the acidic and basic components of the A2B1a subunit of glycinin, the major seed reserve protein of the soybean (Glycine max L. Merr.), were determined. They contain 278 and 180 amino acids, respectively, and have molecular weights of 31,600 +/- 100 and 19,900 +/- 100. The molecular weight of the acidic component is considerably less than that estimated by sodium dodecyl sulfate-gel electrophoresis (37,000). Sequence heterogeneity was detected at several positions scattered throughout the primary structures of both components, indicating that the preparation sequenced was composed of several nearly identical polypeptides. These data, in conjunction with a recently determined nucleotide sequence of the 3'-terminal two-thirds of the analogous glycinin subunit gene, illustrate the complexity of the gene family responsible for synthesis of glycinin subunits.

Amino Acid Sequence↗
PubMed1984

Identification of the cystines which link the acidic and basic components of the glycinin subunits.

The half-cystine residues involved in linking the acidic and basic polypeptides were determined for several glycinin subunits. The cystines were localized with specific cyanogen bromide fragments either by comparing the electrophoretic mobility of nonreduced and reduced fragments, or by co-purifying and then determining the NH2-terminal sequence of the covalently linked fragments. Residues involved in the disulfides were further identified by labeling them with [3H] iodoacetic acid. Only 1 cystine was found to be involved in linking the acidic and basic components of each subunit, and they were in analogous positions in each of the subunits studied. Potential sites for intrapolypeptide cystines were also identified.

Amino Acid Sequence↗
PubMed1983

Characterization of a soybean cultivar lacking certain glycinin subunits.

The 11S storage protein (glycinin) of soybean [Glycine max (L.) Merr., cv. Raiden] was studied by polyacrylamide gel electrophoresis and amino acid sequence analysis. It contained the following subunits composed of acidic (A) and basic (B) polypeptides: A1aB2, A1bB1b, A2B1a, and A3B4. However, it lacked polypeptides A4, A5, and B3 which are present in many other cultivars. A new acidic polypeptide called A6 was present in a low amount and was characterized by amino acid sequence analysis. It was homologous to A4, although of a smaller apparent molecular weight. Since Raiden has an average protein content of about 40% and its glycinin fraction can be purified as a 350,000 D complex which is typical of other cultivars, the results imply polymorphism with respect to glycinin subunit composition. Because there is a wide variation in the methionine content of the various subunits, these findings suggest the possibility of genetically manipulating the nutritional quality of soybean seed protein by altering glycinin subunit composition.

Amino Acid Sequence↗
PubMed1983

Glycinin composition of several perennial species related to soybean.

The 7S and 11S seed storage proteins from four perennials related to soybean (Glycine canescens, G. tomentella, G. tabacina, and G. clandestina) were analyzed by sodium dodecyl sulfate-gel electrophoresis. Each species yielded a unique electrophoretic pattern that varied in the total number of bands and their relative mobilities. In every case, the electrophoretic patterns were substantially different from CX635-1-1-1, the strain of G. max used in this study for comparison. Size heterogeneities among both the 7S and 11S polypeptides of the perennials were evident.Abundant proteins in the 11S fraction from G. tomentella (CSIRO No. 1133) were separated by chromatography on DEAE-Sephadex and then their apparent molecular weights, amino acid compositions, and NH(2)-terminal amino acid sequences were determined. A group of proteins were obtained which resembled the A(1b)-polypeptide components of glycinin from G. max. They had the same size (M(r) approximately 37,000), identical NH(2)-terminal sequences, and similar amino acid compositions to A(1b). A second group of acidic proteins (M(r) approximately 50,000) in G. tomentella had NH(2)-terminal sequences homologous to the A(5) component (M(r) approximately 10,000) of glycinin. The latter group of polypeptides had a substantially higher apparent molecular weight than any acidic polypeptide components of glycinin analyzed previously. A third group of polypeptides purified from G. tomentella were the same size as basic polypeptides of glycinin and had homologus NH(2)-terminal sequences. The results indicated that the perennials exhibit variability in their seed proteins at a level not found among the cultivars of G. max and G. soja and may be useful in studies concerning the origin and organization of genes involved in the synthesis of storage proteins in cultivated soybeans.

Journal Article↗
PubMed1981

Identification of the acidic and basic subunit complexes of glycinin.

Five complexes consisting of one acidic and one basic subunit that were linked via disulfide bonds were purified from unreduced S-alkylated glycinin. The acidic and basic subunits were identified unambiguously using NH2-terminal sequence analysis, sodium dodecyl sulfate (SDS)-electrophoresis, and analytical isoelectric focusing. The subunit pairings are A1aB2, A1bB1b, A2B1a, A3B4, and F2(2)B3. Polypeptide A4 was not linked to a corresponding basic subunit via a disulfide bond. The study shows that pairing between subunits is nonrandom, which is consistent with evidence that glycinin is synthesized as a Mr = 60,000 precursor that undergoes post-translational modification to form the individual linked subunits.

Amino Acid Sequence↗