Isolation of a new member of the ecdysteroid glycoside family: 2-deoxy-20-hydroxyecdysone 22-O-beta-D-glucopyranoside.
A new ecdysteroid glycoside, 2-deoxy-20-hydroxyecdysone 22-O-beta-D-glucopyranoside, is isolated from the herb Silene italica ssp. nemoralis (Waldst. and Kit.) Nyman. The compound is purified with multistep chromatography, such as classical column chromatography on alumina and droplet countercurrent distribution. Also, it is expanded using twice low-pressure reversed-phase liquid column chromatography. Chromatography in four steps results in the purified 2-deoxy-20-hydroxyecdysone 22-O-beta-D-glucopyranoside. Two other ecdysteroids have also been separated, including the formerly identified integristerone A and 24(28)-dehydromakisterone A. (+info)
Introduction of a Na+/H+ antiporter gene from Atriplex gmelini confers salt tolerance to rice.
We engineered a salt-sensitive rice cultivar (Oryza sativa cv. Kinuhikari) to express a vacuolar-type Na+/H+ antiporter gene from a halophytic plant, Atriplex gmelini (AgNHX1). The activity of the vacuolar-type Na+/H+ antiporter in the transgenic rice plants was eight-fold higher than that in wild-type rice plants. Salt tolerance assays followed by non-stress treatments showed that the transgenic plants overexpressing AgNHX1 could survive under conditions of 300 mM NaCl for 3 days while the wild-type rice plants could not. These results indicate that overexpression of the Na+/H+ antiporter gene in rice plants significantly improves their salt tolerance. (+info)
Relationships between seed germinability of Spergularia marina (Caryophyllaceae) and the formation of zonal communities in an inland salt marsh.
BACKGROUND AND AIMS: The formation of zonal communities may be attributed to differences in germination across the community and to timing of germination of seeds present in the seed bank. Our goals were two-fold: (1) to assess the annual germination pattern of Spergularia marina; and (2) to determine whether germination of S. marina differed across zonal communities. METHODS: Fresh seeds were buried in an experimental garden in polyester bags. Bags were harvested monthly for 1 year and exposed to differing 12 h/12 h temperature regimes (5/15 degrees C, 5/25 degrees C, 15/25 degrees C and 20/35 degrees C) with a 12 h dark/12 h light photoperiod. Replicate seeds were exposed to 24 h dark. Seeds were also placed in different zonal communities to assess germinability in the field. KEY RESULTS: Spergularia marina has a primary physiological dormancy. Conditional dormancy occurs from December to May and non-dormancy from June to November. Field germination initiates in the spring when temperatures are cool and salinity is low due to flooding, and ceases in the summer when temperatures exceed germination requirements. Spergularia marina has a light requirement for germination. CONCLUSIONS: If seeds become buried in the field or are light inhibited by Phragmites australis, they will remain dormant until they receive an adequate amount of light for germination. Since S. marina can germinate across all zones in a salt-marsh community, the formation of zonal communities is not determined at the germination stage, but at some later stage of development. (+info)
New triterpenoid saponins from the roots of Sinocrassula asclepiadea.
Five new triterpenoid monodesmosides (sinocrassulosides I-V, 1-5) and six bisdesmosides (sinocrassulosides VI-XI, 6-11), in which 2-11 possess different acyl groups in the glycosidic moieties, were isolated from the roots of Sinocrassula asclepiadea FRANCH. Sinocrassulosides VI (4) and V (5) also contained a novel A-seco aglycone in their structures. All of the structures were determined on the basis of spectroscopic and physicochemical evidence. (+info)
DNA diversity in Hawaiian endemic plant Schiedea globosa.
This is the first report of a study devoted to the population genetics of speciation in the endemic Hawaiian plant genus Schiedea (Caryophyllaceae). Here, we report the estimates of DNA sequence diversity and divergence in a newly isolated nuclear gene from Maui and Oahu Schiedea globosa populations. Overall, the species-wide average heterozygosity per silent site is pi = 0.3%. The silent DNA diversity on the older island of Oahu (pi = 0.24%) is almost twice as high as on the younger Maui (pi = 0.14%). Consistent with this, the haplotype phylogeny suggests a more recent origin of the Maui populations. There is no significant isolation between the two Maui populations (F(st)=0.027), while isolation between the two islands is high (F(st)=0.57, P<0.0001). Pairwise mismatch distributions suggest population growth approximately 660 and 310 thousand generations ago for the Oahu and the Maui populations, respectively, which may be the minimal age for these populations. This is consistent with a fairly neutral frequency spectrum (Tajima's D is 0.34 and -0.94 for the Oahu and the Maui populations, respectively), suggesting that both populations are sufficiently old to have recovered from any initial founder effects. Relatively high nuclear DNA diversity in the S. globosa populations illustrates the usefulness of a DNA sequence-based approach to the population genetics of island plant populations. (+info)
Subcellular distribution of the V-ATPase complex in plant cells, and in vivo localisation of the 100 kDa subunit VHA-a within the complex.
BACKGROUND: Vacuolar H+-ATPases are large protein complexes of more than 700 kDa that acidify endomembrane compartments and are part of the secretory system of eukaryotic cells. They are built from 14 different (VHA)-subunits. The paper addresses the question of sub-cellular localisation and subunit composition of plant V-ATPase in vivo and in vitro mainly by using colocalization and fluorescence resonance energy transfer techniques (FRET). Focus is placed on the examination and function of the 95 kDa membrane spanning subunit VHA-a. Showing similarities to the already described Vph1 and Stv1 vacuolar ATPase subunits from yeast, VHA-a revealed a bipartite structure with (i) a less conserved cytoplasmically orientated N-terminus and (ii) a membrane-spanning C-terminus with a higher extent of conservation including all amino acids shown to be essential for proton translocation in the yeast. On the basis of sequence data VHA-a appears to be an essential structural and functional element of V-ATPase, although previously a sole function in assembly has been proposed. RESULTS: To elucidate the presence and function of VHA-a in the plant complex, three approaches were undertaken: (i) co-immunoprecipitation with antibodies directed to epitopes in the N- and C-terminal part of VHA-a, respectively, (ii) immunocytochemistry approach including co-localisation studies with known plant endomembrane markers, and (iii) in vivo-FRET between subunits fused to variants of green fluorescence protein (CFP, YFP) in transfected cells. CONCLUSIONS: All three sets of results show that V-ATPase contains VHA-a protein that interacts in a specific manner with other subunits. The genomes of plants encode three genes of the 95 kDa subunit (VHA-a) of the vacuolar type H+-ATPase. Immuno-localisation of VHA-a shows that the recognized subunit is exclusively located on the endoplasmic reticulum. This result is in agreement with the hypothesis that the different isoforms of VHA-a may localize on distinct endomembrane compartments, as it was shown for its yeast counterpart Vph1. (+info)
Characterization of antifreeze activity in Antarctic plants.
Deschampsia antarctica and Colobanthus quitensis are the only vascular plants to have colonized the Maritime Antarctic, which is characterized by its permanently low temperature and frequent summer frosts. To understand how the plants survive freezing temperatures year-round, antifreeze activity was assayed in apoplastic extracts obtained from both non-acclimated and cold-acclimated Antarctic plants. By observing the shape of ice crystals grown in dilution series of the extracts, it was found that D. antarctica had antifreeze activity, but C. quitensis did not. D. antarctica exhibited antifreeze activity in the non-acclimated state and this activity increased after cold acclimation. The antifreeze activity in D. antarctica was labile to proteolysis and high temperature, active over a wide pH range, and associated with molecules greater than 10 kDa in molecular weight. These results show that D. antarctica produces antifreeze proteins that are secreted into the apoplast. When examined by SDS-PAGE, the apoplastic extracts from cold-acclimated D. antarctica exhibited 13 polypeptides. It is concluded that D. antarctica accumulates AFPs as part of its mechanism of freezing tolerance. Moreover, this is the first plant in which antifreeze activity has been observed to be constitutive. (+info)
Analysis of promoters recognized by HrpL, an alternative sigma-factor protein from Pantoea agglomerans pv. gypsophilae.
HrpL, an alternative sigma factor, activates the transcription of the Hrp regulon by its binding to a common "hrp box" promoter. Based on computational techniques, the hrp box previously was defined as a consensus bipartite cis element, 5'-GGAACC-N(15-16)-CCACNNA-3'. The present report combines a quantitative in vivo assay for measuring Hrp promoter activity with site-specific mutagenesis to analyze the effect of consensus and nonconsensus nucleotides on promoter activity. The analysis was carried out with Hop effectors of the tumorigenic bacterium Pantoea agglomerans pv. gypsophilae, in which HrpL is indispensable for gall formation. Mutational analysis indicates that the hrp box consensus can be divided into crucial and noncrucial nucleotides. The first 5 nucleotides (nt) of the--35 consensus motif (GGAAC) and the 3 nt of the--10 motif (ACNNA) are crucial, whereas other consensus and adjacent nonconsensus nucleotides exert a significant effect on the promoter's strength. With spacing of 13 or 17 nt between the two motifs, significant activity was still retained. Gel shift assays indicated that deletion of GG from the--35 consensus motif eliminated HrpL binding, whereas mutations in the--10 consensus motif or modification of the spacing, which eliminates promoter activity, did not elicit any effect. The degeneracy in Hrp promoters of four hrp and type III effector genes of P agglomerans pv. gypsophilae indicated significant differences in promoter activity, whereas increasing the promoter strength of the Hop effector, HsvG, resulted in overexpression of gall formation. (+info)